┘ḗ ࢤࣤࣆ࢒ࣝࣤࢪࣈࣞࢡ࣑ࣚ yyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyy Ⓠ⾪ⁿ㢗ཀྵࡦㅦⁿ᫤㛣 yyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyy ≁ืㅦⁿさ᪠ yyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyy ࢨࣤ࣎ࢩ࣑ࢗㅦⁿさ᪠ yyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyy ୌ⯙ㅦⁿさ᪠ yyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyy ࣎ࢪࢰ࣭Ⓠ⾪ㅦⁿさ᪠ yyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyy ெྞ⣬ᘤ yyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyy ⣊≟ⳞฦᏄ⏍∸Ꮥ◂✪ఌఌ์ yyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyy ⣊≟ⳞฦᏄ⏍∸Ꮥ◂✪ఌ㐘ႜጟဤྞ⡑ yyyyyyyyyyyyyyyyyyyyyyyyyyyyy Ở
14
➠㸫ᅂ⣊≟ⳞฦᏄ⏍∸Ꮥࢤ ࣤࣆ ࢒ࣝࣤࢪࣈࣞࢡ࣑ࣚ
᪝᫤㸯ᖲᠺ 18 ᖳ 11 ᭮ 13 ᪝(᭮)-14 ᪝(ℾ)
ఌሔ㸯ኬ㜨ᗋ❟ኬᏕ 㹊࣭࣌ࣜⓉ㮓
㸝ኬ㜨ᗋሗᕰ୯༇Ꮥᅧ⏣㸦㸢㸦㸞
୹ത㸯⣊≟ⳞฦᏄ⏍∸Ꮥ◂✪ఌ
ᚃᥴ㸯⣊≟Ⳟ㐿ఎᏄ◂✪ఌ
11 ᭮ 13 ᪝㸝᭮㸞
12:30-14:30 ཾ㢄Ⓠ⾪㸝O-1㹳O-8㸞
14:30-14:45 ఆ៹
14:45-16:15 ࣎ࢪࢰ࣭Ⓠ⾪㸝ዃᩐ␊ྒ㸞
16:15-17:00 ≁ืㅦⁿձ
Dr. B. Gillian Turgeon
(Department of Plant Pathology,
Cornell University)
17:00-17:45 ≁ืㅦⁿղ
Dr. James R. Kinghorn and Dr. Shiela Unkles
(School of Biology,
University of St. Andrews)
17:45-17:55 ⥪ఌ
18:00- ᠋ばఌ㸝Ꮥ⾙ஹὮఌ㤃㸞
11 ᭮ 14 ᪝㸝ℾ㸞
9:30-12:00 ࢨࣤ࣎ࢩ࣑ࢗ
12:00-13:00 ᫠㣏
13:00-14:30 ࣎ࢪࢰ࣭Ⓠ⾪㸝ഃᩐ␊ྒ㸞
14:30-16:45 ཾ㢄Ⓠ⾪㸝O-9㹳O-17㸞
16:45-16:55 㛚ఌࡡ㎙
2
発表演題および講演 時間
≁ืㅦⁿ ᭮ ᪝㸝᭮㸞
16:15-17:00 ≁ืㅦⁿձ
Secondary metabolites: fundamental and niche-specific roles
in fungal development and cell-cell interactions
Dr. B. Gillian Turgeon (Department of Plant Pathology, Cornell University)
17:00-17:45 ≁ืㅦⁿղ
Fungal Nitrate and Nitrite Transporters: Insights into Structure and Function
Dr. James R. Kinghorn and Dr. Shiela Unkles
(School of Biology, University of St. Andrews)
ࢨࣤ࣎ࢩ࣑ࢗ 11 ᭮ 14 ᪝㸝ℾ㸞9:30-12:00
「バイオマスの有効利 用に向けた糸状菌の役割」
9:30
9:40
10:15
10:50
11:25
「はじめに」
川口 剛司（阪府大院・応生科）
S-1「担 子菌 の生 産 する セ ルロ ー ス分 解系 酵 素 の多 様 性と その 役 割」
天野 良彦 (信州大・物質工学)
S-2「選 択的 白色 腐 朽菌 に よる 木 質バ イオ マ ス のエ ネ ルギ ー・ 化 学資 源 化」
渡辺 隆司（京大 生存圏研）
S-3「バ イオ エタ ノ ール 生 産に 利 用さ れる 糸 状 菌由 来 酵素 」
髙木 忍（ノボザイムズ ジャパン）
S-4「高 機能 型セ ル ラー ゼ 高生 産 Trichoderma reesei 株の 構 築に 向け て 」
森川 康（長岡技科大・生物系）
3
ୌ⯙ㅦⁿ㸝O-1㹳O-8㸞 11 ᭮ 13 ᪝㸝᭮㸞12:30ࠤ14:30
12㸯30 O-1
࠷ࡵࡔ⑋Ⳟ࡞࠽ࡄࡾ஦ࡗࡡ Dicer ᵕ⺦Ⓣ㈻ࡡᶭ⬗ฦ໩
みㆺ├ᶖ㸡ᅰఫᖶ㞕㸡┶ᒜ⁘ᚷ㸡୯ᒁᩔᆍ㸝♼ᡖኬ࣬㎨Ꮥ㒂㸞
12㸯45 O-2
ࢩࣔ࢝࢕࣓ኚ␷⑋Ⳟ Alternaria solani ⏜ᮮ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪ PKSN ࡡⓆ⌟࡛ᶭ⬗
よᯊ
➗ཋᇻ㸦㸡ཀྵᕖⱝ⚽㸧ࠉ⸠஬໌㸦ࠉᾇ⩹ሪ㇇㸦㸝㸦᮶ኬ㝌࣬ⷾࠉ㸧໪ኬ㝌࣬⌦㸞
13㸯00 O-3
㯔Ⳟ A. oryzae ࡡ␏⛸ࢰࣤࣂࢠ㈻㧏⏍⏐ን␏ᰬ ࡡཱི ᚋ
᰷ᮇᓤࠉῳ㎰ὀ♰ࠉ‹୕㇇ࠉ୷ᒜ₮ୌࠉ᭯ᒱᏕࠉ໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬
㎨⏍⛁࣬ᚺ⏍ᕝ㸞
13㸯15 O-4
࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ G ࢰࣤࣂࢠ㈻โᚒᅄ Ꮔ RGS ࡞ࡗ࠷࡙ࡡよᯊ
ཪ㔕⌦⏍ࠉ༞ᰜୌࠉこᮟ㯖㔓Ờ 㸝⏍∸◂㸞
13㸯30 O-5
Aspergillus nidulans ࡞࠽ࡄࡾ His-Asp ᝗ሒఎ㐡⣌ࢰ ࣤࣂࢠ㈻ࡡ⥑⨮Ⓩよᯊ
㔘୷ாᏄࠉᮿᯐⰃ༡ࠉ᮶ಘᏳࠉ㕝ᮄ㯖Ꮔࠉຊ⸠㞖ኃࠉᑚᯐူኰ 㸝ྞኬ࣬
⏍࿤㎨㸞
13㸯45 O-6
⣊≟Ⳟ Aspergillus nidulans ࡡ㎨ⷾᚺ➽࡞࠽ࡄࡾ His-Asp ࣛࣤ㓗࣭ࣛࣝ⣌ࡡよᯊ
ⴏཋኬ♰ 1ࠉᮿᯐⰃ༡ 1ࠉ㔘୷ாᏄ 1 ࠉຊ⸠㞖ኃ 1ࠉ㜷㒂ᩏᝃ 2ࠉᑚᯐူኰ 1ࠉ Ề㔕⊓ 1 㸝1 ྞኬ㝌࣬⏍࿤㎨Ꮥࠉ2 ᮶໪ኬ㝌࣬㎨Ꮥ㸞
14㸯00 O-7
Aspergillus nidulans HOG ⤊㊪≁␏Ⓩ࡝࣭ࣝ࣎ࢰ࣭㐿 ఎᏄⓆ⌟⣌ࡡᵋ⠇
ཿᕖ೸ኯ㑳ࠉཿᕖ㈏Ꮔࠉ᫅⏜⣎Ꮔࠉఫ⸠ዄ὘Ꮔࠉྚぜၤ*ࠉ⸠ᒱᬓ์ࠉ
Ềㆺ἖ࠉ㜷㒂ᩏᝃ
14㸯15 O-8
(᮶໪ኬ㝌㎨࣬ᚺ⏍⛁ࠉ*᮶໪ኬ࣬ᮅᮮ◂)
Aspergillus oryzae ࡞࠽ࡄࡾ steA ࣓࣌ࣞࢡࡡᶭ⬗よᯊ
᲻⏛ᐰெࠉ➁හ㐠㞕 㸝᮶ா㎨ᕝኬ㝌࣬⏍∸ᕝᏕ㸞
ୌ⯙ㅦⁿ㸝O-9㹳O-17㸞 11 ᭮ 14 ᪝㸝ℾ㸞14:30ࠤ16:45
14㸯30 O-9
Aspergillus nidulans ࡡᙟ㈻㌹ᥦ᫤࡞ᚪさ࡛ࡈࡿࡾฦ Ꮔ࠽ࡻࡦ᧧ష࡞ࡗ
࠷࡙ࡡ᳠ゞ
᱋㔕ူ▦ 1ࠉ ౪⏛ᗛᏏ 2ࠉ ௿⸠㟱ኰ 2࣬3 㸝1 ಘኬ࣬⌦ࠉ2 ಘኬ㝌࣬ᕝࠉ
3
14㸯45 O-10
ಘኬ࣬ධᏕᩅ⫩ᶭᵋ㸞
⣊≟Ⳟ Aspergillus nidulans ࡞࠽ࡄࡾ࢞ࢲࢻ࣭ࢭ㐿ఎ Ꮔ⩄ࡡⓆ⌟よᯊ
⏛୯ᙤࠉᒜᓧᬍ୓ࠉᇷහ⿩஄ࠉኯ⏛᪺ᚠ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
4
15㸯00 O-11
》Ⰵ⭁ᮑⳞ Coniophora puteana ࡞ࡻࡾ᳔∸⣵⬂ባ⏜ ᮮኣ⢶ࡡฦよ࣒࢜
ࢼࢫ࣑
ຊ἖ఫᖲࠉ஫༎ᔋᅽ᪝Ꮔࠉ㩢ᓞḿᾀ㸝᮶ኬ㝌࣬㎨⏍⛁࣬⏍ᮞ⛁㸞
15㸯15 O-12
Fusarium oxysporum f. sp. melonis ⏜ᮮࡡβ-1,6-࢝ࣚࢠࢰࢻ࣭ࢭࡡ⢥⿿࡛
ࢠ࣭ࣞࢼࣤࢡ
ㆺཾ㞕သࠉ㜨ᮇ㱗ྒྷࠉᕖᓧ᮶ᙢ㸝ኬ㜨ᗋ❟ኬ࣬⏍࿤⎌ሾ㸞
15㸯30 O-13
ࣛ࣍ࣆࣚࣄࣤ⏍⏐⣊≟Ⳟ Ashbya gossypii ࡡ௥ㅨよᯊ
㔘ᨳ┷ 1 ࠉ⏛ᓞㅅ 1ࠉᮌ㱗ὒ 1,2㸝1 㟴ᒱኬ࣬㎨࣬ᚺ⏍໩ࠉ2 㟴ᒱኬ࣬๭⛁ᢇ㝌࣬⤣ྙࣁ࢕࢛㸞
15㸯45 O-14
》Ⰵ⭁ᮑⳞ࢛࢛ࢗࢫࣚࢰࢢ࡞࠽࠷࡙ࢨࣖࢗ 㓗⪇ᛮ ࡞ᐞ୙ࡌࡾࢰࣤࣂ
ࢠ㈻ࡡᶭ⬗よᯊ
ῳ㑋▩ᶖ 1ࠉኃཬఘ࿰ 1ࠉᱭ⃕ಆ᪺ 1 ࠉ▦ᓧୌྍ 1ࠉᓞ⏛ᖷኰ 2ࠉ᭱㒂Ṃᩝ 1
(1 ாኬ࣬⏍Ꮛ◂ࠉ2 ⚗஬ᕝኬ࣬ᕝ)
16㸯00 O-15
TPP ⤎ྙᆵࣛ࣍ࢪ࢕ࢴࢲࡡᶭ⬗ᨭን
ᒜහ㝧ᐰ 1,2ࠉ୔ይኬ㍔ 2ࠉ❉ᑈ㝧ᩝ 1ࠉఔක༡ 1ࠉこᮟ㢟 1ࠉᮙᮇ├ᕤ 2,3
㸝1 Ⓣ㭧㒿㏸㸝ᰬ㸞࣬◂✪㛜Ⓠᐄࠉ2 ⏝༞ኬ࣬FIBERࠉ3 ⏝༞ኬ࣬⌦ᕝ㸞
16㸯15 O-16
㯔Ⳟ Aspergillus oryzae ࡡࢲ࢓࣐ࣤ㧏⏍⏐ᰬࡡฦᏄ ⫩⛸
ᚠ஬⨶㔓ࠉ❉ᑈ㝧ᩝࠉ஫࿝ົஒ 1ࠉᒜୖఘ㞕ࠉఔක༡ࠉこᮟ㢟 㸝Ⓣ㭧
㒿㏸◂✪㛜Ⓠࠉ1 ᮶໪ኬ㝌㎨࣬⏍∸⏐ᴏ๭ᠺ㸞
16㸯30 O-17
Ềฦྱ㔖ࡡ␏࡝ࡾᅖమᇰ㣬᮪௲ୖ࡞࠽ࡄࡾ 㯔Ⳟ㐿 ఎᏄⓆ⌟ࡡよᯊ
ᑚᯐல⣎Ꮔ㸦ࠉఫ㔕ඔ᫓㸦ࠉ஁⏛༡ඔ㸧ࠉ⛑Ὂ஦㸧ࠉኬ⟺ಘୌ㸦 㸝㸦㔘Ἁᕝᴏ
ኬ࣬ࢣࢿ࣑◂ࠉ㸧᭮᰿෗࣬⥪◂㸞
࣎ࢪࢰ࣭Ⓠ⾪
ዃᩐ␊ྒ : 11 ᭮ 13 ᪝ 14:45-16:15
ഃᩐ␊ྒ : 11 ᭮ 14 ᪝ 13:00-14:30
P-1 㓕ẍ Bni1 ࡡ┞ྜྷࢰࣤࣂࢠ㈻ AoBni1 ࢅ⏕࠷ࡒ㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ Spitzenkörper ࡡྊっ໩
▴ᕖ⤦⌦ࠉḿ㊪ῗஒࠉ᭯ᒱᏕࠉ໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
P-2 ⺧කࢰࣤࣂࢠᵾㆉ࡞ࡻࡾ Aspergillus nidulans ඙❻ᠺ㛏ᶭᵋࡡよᯊ㸯᚜ᑚ⟮࡛࢓ࢠࢲࣤࡡ㛭౿ ࢅ ୯
ᚨ࡞
1
ᇷᑹူஒࠉ2 ⣒ ᭰ᾀࠉ3Naimeh Taheri-Taleshࠉ2 ⳝờ㡝ᙢࠉ 3Berl R. Oakley㸝1 ᚠᓞኬ㝌࣬ࣉࣜࢪࣁ࢕
࢛ࠉ2 ྞཿᒁኬ㝌༈⣌࣬ฦᏄᵾⓏࠉ3 ࢛ࣀ࢕࢛ᕗ❟ኬ࣬ฦᏄ㐿ఎ㸞
P-3 A. oryzae ࡡ Woronin body ᙟᠺ࡞㛭୙ࡌࡾ Aohex1 㐿ఎᏄࡡ◒ቪᰬ࡞࠽ࡄࡾ⾪⌟ᆵよᯊ
୷ᒜ₮ୌࠉ໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
P-4 A. oryzae ࡡ Woronin body ᙟᠺ࡞ᚪさ࡝ Aohex1 㐿ఎᏄ࡞࠽ࡄࡾ㐽ᢝⓏࢪࣈࣚ࢕ࢨࣤࢡࡡណ ⩇࡞㛭
ࡌࡾよᯊ
ᒷᓧ೸ኯ㑳ࠉහ⏛༡ᩅࠉPraveen Rao Juvvadiࠉ୷ᒜ₮ୌࠉ໪ᮇົࡥࡆ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
5
P-5
࢓࢜ࣂࣤ࢜ࣄ࡞࠽ࡄࡾ⺧කࢰࣤࣂࢠ ࡚ᵾㆉ ࡊࡒ NC-RAS-2 㐿ఎᏄࡡྊっ໩
Ⱪἠᖶኯࠉᮟᒜ⫍Ꮔ 㸝㛭᮶Ꮥ㝌ኬ㝌࣬ᕝᏕ◂✪⛁㸞
P-6
N-⤎ྙᆵ⢶㙈ࣄ࣭ࢫࢅ⏕࠷ࡒ A. oryzae ᑚ⬂మහࣝࢠࢲࣤࡡཱིᚋ࡛ᑻᅹよᯊ
ῳ㑋ὀ♰ 1ࠉᮿᑹୌ㑳 1,2ࠉ୷ᒜ₮ୌ 3ࠉ໪ᮇົࡥࡆ 3ࠉ௿⸠ᖶᠺ 1,2㸝⌦◂ 1ࠉCREST2ࠉ᮶ኬ㝌㎨⏍⛁࣬
ᚺ⏍ᕝ 3 㸞
P-7
㯔Ⳟ A. oryzae ࡞࠽ࡄࡾᑚ⬂మࢪࢹࣝࢪࡡྊ っ໩
ኬ㔕⤚Ꮔࠉ୷ᒜ₮ୌࠉ᭯ᒱᏕࠉ໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
P-8
㯔Ⳟ Aspergillus oryzae Ⳟ⣊ᇱ㒂࡞࠽ࡄࡾ࢛ࣜ ࢝ࢾࣚᵋᠺᠺฦࡡ࢛࣭ࢹࣆ࢒ࢩ࣭࡞ࡻࡾᾦ ⬂࡫ࡡ
ཱི㎲ࡲ
ḿ㊪ῗஒࠉⳝ㛣㝧ᚷࠉ᭯ᒱᏕࠉ໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
P-9
ᢰᏄⳞ Coprinus cinereus ⏜ᮮ࢛࣭ࢹࣆ࢒ࢩ࣭ 㛭㏻ࢰࣤࣂࢠ㈻ Atg8 ࣓࣌ࣞࢡࡡᶭ⬗よᯊ
ྚᮟ᪸ᚠࠉῳ㑋 ᙪࠉ㯖⏛᜜ᙢ㸝㤮ᕖኬ࣬㎨㸞
P-10 㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ࢙ࣤࢺࢮ࣭࣑ᵕᵋ㏸ మࡡよᯊ
ᵵཾ⿩ḗ㑳ࠉḿ㊪ῗஒࠉ᭯ᒱ Ꮥࠉ໪ᮇົࡥࡆ (᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ)
P-11
㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ AoVps24 ࡡᾦ⬂ᙟᠺ ࡞㛭ࡌࡾᶭ⬗ࡡよᯊ
㎦ᕦᬏ⣎ࠉⳝ㛣㝧ᚷࠉḿ㊪ῗஒࠉ᭯ᒱᏕࠉ໪ᮇົࡥࡆ (᮶ኬ㝌㹺㎨⏍⛁㹺ᚺ⏍ᕝ)
P-12 ࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ௛╌ჹᙟᠺ࡞ᑊࡌࡾ྘⛸ ᢘ⏍∸㈻ࡡᙫ㡢
㛏ᓞ⏜ኚࠉṂᕖ἖ࠉ᩟⸠ᙤ⳱ࠉ㙂ಲ㧏ᚷ 㸝᮶⌦ኬ⌦ᕝ㸞
P-13 ࢕ࢾ࠷ࡵࡔ⑋Ⳟ࡞࠽ࡄࡾᶭ᲌ๆ⃥ུᐖ࢜ ࣜ ࢨ࣑ࢗ࢕࢛ࣤࢲࣔࢾࣜ MID1 ࣓࣌ࣞࢡࡡᶭ⬗よᯊ
௿⃕┷⃀ࠉ᯵ᮄ▩Ꮔࠉ㙂ಲ㧏ᚷ 㸝᮶⌦ኬ⌦ᕝ㸞
P-14 ࢕ࢾ⛁᳔∸࠷ࡵࡔ⑋Ⳟ(Pyricularia spp.)࡞࠽ࡄࡾ㟸⑋ཋງ㐿ఎᏄ AVR-Pita ࡡᗑ஋᯹Ⰵమንິᶭᵋ
୯㤷࠷ࡘࡲࠉ☶㒂༐ᑖࠉ୯ᒁᩔᆍࠉ┶ᒜ⁘ᚷࠉᅰఫᖶ㞕 㸝♼ᡖኬ࣬㎨㸞
P-15 ࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ㟸⑋ཋᛮ㐿ఎ Ꮔ Avr-pia ࡡࢠ࣭ࣞࢼࣤࢡ
᭧᰷㍜㞕 1ࠉ୔ᮄួ௒ 1ࠉᮿ஬ᖶኯ㑳 1ࠉⰩ⃕Ṃெ 2 ࠉ㨛㢄ⱝᶖ 3 ࠉᖲඳ㔔ୌ஄ 1ࠉ
୯ᓞᨼᙢ 4ࠉෞ⏛ᡛ⏠ 5 㸝1 ໪ኬ㝌㎨࣬ᚺ⏕ⳞᏕࠉ2 ୯ኳ㎨◂ࠉ3 ㎨ᴏ⏍∸㈠″◂ࠉ4 ᮶໪㎨◂ࠉ5 ᨲ㏞ኬ㸞
P-16 Biotinylated NeoechinulinA ໩Ꮥྙᠺ⤊㊪ࡡᨭ ၻ࡛ Aspergillus ᒌࡡ NeoechinulinA ⏍ྙᠺ㐿ఎᏄ⩄ࡡ
᥀⣬
➁㔕ḿὊࠉኬ▴㯖⏜ࠉᇷහ᱀Ꮔࠉ㙂ಲ㧏ᚷ 㸝᮶⌦ኬ⌦ᕝ㸞
P-17 Aspergillus fumigatus ࠿⏍⏐ࡌࡾࢹࣛࣈࣞࢪࢰ ࢲࣤ㢦ࡡ⏍ྙᠺ㐿ఎᏄࢠࣚࢪࢰ࣭
ຊ⸠├ᶖࠉ㧏ᮄᾇࠉὰぜ⾔ᘧࠉ᤻ㆺ⚵᫓ࠉ㛏⏛⿩஄ 㸝⌦◂࣬୯ኳ◂࣬ᢘ⏍∸㈻㸞
P-18 㯔Ⳟ A. oryzae ࡡ III ᆵ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪ 㐿ఎᏄࡡᶭ⬗よᯊ
ເࠎᗛ௥ 1ࠉ⸠஬໌ 1ࠉPraveen Rao Juvvadi2 ࠉ໪ᮇົࡥࡆ 2ࠉᾇ⩹ሪ㇇ 1
㸝1 ᮶ኬ㝌࣬ⷾࠉ2 ᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
P-19 ⧖㏁ࡊᆵࢰ࢕ࣈϨ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪ ࡡ Ⓠ⌟࡛ᶭ⬗よᯊ
᲻ཾᬓ⨶ࠉ⸠஬໌ࠉᾇ⩹ሪ㇇㸝᮶ኬ㝌࣬ⷾ㸞
P-20 㯔Ⳟ A. oryzae ࡡ⧖㏁ࡊᆵࢰ࢕ࣈϨ࣎ࣛࢢࢰ ࢕ࢺྙᠺ㓕⣪㐿ఎᏄࡡⓆ⌟࡛ᶭ⬗よᯊ
Ꮼᒁᬓ༡ 1ࠉເࠎᗛ௥ 1ࠉ໪ᮇົࡥࡆ 2ࠉ⸠஬໌ 1ࠉᾇ⩹ሪ㇇ 1 㸝᮶ኬ㝌࣬ⷾ 1ࠉ
᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ 2㸞
P-21 Penicillium luteo-aurantium ࡡ࣎ࣛࢢࢰ࢕ࢺྙ ᠺ㓕⣪㐿ఎᏄࡡᶭ⬗よᯊ
ఫ㔕⿩ᇱࠉ⸠஬໌ࠉᾇ⩹ሪ㇇ 㸝᮶ኬ㝌࣬ⷾ㸞
P-22 ⣊≟Ⳟⰶ㤮᪐࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪ࡡࢠࣚ ࢕ࢭࣤࢦ࢕ࢠ࣭ࣚࢭࢺ࣒࢕ࣤ
ᐋᓧಆࠉ⸠஬໌ࠉᾇ⩹ሪ㇇ 㸝᮶ኬ㝌࣬ⷾ㸞
6
P-23 㓕ẍࢅ࣌ࢪࢹ࡛ࡊࡒ Aspergillus fumigatus DHN-࣒ࣚࢼࣤ⏍ྙᠺ⣌ࡡ්ᵋᠺ
༞㒂ኚᕵࠉ⸠஬໌ࠉᾇ⩹ሪ㇇ 㸝᮶ኬ㝌࣬ⷾ㸞
P-24 GFP ⼝ྙ⺦Ⓣ㈻ࢅ⏕࠷ࡒ㯔Ⳟࡡࢨࢸࣞࣆ࢚ ࢓⏍⏐ㄢ⟿ᅄᏄ(SreAo)ࡡᑻᅹよᯊ
ῳ㎰஁ᩏࠉఫ⸠ฺḗ㸝ᒷᡥ⏍ᕝ◂㸞
P-25 ⣒㯔Ⳟ㸝Monascus pilosus㸞࡞ࡻࡾ஦ḗ௥ㅨ⏐∸ࡡ⏍⏐
୔Ꮹ๙ྍ 1ࠉᘿ᪺Ễ 1,2ࠉහᐣ஁⨶Ꮔ 3ࠉἑ㔕ຩெ 1 ࠉ㔕ᓧಘ⾔ 1 ࠉ⏐ᮇᘧ஄ 1ࠉ⛼ᇄ㈶஦ 3
㸝1 ᒱᒜ┬ᕝᢇࢬࠉ2JSPSࠉ3 ᒱᒜኬ㝌࣬⏍∸ᶭ⬗໩Ꮥ㸞
P-26 ࢞ࣔ࣊ࢵⴆ㯜⑋Ⳟ Cong:1-1 ࡛ࡐࡡ⑋ཋᛮḖ᥾ᰬ Cong:1-2 ࡞࠽ࡄࡾࢰࣤࣂࢠ㈻Ⓠ⌟ࣂࢰ࣭ࣤࡡ
よᯊ
ᒱ㒂᪺Ꮔࠉྚ⏛㝧ᘇ 1 ࠉ᭯ᮇ⿩ 2ࠉᇸ๑├ 2 ࠉ᲻⩟྿ 3ࠉᑈᒱᚥࠉ᭯Ờງ㸝㎨ᕝኬࠉ1 ㎨⎌◂ࠉ2 ⌦◂ࠉ
3
㎨ኬ㸞
P-27 Colletotrichum acutatum ࡡ࣊ࢿ࣐ࣜ⪇ᛮ㐿ఎᏄ CaBEN1 ࡢș-ࢲ࣭ࣖࣇࣛࣤ㐿ఎᏄࡡ㌹෕࡞㛭ࢂࡖ
࡙࠷ࡾ
୯⏿Ⰳ஦ࠉ୯㔕ḿ᪺ 㸝㎨◂ᶭᵋ ᯕᶖ◂㸞
P-28 ࣛࣤࢥᩤⅤⴘⴝ⑋Ⳟࡡ AM Ẑ⣪⏍ྙᠺ㐿ఎᏄࢠࣚࢪࢰ࣭࡞ࢤ࣭ࢺࡈࡿࡾ Zn(II)2Cys6 ࢰ࢕ࣈ㌹
෕โᚒᅄᏄࡡᶭ⬗
᧓ᮇ౅᪺ࠉඡ⋚ᇱୌ᭹ 1ࠉᒜᮇᖷ༡ 2ࠉᑹㆺ ᾀ 1 ࠉ᯼᳔ᑠᚷ (ྞኬ㝌⏍㎨࣬1 㫵ཱིኬ㎨࣬2 ᒱᒜኬ㎨)
P-29 ࢻࢨ㯦ᩤ⑋Ⳟࡡ㈇✰Ⳟ⣊࡞࠽ࡄࡾὩᛮ㓗 ⣪ ⛸(ROS)ࡡᶭ⬗よᯊ
⋖ᗛὒࠉờ⏛೸ୌࠉ୯ᒁᩔᆍࠉᮌᮠඒ㸝♼ᡖኬ࣬⮤↓⛁Ꮥ㸞
P-30 Fusarium oxysporum ࡡ⬂Ꮔᙟᠺ㛭㏻㌹෕ᅄᏄ Ren1 ࡢல◢㓗㑇ඔ㓕⣪㐿ఎᏄࢅโᚒࡌࡾ
㣜⏛♰ୌ㑳ࠉᑚཋᨼ᪺ࠉ᯼᳔ᑠᚷ 㸝ྞኬ㝌⏍㎨㸞
P-31 Fusarium oxysporum ࡡࣅࣜࣄࣤ㓗ࢸ࢜ࣜ࣍࢞ ࢨ࣭ࣚࢭ㐿ఎᏄࡡྜྷᏽ
ᮇᒜយࠉᑈᒱᚥࠉ᭯Ờງ 㸝᮶ா㎨ᕝኬ࣬㝌㎨⏍∸โᚒ㸞
P-32 ᾈ㏩ᅸ࡞ࡻࡽⓆ⌟࠿ንິࡌࡾ㯔Ⳟࡡ㐿ఎᏄ ⩄࡛ࡐࡡࣈ࣓࣭ࣞࢰ࣭ࡡฺ⏕
⧂⏛೸ࠉ1 ᆊᮇ࿰ಆࠉ2 ᭯㤷ᑋⱝࠉ1 ኬ໪⏜౅ࠉ1 ᰘⷭࢱࣚࣚࢴࢹࠉ1 ᒜ⏛ ಞࠉ1,3 ᒷୖ࿰⿩ࠉ4 ⚽⏛ಞ㸝㔘
Ἁᕝኬࢣࢿ࣑◂ࠉ1 㒿⥪◂ࠉ2 ┬❟ᗀᓞኬࠉ3 ᗀኬ඙❻◂ࠉ4 ᐁ㊰ዥᏄኬ㸞
P-33 㯑Ⳟ Aspergillus oryzae ࡡ bZIP ᆵ㌹෕โᚒᅄᏄ㐿ఎᏄ atfA ࡡよᯊ
ᆊᮇ࿰ಆ 1ࠉᒜ⏛ಞ 1ࠉኬ໪⏜౅ 1ࠉᒷୖ࿰⿩ 1ࠉ⚽⏛ಞ 2ࠉ஫࿝ົஒ 3ࠉ୔୕㔔᪺ 1㸝1 㒿⥪◂ࠉ2 ᐁ㊰ዥ
Ꮔኬࠉ3 ᮶໪ኬ㝌࣬㎨㸞
P-34 ࢓࢜ࣂࣤ࢜ࣄࡡᾈ㏩ᅸࢪࢹࣝࢪ࡚ㄇᑙࡈࡿ ࡾࢡࣛࢬ࣭ࣞࣜྙᠺ࡞࠽ࡄࡾ glycerol dehydrogenase
ࡡᙲ๪
ᒜୖ࿰Ᏻ 1ࠉᆊ㔕┷ᖲ 2ࠉሲ⃕࠵ࡍࡈ 1ࠉ⸠ᮟ┷ 1㸝1 ᮶Ὂኬ࣬⏍࿤ࠉ2 ᮶Ὂኬ᳔࣬ᶭ◂ࢬ㸞
P-35 ࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡࣃࢪࢲࢩࣤ࢞ࢻ࣭ ࢭ Hik1 ࡡ᝗ሒఎ㐡⣌ࡡよᯊ
᲻⏛┷⣟ 1,2ࠉᏫఫ⨶ㄵ 2ࠉᕝ⸠ಆ❮ 1ࠉᮇᒜ㧏ᖶ 1 㸝1 ⌦◂࣬୯ኳ◂ࠉ2 ᮶Ὂኬ࣬ᕝ㸞
P-36 ⣊≟Ⳟ Aspergillus nidulans ࡞࠽ࡄࡾኣᵕ࡝㓗໩ࢪࢹࣝࢪᚺ➽ᶭᵋࡡよᯊ
῞㔕♰ᗀࠉⴏཋኬ♰ࠉ㔘୷ாᏄࠉຊ⸠㞖ኃࠉᑚᯐူኰࠉỀ㔕⊓㸝ྞኬ㝌࣬⏍࿤㎨Ꮥ㸞
P-37 ⣊≟Ⳟ Aspergillus nidulans ࡞࠽ࡄࡾ஦ᠺฦᛮ ᝗ሒఎ㐡⣌ࣛࣤ㓗ᇱ௯௒ᅄᏄ YpdA ࡡよᯊ
ఫ⸠ዄ὘Ꮔࠉཿᕖ೸ኯ㑳ࠉ㜷㒂ᩏᝃ (᮶໪ኬ㝌࣬㎨࣬ᚺ⏍⛁)
P-38 Aspergillus nidulans ࣝࢪ࣎ࣤࢪ࣭ࣝ࢟ࣖࣝࢰ ࣭SskAࠉSrrA ࡡ᫤㛣ⓏⓆ⌟ንິ࡛㎨ⷾវ ུᛮ࡛ࡡ
㛭㏻
ᮿᯐⰃ༡ࠉ㔘୷ாᏄࠉⴏཋኬ♰ࠉ୷஬ῗୌ㑳ࠉỀ㔕⊓ࠉຊ⸠㞖ኃࠉᑚᯐူኰ 㸝ྞኬ㝌⏍࿤㎨࣬⏍∸
ᶭᵋ㸞
7
P-39 In vitro ࣛࣤ㓗࣭ࣛࣝ⣌ࢅ⏕࠷ࡒ Aspergillus nidulans His-Asp ࣛࣤ㓗࣭ࣛࣝࢾࢴࢹ࣭࣠ࢠࡡよᯊ
᮶ಘᏳࠉ㔘୷ாᏄࠉᮿ㮭᫓์ࠉᒜ⠓㈏ྍࠉỀ㔕⊓ࠉຊ⸠㞖ኃࠉᑚᯐူኰ 㸝ྞኬ㝌࣬⏍࿤㎨㸞
P-40 Aspergillus nidulans Histidine Kinase ࡡ㒂న≁␏ⓏⓆ⌟
㕝ᮄ㯖Ꮔࠉ㔘୷ாᏄࠉຊ⸠㞖ኃࠉᑚᯐူኰ 㸝ྞኬ㝌⏍࿤㎨࣬⏍∸ᶭᵋ㸞
P-41 ࢕ࣤࢣ᰷ࣤ⭁⑋Ⳟ࡞࠽ࡄࡾ MAPK 㐿ఎᏄ◒ቪᰬࡡషฝ࡛ࡐࡡ⑋ཋᛮ
໪ᔪຩ඼ࠉ㡪㈙ᬍ஁ࠉⓊ⏣‫‮‬᭹ 㸝ᒪ㜟ኬ㝌࣬㎨㸞
P-42 Neurospora crassa ࡡ grg-1 (glucose-repressible gene)ࡢ OS-2 MAP ࢞ࢻ࣭ࢭ࡞ࡻࡽⓆ⌟ㄢ⟿ࢅུࡄࡾ
ῳ㑌⟿Ꮔ㸦ࠉᆊ㔕┷ᖲ 2 ࠉሲ⃕࠵ࡍࡈ 1ࠉⴘྙ์ᖶ㸨ࠉᮄᮟ┷㸨ࠉ⸠ᮟ┷ 1 (㸦᮶Ὂኬ㹺⏍࿤ࠉ2 ᮶Ὂኬ㹺᳔ᶭ◂ࢬࠉ㸨⌦◂㹺୯ኳ◂)
P-43 Neurospora crassa ࡡ OS-2 MAP ࢞ࢻ࣭ࢭࡡୖὮࡡ㌹෕ㄢ⟿ᅄᏄࡡྜྷᏽ
ሲ⃕࠵ࡍࡈ 1ࠉᆊ㔕┷ᖲ 2ࠉᒜୖ࿰Ᏻ 1ࠉῳ㑌⟿Ꮔ 1 ࠉ⚗᲻ᩝᗛ 1 ࠉ⸠ᮟ┷ 1
㸝1 ᮶Ὂኬ࣬⏍࿤ࠉ 2 ᮶Ὂኬ᳔࣬ᶭ◂ࢬ㸞
P-44 The nsdC gene encoding a novel positive regulator of sexual development of Aspergillus nidulans is
regulated by complicated transcriptional and post-transcriptional control.
Hye-Ryoun Kim and Dong-Min Han (Division of Life Science, Wonkwang University, Korea )
P-45 Molecular analysis of the rapamycin-sensitive and cycloheximide-resistant ActA1 gene in Aspergillus
nidulans
Sun-Hee Noh and Suhn-Kee Chae (Dept. Biochemistry and Biomed RRC, Paichai Univ. Daejeon, 302-735
Korea)
P-46 N. crassa ࡞࠽ࡄࡾࢨࢡࢻࣜ㐿ఎᏄ ncSCD2 ࠉ ncSHK1 ࡡよᯊ
ᕖᮟ┷ᚷࠉୌ▴᫓ᙢ 㸝᮶ὊኬᏕ࣬⏍࿤⛁Ꮥ◂✪⛁㸞
P-47 Neurospora crassa ࡡ NER 㐿ఎᏄ mus-43 ࠉ mus-44 ࡡよᯊ
ఫ⸠ḿொࠉୌ▴᫓ᙢ㸝᮶Ὂኬ࣬⏍࿤⛁Ꮥ㒂㸞
P-48 㯔Ⳟࡡ DNA ligase (lig4) 㐿ఎᏄ◒ቪ࡞ࡻࡾ㧏 㢎ᗐ┞ྜྷ⤄ᥦ࠻ᐙ୹ࡡ㏸ᠺ
Ềㆺ἖ 1 ࠉᕝ⸠Ὂᖲ 1ࠉ ᮿ὾▩ᕤ 1ࠉ ஬୕ᘧୌ 2ࠉ 㜷㒂ᩏᝃ 3 ࠉ ஫࿝ົஒ 1 㸝1 ᮶໪ኬ㝌㎨࣬⏍∸⏐
ᴏ๭ᠺࠉ 2 ᇳ⋚ኬ⌦ࠉ 3 ᮶໪ኬ㝌㎨࣬ᚺ⏍໩㸞
P-49 㯔Ⳟ㸝Aspergillus oryzae㸞࡞࠽ࡄࡾ ORF හ࡫ࡡ poly(A)௛ຊࢨࢡࢻࣜᑙථ࡞ࡻࡾ␏ᖏ poly(A)௛ຊ
⏛୯⍖ᕤ 1ࠉ ᚠᒱ᪸ᩝ 1,2ࠉ 㧏ᮄᚰ 3ࠉ ᩺ㆺᑠᘧ 1ࠉ ஫࿝ົஒ 1㸝1 ᮶໪ኬ࣬㎨࣬⏍∸⏐ᴏ๭ᠺ⛁Ꮥࠉ
2
㔕⏛⏐ᴏ⛁Ꮥ◂✪ᡜࠉ 3 ࢿ࣍ࢧ࢕࣑ࢩࣔࣂࣤ㸞
P-50 㯔 Ⳟ (Aspergillus oryzae)ࡡ DNA ࢹ ࣚ ࣤ ࢪ ࣎ ࢯ ࣤ Crawler ࡡ ㌹ ⛛ Ὡ ᛮ ࡛ transposase 㐿 ఎ Ꮔ ࡡ
3’-RACE よᯊ
ᑚ➗ཋ༡ಘ㸦ࠉᑚ⏷ᾀ㸧ࠉ⛑Ὂ஦㸧ࠉ㧏ᶣ▹⧂㸦ࠉ஫࿝ົஒ㸨
㸝㸦⚽⏛┬⥪㣏◂࣬㓕⣪᚜⏍∸ࠉ㸧᭮᰿෗࣬⥪◂ࠉ㸨᮶໪ኬ㝌㎨࣬⏍∸⏐ᴏ๭ᠺ㸞
P-51 ᰺ᇰ୘Ⰳ⑍≟ࢅ♟ࡌ࢙ࢿ࢞ࢰࢢ࡚᳠ฝࡈ ࡿ ࡒ dsRNA ࢗ࢕ࣜࢪ
ᶋ஬ᑋ㑳ࠉ▹ᕖᨳⱝࠉ㤷᭨⏜⨶ (᲻ᯐ⥪◂)
P-52 ዃᙟ⑍≟ࢅ♟ࡌⳞᗃ᰺ᇰࢨ࢕ࢰࢢ࡚᳠ฝ ࡈ ࡿࡒ㸧ᮇ㙈 RNA ࢗ࢕ࣜࢪ
㤷᭨⏜⨶ࠉ▹ᕖᨳⱝࠉᶋ஬ᑋ㑳 㸝᲻ᯐ⥪◂㸞
P-53 ᪝ᮇ⏐ࠉ࣊ࢹࢻ࣑⏐ࡡல⛸㛣᥃ྙమࢠࣞ ࢓ ࣠ࣄࢰࢢᰬ࡞࠽ࡄࡾ ITS 㒼าࡡዃዼ࡝ୌ⮬
Truong Binh-Nguyen1,2ࠉ ᒱᓧᏏ᫆ 3ࠉLe Xuan-Tham4ࠉ㕝ᮄᙪ 1㸝1 ༐ⴝኬ࣬㝌⮤↓⛁Ꮥࠉ ᩅ⫩ࠉ 2 Tropical
Biology Inst. Vietnamࠉ 3 ࠾ࡍࡈ DNA ◂ࠉ 4 Vietnam Atomic Energy Commission㸞
P-54 Agrobacterium ࢅ⏕࠷ࡒᢰᏄⳞ ࢙ࢿ࢞ࢰࢢ ࡡ ᙟ㈻㌹ᥦἪࡡ㛜Ⓠ
ᒱᮇᬓඖࠉᒜ⏛㞖ெࠉ⛼ෞ⪵ࠉᒱᓧක㞕ࠉୖᆊㄌ 㸝ಘᕗኬ࣬⦼⥌࣬ᚺ⏍⛁ࠉ࣌ࢠࢹࡀࡡࡆ⥪ྙ◂✪
ᡜ㸞
8
P-55 㧏ὩᛮࣛࢡࢼࣤฦよⓉⰅ⭁ᮑⳞ Phanerochaete sordida YK-624 ᰬࡡᙟ㈻㌹ᥦ⣌ᵋ⠇
ᒜᓃ㈶἖ࠉᮄᮟಆ஄ࠉ⩹⏛Ⱪ㸝᮶໪㎨◂ࢬࣤࢰ࣭㸞ࠉᮙ὾❟ᶖࠉᖲ஬ᾀᩝ㸝㟴ᒱኬᏕ㎨Ꮥ㒂㸞
P-56 ࢓ࢡࣞࣁࢠࢷ࣑ࣛࢗࢅ⏕࠷ࡒᮄᮞ⭁ᮑᢰᏄ Ⳟࡡᙟ㈻㌹ᥦ
ຊ἖ఫᖲࠉ஫༎ᔋᅽ᪝Ꮔࠉ㩢ᓞḿᾀ㸝᮶ኬ㝌࣬㎨⏍⛁㸞
P-57 ࢓ࢡࣞࣁࢠࢷ࣑ࣛࢗἪ࡞ࡻࡾࢨ࢕ࢰࢢ (Lentinula edodes) ᙟ㈻㌹ᥦ⣌ࡡ☔❟
ᐋᮇᖶᜠ 1ࠉᖲᒱⰃ㍜ 1ࠉᒜᓃኬ㍔ 1 ࠉ᫤ᮇᬊீ 2ࠉᑹㆺ ᾀ 1ࠉ๑ᕖ஦ኯ㑳 1ࠉඡ⋚ᇱୌ᭹ 1
(1 㫵ཱིኬ࣬㎨ࠉ2 Ⳟ⶷◂✪ᡜ)
P-58 Aspergillus oryzae ⏜ᮮ 㹨 C ࣏࣭࣭࢜ࢅฺ⏕ࡊ ࡒ Aspergillus oryzae ࡡᙟ㈻㌹ᥦ
こᬏᏄࠉᆎᇄ㝧஄ࠉᗀᖏḿெ 㸝ኬ㛭࣬⥪◂㸞
P-59 Aspergillus aculeatus ࡞࠽ࡄࡾ sC 㐽ᢝ࣏࣭࣭࢜ࢅ⏕࠷ࡒᙟ㈻㌹ᥦ⣌ࡡ㛜Ⓠ
㊂❟ᾀ⨶ࠉㆺಞ἖ࠉ⅛ㆺ㡨ୌࠉᕖཾ๙ྒྷ㸝㜨ᗋኬ㝌࣬⏍࿤⎌㸞
ࠉⲠ஬ᇱኰ
(୯㒂ኬ࣬ᚺ⏍)ࠉ㔘ᨳ┷ (㟴ᒱኬ㝌࣬ᚺ⏍໩)
P-60 Aspergillus aculeatus ࡞࠽ࡄࡾ trpC 㐿ఎᏄࢅ⏕࠷ࡒᙟ㈻㌹ᥦ⣌ࡡ㛜Ⓠ
๑⏛ඁ௒ࠉ㔘ᨳ┷ 1 ࠉㆺಞ἖ ࠉ⅛ㆺ㡨ୌ ࠉⲠ஬ᇱኰ 2 ࠉᕖཾ๙ྒྷ㸝㜨ᗋኬ㝌࣬ᚺ⏍⛁ࠉ
1
㟴ᒱኬ࣬ᚺ⏍໩ࠉ 2 ୯㒂ኬ࣬ᚺ⏍㸞
P-61 A. tamarii niaD 㐿ఎᏄࡡⓆ⌟よᯊ
ᮄᮟኣỜ 1ࠉᴃᮇ៿ୌ 1ࠉ໪ᮇ์⾔ 2 ࠉ㕝ᮄ⪵ 1ࠉ᯵ᮄ㇇ 1 㸝1 ㎨◂ᶭᵋ࣬㣏⥪◂ࠉ
2
យ▩⏐ᢇ◂࣬㣏ဗᕝᢇ㸞
P-62 㯑Ⳟ DNAchip ࡞ࡻࡾ㯑Ⳟ Genotyping ࡡ᳠ゞ
ᑚᯐ㟱඼ 1,2ࠉᆊᮇ࿰ಆ㸧ࠉᒜ⏛ಞ㸧ࠉᒷୖ࿰⿩㸦㸡㸧ࠉୖ㣜ொ㸦㸡㸧ࠉ୔୕㔔᪺㸧 㸝㸦ᗀ
ᓞኬ࣬඙❻◂ࠉ㸧㒿⥪◂㸞
P-63 ㈃ᰜ㣬᮪௲ୖ࡚Ⓠ⌟ࡌࡾ㯔ⳞⳞమහࢰࣤ ࣂ ࢠ㈻ࡡよᯊ
໪ᕖ἖ᜠࠉఫ㔕ඔ᫓ࠉᇸᮇකᏄࠉ⏣⏛㞖஄ 1ࠉኬ⟺ಘୌ㸝㔘Ἁᕝኬ࣬ࢣࢿ࣑◂ 1 ⏐
⥪◂㸞
P-64 ⡷ࢽ࢜ᇰᆀ࡚㧏Ⓠ⌟ࡌࡾ㯔Ⳟ㐿ఎᏄ⩄ࡡよ ᯊ
ᑹ㛭೸஦ࠉᒜ୯ᓤ*ࠉఫ㔕ඔ᫓ࠉ⠇㔕༜ኰ*ࠉኬ⟺ಘୌ 㸝㔘Ἁᕝኬ࣬ࢣࢿ࣑◂ࠉ
*⠇㔕㣏ဗᕝᴏ㸞
P-65 㯑Ⳟ DNAchip ࡡ㛜Ⓠ࡛ࢣࢿ࣑よᯊࠉⓆ⌟よ ᯊ࡫ࡡฺ⏕
ᒷୖ࿰⿩㸦㸡㸧ࠉᆊᮇ࿰ಆ㸧ࠉᒜ⏛ಞ㸧ࠉ୔୕㔔᪺㸧 㸝㸦ᗀᓞኬ࣬඙❻◂ࠉ㸧㒿⥪◂㸞
P-66 㯔Ⳟ CCAAT-box ⤎ྙᅄᏄ࡞ࡻࡖ࡙โᚒࡈࡿ ࡾ㐿ఎᏄࡡ⥑⨮Ⓩよᯊ
㧏ᶣ᪺⌌ࠉᮙᒜ⣟ஒࠉྙ⏛⚵▦ࠉఫ㔕ඔ᫓㸟ࠉᑚᯐူኰࠉຊ⸠㞖ኃ㸝ྞኬ㝌⏍࿤㎨࣬
⏍∸ᶭᵋࠉ㸟㔘Ἁᕝᴏኬ࣬ࢣࢿ࣑◂㸞
P-67 㯜㯔Ⳟ⏍⫩Ⳟమහࣈࣞࢷ࢛࣭࣑ࡡよᯊ
♼ኯ㑳ࠉ➁හ㐠㞕 㸝᮶ா㎨ᕝኬ㝌㹺㎨㸞
P-68 㯜㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ CPase 㐿ఎᏄࡡよᯊ
ಚ㔕㐡ᙢࠉ➁හ㐠㞕 㸝᮶ா㎨ᕝኬᏕ࣬㎨࣬ᚺ⏍⛁㸞
P-69 ࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ஦㔔◒ቪᰬࢅ⏕࠷ࡒ ࢬ ࣭ࣜࣞࢪ⢶໩㯔ⳞࡡฦᏄ⫩⛸
୯஬႗ᙢ 1ࠉ᭯ᒱᏕ 1ࠉᕖཾ๙ྒྷ 2ࠉⲠ஬ᇱኰ 3ࠉ໪ᮇົࡥࡆ 1 㸝1 ᮶ኬ㝌࣬㎨⏍⛁࣬
ᚺ⏍ᕝࠉ2 㜨ᗋኬ㝌࣬㎨⏍⛁ࠉ3 ୯㒂ኬ࣬ᚺ⏍㸞
P-70 ஜ⇩ᅰቫ⎌ሾ୯࡞࠽ࡄࡾ㐿ఎᏄ⤄ᥦ࠻㯔Ⳟ ⬂Ꮔࡡ⏍Ꮛᛮࡡよᯊ
ᴃᮇ៿ୌ 1ࠉཿᕖ⫩௥ 1ࠉ໪ᮇ์⾔ 2 ࠉ㕝ᮄ⪵ 1ࠉ᯵ᮄ㇇ 1㸝1 ㎨◂ᶭᵋ࣬㣏⥪◂ࠉ2 យ
▩⏐ᢇ◂࣬㣏ဗᕝᢇ㸞
9
P-71 㯔Ⳟ࡞࠽ࡄࡾ⠾౼ࢤࣞࢼ࣭PCR Ἢ࡛ࡐࡡᚺ⏕
㕝ᮄ⪵ࠉ➁ㆺ༡ᏄࠉᮄᮟኣỜࠉᮿୖ┷⏜⨶ࠉኣ⏛ຉ⏍ࠉᴃᮇ៿ୌࠉ᯵ᮄ㇇㸝㣏⥪◂㸞
P-72 Fusarium oxysporum ⏜ᮮ⢶໩࢓࣐ࢿ㓗࢛࢞ࢨࢱ࣭ࢭࡡ㔘ᒌ࢕࢛ࣤ࡞ࡻࡾὩᛮ໩
⸠ཋ┷⣎ 1ࠉ⅛ㆺ㡨ୌ 1ࠉཿ㈙᫬἖ 2 ࠉⰶ㝘ୌ⏍ 2ࠉ㧏ጏ༜ྒྷ 2ࠉ௑ᮟⱩ⾔ 2ࠉᕖཾ๙ྒྷ 1ࠉ
Ⲡ஬ᇱኰ 3 㸝1 ኬ㜨ᗋኬ㝌࣬ᚺ⏍໩ࠉ2 ᪣໩ᠺࣆ࢒࣭࣏࣬チ᩷ⷾ◂ࠉ3 ୯㒂ኬ࣬⎌ሾ⏍∸㸞
P-73 Rhizopus microsporus ࡡ⣵⬂හභ⏍ࣁࢠࢷࣛ࢓ࡡᙟ㈻㌹ᥦ࡛␏⛸ࢰࣤࣂࢠ㈻Ⓠ⌟
ᮇᒜ㧏ᖶ 1ࠉⴘྙዄ⥬Ꮔ 1ࠉ᲻⏛┷⣟ 1,2ࠉᏫఫ⨶ㄵ 2ࠉᕝ⸠ಆ❮ 1㸝1 ⌦◂࣬୯ኳ◂ࠉ
2
᮶Ὂኬ࣬ᕝ㸞
P-74 㯑Ⳟ(Aspergillus oryzae )ࡡᅖమᇰ㣬ิ᭿࡞Ⓠ⌟ ࡌࡾ㐿ఎᏄ AOS13 ࡡよᯊ
ḯᓞ᜾ 1,2ࠉᒷୖ࿰⿩ 1,2ࠉᆊᮇ࿰ಆ 2ࠉ㉝ᑹ೸ 2ࠉᒜ⏛ಞ 2ࠉ⚽⏛ಞ 3 ࠉୖ㣜ொ 1,2ࠉ
୔୕㔔᪺ 2㸝ᗀኬ࣬඙❻◂ 1ࠉ㒿⥪◂ 2ࠉᐁ㊰ዥᏄኬ 3㸞
P-75 㯜㯔Ⳟ Aspergillus oryzae ࡡ HSP30 㐿ఎᏄࣈ࣓࣭ࣞࢰ࣭ࢅ⏕࠷ࡒᾦమᇰ㣬࡞࠽ࡄࡾ
㓕⣪㐿ఎᏄࡡⓆ⌟࡛ࢰࣤࣂࢠ㈻⏍⏐ࡡ᳠ゞ
ᮿୖ┷⏜⨶ࠉ㕝ᮄ⪵ࠉኣ⏛ຉ⏍ࠉ▴⏛༡ᶖ 1ࠉᴃᮇ៿ୌࠉ᯵ᮄ㇇㸝㣏⥪◂ࠉ1 ᭮᰿෗⥪◂㸞
P-76 A.oryzae ࡡ MAL ࢠࣚࢪࢰ࣭㐿ఎᏄࡢ࢓࣐ࣚ ࣭ࢭ⣌㓕⣪㐿ఎᏄⓆ⌟࡞ࡵ㛭୙ࡌࡾ
㛏ㆺᕖ⚀Ꮔࠉ஫࿝ົஒ 㸝᮶໪ኬ㝌㎨࣬⏍∸⏐ᴏ๭ᠺ㸞
P-77 Aspergillus nidulans ◢㓗㑇ඔ㓕⣪㐿ఎᏄࡡ᎒ Ẵ᮪௲≁␏Ⓩ࡝㌹෕โᚒᶭᵋࡡよ᪺
᱕ᑹಆ௒ࠉ⸠஬㐡ஒࠉ㧏ㆺ├ᶖ 㸝➻ἴኬ㝌࣬⏍࿤⎌ሾ㸞
P-78 Trichoderma reesei ࡞࠽ࡄࡾ࢞ࢨࣚࢻ࣭ࢭ III 㐿ఎᏄࡡㄇᑙⓆ⌟ᶭᵋࡡよᯊ
ཿᕖ㝧⣎ࠉᑚ➗ཋῦࠉᒱ⏛Ᏻᩝࠉ᲻ᕖᗛ 㸝㛏ᒱᢇ⛁ኬ࣬⏍∸㸞
P-79 Trichoderma reesei ࡞࠽ࡄࡾࢬ࣭ࣜࣚࢭ㐿ఎᏄ ࡡㄇᑙᶭᵋࡡよᯊ
ᚷ⏛Ὂ௒ࠉ↻ㆺගᖶࠉᑚ➗ཋῦࠉᒱ⏛Ᏻᩝࠉ᲻ᕖᗛ㸝㛏ᒱᢇ⛁ኬ࣬⏍∸㸞
P-80 㯔Ⳟ Aspergillus oryzae ࡞ࡻࡾ cellobiose phosphorylase ࡡ⏍⏐
ኣ⏛ຉ⏍ࠉ㕝ᮄ⪵ࠉᴃᮇ៿ୌࠉኮ㔕Ⰳᙢ 1ࠉఫࠍᮄሐࠉ᯵ᮄ㇇㸝㣏⥪◂, 1 ಘᕗኬ㝌࣬ᕝ㸞
P-81 ᢰᏄⳞ Phanerochaete chrysosporium ࡡࢬ࣭ࣜࣞࢪฦよ㓕⣪㐿ఎᏄⓆ⌟㔖࡞ࢡࣜࢤ࣭ࢪ
⃨ᗐ࠽ࡻࡦᇰ㣬᫤㛣࠿୙࠻ࡾᙫ㡢
㕝ᮄୌྍࠉ஫༎ᔋᅽ᪝Ꮔࠉ㩢ᓞḿᾀ㸝᮶ኬ㝌࣬㎨⏍⛁㸞
P-82 Cluster analysis of microarray gene expression data during growth of Aspergillus oryzae
in cellulose substrate medium
Avid SHIAU, Praveen Rao JUVVADI, Jun-ichi MARUYAMA, Manabu ARIOKA, Katsuhiko
KITAMOTO (Dept. of Biotechnology, Univ. of Tokyo)
P-83 Aspergillus oryzae ㌹෕ᅄᏄ AoXlnR ᨥ㒼ୖ࡞ ࠵ࡾኣ⢶ฦよ㓕⣪㐿ఎᏄ⩄ࡡྜྷᏽ
㔕ཾ♰ྒྷࠉఫ㔕ඔ᫓1ࠉຊ⸠㞖ኃࠉᑚᯐူኰ㸝ྞኬ㝌⏍࿤㎨ࠉ1㔘Ἁᕝኬ㸞
P-84 Trichoderma reesei ⏜ᮮࢬ࣭ࣜࣚࢭࡡᶭ⬗ᨭⰃ
୯⃕කࠉᒱ⏛࿰ኃࠉⴘྙఘᗽࠉᑚ㔕ᑈ᭰Ꮔࠉᑚ➗ཋῦࠉᒱ⏛Ᏻᩝࠉ᲻ᕖᗛ
㸝㛏ᒱᢇ⛁ኬ࣬⏍∸㸞
P-85 Aspergillus aculeatus ⏜ᮮ aviIII 㐿ఎᏄࡡ A.aculeatus ࢅᐙ୹࡛ࡊࡒ㧏Ⓠ⌟
ᯀ᮶ኚᾇࠉ㔘ᨳ┷ 1ࠉㆺಞ἖ࠉ⅛ㆺ㡨ୌࠉⲠ஬ᇱኰ 2ࠉᕖཾ๙ྒྷ㸝㜨ᗋኬ㝌࣬ᚺ⏍
⛁ࠉ1 㟴ᒱኬ㎨࣬ᚺ⏍⛁ࠉ2 ୯㒂ኬ࣬ᚺ⏍㸞
P-86 Aspergillus aculeatus ⏜ᮮ β-࢞ࢨࣞࢨࢱ࣭ࢭ㐿 ఎᏄࡡ A. oryzae ࡞࠽ࡄࡾⓆ⌟
ᑚこ 㐡ஒࠉ⏛ᑹ⸓Ꮔࠉㆺ ಞ἖ࠉ⅛ㆺ㡨ୌࠉⲠ஬ᇱኰ 1 ࠉᕖཾ๙ྒྷ㸝㜨ᗋኬ㝌࣬
ᚺ⏍⛁ࠉ1 ୯㒂ኬ࣬ᚺ⏍㸞
10
P-87 ࣃࢣ࢜ࣄ࢞ࢲࣤࢸ࢓ࢬࢲ࣭ࣚࢭ㐿ఎᏄࡡ ᕝ ᏕⓏฺ⏕
⡷ᮟ᫥Ꮔࠉᮟᒜ⫍Ꮔ㸝㛭᮶Ꮥ㝌ኬ࣬ᕝ࣬∸㈻⏍࿤⛁Ꮥ㸞
P-88 ⣊≟Ⳟ Aspergillus nidulans ࡡ࣐࢛ࢨࣤᵕࢺ࣒࢕ࣤࢅᣚࡗ࢞ࢲࣤྙᠺ㓕⣪
CsmAࠉCsmB ࡡࢺ࣒࢕ࣤ㛣ࡡᶭ⬗Ⓩᕣ␏ࡡよᯊ
ᑘᓧ┷ᴃࠉ➁ୖ඼⏠ࠉᇷහ⿩஄ࠉኯ⏛᪺ᚠ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
P-89 ⣊≟Ⳟ Aspergillus nidulans ࡡࢠࣚࢪ III ࢞ࢲࣤྙᠺ㓕⣪㸝ChsB㸞ࡡ⣵⬂හᑻᅹ
㒂నࡡよᯊ
⚗⏛ୌ἖ 1ࠉᒜୖಞୌ 2ࠉᇷහ⿩஄ 1 ࠉኯ⏛᪺ᚠ 1㸝᮶ኬ࣬㎨⏍⛁࣬1 ᚺ⏍ᕝ࣬2 ⏍⎌⏍㸞
P-90 ⣊≟Ⳟ Aspergillus nidulans ࡡ࢞ࢲࢻ࣭ࢭ㐿ఎᏄ chiHࠉchiR ࡡᶭ⬗よᯊ
ᒜᓧᬍ୓ࠉ⏛୯ᙤࠉᇷහ⿩஄ࠉኯ⏛᪺ᚠ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
P-91 Rhizopus oryzae ࡡ࢞ࢲࣤྙᠺ㓕⣪㐿ఎᏄࡡⓆ ⌟よᯊ
㜷㒂Ṅࠉὰ㔕⾔ⶮࠉ᭧᰷㍜㞕㸝໪ኬ㝌㎨࣬ᚺ⏕ⳞᏕ㸞
11
Special Lecture I
Secondary metabolites: fundamental and niche-specific roles in fungal development and
cell-cell interactions
B. Gillian Turgeon and Shinichi Oide
Department of Plant Pathology
334 Plant Science Bldg.
Cornell University
Ithaca, NY, 14853
An exhaustive characterization of the set of non-ribosomal peptide synthetase (NPS) genes of the corn
pathogen, Cochliobolus heterostrophus, and the small molecule peptides produced by the enzymes they
encode, has been undertaken to ascertain what peptide secondary metabolite products are doing for the
fungal cell. Non-ribosomal peptide synthetases (NRPSs) are multimodular enzymes that make small
non-ribosomal peptides (NRPs) through a mechanism, independent of organelles best known for protein
synthesis, the ribosomes. To date, the NRPS method of peptide biosynthesis has been described for
filamentous ascomycete fungi (and to a limited extent, for basidiomycete fungi) and for bacteria, only. The
diversity of possible NRP products is potentially limitless as NRPs can be composed of D- and L-amino
acids, protein and non-protein amino acids, hydroxy acids, ornithine, b-amino acids, and other unusual
constituents. Furthermore, NRPs can be linear, cyclic, or branched cyclic, and may be modified by
glycosylation, N-methylation or acylation. In addition to structural diversity, NRPs have an astonishingly
broad spectrum of biological activities, many of which have been useful in medicine, agriculture, industry,
and biological research. However, despite the fact that activities of the peptide products with respect to
interactions with other organisms are well-documented, and certainly remarkable, to suggest that this is their
primary function is likely incorrect. For example, only one of the 12 C. heterostrophus NPS genes is required
for virulence on the host, maize. In fact, the physiological significance of these small peptides to the
producing fungi is largely unknown. We document that NRPS enzymes are purveyors of small molecules for
both basal metabolism and for specialized environmental niches. Certain NPSs are required for sexual
reproduction, asexual reproduction, virulence, as nutrient ‘gatherers’ (such as iron-gathering siderophores),
for responses to oxidative, nitrosative, nutrient, pH stress etc, for growth, and for hydrophobicity. Our
results indicate that 8 of the 12 C. heterostrophus NPS genes are not conserved, while four are conserved in
other ascomycetes, and we have examined functional conservation of conserved genes in the wheat pathogen,
Fusarium graminearum, and the Arabidopsis pathogen, Alternaria brassicicola. Comprehension of
secondary metabolite function, from the perspective of the fungal cell itself, impacts our understanding of
evolution of fungal pathogenicity mechanisms by addressing how pathogens become pathogens.
12
Special Lecture II
Fungal Nitrate and Nitrite Transporters: Insights into Structure and Function
Shiela E Unkles and James R Kinghorn,
University of St Andrews, United Kingdom
Nitrate is a major source of nitrogen for microbes and plants and it is a nutrient that most frequently
limits their growth. In the natural environment, nitrate is the most ephemeral of inorganic nutrients,
concentrations varying by as much as five orders of magnitude. Nitrate is readily leached from soils,
contributing to eutrophication of natural water systems and simultaneously lowering nitrogen availability to
land organisms. In addition, nitrate based fertilisers, widely used at around 1011 kg per annum to improve
crop yield, have the inherent disadvantage of cost, globally costing roughly US$50 billion. There are also
health concerns as nitrogen oxides derived from nitrate may be generated in the human gut.
Nitrate is converted to nitrite by the enzyme nitrate reductase and then to ammonium by nitrite
reductase activity. Ammonium is used for the production of organic nitrogen molecules including amino
acids. Whilst there is considerable information regarding nitrate and nitrite reductases, there is significantly
less on the transport of nitrate and nitrite into cells. This first step in the assimilation of nitrate to ammonium,
involves the influx of nitrate into cells, an active process that may require up to 25 kJ mol-1, depending upon
external nitrate concentration. Two nitrate transporter proteins are present in Aspergillus nidulans, NrtA
(formerly CrnA) and NrtB, and both are high-affinity systems. The A. nidulans nrtA gene was the first
eukaryotic gene encoding high-affinity nitrate transporter to be isolated and has acted as a paradigm for such
high-affinity systems, leading to the cloning and identity of orthologous genes from a number of species,
including A. oryzae (which has a single nitrate transporter). A secondary structure model was proposed for
the 57 kDa (507 amino acids) NrtA protein in which 12 hydrophobic transmembrane domains (Tm) in
alpha-helical conformation, pass through the membrane connected by hydrophilic loops. The NrtA
homologues belong to a distinct cluster (nitrate-nitrite porter family, TC 2.A.1.8) of the largest secondary
transporter family known, the major facilitator superfamily (MFS: TC 2.A.1) that comprises a vast range of
functionally diverse transporters, each containing motifs characteristic of the superfamily (Fig. 1, positions
marked by MFS). Recently, the three dimensional structures of three bacterial MFS members have been
determined, the oxalate (OxlT: TC 2.A.1.11.1), glycerol-3-phosphate (GlpT: TC 2.A.1.4.3) transporters and
lactose permease (LacY: TC 2.A.1.5.1). Although the primary sequence similarity between these and NrtA is
low, comparative modelling to these structures has allowed us to develop a three dimensional representation
of NrtA (Fig. 1).
Fig. 1. Three dimensional representation of
NrtA as viewed from the side within the
plane of the membrane.
13
Several Tms of NrtA are predicted by helical wheel analysis to form amphipathic α-helices
suggesting that the hydrophilic faces of such Tms could form the lining of an aqueous pore through which
nitrate is channelled. Among the amino acids of these helices, are a number of residues that are highly
conserved in all nitrate transporters, with at least 95% conservation in 52 proteins from archaebacteria,
eubacteria, fungi, algae and plants. Some of these amino acids are found within a motif present in all nitrate
transporters,
F/Y/K-x3-I/L/Q/R/K-x-G/A-x-V/A/S/K-x-G/A/S/N-L/I/V/F/Q-x1,2-G-x-G-N/I/M-x-G-G/V/T/A which is
not observed in any other classes of MFS proteins. This residue stretch has been referred to as the nitrate
signature of which there are two copies within NrtA, located in Tm 5 and Tm 11. The first copy contains a
highly conserved (48/52) polar asparagine residue, N168, while in the second motif, N459 is conserved in
eukaryotes. Also prominent are two charged polar arginine residues, R87 and R368, which are completely
conserved in both prokaryotes and eukaryotes, within helices Tm 2 and Tm 8, respectively. Towards gaining
an understanding of the importance of residues and their role in the structure and function of this important
anion transporter, we have altered these amino acids and characterised the resulting mutant strains using the
short-lived tracer 13NO3-. Possible roles of such residues in the function of NrtA will be discussed as well as
progress towards crystallographic structure determination.
The anion nitrite can also be used as a source of nitrogen by many bacteria, algae, fungi and plants.
Certain nitrate uptake systems such as A. nidulans NrtA and NrtB have been shown to transport nitrite as
well as nitrate. In addition, there are transporters that appear to transport nitrite, but not nitrate. These include
Escherichia coli NirC, Chlamydomonas reinhardtii NaR1 and A. nidulans NitA (and A. oryzae NitA and
NitB). Such proteins as belong to a family, quite separate from the MFS, namely the formate/nitrite
transporter family (FNT: TC 2.A.44), and many are thought to transport nitrite and/or the structurally related
formate. In contrast to the nitrate transporter group, members of the FNT family probably possess 6 Tms and
are therefore much smaller (at the primary protein sequence level) than NNP proteins, the A. nidulans NitA
protein possessing 310 residues. A secondary structure model of the A. nidulans 33.6 kDa NitA protein is
given in Fig. 2. Multiple sequence alignments of 186 proteins have indicated several residues and motifs that
are conserved within this family and therefore are likely to be involved in the function of these proteins.
These are clear targets for analysis by mutagenesis followed by biochemical characterisation and this will be
discussed. Unlike the MFS family, there are no crystal structures as yet for the FNT family and our progress
towards this goal will be reported.
Fig. 2. Secondary structure model of NitA.
Conserved formate/nitrate motifs are outlined and
conserved residues are shaded in red (>95%), green
(charge conservation) or grey (>60%)
14
S-1
ᢰᏄⳞࡡ⏍⏐ࡌࡾࢬ࣭ࣜࣞࢪฦよ⣌㓕⣪ࡡኣᵕᛮ࡛ࡐࡡᙲ๪
ಘᕗኬᏕᕝᏕ㒂∸㈻ᕝᏕ⛁
ኮ㔕Ⰳᙢࠉ㔕ᓧຉୌࠉỀ㔕ḿᾀࠉ♼⏛㮒஁
㸦㸣
ࡢࡋࡴ࡞
ࢬ࣭ࣜࣞࢪࡢ⏍∸ᅥ࡞࠽࠷࡙᭩ࡵ㇇ᐣ࡞Ꮛᅹࡊࠉ࠾ࡗ්⏍ྊ⬗࡝ࣁ࢕࢛࣏ࢪ࡚࠵ࡾࠊࡆࡿࡼࡢᮅ
ฺ⏕࡚࠵ࡽࠉࡱࡒ༎ฦ࡝㔖࠿Ꮛᅹࡌࡾࡆ࡛࠾ࡼࠉ㣏ᩩࡷ࢙ࢾ࣭ࣜ࢟ࡐࡊ࡙⎌ሾၡ㢗ࢅよỬࡌࡾࡒࡴ
ࡡ㈠″࡛ࡊ࡙᭿ᙽࡈࡿ࡙࠷ࡾࠊࡐࡆ࡚ࠉኣࡂࡡ◂✪⩽࠿ࢬ࣭ࣜࣞࢪࡡࢡࣜࢤ࣭ࢪ࡫ࡡฦよ࡞ࡗ࠷࡙
◂✪ࡊ࡙ࡀ࡙࠷ࡾࠊ࡝ࡏ࡝ࡼࠉࢡࣜࢤ࣭ࢪࡢᐖ᪾࡞ኣࡂࡡ໩ᠺဗ࡞ንᥦ࡚ࡀࡾ࠾ࡼ࡚࠵ࡾࠊࡊ࠾ࡊ
࡝࠿ࡼࠉࣁ࢕࢛࣏ࢪࡢ├᥃⇖↕ࡌࡾ௧አ࡞ࠉࡆࡿࡼࡡ┘Ⓩ࡞ࡢฺ⏕ࡈࡿ࡙࠷࡝࠷ࡡ࠿⌟≟࡚࠵ࡾࠊ
ࡆࡿ࠿ฺ⏕ࡈࡿ࡝࠷⌦⏜ࡡ࠷ࡂࡗ࠾ࡢࠉࡐࡡንᥦ࡞࠾࠾ࡾࢤࢪࢹࡷฦよࡌࡾ㓕⣪ࡡり፳ຝ⋙ࡡ఩ࡈ
࡞࠵ࡾ࡛⩻࠻ࡼࡿࡾࠊ◂✪⩽ࡡከࡢࠉ▯᫤㛣࡞ࡆࡿࡼࡡࣁ࢕࢛࣏ࢪࢅฦよ࡚ࡀࡾࢪ࣭ࣂ࣭㓕⣪ࢅష
ࡾࡆ࡛࡚࠵ࢀ࠹ࠊᠻࠍࡢ㓕⣪ࡡຝ⋙ࢅᨭၻࡌࡾᡥṹ࡛ࡊ࡙㐿ఎᏄᕝᏕࡷࣁ࢕࢛࢕ࣤࣆ࢚࣏ࢷ࢔ࢠࢪ
ࡡࡻ࠹࡝ᙁງ࡝ᢇ⾙ࢅᡥ࡞ථࡿࡒ࠿ࠉࣁ࢕࢛࣏ࢪฦよࡡࡒࡴࡡࢪ࣭ࣂ࣭㓕⣪ࡡ࢕࣒࣭ࢩࡢࡱࡓᏽ࠾
࡚ࡢ࡝࠷ࠊࡳࡊࢀࡆࡿࡼࡡከࢅ㏛࠹ࡻࡽࡵࠉᆀ㐠࡚ࡢ࠵ࡾ࠿ࢬ࣭ࣜࣚࢭࡡᵋ㏸࡛ᶭ⬗ࡡ㛭౿ࡷࠉ┞
஋ష⏕࡝࡜ࡡฦよ࣒࢜ࢼࢫ࣑࡞㛭ࡊ࡙ᇱ♇Ⓩ▩ぜࢅᚋࡾࡆ࡛࡞ࡻࡽࠉ๑㏑ࡡၡ㢗ࢅ᰷ẴࡻࡂよỬࡊ
࡙࠷ࡂᚪさ࠿࠵ࢀ࠹ࠊ
㸧㸣
ᢰᏄⳞࡡ⏍⏐ࡌࡾ㓕⣪ࡡኣᵕᛮ
ᙔ◂✪ᐄ࡚ࡢࠉཿࡂ࠾ࡼᢰᏄⳞࡡ⏍⏐ࡌࡾࢬ࣭ࣜࣞࢪฦよ⣌ࡡ㓕⣪࡞㛭ࡌࡾ◂✪ࢅ⾔ࡖ࡙ࡀࡒࠊ
≁࡞ࣃࢱࢻࢨࢰࢢ┘ࡡࢗࢪࣁࢰࢢ㸝Irpex lacteus㸞ࡢࠉこ⃕඙⏍㸝᮶ாᩅ⫩ኬᏕྞヽᩅ᤭㸞࠿᫓࿰㸦
㸩ᖳࡻࡽ◂✪ࢅ㛜ጙࡊࡒⳞ࡚࠵ࡽࠉ㛏ᖳࡡṌྍ࠿࠵ࡾࠊࡆࡡ◂✪ࡢࠉᙔ᫤ࡡ୯ᅗ࡚ࡡ㌱㤷ࡡ㣣ᩩ୘
㊂࠾ࡼࠉᮄ㈻⣌㣣ᩩࢅ㛜Ⓠ࡚ࡀ࡝࠷࠾࡛ࡡ㝛㌱࠾ࡼࡡ౪㢏࡞ࡻࡾࡵࡡ࡚࠵ࡖࡒࡻ࠹ࡓ㸝㸦㸞ࠊᮇⳞ
ࡢࠉࢬ࣭ࣜࣚࢭࡢࡵ࡛ࡻࡽࠉࡐࡡ௙ࡡኣ⢶㢦ࡷࢰࣤࣂࢠ㈻ࢅฦよࡌࡾ⬗ງ࠿㧏ࡂࠉࡐࡿࡼࡡ㓕⣪࡞
㛭ࡌࡾ⢥⿿ࡷࡐࡡᛮ㈻࡞㛭ࡌࡾᩝ⊡ࡢᩐኣࡂሒ࿈ࡈࡿ࡙࠷ࡾࠊ᭩㎾࡚ࡢࠉ㐿ఎᏄᕝᏕⓏᡥἪ࠿Ⓠ㐡
ࡊࠉᮇⳞࡡ୹ᠺฦ࡚࠵ࡾ㓕⣪⩄ࡡ㐿ఎᏄ࠿ࢠ࣭ࣞࢼࣤࢡࡈࡿࡒࠊ⌟ᅹࡱ࡚࡞᪺ࡼ࠾࡞࡝ࡖ࡙࠷ࡾࡡ
ࡢࠉGH ࣆ࢒࣐࣭ࣛ㸪ࡡ࢙ࣤࢺࢡࣜ࢜ࢻ࣭ࢭ㐿ఎᏄ㸦಴ࠉྜྷ㸫ࡡࢬࣞࣄ࢛ࣃࢺ࣭ࣞࣚࢭ㐿ఎᏄ㸦಴ࠉ
ྜྷ㸬ࡡࢬࣞࣄ࢛ࣃࢺ࣭ࣞࣚࢭ㐿ఎᏄ㸨಴ࠉ࠽ࡻࡦ㓗໩㑇ඔ㓕⣪࡚࠵ࡾࢬࣞࣄ࢛࣭ࢪ⬲Ề⣪㓕⣪㸦಴
࡚࠵ࡾ㸝㸧㸞ࠊ᭩㎾࡞࡝ࡖ࡙ࠉᢰᏄⳞ Phanerochaete crysosporium ࡡࢣࢿ࣑᝗ሒ࠿᪺ࡼ࠾࡛࡝ࡽࠉᢰ
ᏄⳞࡵኣࡂࡡࢬ࣭ࣜࣚࢭ㐿ఎᏄࢅᣚࡗࡆ࡛࠿♟ြࡈࡿࡒࠊ࠽ࡐࡼࡂࠉᮇⳞࡵᩐኣࡂࡡࢬ࣭ࣜࣚࢭ㐿
ఎᏄ࠿Ꮛᅹࡌࡾ࡛⩻࠻ࡼࡿࡾ࠿ࠉⳞమአ࡞⏍⏐ࡈࡿࡾࢰࣤࣂࢠ㈻ࡡ࠹ࡔ࡚ࡢࠉ୕オࡡࡵࡡࡡ༥ࡴࡾ
๪ྙ࠿ᅸ಻Ⓩ࡞㧏࠷ࠊ
㸨㸣
ࢬ࣭ࣜࣚࢭᠺฦ㛣ࡡ┞஋ష⏕
ᇱ㈻࡚࠵ࡾኮ↓ࡡࢬ࣭ࣜࣞࢪࡢ⛸ࠍࡡᙟឺࢅࡵࡔࠉࡐࡡኣᵕᛮࡢ⤎ᬏᵋ㏸ࡣ࠾ࡽ࡚ࡢ࡝ࡂࠉࡐࡡ
ฦᕱ࡞࠽࠷࡙ࡵヾࡴࡼࡿࡾ(㸨)ࠊࡆࡡᇱ㈻ᵋ㏸ࡡ々㞟ࡈࡢࢬ࣭ࣜࣚࢭࡡᠺฦࡡኣᵕᛮࢅ⏍ࡲࡓࡊ࡙
࠷ࡾࡡ࠾ࡵࡊࡿ࡝࠷ࠊኣࡂࡡ᚜⏍∸࠿ᑛ࡝ࡂ࡛ࡵ 2 ⛸௧୕ࡡᠺฦࢅ⏍⏐ࡊ࡙࠽ࡽࠉࡐࡿࡼࡢ┞஋Ⓩ
࡞ࢬ࣭ࣜࣞࢪฦよࢅり፳ࡊ࡙࠷ࡾࡆ࡛࠿ሒ࿈ࡈࡿ࡙࠷ࡾ(㸩)ࠊࡱࡒࠉࡐࡿࡼࡡ㓕⣪ࡢୌḗᵋ㏸ࡡẒ
㍉࠽ࡻࡦࡐࡡ␧Ềᛮࢠࣚࢪࢰ࣭よᯊ࡞ࡻࡽ 12 ௧୕ࡡࢡ࣭ࣜࣈ࡞ฦ㢦ࡈࡿ࡙࠷ࡾ(㸪)ࠊྜྷࡋࢡࣜࢤࢨ
ࣜࣀ࢕ࢺ࣭ࣞࣚࢭ㸝GH㸞ࣆ࢒࣐࣭ࣛ࡞ᒌࡌࡾࡌ࡬࡙ࡡᠺฦࡢࠉࢡࣜࢤࢨࢺ⤎ྙࡡຊỀฦよ࡞㝷ࡊ
࡙ࠉྜྷࡋ❟మ㐽ᢝᛮࢅ♟ࡌࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࠊᠻࠍࡢࠉࢬ࣭ࣜࣞࢪฦよࡡ┞஋ష⏕࡞ࡗ࠷࡙ࠉࢬ
࣭ࣜࣞࢪ⦼⥌ࡡᅂ㌹࡛࠷࠹ふⅤ࠾ࡼ⩻ᐳࡊࡒࠊࡌ࡝ࢂࡔࠉ᭩㐲࡝ฦよࡡ᫤࡞ࡢࠉࢬ࣭ࣜࣞࢪ⦼⥌ࡡ
ᧅࡽ࠿よࡿࡾࡆ࡛࠾ࡼࠉ⦼⥌ࡡᮆ❻࠿ࡂࡾࡂࡾ࡛ᅂ㌹ࡌࡾ⌟㇗ࢅぜฝࡊࡒࠊࡆࡡ⌟㇗࡞ࡗ࠷࡙ࠉ୕
オࡡ㸨ࡗࡡࣆ࢒࣐࣭ࣛ࡞ᒌࡌࡾ㓕⣪ࡡ┞஋ష⏕ࢅ᳠ゞࡊࠉࣆ࢒࣐࣭ࣛ㸬࡛㸪࡞ᒌࡌࡾ࢙ࣤࢺࢡࣜ࢜
ࢻ࣭ࢭ࡛ࢬࣞࣄ࢛ࣃࢺ࣭ࣞࣚࢭࡡ┞஋ష⏕࠿ᙁ࠷ࡆ࡛ࢅሒ࿈ࡊ࡙࠷ࡾ㸝㸫㸞ࠊࡊ࠾ࡵࠉᅂ㌹⌟㇗࡞
15
ࡢࡐࡡ㔖Ẓ࠿㔔さ࡚࠵ࡾࡆ࡛࠿ฦ࠾ࡖ࡙࠷ࡾࠊ
㸩㸣
ࢬ࣭ࣜࣞࢪฦよ࡞࠽ࡄࡾ௙ࡡ㓕⣪ࡡ㛭୙
ࢬ࣭ࣜࣞࢪฦよࢅ⾔࠹඼ᆵⓏ࡝⏍ឺ⣌࡚ࡢࠉࢬ࣭ࣜࣞࢪࢅฦよࡌࡾ⣵Ⳟࡷ⣊≟Ⳟ࠿୘⁈ᛮࢬࣜࣞ
࣭ࢪࢅ⣵⬂࠿ฺ⏕࡚ࡀࡾࢡࣜࢤ࣭ࢪࡷࢬࣞࣄ࢛࣭ࢪࡡࡻ࠹࡝ྊ⁈ᛮࡡ⢶࡞ንᥦࡌࡾࡒࡴࠉ┞஋Ⓩ࡞
഼ࡂ㓕⣪⩄ࢅฦἢࡊ࡙࠷ࡾࠊࢬ࣭ࣜࣞࢪࢅฦよࡌࡾ᚜⏍∸ࡡ୯࡚ࡵࠉⓉⰅ⭁ᮑⳞࡷ㌶⭁Ⳟ࠽ࡻࡦᑛ
࡝ࡂ࡛ࡵ ⛸ࡡ》Ⰵ⭁ᮑⳞࡢࠉᇰᆀ୯࡞௛㝮Ⓩ࡞ࢬࣞࣄ࢛࣭ࢪ⬲Ề⣪㓕⣪㸝()-㸞ࢅ⏍⏐ࡌࡾࠊ()ࡡ⏍∸Ⓩ࡝ᶭ⬗࡞ࡗ࠷࡙ࡢᮅよ᪺࡚࠵ࡾ࠿ࠉ᭩㎾Ề㓗໩ࣚࢩ࢜ࣜ⏍ᠺ⣌࡞ࡗ࠷࡙ᨥᣚࡌࡾ⤎ᯕ࠿ሒ
࿈ࡈࡿ࡙࠷ࡾࠊࡆࡡ⏍ᠺࣚࢩ࢜ࣜࡢࠉࢬ࣭ࣜࣞࢪࠉࣉ࣐ࢬ࣭ࣜࣞࢪ࠽ࡻࡦࣛࢡࢼࣤࢅಞ㣥ࡊฦよࡌ
ࡾࡆ࡛࠿࡚ࡀࡾ࡚࠵ࢀ࠹ࠊᠻࠍࡢ ()- ࠿ .OFHWHXV ࡡᇰ㣬୯࡞ࢬ࣭ࣜࣚࢭ࡛භ࡞ฦἢࡈࡿࡾࡆ࡛
ࢅ☔ヾࡊ࡙࠷ࡾࡡ࡚ࠉșࢡࣜ࢜ࢻ࣭ࢭࢅྱࡴࡒࢬ࣭ࣜࣞࢪฦよ㓕⣪ࢨࢪࢷ࣑ࡡ┞஋Ⓩ࡝ష⏕
࡞࠽ࡄࡾ ()- ࡡ⏍∸Ⓩ࡝ᶭ⬗ࢅ◂✪ࡌ࡬ࡀ࡛⩻࠻ࡒࠊᠻࠍࡡᐁ㥺࡚ࡢࠉ()- ࡛㕪࢕࢛ࣤࡡ㑇ඔࢨࢪ
ࢷ࣑࡚๑ฌ⌦ࡊࡒࢬ࣭ࣜࣞࢪࡢࠉࢬ࣭ࣜࣚࢭ࡞࡛ࡖ࡙ࡢⰃ࠷ᇱ㈻࡛࡝ࡼ࡝࠾ࡖࡒࠊᑊ↯Ⓩ࡞ࠉ()࠿භᏋࡌࡾ⣌࡚ࡢࠉࢬ࣭ࣜࣞࢪฦよ㓕⣪ࡡ⬗ງࡢቌኬࡊࡒࠊࡈࡼ࡞ࠉ()- ࡛șࢡࣜࢤࢨࢱ࣭ࢭ࠽ࡻ
ࡦࢬ࣭ࣜࣚࢭࢅྱࡳࢨࢪࢷ࣑࡚ࡢࠉࢬ࣭ࣜࣚࢭࡡࡲࡡሔྙࡻࡽࠉ␏࡝ࡾ⤎ᬏᵋ㏸ࢅࡵࡗࢬ࣭ࣜࣞࢪ
࡞ᑊࡊ࡙ࠉࡻࡽຝᯕⓏ࡞ฦよࡌࡾࡆ࡛࠿ࢂ࠾ࡖࡒࠊ⌟ᅹࠉࢬ࣭ࣜࣞࢪฦよ࡞ᯕࡒࡌ ()- ࡡᙲ๪ࢅ☔
ヾࡌࡾࡒࡴࠉ()- 㑇ඔࢨࢪࢷ࣑ࢅྱࡳࢬ࣭ࣜࣞࢪฦよ⣌࡚ฌ⌦ࡊࡒࢬ࣭ࣜࣞࢪࡡᙟឺࡷ∸⌦໩ᏕⓏ
ᛮ㈻ࡡን໩࡞ࡗ࠷࡙᳠ゞࡊ࡙࠷ࡾࠊ
㸪㸣
ࢬ࣭ࣜࣞࢪ㓕⣪ฦよࢅຐࡄࡾ௙ࡡᅄᏄࡡよ᪺ࡡ㔔さᛮ
ኣࡂࡡ◂✪⩽࠿ࠉࢺ࣒࢕ࣤࢨࣔࢴࣆࣛࣤࢡࡷ㒂న≁␏Ⓩንᥦ࡝࡜࡞ࡻࡽࠉ㓕⣪ࡡり፳ὩᛮࢅᨭⰃ
ࡌࡾࡒࡴ࡞◂✪ࡊ࡙࠷ࡾࠊࡱࡒࠉ⮤↓⏲࠾ࡼ⪇⇍ᛮࡡࢬ࣭ࣜࣚࢭࡡࡻ࠹࡝≁␏Ⓩ࡝㓕⣪ࢅ⏍⏐ࡌࡾ
᚜⏍∸ࡵࢪࢠ࣭ࣛࢼࣤࢡࡈࡿ࡙ࡀ࡙࠷ࡾࠊࡊ࠾ࡊ࡝࠿ࡼࠉࢬ࣭ࣜࣚࢭࡡࡲ࡚ࡡฦよ࡚ࡢࠉ⤎ᬏᛮࢬ
࣭ࣜࣞࢪࡢ࠵ࡱࡽ࡞ࡵᇻᅖ࡝ᵋ㏸ࢅࡊ࡙࠷ࡾࡡ࡚ࠉ๑ฌ⌦࠿ᚪさ࡚࠵ࢀ࠹ࠊୌ᪁ࠉᢰᏄⳞࡷ㧏⇍ᛮ
⣵Ⳟ࡝࡜ࡡ᚜⏍∸ࡡⳞమࡐࡡࡵࡡࢅ⏕࠷ࡒ᫤࡞ࡢࠉࢬ࣭ࣜࣞࢪࡢᏰධ࡞ྊ⁈໩࡚ࡀࡾࠊࡐࡿࡼࡡ⣵
⬂ࡢࠉࢬ࣭ࣜࣚࢭ௧አ࡞௙ࡡ㓕⣪ࡷ∸㈻ࢅ⏍⏐ࡊࠉࡆࡿࡼ࠿ࢬ࣭ࣜࣞࢪฦよ࡞㛭୙ࡊ࡙࠷ࡾ࠾ࡵࡊ
ࡿ࡝࠷ࠊ᭩⤂Ⓩ࡞ࡢࠉࢬ࣭ࣜࣚࢭࢅྱࡴࡒධమࡡ㓕⣪ฦよ⣌࡛ࡊ࡙ࢬ࣭ࣜࣞࢪฦよࢅ⩻ᐳࡌ࡬ࡀ࡛
⩻࠻ࡾࠊ
㸝㸦㸞 こ⃕ୌಆ㸯Irpex lacteus ࢬ࣭ࣜࣚࢭࡡิ᭿㓕⣪ᏕⓏ◂✪ࡡṄࡲ㸝ࡐࡡ㸦㸞ࠊ
㸝2004㸞
㸝ᰬ㸞ᐣኃ
⤊ῥ௛ᒌ㜷㒂◂✪ᡜ◂✪ሒ࿈㸡97㸡1㸢14.
㸝㸧㸞 ኮ㔕Ⰳᙢࠉ♼⏛㮒஁㸯ⓉⰅ⭁ᮑⳞ Irpex lacteus ࡡࣁ࢕࢛࣏ࢪฦよ࡞㛭ࢂࡾ㓕⣪⣌㸝2004㸞
Cellulose communication, 11(4), 175-180.
㸝㸨㸞 Hackney, J.M., Atalla, R.H., VanderHart, D.L. (1994) Int. J. Biol. Macromol. 16, 215-218
㸝㸩㸞 Nisizawa, K., Tomita, Y., Kanda, T., Suzuki, H., Wakabayashi, K. (1972) in Fermentation
technology today. Terui G, (ed), Society of Fermentation Technology, Osaka, pp. 719-725.
㸝㸪㸞 http://afmb.cnrs-mrs.fr/~pedro/CAZY/db.html.
㸝㸫㸞 Amano, Y., Kanda, T. (2002) Trends in Glycoscience and Glycotechnology, 14(75), 27-34.
Variations of cellulases and related enzymes from Basidiomycetes and their roles in cellulose
degradation
Yoshihiko Amano, Kouichi Nozaki, Masahiro Mizuno, Takahisa Kanda
Department of Chemistry and Material Engineering Faculty of Engineering, Shinshu University
16
S-2
㐽ᢝⓏⓉⰅ⭁ᮑⳞ࡞ࡻࡾᮄ㈻ࣁ࢕࢛ ࣏ࢪࡡ࢙ࢾ࣭ࣜ࢟࣬໩Ꮥ㈠″໩
ா㒌ኬᏕ ⏍Ꮛᅥ◂✪ᡜ
ῳ㎰ 㝧ྒྷࠉῳ㑌 ᓤெࠉᮇ⏛ ୙ୌ
1. ࡢࡋ ࡴ ࡞
᛬⃥࡝ெཾቌຊ࡛໩▴㈠″ࡡᾐ㈕࡞ఔ࠹ CO2 ⃨ᗐࡡቌຊࢅ⫴ᬊ࡛ࡊ࡙ࠉ්⏍⏐ྊ⬗࡝㈠″࡚࠵ࡾ
ࣁ࢕࢛࣏ࢪ࠾ࡼ᭯⏕࡝࢙ࢾ࣭ࣜ࢟࣬໩Ꮥཋᩩࢅ⏍⏐ࡌࡾࡆ࡛࠿ịࡴࡼࡿ࡙࠷ࡾࠊࡊ࠾ࡊ࡝࠿ࡼࠉᮄ
ᮞ࡝࡜ࡡࣛࢡࢿࢬ࣭ࣜࣞࢪࡡ㓕⣪⢶໩࣬Ⓠ㓕࡞ࡢࠉࣛࢡࢼࣤ࡞ࡻࡽ⿍しࡈࡿࡒ⣵⬂ባኣ⢶ࢅ㟚ฝࡈ
ࡎࡾ๑ฌ⌦࠿ᚪさ࡚࠵ࡽࠉࡆࡡࣈࣞࢬࢪ࠿ᠺฦฺ⏕ࡡኬࡀ࡝㝸ᐐ࡛࡝ࡖ࡙࠷ࡾࠊࡆ࠹ࡊࡒⅤࢅ⫴ᬊ
࡛ࡊ࡙ࠉᠻࠍࡢᮄᮞ୯ࡡࣛࢡࢼࣤࢅ㧏㐽ᢝⓏ࡞ฦよࡌࡾ㐽ᢝⓏⓉⰅ⭁ᮑⳞ㸝ࣛࢡࢼࣤฦよᛮᢰᏄⳞ㸞
ࡡ⏍∸ᶭ⬗ࢅฺ⏕ࡊࡒᮄ㈻ࣁ࢕࢛࣏ࢪንᥦἪࢅ㛜Ⓠࡌࡾ࡛࡛ࡵ࡞ࠉᶭ⬗ቌᙁ࡞ྡྷࡄࡒ◂✪ࢅ⾔ࡖ࡙
࠷ࡾࠊࡆࡆ࡚ࡢࠉ㐽ᢝⓏⓉⰅ⭁ᮑⳞࡡᮄ㈻ࣁ࢕࢛࣏ࢪንᥦ࡫ࡡᚺ⏕➴࡞ࡗ࠷࡙㏑࡬ࡾࠊ
2. ᮄ㈻ ࣁ ࢕࢛ ࣏ࢪ ࡡ㓕 ⣪ ⢶໩ ๑ฌ ⌦
ᮄᮞ⤄⧂ࡡ୯࡚ࢬ࣭ࣜࣞࢪࠉࣉ࣐ࢬ࣭ࣜࣞࢪࡢࠉࣛࢡࢼࣤ࡞ࡻࡽ⿍しࡈࡿ࡙࠷ࡾࡒࡴࠉࡆࡿࡼࡡ
⣵⬂ባኣ⢶ࢅࢬ࣭ࣜࣚࢭࠉࣉ࣐ࢬ࣭ࣜࣚࢭ࡚ຊỀฦよࡌࡾࡒࡴ࡞ࡢࠉᮄᮞ⣵⬂ባࡡᐠ࡝ࣂࢴ࢞ࣤࢡ
ࢅ◒ቪࡊ࡙⣵⬂ባኣ⢶ࢅ㟚ฝࡈࡎࡾ๑ฌ⌦࠿ᚪさ࡛࡝ࡾࠊⵠ↳ࠉỀⵠẴ⇷◃ࠉ࢓࣓ࣤࢼ࢓⇷◃(AFEX)ࠉ
CO2 ⇷◃ࠉⵠ↳ࠉ⢂◃ࠉࢮࣜ࣍ࣛࢨࢪࠉ࢛ࢯࣤ㓗໩ࠉ㓗ฌ⌦ࠉ࢓ࣜ࢜ࣛฌ⌦ࠉ࣏࢕ࢠࣞἴ↯ᑏࠉ㞹
Ꮔ⥲↯ᑏࠉȚ⥲↯ᑏࠉᮄᮞ⭁ᮑⳞฌ⌦ࠉ࡝࡜ᵕࠍ࡝๑ฌ⌦Ἢ࠿ࡆࡿࡱ࡚᳠ゞࡈࡿ࡙ࡀࡒࠊᮄᮞ㓕⣪
⢶໩๑ฌ⌦Ἢࡡ୯࡚ࠉ⇷◃ࠉⵠ↳ࠉ࣏࢕ࢠࣞἴ↯ᑏࠉࢮࣜ࣍ࣛࢨࢪ➴⇍໩ᏕⓏᡥἪࡡኣࡂࡢࠉୌ⯙
࡞ᗀⴝᶖᮞ࡞Ẓ㍉ࡊ࡙㔢ⴝᶖᮞ࡞ᑊࡌࡾ๑ฌ⌦ຝᯕ࠿఩࠷ࠊ㔢ⴝᶖࡡ୯࡚ࡵࠉᠻ࠿ᅗࡡெཾᯐࡡ⣑
6 ๪ࢅ༥ࡴࡾࢪ࢟ᮞࡢ≁࡞๑ฌ⌦ຝᯕࢅᚋࡾࡆ࡛࠿㞬ࡊ࠷ࠊౚ࠻ࡣࠉ࣏࢕ࢠࣞἴỀ⇍ཬᚺࡢࠉᗀⴝ
ᶖࣇࢻᮞ࡞ᑊࡊ࡙ࡢ 93㸚(ኣ⢶ᙔࡒࡽࡡ㑇ඔ⢶཭⋙)࡛࠷࠹㧏࠷㓕⣪⢶໩⋙ࢅ୙࠻ࡾ࠿ࠉ㔢ⴝᶖࣃࢿ
࢞࡞ᑊࡊ࡙ࡢ 61%ࠉࢪ࢟ᮞ࡞ᑊࡊ࡙ࡢࠉ᭩ኬ࡚ࡵ 36㸚ࡡ㓕⣪⢶໩⋙ࡊ࠾୙࠻࡝࠷ 1)ࠊࡆ࠹ࡊࡒၡ㢗
Ⅴࢅᡬ㛜ࡌࡾࡒࡴࠉ㔢ⴝᶖᮞࡡ⇷◃๑ฌ⌦࡚ࡢࠉ◪㓗ࡷ SO2 ࠉ᭯ᶭ㓗ࠉࣜ࢕ࢪ㓗ࠉ࢓ࣜ࢜ࣛ㐛㓗໩
Ề⣪➴ࢅり፳࡛ࡊ࡙౐⏕ࡌࡾ᪁Ἢ࠿᳠ゞࡈࡿ࡙ࡀࡒࠊࡊ࠾ࡊ࡝࠿ࡼࠉ᭯ᐐ࡝ⷾဗࢅ౐⏕ࡌࡾࡆ࡛ࡢ
㓕⣪⢶໩Ἢࡡ࣒ࣛࢴࢹࢅ᥾࡝࠹ࡆ࡛࡞࡝ࡾࠊࡱࡒࠉ⇍໩ᏕⓏฌ⌦࡞࠽࠷࡙ࡢࠉ⢶㦭᰹ࡡ⇍ฦよ࠿㉫
ࡆࡾῺᗐ࡛๑ฌ⌦ຝᯕ࠿ᚋࡼࡿࡾῺᗐᇡ࠿㎾᥃ࡊ࡙࠷ࡾࡒࡴࠉ๑ฌ⌦Ὼᗐࢅୖࡅ࡙Ⓠ㓕㜴ᐐ∸㈻ࡡ
⏍ᠺࢅ᭩ᑚ㝀࡞ᢒ࠻ࡾࡆ࡛࠿᭻ࡱࡊ࠷ࠊࡆ࠹ࡊࡒⅤࢅ⫴ᬊ࡛ࡊ࡙ࠉⓉⰅ⭁ᮑⳞ࡞ࡻࡾࣛࢡࢼࣤฦよ
ཬᚺࢅࢪ࢟࡝࡜ࡡᮄ㈻ࣁ࢕࢛࣏ࢪࡡ㓕⣪⢶໩๑ฌ⌦Ἢ࡛ࡊฺ࡙⏕ࡌࡾࡆ࡛࠿Ἰ┘ࡈࡿ࡙࠷ࡾࠊࡆࡆ
࡚ࡢࠉⓉⰅ⭁ᮑⳞࢅ⏕࠷ࡒࣁ࢕࢛࣏ࢪ⢶໩๑ฌ⌦Ἢ࡛ࡐࡡ㛭㏻◂✪ࢅ⣺௒ࡌࡾࠊ
3. ࣛ ࢡ ࢿ ࢬ ࣜ ࣞ ࣭ ࢪ ࡡ 㓕 ⣪ ⢶ ໩ ࣬
࢙ࢰࢿ࣭ࣜⓆ㓕ࡡࡒࡴࡡⓉⰅ⭁ᮑ
Ⳟ๑ ฌ⌦
ᅒ 㐽ᢝⓏⓉⰅ⭁ᮑⳞࢅ⏕࠷ࡒࢪ࢟࡝࡜ࡡᮄᮞࡡንᥦ
17
ࣛࢡࢼࣤࢅᖏῺ࡚ฦよࡌࡾⓉⰅ⭁ᮑⳞ
㸝ࣛࢡࢼࣤฦよᛮᢰᏄⳞ㸞࡞ࡻࡾ๑ฌ⌦
ࡢࠉฌ⌦᫤㛣࠿㛏࠷࡛࠷࠹ḖⅤࡢ࠵ࡾࡵ
ࡡࡡࠉ᭯ᐐ࡝ⷾဗࢅ౐⏕ࡊ࡝࠷ࡆ࡛ࠉ࢙
ࢾ࣭ࣜ࢟࢕ࣤࣈࢴࢹ࠿ᑚࡈ࠷ࡆ࡛ࠉⳞᰬ
࡞ࡻࡖ࡙ࡢ㔢ⴝᶖᮞ࡞ᑊࡊ࡙ࡵ㧏࠷ຝ
ᯕࢅ୙࠻ࡾࡆ࡛࠾ࡼἸ┘ࡈࡿࡾࠊⓉⰅ⭁
ᮑⳞࡢࠉୌ⯙࡞ࣛࢡࢼ࡛ࣤࢬ࣭ࣜࣞࢪࢅ
ྜྷ᫤࡞ฦよࡌࡾࠊࡆࡡࡻ࠹࡝㟸㐽ᢝⓏⓉ
Ⰵ⭁ᮑⳞ࡞ࡻࡾⳞฌ⌦ࡢࠉࣁ࢕࢛࣏ࢪን
ᥦࡡ๑ฌ⌦࡛ࡊ࡙ࡢฺ⏕౮ೋ࠿఩࠷ࠊࡆ
ࡿ࡞ᑊࡊࠉCeriporiopsis subvermispora ➴
ࡡ㐽ᢝⓏⓉⰅ⭁ᮑⳞ࡛࿣ࡣࡿࡾⳞࡢࠉࢬ
࣭ࣜࣞࢪࡡ᥾ഭࢅ᭩ᑚ㝀࡞ᢒ࠻ࡗࡗࠉࣛ
ࢡࢼࣤࢅ㧏㐽ᢝⓏ࡞ฦよࡌࡾ⬗ງࢅ᭯
ࡊ࡙࠽ࡽࠉⳞฌ⌦ࢅ⤄ࡲ㎲ࢆࡓࣂࣜࣈ໩
㸝ࣁ࢕࢛ࣂࣜࣅࣤࢡ㸞࡞㛭㏻ࡊ࡙ࠉ◂✪
࠿㐅ࡴࡼࡿ࡙ࡀࡒ 2,3)ࠊ㐽ᢝⓏⓉⰅ⭁ᮑⳞ
࡚ࡢࠉᮄᮞࡡ⣵⬂ྜྷኃࢅ᥃╌ࡊ࡙࠷ࡾ⣵
⬂㛣ᒒࡡࣛࢡࢼࣤ࠿⭁ᮑิ᭿࠾ࡼฦよ
ࡈࡿࡾࡒࡴࠉࡆࡡⳞ࡚ 2-4 㐄㛣ฌ⌦ࡊࡒ
ᚃ࡞ᶭ᲌Ⓩ࡝ࣂࣜࣈ໩ࢅ⾔࠹࡛㔔㔖΅
ᑛࢅࡐࡿ࡮࡜ఔ࠹ࡆ࡛࡝ࡂ᭩ኬ 47㸚ࡵ
ࡡ࢙ࢾ࣭ࣜ࢟๎΅ຝᯕ࠿ᚋࡼࡿࡾࠊ㐽ᢝ
ᅒ 2 ⓉⰅ⭁ᮑⳞฌ⌦ࢪ࢟ࢲࢴࣈࡡ࣭࣒ࣜ
ⓏⓉⰅ⭁ᮑⳞฌ⌦ࢅࠉࢠࣚࣆࢹἪࡷࢦࣜ
ࣤ ᾦ ࡞ ࡻ ࡾ in vitro ᭯ ᶭ ∸ ᾐ ໩ ⋙
(IVOMD)࡛࢝ࢪⓆ⏍㔖(IVGP)13)
ࣆ࢒࢕ࢹἪ࡝࡜ࢢ࣐࢜ࣜࣂࣜࣈ໩࡛⤄
ࡲྙࢂࡎࡾ࡛ࠉࣂࣜࣈ໩࡞ᚪさ࡝ⷾဗᢖ
ථ㔖ࢅ΅ᑛࡈࡎࡾࡆ࡛࠿࡚ࡀࡾࠊ
ⓉⰅ⭁ᮑⳞฌ⌦ࡢࠉࣁ࢕࢛ࣂࣜࣅࣤࢡࡡ௙ࠉᮄᮞ࠽ࡻࡦ㟸ᮄᮞࣛࢡࢿࢬ࣭ࣜࣞࢪࡡ㓕⣪⢶໩࣬Ⓠ
㓕๑ฌ⌦࡞ฺ⏕࡚ࡀࡾ㸝ᅒ 1㸞ࠊ➱⩽ࡼࡢࠉC. subvermispora ࡝࡜ࡡ㐽ᢝⓏⓉⰅ⭁ᮑⳞ࡞ࡻࡾᮄᮞ⭁
ᮑ࡛ࢮࣜ࣍ࣛࢨࢪࢅ⤄ࡲྙࢂࡎࡒᮄᮞࡡ⢶໩࢙࣬ࢰࢿ࣭ࣜⓆ㓕ࡡࡒࡴࡡ๑ฌ⌦Ἢࢅ᳠ゞࡊ࡙࠷ࡾ 4-8)ࠊ
ⓉⰅ⭁ᮑⳞ࡚ࣇࢻᮞࢲࢴࣈࢅ 2 㐄㛣࠾ࡼ 8 㐄㛣⭁ᮑࡈࡎࠉ⭁ᮑᮞࢅ 180Υ࡚࢙ࢰࢿࣛࢨࢪࡊࠉᚋࡼ
ࡿࡒ୘⁈ᛮࣂࣜࣈ⏤ฦࢅࢬ࣭ࣜࣚࢭ࡛㓕ẍ Saccharomyces cerevisiae ࡚ె⾔々Ⓠ㓕ࡊࡒࠊ౐⏕ࡊࡒⓉ
Ⰵ⭁ᮑⳞࡡ୯࡚ࡢࠉࣛࢡࢼ࡛ࣤࢬ࣭ࣜࣞࢪࢅྜྷ᫤࡞ฦよࡌࡾⓉⰅ⭁ᮑⳞ Coriolus versicolor ࡷ
Pleurotus ostreatus ࡞Ẓ࡬࡙ࠉࣛࢡࢼࣤฦよࡡ㐽ᢝᛮ࠿㧏࠷ C. subvermispora ࠿㧏࠷Ⓠ㓕཭⋙ࡡቌኬ
ຝᯕࢅ୙࠻ࡒࠊ㐽ᢝⓏⓉⰅ⭁ᮑⳞ C. subvermispora ࡚ 8 㐄㛣ฌ⌦ࡊࡒࡵࡡࡢࠉⳞᮅฌ⌦ࡡࡵࡡ࡞Ẓ
㍉ࡊࠉ1.6 ಶ࢙ࢰࢿ࣭ࣜ཭⋙ࢅቌኬࡈࡎࡒ 4)ࠊࡆ࠹ࡊࡒⳞฌ⌦ຝᯕࡢࠉ㔢ⴝᶖ࡞࠽࠷࡙㢟ⴥ࡞⌟ࡿࡾࠊ
㔢ⴝᶖࢪ࢟ᮞࢲࢴࣈࢅ 200Υࠉ1 ᫤㛣ࡡ᮪௲࡚࢙ࢰࢿࣛࢨࢪฌ⌦ࡊࠉᚋࡼࡿࡒࢲࢴࣈࢅࢬ࣭ࣜࣚࢭ࡚
㓕⣪⢶໩ࡌࡾ࡛ࠉᗀⴝᶖࣇࢻᮞࡡሔྙ࡛ࡢᑊ㇗Ⓩ࡞ࠉ7㸚ࡡ㓕⣪⢶໩⋙ࡊ࠾୙࠻࡝࠷ࠊࡊ࠾ࡊ࡝࠿ࡼࠉ
ࢪ࢟ᮞࢅ C. subvermispora ࡚࠵ࡼ࠾ࡋࡴ 8 㐄㛣⭁ᮑࡈࡎࡒᚃ࡞࢙ࢰࢿࣛࢨࢪฌ⌦ࢅࡌࡾ࡛ࠉ㓕⣪⢶
໩⋙ࡢⳞᮅฌ⌦ࡡ 7 ಶ௧୕࡞୕᪴ࡊࡒࠊ㐽ᢝⓏⓉⰅ⭁ᮑⳞฌ⌦ࢅ⤄ࡲ㎲ࡳࡆ࡛࡞ࡻࡽࠉ◪㓗࡝࡜ࡡ
໩Ꮥり፳ࢅ౐⏕ࡌࡾࡆ࡛࡝ࡂ㔢ⴝᶖࡡ࢙ࢰࢿࣛࢨࢪࣁ࢕࢛ࣛࣆ࢒࢕ࢻ࣭ࣛ࠿ྊ⬗࡛࡝ࡾࠊ
ᅗහ࡚᲻ᯐࣁ࢕࢛࣏ࢪࢅⓉⰅ⭁ᮑⳞ࡚ฌ⌦ࡌࡾࡒࡴ࡞ࡢࠉᅗ⏐ࡡⳞࢅฺ⏕ࡌࡾࡆ࡛࠿᭻ࡱࡊ࠷ࠊ
ࡆࡡࡒࡴࠉࢪ࢟ᮞࡡ⢶໩๑ฌ⌦࡞ຝᯕࡡ࠵ࡾⓉⰅ⭁ᮑⳞࢅฦ㞫࣬㐽ᢜࡊࠉࢪ࢟ᮞ࠾ࡼࡡ࢙ࢰࢿ࣭ࣜ
⏍⏐࡞㛭ࡌࡾ◂✪ࢅ㐅ࡴ࡙࠷ࡾ 9)ࠊฦ㞫ࡊࡒ᩺ぞᰬࡢࠉࢪ࢟ᮞ࡞ᑊࡌࡾ㔔㔖΅ᑛ⋙࠿ C. subvermispora
ࡻࡽᑛ࡝࠷࡞ࡵ࠾࠾ࢂࡼࡍࠉ࢙ࢰࢿࣛࢨࢪࡷ㧏ἓⅤ࢓ࣜࢤ࣭ࣜࢅ⏕࠷ࡾࢮࣜ࣍ࣛࢨࢪ࡚ࡢࠉC.
subvermispora ࢅ୕ᅂࡾ๑ฌ⌦ຝᯕࢅ♟ࡊࡒࠊᮇⳞࡢࠉ࣏࢛ࣤ࢝ࣤ࣋ࣜ࢞ࢨࢱ࣭ࢭࢅ⏍⏐ࡊࠉᮄᮞ୯
ࡡࣛࣅࢴࢺࢅ΅ᑛࡈࡎࡾࠊ⌟ᅹࠉ᩺ぞᰬ࡛࣏࢕ࢠࣞἴࢮࣜ࣍ࣛࢨࢪࢅ⤄ࡲྙࢂࡎࡒ๑ฌ⌦Ἢࡡ㛜Ⓠ
18
◂✪ࢅᐁ᪃୯࡚࠵ࡾࠊ࣏࢕ࢠࣞἴ↯ᑏ⿞⨠ࡢࠉ2.45GHz ࡛ 5.8GHz ࡡ↯ᑏ㒂ࢅങ࠻ࠉ3 ḗඔ㞹☚⏲ࢨ
࣐࣭ࣖࣝࢨ࡚ࣘࣤ࢞ࣔࣄࢷ࢔࣭ࢅスゝࡊࡒ⿞⨠ࢅ㛜Ⓠࡊࡒࠊᮇ⿞⨠ࡢ↯ᑏ㐛⛤ࢅࢤࣤࣅ࣭ࣖࢰࣈࣞ
ࢡ࣑࡚ࣚโᚒࡌࡾࠊ
ⓉⰅ⭁ᮑⳞฌ⌦ࡢࠉᮄᮞࡡ
௙ࠉ㟸ᮄᮞ⣌ࣛࢡࢿࢬ࣭ࣜࣞࢪ
࡚࠵ࡾ࢛࢕ࣜࣂ࣭࣑ࡡ✭ᯕᡛ
(EFB)࡞ᑊࡊ࡙ࡵ⬲ࣛࢡࢼࣤ࡞
ࡻࡾ⢶໩࣬Ⓠ㓕ಀ㐅ຝᯕࢅ♟ࡌ
10)
ࠊⓉⰅ⭁ᮑⳞฌ⌦ࡢࠉࣁ࢝ࢪ
ࡡ⢶໩࣬Ⓠ㓕๑ฌ⌦࡞ࡵຝᯕࢅ
♟ࡌ 11)ࠊ
ᮄ㈻ࣁ࢕࢛࣏ࢪࡡ᚜⏍∸Ⓠ
㓕࡞࠽࠷࡙ࡢࠉⓆ㓕㜴ᐐࡡ఩΅
ࡵኬࡀ࡝ㄚ㢗࡚࠵ࡾࠊⓆ㓕㜴ᐐ
∸㈻ࡡ㝎ཡ࡞ࡢ࢛࣭ࣁ࣭ࣚ࢕
࣐ࣤࢡࠉὩᛮ⅛ࡷ࢕࢛ࣤஹᥦᶖ
⬙࡞ࡻࡾྺ╌ࠉ΅ᅸ㝎ཡࠉ㓗໩
㓕⣪࡞ࡻࡾฌ⌦࡝࡜ᵕࠍ࡝᪁
Ἢ ࠿ ▩ ࡼ ࡿ ࡙ ࠷ ࡾ ࠊ C.
subvermispora ࡢᮄᮞ࡞ࡵ࡛ࡵ
࡛ྱࡱࡿ࡙࠷ࡒ⬙⫣㓗࡝࡜ࡡ
ᅒ 3 ⓉⰅ⭁ᮑⳞฌ⌦ࢪ࢟ࢲࢴࣈࡡ࣒
Ⓠ㓕㜴ᐐ∸㈻ࢅฦよ㝎ཡࡌࡾࠊ
ࢰࣤⓆ㓕 14)
ࡱࡒࠉⓉⰅ⭁ᮑⳞฌ⌦ࡢࠉⳞฌ
⌦࡛⤄ࡲྙࢂࡎࡾ⇷◃ࡷࢮࣜ
࣍ࣛࢨࢪ࡝࡜ࡡ๑ฌ⌦ࡡῺᗐࢅ
ୖࡅࡾࡆ࡛࡞ࡻࡖ࡙ࠉ๑ฌ
⌦࡞ࡻࡖ࡙⏍ᠺࡌࡾⓆ㓕
㜴ᐐ∸㈻ࡡ㔖ࢅ఩΅ࡈࡎ
ࡾࠊⓆ㓕㜴ᐐࡡ఩΅࡞ࡢࠉ
ⓉⰅ⭁ᮑⳞ࠿⏍⏐ࡌࡾࣚ
ࢴ࣭࢜ࢭࡡฺ⏕ࡵ᳠ゞࡈ
ࡿ࡙࠷ࡾࠊ42Υ࡚⏍⫩ྊ⬗
࡝ ⪇ ⇍ᛮⓉ Ⰵ⭁ᮑⳞ
Trametes versicolor RC3 ᰬ
⏜ᮮࡡࣚࢴ࣭࢜ࢭࡢࠉⓆ㓕
㜴ᐐࡡ఩΅࡞㧏࠷ຝᯕࢅ
♟ࡌࠊ࣋ࣤࢹ࣭ࢪⓆ㓕ᛮ㓕
ẍ Pichia stipitis ࡞ࡻࡾࣇࢻ
ᮞ࢙ࢰࢿࣛࢨࢪྊ⁈ᛮ⏤
ฦࡡ࢙ࢰࢿ࣭ࣜⓆ㓕࡞࠽
࠷࡙ࠉRC3 ⏜ᮮࡡࣚࢴ࣭࢜
ᅒ 4 㐽ᢝⓏ࠽ࡻࡦ㟸㐽ᢝⓏⓉⰅ⭁ᮑⳞࡡᮄᮞ⭁ᮑᵕᘟ 7,8,19)
ࢭฌ⌦ࡢࠉ࢙ࢰࢿ࣭ࣜ཭⋙
㓕⣪ࡢฦᏄ㔖࠿ኬࡀ࠷ࡒࡴࠉᮄᮞ⣵⬂ባහ࡞౴ථ࡚ࡀ࡝࠷ࠊ㟸㐽ᢝⓏⓉⰅ⭁ᮑ
ࢅ 15㸚࠾ࡼ 78㸚࡫ྡྷ୕ࡈ
Ⳟࡢࠉࣆ࢘ࣤࢹࣤཬᚺ࡞ࡻࡽࣃࢺࣞ࢞ࢨࣜࣚࢩ࢜ࣜ(•OH)ࢅ⏍ᠺࡈࡎ࡙ᮄᮞ⣵⬂
ባࢅ౴㣏ࡊࠉኬࡀࡂ㛜࠷ࡒᏇ࡞㓕⣪࠿౴ථࡊ࡙ࣛࢡࢼ࡛ࣤࢬ࣭ࣜࣞࢪࢅྜྷ᫤ฦ
ࡎࡒࠊRC3 ⏜ᮮࡡࣚࢴ࣭࢜
よࡌࡾࠊ㐽ᢝⓏⓉⰅ⭁ᮑⳞࡢࠉᮄᮞ⣵⬂ባ࡞㓕⣪ࡷⳞ⣊ࢅ㐅ථࡈࡎࡾࡆ࡛࡝ࡂࠉ
ࢭࡡ୯ࡡ㓗ᛮ࢓࢕ࢮࢧ࢕
㓕⣪࠾ࡼ㞫ࡿࡒሔᡜࡡࣚࢩ࢜ࣜⓆ⏍⣌࡚ࣛࢡࢼࣤࢅ㧏㐽ᢝⓏ࡞ฦよࡌࡾࠊFe3+
࣑࡞ࡗ࠷࡙ࡢࠉ㐿ఎᏄࢠࣞ
ࡡ㑇ඔࡡ௥ㅨ∸࡞ࡻࡾโᚒ࠿㐽ᢝᛮࢅỬࡴࡾኬࡀ࡝㘵࡛࡝ࡾࠊ
࣭ࢼࣤࢡ࡛ࣃࣚࢰࢢᙟ㈻
㌹ᥦ⣌ࢅฺ⏕ࡊࡒ␏⛸Ⓠ
19
⌟࡞ᠺຉࡊ࡙࠷ࡾ 12)ࠊᮇࣚࢴ࣭࢜ࢭࡢࠉT㸣versicolor ࡡ LAC4 ࡛ 81%ࡡ┞ྜྷᛮࢅࡵࡗ᩺ぞ࡝㓕⣪࡚
࠵ࡾࠊ
5.Ⓣ Ⰵ⭁ ᮑⳞ ฌ⌦ ᮄ ᮞࡡ ᐓ⏾ 㣣 ᩩ໩ ࡛࣒ ࢰࣤ Ⓠ㓕 ๑ฌ ⌦
㐽ᢝⓏⓉⰅ⭁ᮑⳞฌ⌦ࡢࠉᮄᮞࡡᐓ⏾㣣ᩩ໩ࡷ࣒ࢰࣤⓆ㓕๑ฌ⌦࡞㧏࠷ຝᯕࢅ♟ࡌࠊࡆࡿࡱ࡚ࠉ
ᮄᮞࢅ⢊㣣ᩩ໩ࡌࡾࡒࡴ࡞⇷◃ฌ⌦ࠉ⢂◃ฌ⌦࠿᳠ゞࡈࡿ࡙ࡀࡒ࠿ࠉ⇷◃ฌ⌦࠽ࡻࡦ⢂◃ฌ⌦ᮄᮞ
ࡡཬⰼᐓ⏾࡞ࡻࡾᾐ໩⋙ࡢ఩࠷ࠊ≁࡞ࠉ⇷◃࡚ࡢ㔢ⴝᶖᮞ࡞ᑊࡌࡾฌ⌦ຝᯕ࠿఩ࡂࠉࡱࡒᐓ⏾ࡡႰ
ይᛮࢅ᥾࡝࠹࢓ࣜࢸࣃࢺ㢦࠿⏍ᠺࡌࡾࠊࡐࡆ࡚ࠉ㔢ⴝᶖࢪ࢟ᮞ࠾ࡼᰜ㣬౮ࡡ㧏࠷ᐓ⏾⢊㣣ᩩࢅ⏍⏐
ࡌࡾࡒࡴ࡞ⓉⰅ⭁ᮑⳞฌ⌦ࢪ࢟ᮞࡡཬⰼᐓ⏾࣭࣒ࣜࣤᾦ࡞ࡻࡾᾐ໩ᛮモ㥺ࢅ⾔ࡖࡒࠊ㣣ᩩ໩ࡢࣛࢡ
ࢼࣤ࡞ࡻࡾ⣵⬂ባኣ⢶ࡡ⿍しࢅ◒ቪࡊ࡙ࠉཬⰼᐓ⏾࠿ࣁ࢕࢛࣏ࢪ୯ࡡࢬ࣭ࣜࣞࢪࡷࣉ࣐ࢬ࣭ࣜࣞࢪ
ࢅฦよฺ࣬⏕ࡊࡷࡌࡂࡌࡾฌ⌦࡚࠵ࡽࠉ⢶໩๑ฌ⌦࡛ࡲ࡝ࡌࡆ࡛ࡵ࡚ࡀࡾࠊཬⰼᐓ⏾ࡢ㸩ࡗࡡ⫮ࢅ
ࡵࡔࠉࡐࡡ࠹ࡔࡡ➠ୌ⫮㸝࣭࣒ࣜࣤ㸞୯࡞ࡢ᳔∸⣵⬂ባኣ⢶ࡡฦよງࢅࡵࡗ᎒Ẵᛮ᚜⏍∸⩄࠿Ჿ᜝
ࡌࡾࠊ㣣ᩩ໩ࡡフ౮Ἢࡡ୯࡚ࠉཬⰼᐓ⏾ࡡ➠ୌ⫮࠾ࡼ࣭࣒ࣜࣤᾦࢅమአ࡞ཱིࡽฝࡊࠉࡐࡿࢅ⏕࠷࡙
in vitro ࡚᭯ᶭ∸ࡡྊ⁈໩⋙ࡷ࢝ࢪⓆ⏍㔖ࢅῼ
ᏽࡌࡾ᪁Ἢࡢࠉᑛ㔖ࡡࢦࣤࣈ࡚ࣜ⠾౼࡞モ㥺
࡚ࡀࡾࡆ࡛࠾ࡼࠉ⢊㣣ᩩࡡᾐ໩ᛮフ౮Ἢ࡛ࡊ
࡙ᗀࡂฺ⏕ࡈࡿ࡙࠷ࡾࠊࡆࡡ᪁Ἢ࡚ࡢࠉྊ⁈
໩ࡊࡒ⢶࠿ᰜ㣬″࡛ࡊฺ࡙⏕ࡈࡿࡾ࡛ࡢ㝀ࡼ
࡝࠷ࡒࡴࠉ᭯ᶭ∸ࡡྊ⁈໩⋙㸝࠵ࡾ࠷ࡢ୘⁈
ᛮṟ⁞ࡡṟᏋ⋙㸞࡛ྜྷ᫤࡞ࠉ࢝ࢪⓆ⏍㔖ࢅῼ
ᏽࡌࡾࡆ࡛࠿᭻ࡱࡊ࠷ࠊࢪ࢟ᮞࢅࠉⓉⰅ⭁ᮑ
Ⳟ C. subvermispora ࠉ ࢨ ࢕ ࢰ ࢢ (Lentinula
edodes)ࠉࣃࣚࢰࢢ(Pleurotus ostreatus)ࠉࢻ࣒ࢤ
(Pholiota nameko)࡚Ⳟฌ⌦ࡊࠉ⭁ᮑᮞࡡཬⰼᐓ
⏾࣭࣒ࣜࣤᾦ࡞ࡻࡾ in vitro ᾐ໩ᛮモ㥺ࢅ⾔ࡖ
ࡒ 13)ࠊࡐࡡ⤎ᯕࠉⓉⰅ⭁ᮑⳞࢅ᥃⛸ࡊ࡙࠷࡝
࠷ࢪ࢟ᮞࡡ᭯ᶭ∸ᾐ໩⋙ࡢࠉ
4.7-6.8%ࡡ㛣࡚࠵
ࡖࡒࡡ࡞ᑊࡊࠉC. subvermispora ࢅᇰ㣬ࡊࡒࢪ
ᅒ 5 㐽ᢝⓏⓉⰅ⭁ᮑⳞⳞమአ௥ㅨ∸ ceriporic
࢟ᮞࡡᾐ໩⋙ࡢࠉ44.6%ࡱ࡚୕᪴ࡊࡒ㸝ᅒ 2㸞ࠊ
acid ࡞ࡻࡾࣆ࢘ࣤࢹࣤཬᚺ⣌࡞࠽ࡄࡾࢬࣜ
࣭ࣞࢪฦよࡡᢒโ 19)
ࡆࡿࡢࠉ᪝ᮇᵾ‵㣣ᩩᠺฦ⾪࡞オ㍍ࡈࡿ࡙࠷
ࡾⵠ↳⇷◃ฌ⌦ࢪ࢟ᮞࡡ TDN㸝ᾐ໩࡚ࡀ
ࡾᰜ㣬ฦ⥪㔖㸞11.0%ࡡ 4 ಶ࡞ᙔࡒࡾࠊࡆ
ࡡᐁ㥺⤎ᯕࡢࠉࢪ࢟ᮞ࠿ C. subvermispora
ฌ⌦࡞ࡻࡖ࡙ࠉ࣭࣒ࣜࣤ᚜⏍∸ࡡ᳔∸⣵⬂
ባฦよ㓕⣪⩄ࡡష⏕ࢅུࡄࡷࡌ࠷≟ឺ࡞
ን໩ࡊࡒࡆ࡛ࢅ♟ࡊ࡙࠽ࡽࠉ㐽ᢝⓏⓉⰅ⭁
ᮑⳞฌ⌦ࢅࢪ࢟ᮞࡡᵕࠍ࡝Ⓠ㓕⏍⏐࡞ฺ
⏕࡚ࡀࡾࡆ࡛ࢅ♟ࡌࠊ
ᐁ㝷ࠉC. subvermispora ࡞ࡻࡾࢪ࢟ᮞฌ⌦
ࡢࠉୖỀỗἶࢅ⏕࠷ࡒࢪ࢟ᮞࡡ࣒ࢰࣤⓆ㓕
๑ฌ⌦࡞ࡵຝᯕࢅ♟ࡊࡒ 14)ࠊᑚ㯇ࣆࢪ࣏ࢅ
ྱࡳࢪ࢟ᮞࢲࢴࣈ࡞ C. subvermispora ࢅ᳔
Ⳟࡊࠉ8 㐄㛣ᇰ㣬ࡌࡾ࡛ࠉ⭁ᮑࢪ࢟ᮞࡡኣ
⢶㸝࣌ࣞࢬ࣭ࣜࣞࢪ㸞࠵ࡒࡽ 35㸚ࠉཋᩩ
ࢪ࢟ᮞ࠵ࡒࡽ 25%ࡡ㌹ᥦຝ⋙࡚ࣁ࢕࢛࢝
ࢪ㸝࣒ࢰ⃨ࣤᗐ 55-60㸚㸞࠿⏍ᠺࡊࡒ(ᅒ
ᅒ 6 㐽ᢝⓏⓉⰅ⭁ᮑⳞࡡⳞమአ௥ㅨ∸ ceriporic
3)ࠊ⭁ᮑฌ⌦࡞࠽࠷࡙ࡢࠉᑚ㯇ࣆࢪ࣏ࡡ῟
acid ࡞ࡻࡾࢬ࣭ࣜࣞࢪฦよࡡᢒโᶭᵋ 20)
ຊ࠿๑ฌ⌦ຝᯕࢅ㧏ࡴࡒࠊࡆࡡ⭁ᮑຝᯕ࡛
20
ș-O-4 ࢓࢙࣭ࣛࣜࢷࣜ⤎ྙࡡว᩷⋙࡞ࡢ┞㛭࠿ヾࡴࡼࡿࡒࠊⓉⰅ⭁ᮑⳞ C. subvermispora ࡷࢨ࢕ࢰ
ࢢࡢࠉࣁ࢝ࢪࡡ㣣ᩩ໩࡞ࡵຝᯕࢅ♟ࡌ 15)ࠊ
6㸣㐽 ᢝⓏ ⓉⰅ ⭁ ᮑⳞ ࡞ࡻ ࡾ ࣛࢡ ࢼࣤ ฦよ ᶭ ᵋ
ⓉⰅ⭁ᮑⳞࡢࠉⳞమአ࡞ࣛࢡࢼࣤࡷࢬ࣭ࣜࣞࢪࡡฦよ㓕⣪ࢅฦἢࡌࡾࠊ࡛ࡆࢀ࠿ࠉࡆࡿࡼࡡ㓕⣪
ࡡฦᏄࢦ࢕ࢫࡢᮄᮞ⣵⬂ባ࡞ඔࠍᏋᅹࡊ࡙࠷ࡾ⣵Ꮗࡡ├ᙼࡻࡽኬࡀ࠷ࡒࡴ࡞ࠉฦἢࡈࡿࡒⳞమአ㓕
⣪ࡢࡐࡡࡱࡱ࡚ࡢᮄᮞ⣵⬂ባ୯࡞㐅ථ࡚ࡀ࡝࠷ࠊࡆࡡࡒࡴࠉኣࡂࡡⓉⰅ⭁ᮑⳞࡢࠉὩᛮ㓗⣪࡚࠵ࡾ
ࣃࢺࣞ࢞ࢨࣜࣚࢩ࢜ࣜ(·OH)ࢅ㐼⛛㔘ᒌࡡࣝࢺࢴࢠࢪཬᚺࢅ௒ࡊ࡙Ⓠ⏍ࡈࡎࡾࡆ࡛࡞ࡻࡽᮄᮞ⣵⬂
ባ୯ࡡኣ⢶ࢅ࡯ࢀ࡯ࢀ࡞ࡊࠉ⤎ᯕ࡛ࡊ࡙㛜࠷ࡒᮄᮞ⣵⬂ባࡡኬࡀ࡝Ꮗ࡞⮤ฦࡡฝࡌⳞమአ㓕⣪ࢅ㐅
ථࡈࡎࡾ㸝ᅒ 4㸞ࠊࡆࡿ࡞ᑊࡊࠉC. subvermispora ➴ࡡ㐽ᢝⓏⓉⰅ⭁ᮑⳞࡢࠉᮄᮞ⭁ᮑ࠿࠾࡝ࡽ㐅⾔
ࡊࡒṹ㝭࡞࡝ࡖ࡙ࡵࠉ⮤ฦࡡฦἢࡊࡒⳞమአ㓕⣪ࢅᮄᮞ⣵⬂ባහ࡞㐅ථࡈࡎࡾࡆ࡛࡝ࡂࠉ㓕⣪࠾ࡼ
㐪ࡂ㞫ࡿࡒ⣵⬂㛣ᒒࡷ⣵⬂ባ῕ᒒࡡࣛࢡࢼࣤࢅ఩ฦᏄ௥ㅨ∸ࢅฺ⏕ࡊ࡙㧏㐽ᢝⓏ࡞ฦよࡌࡾ 3)ࠊ༳
ࡔࠉ㐽ᢝⓏⓉⰅ⭁ᮑⳞࡢࣛࢡࢼࣤࡡࣚࢩ࢜ࣜฦよࢅḾࡴࡾࡆ࡛࡝ࡂࠉ㓗⣪࠽ࡻࡦ㕪࢕࢛ࣤᏋᅹୖ࡚
·
OH ࡡ⏍ᠺࢅᢒโࡌࡾᶭᵋࢅࡵࡗࠊᠻࠍࡢࠉ㐽ᢝⓏⓉⰅ⭁ᮑⳞࡡᇰ㣬∸࠾ࡼ㕪ࡡࣝࢺࢴࢠࢪཬᚺࢅ
ᢒโࡌࡾࡆ࡛࡞ࡻࡽ·OH ࡡ⏍ᠺࢅ㜴Ḿࡌࡾ᩺ぞ௥ㅨ∸ࢅ༟㞫ࡊࡒ㸝ᅒ 5,6㸞15-18)ࠊࡆࡡ∸㈻ࡢࠉ㕪࢕
࢛ࣤࠉH2 O2ࠉࣃࢺࣞ࢞ࢿࣤ࡝࡜ Fe3+㑇ඔ๠Ꮛᅹୖ࡞࠽࠷࡙ࠉࣆ࢘ࣤࢹࣤཬᚺ࡞ࡻࡾ·OH ࡡ⏍ᠺ࡛ࢬ
࣭ࣜࣞࢪࡡよ㔔ྙࢅᙁງ࡞ᢒโࡌࡾ 19)ࠊ
㐽ᢝⓏⓉⰅ⭁ᮑⳞ C. subvermispora ࡢࠉᮄᮞ⭁ᮑࡡิ᭿࡞㣤࿰࠽ࡻࡦ୘㣤࿰⬙⫣㓗࡛࣏ࣤ࢝ࣤ࣋
࢛ࣜ࢞ࢨࢱ࣭ࢭ(MnP)ࢅ⏐⏍ࡊ 20)ࠉMnP ࡷᣉᩋྊ⬗࡝ Mn3+㘊మࢅ㛜ጙ๠࡛ࡌࡾ⬙㈻㐛㓗໩࡞ࡻࡽ࢓
ࢨࣜࣚࢩ࢜ࣜࢅ chain carrying ࣚࢩ࡛࢜ࣜࡌࡾࣚࢩ࢜ࣜ㏻㙈ཬᚺࢅ㉫ࡆࡌ 21,22)ࠊMnP ࡞ࡻࡾ⬙㈻㐛
㓗໩ࡢ㟸ࣆ࢘ࢿ࣭ࣜᛮࣛࢡࢼ࣓ࣤࢸࣜࢅฦよࡌࡾࡆ࡛࠾ࡼ 23)ࠉᮇⳞࡡࣛࢡࢼࣤฦよᶭᵋࡡୌࡗ࡛ࡊ
࡙Ἰ┘ࡈࡿ࡙࠷ࡾࠊࡱࡒࠉ⬙㈻㐛㓗໩୯㛣మࡡ࣓ࢸ࡚ࣜ࠵ࡾ᭯ᶭࣃࢺ࢛ࣞ࣋ࣜ࢞ࢨࢺࢅ㔘ᒌ㘊మ࡛
ཬᚺࡈࡎ࡙࣭࢜࣍ࣤࢬࣤࢰ࣭ࣚࢩ࢜ࣜࢅ⏍ᠺࡈࡎࡾ࡛ࠉ㟸ࣆ࢘ࢿ࣭ࣜᛮྙᠺࣛࢡࢼࣤ࠿఩ฦᏄ໩ࡌ
ࡾࡲ࡚࡝ࡂ 24)ࠉᮄᮞ⣵⬂ባ࠽ࡻࡦ⣵⬂㛣ᒒ୯ࡡࣛࢡࢼࣤ࠿ฦよࡊ࡙ࣂࣜࣈ໩࠿㉫ࡀࠉᮄᮞ⣵⬂࠿๡
㞫ࡌࡾ 25)ࠊ⬙㈻㐛㓗໩࡝࡜ࡡ㐽ᢝⓏⓉⰅ⭁ᮑⳞࡡᶭ⬗よᯊ࡛⬗ງቌᙁࡡࡒࡴࠉC. subvermispora ࡡ
୘㣤࿰⬙⫣㓗⏍ྙᠺ㓕⣪࡚࠵ࡾǼ9-ཀྵࡦǼ12 ࢸࢦࢲ࣭ࣖࣚࢭࡡ㐿ఎᏄࢠ࣭ࣞࢼࣤࢡ࡛ྜྷⳞࡡᙟ㈻
㌹ᥦ⣌ࡡ㛜Ⓠࠉࣛࢡࢼࣤฦよ࡞├᥃㛭୙ࡌࡾࣚࢩ࢜ࣜ⛸ࡡཬᚺよᯊࢅ⾔ࡖ࡙࠷ࡾࠊ
7. ࠽ࢂ ࡽ ࡞
㐽ᢝⓏⓉⰅ⭁ᮑⳞ C. subvermispora ࡢࠉࢣࢿ࣑࠿ᮅよㄖ࡚࠵ࡽࠉᙟ㈻㌹ᥦ⣌ࡡᵋ⠇࠿㞬ࡊࡂࠉࣛ
ࢡࢼࣤฦよ࡞ࡢ㓕⣪ࡡ├᥃ཬᚺ࡚ࡢ࡝ࡂࠉ௥ㅨ∸ࡡཬᚺ࠿㛭୙ࡌࡾࡆ࡛࠾ࡼࠉࡐࡡᶭ⬗よᯊ࡛ࣛࢡ
ࢼࣤฦよ⬗ࡡቌᙁࡢᐖ᪾࡚ࡢ࡝࠷ࠊࡊ࠾ࡊ࡝ࡼ࠿ࠉࡆࡡⳞࡢࠉࣛࢡࢼࣤࢅ㓕⣪࠾ࡼ㞫ࡿࡒሔ࡚㧏㐽
ᢝⓏ࡞ฦよࡌࡾࣗࢼ࣭ࢠ࡝Ⳟమአࡡࣚࢩ࢜ࣜโᚒࢨࢪࢷ࣑ࢅࡵࡖ࡙࠽ࡽࠉࡱࡒ㟸㐽ᢝⓏⓉⰅ⭁ᮑⳞ
࡛␏࡝ࡽࠉᮄ㈻ࣁ࢕࢛࣏ࢪንᥦ࡫ࡡᖕᗀ࠷ᚺ⏕ᒈ㛜࠿ྊ⬗࡚࠵ࡾࠊᇱ♇࣬ᚺ⏕ࡡ୦㟻࠾ࡼ㐽ᢝⓏⓉ
Ⰵ⭁ᮑⳞ࡞㛭ࡌࡾ◂✪ࢅ╌ᐁ࡞㐅ࡴࡾࡆ࡛࠿ᚪさ࡛ᛦࢂࡿࡾࠊ
参考文献
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6) ῳ㎰㝧ྒྷ㸯ࢢ࣐࢙࢜ࣜࣤࢩࢼ࢓ࣛࣤࢡ, 48, 30-35 (2003)
7) ῳ㎰㝧ྒྷ㸯࢙ࢤࣁ࢕࢛࢙ࢾ࣭ࣜ࢟ࡡ᭩๑⥲̺ࢭ࢙࣐ࣞࢴࢨࣘࣤᆵ♣ఌࢅ┘ᣞࡊ࡙̺ࠉࢨ࣭࢙࣑ࢨ࣭ฝ∟ࠉ
᮶ாࠉpp. 68-78 (2005).
8) ῳ㎰㝧ྒྷ㸯ᮄ㈻⣌᭯ᶭ㈠″ࡡ᩺ᒈ㛜ࠉࢨ࣭࢙࣑ࢨ࣭ฝ∟ࠉ᮶ாࠉpp. 68-79 (2005).
9) ῳ㎰㝧ྒྷ㸯⏛㑌ಆ᭹ࠉ㤷ሔಕᚠࠉ▦㔕೸ኯࠉῳ㑌ᓤெࠉᮇ⏛୙ୌࠉᒱ⏛ಆᶖࠉⓉ஬ఘ᪺㸯ᖲᠺ 17 ᖳᗐ᪝
21
ᮇ⏍∸ᕝᏕఌㅦⁿさ᪠㞗ࠉ183 (2005).
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313-316 (2004).
11) Samsuri, M., Prasetya, B., Hermiati, E., Idiyanti, T., Okano, K., Syafwina,, Honda, Y., Watanabe, T. : Proc. The
Fifth Intern. Wood Sci. Symp., 317-323 (2004).
12) こᅽ௒ࠉ୯ᮇකᯣࠉKhanongnuch, C.ࠉῳ㑌ᓤெࠉᮇ⏛୙ୌࠉῳ㎰㝧ྒྷ㸯2006 ᖳᗐ㎨ⰹ໩Ꮥఌኬఌㅦⁿさ
᪠㞗ࠉ275 (2006).
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180, 205-211 (1999).
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(2003)
Conversion of wood biomass into energy and chemical resources by selective white rot fungi
Takashi Watanabe, Takahito Watanabe, Yoichi Honda
Research Institute for Sustainable Humanosphere, Kyoto University
22
S-3
ࣁ࢕࢛࢙ࢰࢿ࣭ࣜ⏍⏐࡞ฺ⏕ࡈࡿࡾ ⣊≟Ⳟ⏜ᮮ㓕⣪㻃
ࢿ࣍ࢧ࢕࣑ࢫ ࢩࣔࣂࣤᰬᘟఌ♣ ◂✪㛜Ⓠ㒂 㧐ᮄ ᚰ㻃
㻃
ࡢࡋࡴ࡞㻃
ୠ⏲Ⓩ࡞ࡲ࡙ࠉࣁ࢕࢛࣏ࢪ⇖ᩩࡡฺ⏕ࡢ⌟ᅹ᛬㏷࡞ᣉኬࡊࡗࡗ࠵ࡽࠉࣁ࢕࢛࢙ࢰࢿ࣭ࣜࡷ
ࣁ࢕࢛ࢩ࣭ࢭࣜࡡ⏍⏐㔖ࡢᖳ㛣⣑㸦㸥㹳㸧㸥㸚ࡡఘࡦࢅ♟ࡊ࡙࠷ࡾࠊ⌟ᅹࡆࡿࡼࡡࣁ࢕࢛
࣏ࢪ⇖ᩩࡢࠉ㣏⣂࣬㣣ᩩ࡛࡝ࡾ᳔∸ཋᩩ࠾ࡼ⏍⏐ࡈࡿ࡙࠷ࡾ࠿ࠉᑑᮮࡢ㟸㣏⣂ཋᩩࡡฺ⏕
࠿㔔さっࡈࡿࡾ࡛⩻࠻ࡼࡿࡾࠊ௑ᅂࡢࠉ⌟ᅹࡱࡒࡢᑑᮮࡡࣁ࢕࢛࢙ࢰࢿ࣭ࣜ⏍⏐࡞ฺ⏕ࡈ
ࡿࡾ⣊≟Ⳟ⏜ᮮࡡ㓕⣪ࢅ⣺௒ࡌࡾࠊ㻃
㻃
㸦㸞 ⃞⢂ฦよ㓕⣪㻃
⃞⢂ཋᩩࢅⵠ↳࣬⢮໩ᚃ࢓࣐࣭ࣚࢭ࡞ࡻࡽ⢶໩ࡊࣇࢺࢗ⢶࣬ᯕ⢶➴ࢅ⿿㏸ࡌࡾࣈࣞࢬࢪࡢࠉ
㸦㸮㸬㸥ᖳ௥࡞☔❟ࡊ௧ᮮ࡮࡯ንࢂࡾࡆ࡛࡝ࡂฺ⏕ࡈࡿ࡙࠷ࡾࠊ⌟ᅹ⡷ᅗ࡚ࡢࠉࡆࡡࣈࣞ
ࢬࢪࢅᚺ⏕ࡊ࡙ࠉ࡛࠹ࡵࢀࡆࡊཋᩩࡻࡽࣁ࢕࢛࢙ࢰࢿ࣭ࣜࢅ⏍⏐ࡊ࡙࠷ࡾࠊ㻃
㻃
㻃
!"#$%&'(
)"#$%&'(
㻃
Enzymes
CO 2
㻃
Hammer
Jet
㻃
mill
cooker
Whole
LiqueSlurry
Mash
㻃
grain
faction
tank
cooling
Ethanol
㻃
Fermentation
Yeast
Fresh water &
㻃
recycled water
㻃
㻃
Ethanol production process - Dry milling
㻃
㻃
㻃
ࡆࡆฺ࡚⏕ࡈࡿࡾᾦ໩ᆵ࢓࣐࣭ࣚࢭࡷᯖวࡽ㓕⣪ࡢ⣵Ⳟ⏜ᮮ࡚࠵ࡾ࠿ࠉ⢶໩ᆵ࢓࣐࣭ࣚࢭ
㻋ࢡࣜࢤ࢓࣐࣭ࣚࢭ㻌ࡢ 㻤㼖㼓㼈㼕㼊㼌㼏㼏㼘㼖㻃㼑㼌㼊㼈㼕 ࡷ 㻵㼋㼌㼝㼒㼓㼘㼖 ᒌⳞ࡝࡜⣊≟Ⳟ⏜ᮮࡡ㓕⣪࡚࠵ࡾࠊ㻃
㎾ᖳࠉ↋ⵠ↳⃞⢂ࡻࡽ࢙ࢰࢿ࣭ࣜࢅ⏍⏐ࡌࡾࣈࣞࢬࢪ࠿㛜Ⓠࡈࡿࡒࠊᐁ⏕໩࡞ᠺຉࡊࡒࡡ
ࡢ⡷ᅗࡡ࢙ࢰࢿ࣭࣒࣭࣭࡚ࣜ࢜ࠉࡆࡡᐁ⏕໩࡞ᐞ୙ࡊࡒࡡ࠿ᘚ♣㛜ⓆࡡᨭⰃᆵ⣊≟Ⳟ⏜ᮮ
࢓࣐࣭ࣚࢭ࡚࠵ࡾࠊࡆࡿࡢࠉࡱࡍ⃞⢂ྺ╌ࢺ࣒࢕ࣤ㻋㻶㼗㼄㼕㼆㼋㻃㼅㼌㼑㼇㼌㼑㼊㻃㼇㼒㼐㼄㼌㼑㻌㻃 ࢅᨭⰃࡊࠉࡈࡼ
࡞り፳ࢺ࣒࢕ࣤ࡞ࡵᨭⰃࢅຊ࠻࡙ᚋࡒ㓕⣪࡚࠵ࡾࠊࡆࡡ᩺ぞ㛜Ⓠ㓕⣪࡞ࡻࡽࠉࣈࣞࢬࢪ࡞
ᚪさ࡝㓕⣪῟ຊ㔖ࢅᐁ⏕໩ࣝ࣊ࣜࡱ࡚఩΅ࡌࡾࡆ࡛࡞ᠺຉࡊࡒࠊࡆࡡ᩺ࡊ࠷࢙ࢰࢿ࣭ࣜ⿿
㏸ࣈࣞࢬࢪࡢ୹࡞⡷ᅗහ࡚ᣉኬࡊࡗࡗ࠵ࡾࠊ㻃
㻃
㸧㸞 ࣛࢡࢿࢬ࣭ࣜࣞࢪฦよ㓕⣪㻃
⌟ᅹ⡷ᅗ࡚⾔ࢂࡿ࡙࠷ࡾ࡛࠹ࡵࢀࡆࡊཋᩩ࠾ࡼࡡ࢙ࢰࢿ࣭ࣜ⏍⏐ࡢࠉ㣣ᩩ⏕ཋᩩࡡ☔ಕࢅ
⩻៎ࡌࡾ࡛ 㻕㻓㻔㻕 ᖳ࡞ࡢ㝀⏲࡞㐡ࡌࡾ࡛ぜࡼࡿ࡙࠷ࡾࠊࡐࡡࡒࡴ⡷ᅗᨳᗋࡢࠉᮅฺ⏕㈠″࡚
࠵ࡾࣛࢡࢿࢬ࣭ࣜࣞࢪ⣌ࣁ࢕࢛࣏ࢪࢅཋᩩ࡛ࡊࡒࣁ࢕࢛࢙ࢰࢿ࣭ࣜ⏍⏐ࢅ 㻕㻓㻔㻖 ᖳࡱ࡚࡞
ᐁ⏕໩ࡌࡾࡆ࡛ࢅ┘ᣞࡊ࡙ࠉ⌟ᅹ┊ࢆ࡞◂✪ຐᠺࢅ⾔ࡖ࡙࠷ࡾࠊ⏐ᴏ⏕㓕⣪࣒࣭࣭࡛࢜ࡊ
࡙ࡡᘚ♣⡷ᅗᨥ♣ࡢࠉ࢙ࢾ࣭ࣜ࢟┤ 㻧㻲㻨 ࡻࡽ◂✪ጟォຐᠺࢅུࡄࠉࣛࢡࢿࢬ࣭ࣜࣞࢪฦよ
⏕ࡡ 㻷㼕㼌㼆㼋㼒㼇㼈㼕㼐㼄㻃㼕㼈㼈㼖㼈㼌 ⏜ᮮࢬ࣭ࣜࣚࢭࡡᨭⰃࢅ⾔ࡖࡒࠊ౐⏕ࡊࡒᇱ㈻ࡢࠉභྜྷ◂✪඙࡚࠵
ࡾᅗ❟࢙ࢾ්࣭ࣜ࢟⏍◂✪ᡜ 㻱㻵㻨㻯 ࠾ࡼ౩⤝ࡈࡿࡒ๑ฌ⌦ῥࡲࡡ࡛࠹ࡵࢀࡆࡊᗣᲘ∸㻃
㻋㼆㼒㼕㼑㻃 㼖㼗㼒㼕㼙㼈㼕㻌㻃 ࡚࠵ࡾࠊ㻗 ᖳ㛣࡞ࢂࡒࡾ◂✪ࡡᠺᯕ࡛ࡊ࡙ࠉᙔิࡡ┘ᵾ࡚࠵ࡖࡒ࢙ࢰࢿ࣭ࣜ
23
⿿㏸࡞࠾࠾ࡾ㓕⣪ࢤࢪࢹࡡ 㻔㻓 ಶ๎΅㸝㸦㸥ฦࡡ㸦࡞ࡌࡾࡆ࡛㸞ࢅኬࡀࡂ୕ᅂࡽࠉ㻱㻵㻨㻯 ࡞
ࡻࡾ๑ฌ⌦ᢇ⾙ࡡᨭⰃ࡛ྙࢂࡎ࡙᭩⤂Ⓩ࡞㓕⣪࡞࠾࠾ࡾࢤࢪࢹࢅ 㻖㻓 ฦࡡ 㻔 ࡞఩΅ࡌࡾࡆ࡛
࡞ᠺຉࡊࡒࠊ㓕⣪ഁࡡᨭⰃⅤࡢࠉ࠽ࡵ࡞ࢬ࣭ࣜࣚࢭ࡞ྱࡱࡿࡾ㓕⣪ᠺฦࡡᵋᠺẒࡡᨭⰃ㻋ș
㻐ࢡࣜࢤࢨࢱ࣭ࢭࡡẒ⋙ࢅྡྷ୕㻌ࠉ௙ࡡࣁ࢕࢛࣏ࢪฦよ⣊≟ⳞࡻࡽⓆぜࡈࡿࡒฦよὩᛮྡྷ୕
ᅄᏄࠉ㓕⣪⿿㏸ࣈࣞࢬࢪࡡᨭⰃ࡞ࡻࡾ⿿㏸ࢤࢪࢹࡡ๎΅࡝࡜࡞ࡻࡾࠊ㻃
⌟ᅹࡢࠉ㻘 ᖳᚃࡡᐁ⏕໩ࢅ┘ᣞࡊࠉ྘ᅗࡡභྜྷ◂✪඙࡛࡛ࡵ࡞ᐁッモ㥺ࢅ⾔ࡖ࡙࠷ࡾࠊ㻃
㻃
㻃
㻃
Enzyme is one of the keys to the process
Biomass
Collection
PreEnzyme
treatment Hydrolysis
Flow of biomass to fuel ethanol
Fungal enzymes used for Bio-ethanol production
Shinobu Takagi
Novozymes Japan Ltd. R&D
24
Fermentation
Recovery/
distillation
Fuel
Ethanol
S-4
㧏ᶭ⬗ᆵࢬ࣭ࣜࣚࢭ㧏⏍⏐ Trichoderma reesei ᰬࡡᵋ⠇࡞ྡྷࡄ࡙
᲻ᕖ ᗛࠉᒱ⏛Ᏻᩝࠉᑚ➗ཋ ῦ㸝㛏ᒱᢇ⾙⛁ᏕኬᏕ⏍∸⣌㸞
ࠜࡢࡋࡴ࡞ࠝ㻃
ࢬ࣭ࣜࣞࢪ⣌ࣁ࢕࢛࣏ࢪ㸝ࣛࢡࢿࢬ࣭ࣜࣞࢪ㸞࠾ࡼࡡ⢶⏍⏐ࡢࠉ⌟ᅹ⅛㓗࢝ࢪቌຊᢒโࢅ┘ᣞࡊ
ࡒ⇖ᩩ࢙ࢰࢿ࣭ࣜ࡝࡜ࡡࣁ࢕࢛࣏ࢪ࢙ࢾ࣭ࣜ࢟࡞↌Ⅴ࠿ᙔ࡙ࡼࡿ࡙࠷ࡾ࠿ࠉ▴ἔᯜῤࢅ⩻៎ࡊࡒ୯
㛏᭿Ⓩ࡞ࡢࠉᆀ⌣ぞᶅ࡚ࡡ㣏⣂࣬㣏ဗࡷ໩Ꮥᕝᴏཋᩩࡡ⏍⏐࡞ᚪさ࡝ᢇ⾙࡛ࡊ࡙㟸ᖏ࡞㔔さ࡛࡝ࡾ
㸝ࣁ࢕࢛ࣛࣆ࢒࢕ࢻ࣭ࣛ㸞ࠊࢬ࣭ࣜࣞࢪࡡ⢶໩ࡢ㛏࠷Ṍྍࡡ࠵ࡾᠺ⇅ࡊࡒ㓗⢶໩࡞௥ࢂࡖ࡙ࠉ௑ᚃ
ࡢᨭⰃࡡఴᆀ࠿ṟࡖ࡙࠷ࡾࢬ࣭ࣜࣚࢭ࡞ࡻࡾ㓕⣪⢶໩ࡡฝ␊࠿᭿ᙽࡈࡿ࡙࠷ࡾࠊ
ࢬ࣭ࣜࣞࢪ⣌ࣁ࢕࢛࣏ࢪࡢࢬ࣭ࣜࣞࢪ࠿⤎ᬏᵋ㏸ࢅཱིࡖ࡙࠷ࡾࡆ࡛࡞ຊ࠻࡙ࠉࣉ࣐ࢬ࣭ࣜࣞࢪࡷ
ࣛࢡࢼࣤ࠿々㞟࡞⤙ࡲྙࡖࡒᵋ㏸ࢅ᭯ࡊ࡙࠷ࡾࡒࡴࠉࢬ࣭ࣜࣚࢭ࡞ࡻࡾ㓕⣪⢶໩࡚ࡢࠉ๑ฌ⌦࠽ࡻ
ࡦኬ㔖ࡡࢬ࣭ࣜࣚࢭ࠿ᚪさ࡛࠷࠹ㄚ㢗࠿ᕝᴏ໩ࢅ㜴ࢆ࡚࠷ࡾࠊ
9ULHKRIHUPFUHHVHL㸝ࡆࡿࡱ࡚୘ᏰධⳞ࡞ฦ㢦ࡈࡿ࡙࠷ࡒ࠿ࠉ᭯ᛮୠ௥ -^SRHUHFMHHRULQF ࠿ぜ
ฝࡈࡿࡒ㸞ࡢ㔕⏍ᰬࡡⓆぜ࠾ࡼ㸫㸥ᖳࡡṌྍ࠿࠵ࡽࠉ⤎ᬏࢬ࣭ࣜࣞࢪࢅຝ⋙ࡻࡂฦよྊ⬗࡝ኬ㔖ࡡ
ࢬ࣭ࣜࣚࢭࢅฦἢࡌࡾࡆ࡛࠾ࡼࠉࢬ࣭ࣜࣞࢪࡡ㓕⣪⢶໩࡞ࡢḖ࠾ࡎ࡝࠷⣊≟Ⳟ࡚࠵ࡾࠊ9UHHVHL
ࡡࢬ࣭ࣜࣚࢭ࡞㛭ࡊ࡙ࠉ಴ࠍࡡࢬ࣭ࣜࣚࢭᠺฦࡡ㓕⣪໩ᏕⓏᛮ㈻ࠉᵋ㏸ᶭ⬗┞㛭ࠉ㓕⣪࠽ࡻࡦࡐࡡ
㐿ఎᏄࡡⓆ⌟โᚒᶭᵋ࡝࡜࡞㛭ࡊ࡙↋ᩐࡡሒ࿈࠿࠵ࡾࠊࡊ࠾ࡊࠉᫎᖳࢣࢿ࣑㒼า࠿ප⾪ࡈࡿࡒࡆ࡛
࠾ࡼࠉࡆࡿ࠾ࡼࡡ◂✪ࡡ᪁ྡྷᛮࡷ㐅⾔㏷ᗐࡢ᩺ࡊ࠷ṹ㝭࡞ථࡖࡒ࡛⩻࠻ࡼࡿࡾࠊ
➱⩽➴ࡡ◂✪ࢡ࣭ࣜࣈ࡚ࡢࠉ9UHHVHL ࡡ⏍⏐ࡌࡾኣᩐࡡࢬ࣭ࣜࣚࢭ࣬࢞ࢨࣚࢻ࣭ࢭࡡᛮ㈻ࡷࡐ
ࡿࡼ㐿ఎᏄࡡ␏⛸ᐙ୹Ⓠ⌟࡛ࢰࣤࣂࢠ㈻ᕝᏕⓏᨭንࢅ㐅ࡴࡾୌ᪁ࠉࢬ࣭ࣜࣚࢭ㸝࠽ࡻࡦ࢞ࢨࣚࢻ࣭
ࢭ㸞ࡡㄇᑙⓆ⌟ᶭᵋࡡよ᪺࡞ཱིࡽ⤄ࢆ࡚ࡀࡒࠊࡆࡆ࡚ࡢࠉࢬ࣭ࣜࣞࢪ⣌ࣁ࢕࢛࣏ࢪࡡ㓕⣪⢶໩ࡡᐁ
⌟࡞ྡྷࡄࡒཱིࡽ⤄ࡲ࡞⤘ࡖ࡙ࠉࡆࡿࡱ࡚ࡡ⤎ᯕ࡛௑ᚃࡡゝ⏤ࢅ㏑࡬ࡒ࠷ࠊ
䛀಴䚱䛴䜿䝯䝭䞀䝀㐿ఎᏄ䛴␏⛸ᐙ୹Ⓠ⌟䛮㐅໩ฦᏄᕝᏕⓏᨭን䛁㻃
㻔㻌9UHHVHLࡢኣᩐࡡࢬ࣭ࣜࣚࢭࢅฦἢࡌࡾࡡ࡚ࠉ಴ࠍࡡࢬ࣭ࣜࣚࢭࡡᛮ㈻ࢅよ᪺ࡌࡾࡒࡴ࡞ࡢࠉ
ࢬ࣭ࣜࣚࢭࢅ⏍⏐ࡊ࡝࠷␏⛸ᐙ୹㸝࠵ࡾ࠷ࡢ⏍⏐ࡊ࡝࠷᮪௲㸞࡚Ⓠ⌟ࡈࡎࡾᚪさ࠿࠵ࡾࠊ⾪࡞♟ࡊ
ࡒࡻ࠹࡞ࠉࡆࡿࡱ࡚࡞ 9UHHVHL ࡡ⏍⏐ࡌࡾ୹さ࡝ࢬ࣭ࣜࣚࢭ࣬࢞ࢨࣚࢻ࣭ࢭࡡ␏⛸ᐙ୹࡚ࡡⓆ⌟
ࢅ⾔࠷ࠉ⤄ᥦ࠻㓕⣪ࡡᛮ㈻࠵ࡾ࠷ࡢࢬ࣭ࣜࣞࢪฦよ࡞࠽ࡄࡾ༝዇ష⏕ࢅ᳠ゞࡊ࡙ࡀࡒࠊ᭩⤂Ⓩ࡝┘
ᵾࡢࢬ࣭ࣜࣞࢪฦよ࡞࠽ࡄࡾ୹さࢬ࣭ࣜࣚࢭᠺฦࡡ᭩㐲Ẓ⋙ࢅỬᏽࡌࡾࡆ࡛࡚࠵ࡾࠊ
㻕㻌ࡆࡿࡼࡡ㐿ఎᏄࢅ⏕࠷࡙ࢰࣤࣂࢠ㈻ᕝᏕⓏ࡞ᨭንࢅモࡲࠉⱕᖱࡡᨭⰃ㓕⣪ࢅㄢ⿿ࡊࡒࠊࡊ࠾ࡊࠉ
ࡆࡡ᪁Ἢࡢᵋ㏸
ᶭ⬗┞㛭➴ࡡ◂
✪࡞ࡢ᭯ຝ࡚࠵
ࡾ࠿ࠉẒὩᛮࡡ
ቌᙁ➴࡞ᑊࡌࡾ
ᣞ㔢ࢅᚋࡾࡆ࡛
ࡢᅏ㞬࡚࠵ࡖࡒ
ࡒࡴࠉ⌟ᅹࡢ㐅
໩ฦᏄᕝᏕⓏᨭ
ንࢅモࡲ࡙࠷ࡾࠊ
ᨭን࡞ࡢኣᩐࡡ
ን␏ࣚ࢕ࣇࣚࣛ
࣭ࡡᵋ⠇࠿ྊ⬗
࡝ኬ⭘Ⳟࡷ㓕ẍ
㸝 8FHHKFURP^HH
VHHUH[LVLFH 㸞
࡚ࡡⓆ⌟࠿ᚪさ
࡚࠵ࡾࡒࡴࠉ*,
25
.㸝8HHUH[LVLFH㸞
ࠉ*,.. ࠽ࡻࡦ *,...㸝ኬ⭘Ⳟ㸞࡞ᑊࡊ࡙ࠉ࢙࣭ࣚࣈ࣭ࣞࣤ 5(7 Ἢ࡞ࡻࡾᨭን
ࢅ㐅ࡴ࡙࠷ࡾࠊᨭንࡡ࣓ࢸ࡛ࣜࡊ࡙඙⾔ࡊ࡙࠷ࡾ *,... ࡚ࡢࠉⓆ⌟㔖ቌኬࠉẒὩᛮྡྷ୕࠽ࡻࡦ⪇
⇍ᛮྡྷ୕࡝࡜ࡡ⮾࿝῕࠷ን␏మࡵཱིᚋࡊ࡙࠷ࡾࠊ
䛀㻷㻑㻃㼕㼈㼈㼖㼈㼌 䛴┞ྜྷ⤄ᥦ䛎䜘ฺ⏕䛝䛥䜿䝯䝭䞀䝀ㄇᑙโᚒᶭᵋ䛴よ᪺䛁㻃
ࡵ࠹ୌࡗࡡ◂✪ࡡ୯ᚨࡢࢬ࣭ࣜࣚࢭࡡㄇᑙโᚒᶭᵋ࡞㛭ࡌࡾ◂✪࡚࠵ࡾࠊ9UHHVHL ࡢࠉࢬࣜࣞ
࣭ࢪ࡛ࡐࡡㄇᑙమ࠽ࡻࡦࣚࢠࢹ࣭ࢪࡷࢮ࣭ࣜ࣍ࢪࡡᏋᅹୖ࡚ࠉࢬ࣭ࣜࣚࢭࢅㄇᑙࡌࡾࠊࡆࡡㄇᑙᶭ
ᵋࡡレ⣵ࡢ࡮࡛ࢆ࡜よ᪺ࡈࡿ࡙࠽ࡼࡍࠉ&(*.. ࠿။ୌࡡ≁␏Ⓩ࡝㌹෕Ὡᛮ໩ᅄᏄ࡛ࡊ࡙ሒ࿈ࡈࡿ࡙
࠷ࡾ࠿ࠉᑛ࡝ࡂ࡛ࡵ㸬⛸ࡡࢬ࣭ࣜࣚࢭ㐿ఎᏄࡡ㌹෕࡞㛭ࡊ࡙ࠉձ࡝ࡏྜྷ᫤࡞ㄇᑙ࠿㛜ጙࡈࡿࡾࡡ࠾ࠉ
ղ࡝ࡏ㌹෕㔖࠿ኬࡀࡂ␏࡝ࡾࡡ࠾ࠉ࡝࡜ࡡ⌦⏜ࡢධࡂ୘᪺࡚࠵ࡾࠊ
ࡐࡆ࡚ࠉJXV ࢅ࣭ࣝ࣎ࢰ࣭㐿ఎᏄ࡛ࡊ࡙ࠉ㌹෕㔖ࡡᑛ࡝࠷㸝࢓ࢠࢲࣤࡡⓆ⌟㔖ࡻࡽࡵኣ࠷㸞*,...ࠉ
࠽ࡻࡦࢬ࣭ࣜࣚࢭ࡛ྜྷᣪິࢅࡌࡾ࢞ࢨࣚࢻ࣭ࢭ... ࡡ୦㐿ఎᏄࡡ୕Ὦ㒼าࢅḖ኶ን␏࠵ࡾ࠷ࡢ㡷
ᇡࡷⅤን␏ࢅ⾔࠷ࠉ┞ྜྷ⤄ᥦ࠻࡞ࡻࡖ࡙よᯊࡊ࡙࠷ࡾࠊ⌟ᅹㄇᑙ㌹෕࡞㔔さ࡝㡷ᇡࡡ≁ᏽࡷὩᛮ໩
ᅄᏄ &(*.. ࡓࡄ࡚ㄕ᪺୘ྊ⬗࡝⌟㇗ࢅぜฝࡊ࡙࠷ࡾࠊ௑ᚃࠉࡆࡿࡼࡡ㡷ᇡࢅྱࡴ࡙ࠉ┘Ⓩࡡࢬࣜ
࣭ࣚࢭẒ⋙࡞ぜྙࡖࡒⓆ⌟㔖ࢅ㐡ᠺࡈࡎࡾࡆ࡛ࢅ┘ᵾ࡞᳠ゞࡊ࡙࠷ࡂ஢ᏽ࡚࠵ࡾࠊ
䛀㧏ᶭ⬗ᛮ䜿䝯䝭䞀䝀㧏⏍⏐ 㻷㻑㻃㼕㼈㼈㼖㼈㼌 ᰬ䛴ᵋ⠇䛱ྡྷ䛗䛬䛁㻃
௧୕ࡡࢬ࣭ࣜࣞࢪฦよ࡞࠽ࡄࡾ྘ࢬ࣭ࣜࣚࢭᠺฦࡡ᭩㐲Ẓ⋙ࡡ᳠ゞࠉ྘ᠺฦࡡᨭንࠉㄇᑙㄢ⟿ᶭ
ᵋࡡよ᪺ࢅ⤄ࡲྙࢂࡎࡾࡆ࡛࡞ࡻࡖ࡙ࠉ┞ྜྷ⤄ᥦ࠻࡞ࡻࡾ㧏ᶭ⬗ᆵࢬ࣭ࣜࣚࢭ㧏⏍⏐ን␏ᰬࡡᵋ⠇
࠿ྊ⬗࡞࡝ࡾ࡛⩻࠻࡙࠷ࡾࠊ
ᚉᮮࠉኬ㔖࡞Ⓠ⌟ࡊ࡙࠷ࡾ ('-. ࡡࣈ࣓࣭ࣞࢰ࣭ࢅฺ⏕ࡊࡒ␏⛸㓕⣪ࢰࣤࣂࢠ㈻Ⓠ⌟࠿モࡲࡼࡿ
࡙࠷ࡾ࠿ࠉ࠷ࡍࡿࡵ ('-. ࡮࡜ࡡኬ㔖Ⓠ⌟࡞ᠺຉࡊ࡙࠷࡝࠷ࠊ୕オࡡᢇ⾙ࡢ␏⛸㓕⣪ࢰࣤࣂࢠ㈻ࡡ
ኬ㔖Ⓠ⌟ࢅ⾔࠹㝷࡞ࡵ㐲⏕࡚ࡀࡾࡵࡡ࡛᭿ᙽࡊ࡙࠷ࡾࠊ
ࡱࡒࠉ9UHHVHL ࢬ࣭ࣜࣚࢭ࡞㛭ࡌࡾ◂✪࠽ࡻࡦࢬ࣭ࣜࣚࢭ⏍⏐࡞ࡗ࠷࡙ࡢ࡮࡛ࢆ࡜࠿ᾦమᇰ㣬
࡚⾔ࢂࡿ࡙ࡀࡒࠊࡆࡿࡢࢬ࣭ࣜࣞࢪ⣌ࣁ࢕࢛࣏ࢪ࠾ࡼࡡ⢶⏍⏐ࢅ⩻࠻ࡒሔྙࡢᾦమᇰ㣬࠿ᚪ↓࡚࠵
ࡾࡒࡴ࡚࠵ࢀ࠹ࠊࡆࡡࡻ࠹࡝ᾦమᇰ㣬࡚ࡡᠺᯕ࡞ຊ࠻࡙ࠉ&VSHUJLOOXV ᒌ࡚᪺ࡼ࠾࡞࡝ࡽࡗࡗ࠵ࡾ
ᅖమᇰ㣬࡛ᾦమᇰ㣬࡚ࡡⓆ⌟ㄢ⟿ᶭᵋࡡᕣ␏ࢅ 9UHHVHL ࡞㐲⏕ࡈࡎࡾࡆ࡛࡞ࡻࡖ࡙ࡵࠉ㧏ᶭ⬗ᆵ
9UHHVHL ን␏ᰬࡡᵋ⠇ࢅ㐅ࡴࡾࡆ࡛࠿ྊ⬗࡛⩻࠻ࠉ⌟ᅹ◂✪ࢅࢪࢰ࣭ࢹࡈࡎࡒࠊ
Construction of highly functional cellulase-highly producing Trichoderma reesei mutants
Yasushi Morikawa
Dept. of BioEngineering, Nagaoka University of Technology
26
O-1
࠷ࡵࡔ⑋Ⳟ࡞࠽ࡄࡾ஦ࡗࡡ Dicer ᵕ⺦Ⓣ㈻ࡡᶭ⬗ฦ໩
みㆺ├ᶖ㸡ᅰఫᖶ㞕㸡┶ᒜ⁘ᚷ㸡୯ᒁᩔᆍ㸝♼ᡖኬ࣬㎨Ꮥ㒂㸞
Dicer ⺦Ⓣ㈻ࡢࠉRNA ࢦ࢕ࣝࣤࢨࣤࢡ(RNAi)⤊㊪ࡡ㘵㓕⣪࡚࠵ࡽࠉ㛏࠷஦ᮇ㙈 RNA ࡡ๑㥉మࢅ 20-30bp ⛤
ᗐࡡᑚࡈ࡝ RNA ฦᏄ࡞ว᩷ࡌࡾり፳ష⏕ࢅᣚࡗࠊ࢕ࢾ⛁᳔∸࠷ࡵࡔ⑋Ⳟࢣࢿ࣑࡞ࡢࠉ㸧⛸ࡡ Dicer ᵕ㐿ఎᏄࠉ
Mdl-1 ࠽ࡻࡦ MDL-2 ࠿ヾࡴࡼࡿࡾ࠿ࠉࡆࡿࡼࡡ◒ቪᰬ(Knock-Out, KO)ࢅష⿿ࡊࡒᡜࠉMDL-2 KO ᰬࡡࡲ࠿ࣉ
࢓ࣅࣤ RNA ࡞ࡻࡽㄇᑙࡈࡿࡾ RNA ࢦ࢕ࣝࣤࢨࣤࢡ࡞Ḗ᥾࠿࠵ࡾࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊࡱࡒࠉහᅹᛮࡡࢹ
ࣚࣤࢪ࣎ࢯࣤ࡞ᑊࡌࡾ siRNA ࡡ⵫✒ࡵ MDL-2 KO ᰬ࡚ᾐ኶ࡊ࡙࠽ࡽࠉ⮤↓᮪௲ୖ࡚ࡢ MDL-2 ࡡࡲ࠿ RNA
ࢦ࢕ࣝࣤࢨࣤࢡ⤊㊪࡞㛭୙ࡌࡾࡆ࡛࠿♟ြࡈࡿࡒࠊୌ᪁ࠉ
Ⳟ⣊࡚ࡡ MDL-1 ࠽ࡻࡦ MDL-2 ࡡⓆ⌟㔖ࢅ Real Time
PCR ࡚ᏽ㔖ࡊࡒᡜࠉMDL-2 mRNA ࡢ MDL-1 mRNA ࡞Ẓ࡬࡙⣑ 15 ಶࡡⓆ⌟㔖࠿࠵ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊ
ࡆࡡ㌹෕㔖ࡡᕣ࠿ Dicer ᵕ⺦Ⓣ㈻ࡡᶭ⬗ฦ໩࡞୙࠻ࡾᙫ㡢ࢅㄢ࡬ࡾࡒࡴࠉAspergillus nidulans ࡡ gpd ࣈ࣓࣭ࣞ
ࢰ࣭ࢅ⏕࠷ࡒᙁⓆ⌟⣌ࢅฺ⏕ࡊࡒࠊMDL-2 KO ᰬ࡚ࠉMDL-1 ࠽ࡻࡦ MDL-2 mRNA ࢅᙁⓆ⌟ࡈࡎࡒᡜࠉྜྷ⛤
ᗐࡡⓆ⌟㔖ࢅ♟ࡊࡒ࠿ࠉMDL-2 ᙁⓆ⌟ᰬ࡚ࡢࠉ㔕⏍ᰬ࡛ྜྷ⛤ᗐࡡࢦ࢕ࣝࣤࢨࣤࢡຝ⋙ࢅ♟ࡊࠉMDL-1 ᙁⓆ⌟
ᰬ࡚ࡢࠉ఩ຝ⋙ࡡ RNA ࢦ࢕ࣝࣤࢨࣤࢡ⬗ࢅ♟ࡊࡒࠊࡆࡿࡼࡡࡆ࡛ࡢ௧ୖ࡞♟ࡌ࠷ࡂࡗ࠾ࡡ♟ြࢅ୙࠻ࡾࠊ㸦㸣
⮤↓᮪௲ୖ࡚ࡢ RNA ࢦ࢕ࣝࣤࢨࣤࢡ⤊㊪࡚ᶭ⬗ࡊ࡙࠷࡝࠷ MDL-1 ⺦Ⓣ㈻ࡵࠉRNA ࢦ࢕ࣝࣤࢨࣤࢡ⤊㊪࡚ᶭ
⬗ࡊᚋࡾ⺦Ⓣ㈻࡚࠵ࡾࠉ㸧㸣MDL-1 ࡛ MDL-2 ࡢࡌ࡚࡞⺦Ⓣ㈻࡚ࣝ࣊ࣜฦ໩ࡊ࡙࠽ࡽࠉMDL-2 ࠿ࡻࡽຝ⋙Ⓩ
࡞഼ࡂࠉ㸨㸣⮤↓᮪௲ୖ࡚ࡡ MDL-1 ࡛ MDL-2 ࡡᶭ⬗ฦ໩࡞ࡢ㌹෕ࢤࣤࢹ࣭ࣞࣜ࠿㛭୙ࡊ࡙࠷ࡾ㸣㸩㸣MDL-2
ࡡⓆ⌟㔖ࡢࠉ࠷ࡵࡔ⑋Ⳟࡡ RNA ࢦ࢕ࣝࣤࢨࣤࢡຝ⋙ࡡโ㝀さᅄ࡚ࡢ࡝࠷ࠊ
Two dicer-like proteins in Magnaporthe oryzae
Naoki Kadotani, Yukio Tosa, Shigeyuki Mayama, Hitoshi Nakayashiki
(Fac. of Agri., Kobe Univ.)
O-2
ࢩࣔ࢝࢕࣓ኚ␷⑋Ⳟ Alternaria solani ⏜ᮮ ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪ PKSN ࡡⓆ⌟࡛ᶭ⬗
よᯊ
➗ཋ ᇻ㸦ࠉཀྵᕖⱝ⚽㸧ࠉ⸠஬ ໌㸦ࠉᾇ⩹ሪ㇇㸦 㸝㸦᮶ኬ㝌࣬ⷾࠉ㸧໪ኬ㝌࣬⌦㸞
࠘┘Ⓩ࠙ࢩࣔ࢝࢕࣓ኚ␷⑋Ⳟ Alternaria solani ࡡ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪ PKSN ࡢࠉ8 ಴ࡡ S-adenosyl methionine
(SAM)⏜ᮮࡡഁ࣒ࢲࣜᇱࢅᣚࡗࢸ࢜ࢢࢰ࢕ࢺ alternapyrone ࢅ⏍ᠺࡌࡾࠊ1) ࡆࡿࡼࡡഁ࣒ࢲࣜᇱࡢࠉPKSN ୕ࡡ
࣒ࢲࣜᇱ㌹⛛㓕⣪㸝MeT㸞ࢺ࣒࢕ࣤࡡり፳ཬᚺ࡞ࡻࡽᑙථࡈࡿࡾ࡛⩻࠻ࡼࡿࡾ࠿ࠉ௙ࡡ⣊≟Ⳟ࣎ࣛࢢࢰ࢕ࢺ
ྙᠺ㓕⣪ࡡ MeT ࢺ࣒࢕࡛ࣤྜྷᵕࠉࡐࡡᶭ⬗ࡡレ⣵ࡢ୘᪺࡚࠵ࡾࠊࡐࡆ࡚ࠉ௑ᅂ PKSN ࡡ㓕ẍⓆ⌟⣌ࢅ☔❟ࡊࠉ
MeT ࢺ࣒࢕ࣤࡡᶭ⬗よᯊࢅモࡲࡒࡡ࡚ሒ࿈ࡌࡾࠊ
࠘ᐁ㥺࡛⤎ᯕ࠙MeT Ὡᛮ࡞ᚪ㡪࡝࢓࣐ࢿ㓗ṟᇱࡡ≁ᏽࢅ┘Ⓩ࡛ࡊ࡙Ⅴን␏ᑙථᐁ㥺ࢅ㐅ࡴࡾ࡞࠵ࡒࡽࠉฝⰾ
㓕ẍ Saccharomyces cerevisiae ࢅᐙ୹࡛ࡊࡒࡻࡽຝ⋙Ⓩ࡝Ⓠ⌟⣌ࡡᵋ⠇ࢅモࡲࡒࠊ㓕ẍ⏕Ⓠ⌟࣊ࢠࢰ࣭
pYES-DEST52 ࡞ pksN cDNA ࢅ⤄ࡲ㎲ࡲࠉBacillus subtilis ⏜ᮮ࣌ࢪ࣌ࣂࣤࢷࢷ࢕ࢼࣜ໩㓕⣪㐿ఎᏄ sfp ࡛࡛ࡵ
࡞ࠉฝⰾ㓕ẍ࡞ᑙථࡊࡒࠊᇰ㣬࣬ㄇᑙ᮪௲ࢅ⛸ࠍ᳠ゞࡊࡒ⤎ᯕࠉPKSN ᙟ㈻㌹ᥦ㓕ẍ࡞ࡻࡾ alternapyrone ࡡ
⏍⏐࡞ᠺຉࡊࡒࠊࡆࡿࡢࠉ⣊≟Ⳟ㑇ඔᆵ PKS ࡛ࡊ࡙㓕ẍ࡚ᶭ⬗Ⓠ⌟ࡊࡒ᭩ิࡡౚ࡚࠵ࡾࠊࡆࡡ PKSN 㓕ẍⓆ
⌟⣌ࢅ⏕࠷ࠉMeT ࢺ࣒࢕ࣤ࡫ࡡን␏ᑙථᐁ㥺ࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉPKSN ࡡ MeT ࢺ࣒࢕ࣤ୯ࠉalternapyrone
⏍ᠺ࡞ᚪ㡪ࡡ࢓࣐ࢿ㓗ṟᇱࢅྜྷᏽࡊࡒࠊࡱࡒࠉMeT ን␏మ࡞ࡻࡾഁ࣒ࢲࣜᇱࢅᣚࡒ࡝࠷ alternapyrone ㄇᑙమ
ࡡ⏍ᠺࡢヾࡴࡼࡿ࡝࠾ࡖࡒࠊ
1) I. Fujii et al., Chem. Biol., 12, 1301-1309 (2005)
Expression and functional analysis of polyketide synthase PKSN from Alternaria solani
Ken Kasahara1, Hideaki Oikawa2, Isao Fujii1, Yutaka Ebizuka1
(1Graduate School of Pharm. Sciences, Univ. of Tokyo, 2Graduate School of Science, Hokkaido Univ.)
27
O-3
㯔Ⳟ A. oryzae ࡡ␏⛸ࢰࣤࣂࢠ㈻㧏⏍⏐ን␏ᰬࡡཱིᚋ
᰷ᮇᓤ㸡ῳ㎰ὀ♰ࠉ‹୕㇇㸡୷ᒜ₮ୌ㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
࠘┘Ⓩ࠙ඁࡿࡒࢰࣤࣂࢠ㈻ฦἢ⬗ࢅ᭯ࡌࡾ㯔Ⳟ A. oryzae ࡢࠉ␏⛸ࢰࣤࣂࢠ㈻⏍⏐ࡡᐙ୹࡛ࡊ࡙᭿ᙽࡈࡿ࡙࠷
ࡾࠊࡊ࠾ࡊࠉ㧏➴⏍∸⏜ᮮࢰࣤࣂࢠ㈻ࡡኬ㒂ฦࡢ⏍⏐ᛮ࠿఩࠷ࡡ࡞ࡵ㛭ࢂࡼࡍࠉA. oryzae ࡞࠽࠷࡙ࡆࡿࡼࢅ
㧏⏍⏐ࡌࡾࡒࡴࡡᐙ୹ࢅ๭⿿ࡌࡾモࡲࡢ࡮࡛ࢆ࡜⾔ࢂࡿ࡙࠷࡝࠷ࠊࡆࡿࡱ࡚࡞ᠻࠍࡢࠉⳞమහ㓗ᛮࣈࣞࢷ࢓
࣭ࢭ㐿ఎᏄ(pepE)࡛ࢹࣛ࣋ࣈࢲࢩࣜ࣋ࣈࢲࢱ࣭ࢭ㐿ఎᏄ(tppA)ࡡ㸧㔔◒ቪ࡞ࡻࡽࠉࣃࢹࣛࢯࢲ࣭࣑ࡡ⏍⏐㔖࠿
⣑ 1.6 ಶ࡞࡝ࡾࡆ࡛ࢅሒ࿈ࡊ࡙࠷ࡾࠊᮇ◂✪࡚ࡢࠉ୕オ㐿ఎᏄࡡ㸧㔔◒ቪᰬࢅࡵ࡛࡞ࡊ࡙ࠉࣛࢯࢲ࣭࣑Ὡᛮ
ࢅᣞᵾ࡞␏⛸ࢰࣤࣂࢠ㈻㧏⏍⏐ን␏ᰬࡡ⫩⛸ࢅ⾔ࡖࡒࠊ
࠘᪁Ἢ࡛⤎ᯕ࠙᭩ิ࡞ࠉ㸩㔔ᰜ㣬さịᛮᰬࢅ⏕࠷࡙ pepE, tppA㸧㔔◒ቪᰬ(NS-tApE)ࢅష⿿ࡊࡒࠊࡆࡡᰬࡡௗ
ࢗࢨ࣓࢞ࢨࣤ⏍⏐㔖ࢅࢤࣤࢹ࣭ࣞࣜᰬ࡛Ẓ㍉ࡊࡒ࡛ࡆࢀࠉ⣑ 1.8 ಶࡡቌຊ࠿ヾࡴࡼࡿࡒࠊࡆࡡࡆ࡛࠾ࡼࠉpepE,
tppA㸧㔔◒ቪ࠿௙ࡡ␏⛸ࢰࣤࣂࢠ㈻࡞ࡵ᭯ຝ࡚࠵ࡾࡆ࡛࠿♟ࡈࡿࡒࠊࡈࡼ࡞ࠉNS-tApE ᰬ࡞ࣃࢹࣛࢯࢲ࣭࣑
Ⓠ⌟ࣈࣚࢪ࣐ࢺࢅ niaD 㐿ఎᏄᗑ࡞༟ࢤࣅ࣭࡚ᑙථࡊࠉࡆࡡᰬࡡฦ⏍Ꮔ࡞ᑊࡊ࡙⣰አ⥲↯ᑏ࡞ࡻࡾን␏ฌ⌦ࢅ
⾔ࡖࡒࠊን␏ฌ⌦ᚃࡡฦ⏍Ꮔࢅ Micrococcus Ⳟమථࡽᐨኮᇰᆀ࡞ሧἋࡊࠉࣛࢯࢲ࣭࣑Ὡᛮ࡞ࡻࡾ⁈Ⳟࡡࣀ࣭ࣞ
࠿ኬࡀࡂ࡝ࡖࡒᰬࢅࢪࢠ࣭ࣛࢼࣤࢡࡊࡒࠊ᭩ࡵ㧏࠷Ὡᛮࢅ♟ࡊࡒᰬ࠾ࡼⓆ⌟ࣈࣚࢪ࣐ࢺࢅ࢞ࣖ࢓ࣛࣤࢡ࡞ࡻ
ࡽ⬲ⴘࡈࡎࠉ㧏⏍⏐ን␏ᰬࢅ㸬ᰬཱིᚋࡊࡒࠊࡐࡡ㸬ᰬࢅ AUT1㹳7 ᰬ࡛ྞࡘࡄࠉ⌟ᅹ AUT1 ᰬ࡞࣓࢞ࢨࣤⓆ⌟
ࣈࣚࢪ࣐ࢺࢅᙟ㈻㌹ᥦࡊࠉࡐࡡ⏍⏐㔖࡞ࡗ࠷࡙᳠ゞࡊ࡙࠷ࡾࠊ
Breeding of the A. oryzae mutants for higher-level production of heterologous proteins
Takashi Nemoto, Taisuke Watanabe, Yutaka Mizogami, Jun-ichi Maruyama, Manabu Arioka, Katsuhiko Kitamoto
(Dept. of Biotechnol., Univ. of Tokyo)
O-4
࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ G ࢰࣤࣂࢠ㈻โᚒᅄᏄ RGS ࡞ࡗ࠷࡙ࡡよᯊ
ཪ㔕⌦⏍㸡༞ᰜୌ㸡こᮟ㯖㔓Ờ 㸝⏍∸◂㸞
࢕ࢾ࠷ࡵࡔ⑋Ⳟࡢࠉ3 㔖మ G ࢰࣤࣂࢠ㈻࡞ࡻࡽአ⏲ࡡ␧Ềᛮࡷ᳔∸ᠺฦ࡝࡜ࢅヾㆉࡊࠉវ᯹ჹᏻ࡚࠵ࡾ௛
╌ჹࢅᙟᠺࡌࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊ3 㔖మ G ࢰࣤࣂࢠ㈻șࢦࣇࣗࢼࢴࢹ㸝Mgb1㸞ࡡḖ᥾ᰬࡢ௛╌ჹᙟᠺࢅ⾔
࠻ࡍࠉ᳔∸⣵⬂හ࡫ࡡ౴ථࠉវ᯹ࡵ࡚ࡀ࡝࠷ࡆ࡛࠾ࡼࠉG ࢰࣤࣂࢠ㈻࠿វ᯹࡞㔔さ࡝഼ࡀࢅᢰࡖ࡙࠷ࡾࡆ࡛
࠿᪺ࡼ࠾࡚࠵ࡾࠊ3 㔖మ G ࢰࣤࣂࢠ㈻ࡡโᚒᅄᏄ࡛ࡊ࡙▩ࡼࡿࡾ RGS ࢰࣤࣂࢠ㈻ࡢࠉȘࢦࣇࣗࢼࢴࢹࡡ୘Ὡ
ᛮ໩ࢅಀ㐅ࡌࡾࡆ࡛࡚ 3 㔖మ G ࢰࣤࣂࢠ㈻ࢅㇿ࡞โᚒࡌࡾࡆ࡛࠿ฦ࠾ࡖ࡙࠷ࡾࠊ࠷ࡵࡔ⑋Ⳟ࡞ࡢ RGS ࢰࣤࣂ
ࢠ㈻࡛┞ྜྷᛮࡡ࠵ࡾࢰࣤࣂࢠ㈻ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ࠿ࢣࢿ࣑୯࡞ 4 ಴Ꮛᅹࡊࠉࡐࡡ࠹ࡔࡡ 1 ࡗ MRGS2 ࡡ㌹෕
㔖࠿ G ࢰࣤࣂࢠ㈻șࢦࣇࣗࢼࢴࢹࡡḖ᥾ᰬ࡞࠽࠷࡙ኬࡀࡂ΅ᑛࡊ࡙࠷ࡒࠊࡐࡆ࡚ MRGS2 ࡡḖ᥾ᰬ mrgs2 ࢅ
ష⿿ࡊࡒࠊば࿰ᛮࡡ࢕ࢾࡡⴝ㠟࡞⬂Ꮔࢅ᥃⛸ࡊ࡙ 24 ᫤㛣ᚃࠉ㔕⏍ᰬ Guy11 ࡚ࡢ࡮࡛ࢆ࡜࠿ୌ⣵⬂࡫ࡡࡲࡡ
౴ථ࠿ぜࡼࡿࡒࡡ࡞ᑊࡊࠉmrgs2 ࡚ࡢኣ⣵⬂࡫ࡡ౴ථ࠿ኣࡂぜࡼࡿࡒࠊࡊ࠾ࡊࠉmrgs2 ࡡ⬂Ꮔࢅば࿰ᛮࡡ࢕ࢾ
࡞ᄂ㟕᥃⛸ࡊࡒ࡛ࡆࢀࠉᢤᢘᛮ⑋ᩤ࠿ぜࡼࡿࡒࠊࡆࡿࡼࡡ⤎ᯕࡻࡽࠉMrgs2 ࡢ࠷ࡵࡔ⑋Ⳟࡡ౴ථⳞ⣊ࡡᙟᠺ
࠽ࡻࡦ࢕ࢾࡡᢤᢘᛮཬᚺㄇᑙ࡞㛭ࢂࡖ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡾࠊ
Mrgs2, a regulator of G-protein signalling is involved in infectious hyphal development in Magnaporthe grisea.
Ayami Tomono, Eiichi Minami, Marie Nishimura (NIAS)
28
O-5
Aspergillus nidulans ࡞࠽ࡄࡾ His-Asp ᝗ሒఎ㐡⣌ࢰࣤࣂࢠ㈻ࡡ⥑⨮Ⓩよᯊ
㔘୷ாᏄ㸡ᮿᯐⰃ༡ࠉ᮶ಘᏳࠉ㕝ᮄ㯖Ꮔࠉຊ⸠㞖ኃࠉᑚᯐူኰ 㸝ྞኬ࣬⏍࿤㎨㸞
⣊≟Ⳟ Aspergillus nidulans ࡞ࡢࠉHis-Asp ࣛࣤ㓗࣭ࣛࣝᆵ᝗ሒఎ㐡ᶭᵋ࡞㛭୙ࡌࡾᅄᏄ࡛ࡊ࡙ࠉhybrid ᆵ
histidine kinase (HK) 15 ⛸㢦ࠉresponse regulator (RR) 4 ⛸㢦ࠉHK ࡛ RR 㛣ࡡࣛࣤ㓗ᇱ㌹⛛୯㛣ᅄᏄ࡛ࡊ࡙㸦⛸
㢦ࡡ HPt ࠿Ꮛᅹࡌࡾࠊ⣊≟Ⳟ࡚ࡢࡆࡿࡼࡡᅄᏄ࠿々㞟࡝ࣛࣤ㓗㌹⛛ࡡࢾࢴࢹ࣭࣠ࢠࢅᙟᠺࡊࠉᾈ㏩ᅸࡷ㎨ⷾࠉ
㓗໩ࢪࢹࣝࢪ࡝࡜ࡡᚺ➽ࡷ↋ᛮ⏍Ṣࠉ᭯ᛮ⏍Ṣࡡโᚒ࡞㔔さ࡝ᙲ๪ࢅᢰ࠹࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊᮇ◂✪ࡢ⣊≟
Ⳟ᝗ሒఎ㐡⣌ࢰࣤࣂࢠ㈻ࢅฦᏄ⏍∸ᏕⓏᡥἪ࡞ࡻࡽ⥑⨮Ⓩ࡞よᯊࡌࡾࡆ࡛࡚ࠉ々㞟࡝᝗ሒఎ㐡ࢾࢴࢹ࣭࣠ࢠ
ࡡධㇲࢅ᪺ࡼ࠾࡞ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙࠷ࡾࠊࡱࡍࠉReal Time PCR ࡞ࡻࡽධᅄᏄࡡⓆ⌟᫤᭿࡛㔖ࢅྜྷᏽࡊࠉ
ࡈࡼ࡞ GFP ࡛ࡡ⼝ྙ㐿ఎᏄࢅ⏕࠷࡙Ⓠ⌟㒂నࢅふᐳࡊࡒࠊࡱࡒࠉHK, HPt, RR ࡡࣛࣤ㓗㌹⛛ཬᚺ࡞㔔さ࡝࢓
࣐ࢿ㓗ࢅ⨠ᥦࡊࡒን␏ᰬࡡష⿿࠽ࡻࡦᶭ⬗ን໩ࡡ in vivo ふᐳ࡛⢥⿿ࢰࣤࣂࢠ㈻ࢅ⏕࠷ࡒ in vitro ࣛࣤ㓗㌹⛛ᐁ
㥺ࢅ⾔ࡖࡒࠊࡆࡿࡼࡡ⤎ᯕࡻࡽ A. nidulans ࡞࠽ࡄࡾ His-Asp ࣛࣤ㓗࣭ࣛࣝࢅ௒ࡊࡒ᝗ሒఎ㐡⣌ࡡᶭ⬗࡞ࡗ࠷
࡙⩻ᐳࡌࡾࠊ
Comprehensive analysis of signal transduction factors for His-Asp phosphorelay in Aspergillus nidulans
Kyoko Kanamaru, Yoshihiro Matsubayashi, Nobuhiro Azuma, Asako Suzuki, Masashi Kato, Tetsuo Kobayashi
(Dept of Biological Mechanisms and Functions, Nagoya Univ.)
O-6
⣊≟Ⳟ Aspergillus nidulans ࡡ㎨ⷾᚺ➽࡞࠽ࡄࡾ His-Asp ࣛࣤ㓗࣭ࣛࣝ⣌ࡡよᯊ
ⴏཋኬ♰ 1ࠉᮿᯐⰃ༡ 1ࠉ㔘୷ாᏄ 1 ࠉຊ⸠㞖ኃ 1ࠉ㜷㒂ᩏᝃ 2ࠉᑚᯐူኰ 1ࠉỀ㔕⊓ 1 㸝1 ྞኬ㝌࣬⏍࿤㎨Ꮥࠉ
2
᮶໪ኬ㝌࣬㎨Ꮥ㸞
His-Asp ࣛࣤ㓗࣭ࣛࣝ⣌ࡢࣁࢠࢷࣛ࢓࠾ࡼ㧏➴᳔∸࡞ࡱ࡚ᬉ㐚Ⓩ࡞Ꮛᅹࡌࡾ⎌ሾᚺ➽᝗ሒఎ㐡ᶭᵋ࡚࠵ࡾࠊ
⣊≟Ⳟ࡞࠽࠷࡙ࡵ࠷ࡂࡗ࠾ࡡᅄᏄࡢሒ࿈ࡈࡿ࡙࠷ࡾ࠿ࠉࡐࡡ⎌ሾᚺ➽࣒࢜ࢼࢫ࣑ࡡమ⣌Ⓩ࡝⌦よࡢࡱࡓὰ࠷ࠊ
ᠻࠍࡢ A. nidulans ࡡࢣࢿ࣑㒼า࠾ࡼࠉHis-Asp ࣛࣤ㓗࣭ࣛࣝ⣌ࡡᵋᠺᅄᏄ࡚࠵ࡾ Histidine kinase (HK)ࠉHPt
ᅄᏄࠉResponse regulator (RR)ࢅࡐࡿࡑࡿ 15 ⛸㢦ࠉ1 ⛸㢦ࠉ4 ⛸㢦ྜྷᏽࡊࠉࡆࡿࡼࡡᅄᏄࢅ⥑⨮Ⓩ࡞よᯊࡌࡾ
ࡆ࡛࡚ His-Asp ࣛࣤ㓗࣭ࣛࣝ⣌ࡡమ⣌Ⓩ࡝⌦よࢅ┘ᣞࡊ࡙࠷ࡾࠊࡆࡿࡱ࡚ࡡよᯊ࡚ࠉNik1-type ࡡ HK ࡚࠵ࡾ
NikA ࠿ࢩ࢜ࣜ࣍࢞ࢨ࢕࣐ࢺ⣌࠽ࡻࡦࣆ࢘ࢼࣜࣅ࣭ࣞࣜ⣌ࡡ㎨ⷾᚺ➽࡞㛭୙ࡌࡾࡆ࡛ࢅ᪺ࡼ࠾࡞ࡊࡒࠊࡗࡱࡽࠉ
NikA ◒ቪᰬࡢ㎨ⷾ࡞ᑊࡊ࡙⪇ᛮࢅ♟ࡊ࡙࠷ࡒࠊࡆࡡ⌟㇗ࢅ His-Asp ࣛࣤ㓗࣭ࣛࣝ⣌ࡡࢨࢡࢻࣜఎ㐡࣓ࢸࣜࢅ
ࡵ࡛࡞⩻࠻ࡾ࡛ࠉୖὮᅄᏄ࡚࠵ࡾ RR ࠿㎨ⷾᚺ➽࡞㛭୙ࡌࡾࡆ࡛࠿஢᝷ࡈࡿࡾࠊࡐࡆ࡚ࠉࡆࡿࡱ࡚࡞ཱིᚋࡊ
ࡒ㸩⛸㢦ࡡ RR ◒ቪᰬ࠽ࡻࡦኣ㔔 RR ◒ቪᰬࢅ⏕࠷࡙㎨ⷾវུᛮࢅよᯊࡊࡒ࡛ࡆࢀࠉSskA ࡛ SrrA ࡡ஦㔔◒ቪ
ᰬ࠿㎨ⷾ⪇ᛮࢅ♟ࡌࡆ࡛࠿ࢂ࠾ࡖࡒࠊࡆࡡࡆ࡛࠾ࡼ㎨ⷾវུᛮ࡞ࡢ SskA ౪ᏋⓏ⤊㊪࡛ SrrA ౪ᏋⓏ⤊㊪ࡡ୦
᪁࠿ᐞ୙ࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡒࠊࡆࡿࡱ࡚࡞ SskA ࡢ Hog1-type MAPK (HogA)࢜ࢪࢢ࣭ࢺࢅโᚒࡊ࡙࠷ࡾࡆ
࡛࠿ࢂ࠾ࡖ࡙࠽ࡽࠉ⌟ᅹࠉ㎨ⷾࡡ῟ຊ࡞ࡻࡾ HogA MAPK ࡡὩᛮࡷࠉࡐࡡୖὮโᚒᅄᏄࡡⓆ⌟ࣂࢰ࣭ࣤ࡝࡜
ࢅよᯊ୯࡚࠵ࡾࠊࡆࡿࡼࡡ⤎ᯕࢅ㊻ࡱ࠻࡙ࠉ⣊≟Ⳟࡡ㎨ⷾᚺ➽ᶭᵋࡡධమാ࡞ࡗ࠷࡙⩻ᐳࡊ࡙ࡲࡾࠊ
Analysis of the His-Asp phosphorelay signaling pathway in response to fungicide in Aspergillus nidulans
Daisuke Hagiwara, Yoshihiro Matsubayashi, Kyoko Kanamaru, Masashi Kato, Tetsuo Kobayashi, Takeshi Mizuno
(Grad. Sch. Bioagricultural Sci. Nagoya Univ.)
29
O-7
Aspergillus nidulans HOG ⤊㊪≁␏Ⓩ࡝࣭ࣝ࣎ࢰ࣭㐿ఎᏄⓆ⌟⣌ࡡᵋ⠇
ཿᕖ೸ኯ㑳ࠉཿᕖ㈏Ꮔࠉ᫅⏜⣎Ꮔࠉఫ⸠ዄ὘Ꮔࠉྚぜၤ*ࠉ⸠ᒱᬓ์ࠉỀㆺ἖ࠉ㜷㒂ᩏᝃ
(᮶໪ኬ㝌㎨࣬ᚺ⏍⛁ࠉ*᮶໪ኬ࣬ᮅᮮ◂)
ᠻࠍࡢࡆࡿࡱ࡚࡞ A. nidulans ࡡ㧏ᾈ㏩ᅸᚺ➽ HOG (high-osmolarity glycerol response) ⤊㊪ (AnHOG ⤊
㊪) ࡡධᐖよ᪺ࡡࡒࡴࠉ⤊㊪ᵋᠺ㐿ఎᏄࡡ༟㞫ࠉ㓕ẍ┞ྜྷ㐿ఎᏄ࡛ࡡẒ㍉よᯊࠉ㐿ఎᏄ◒ቪᰬࢅ⏕࠷ࡒ㐿ఎ
ᏕⓏよᯊࢅ⾔࠷ࠉAnHOG ⤊㊪ࡡ㐵᩷ࡷᜇᖏⓏὩᛮ໩࠿Ⳟమࡡ⏍⫩࡞ᝇᙫ㡢ࢅཀྵ࡯ࡌࡆ࡛ࢅぜฝࡊࡒࠊິ᳔
∸វ᯹⣊≟Ⳟ (A. fumigatus ࡷ Magnaporthe grisea ࡝࡜) ࡵ┞ྜྷ⤊㊪ࢅ᭯ࡌࡾࡆ࡛࠾ࡼࠉࡆࡡ⤊㊪ࡡິឺࢅ◂
✪ࡌࡾࡒࡴࡡ࣭ࣝ࣎ࢰ࣭ࢅ㛜Ⓠࡌࡾࡆ࡛࡛ࡊࡒࠊA. nidulans 㔕⏍ᰬཀྵࡦࠉAnhogA ◒ቪᰬࡡᾈ㏩ᅸๆ⃥᫤ࡡ㐿
ఎᏄⓆ⌟ᚺ➽よᯊ࠾ࡼࠉ࣭ࣝ࣎ࢰ࣭ು⿭㐿ఎᏄࢅ々ᩐ㐽ᢝࡊࡒࠊAnHOG ⤊㊪࡞ᨥ㒼ࡈࡿࡾ࡛஢᝷ࡈࡿࡾ㐿
ఎᏄࡡࣈ࣓࣭ࣞࢰ࣭㡷ᇡࢅ࣭ࣝ࣎ࢰ࣭㐿ఎᏄ࡞㏻⤎ࡊࠉ⤊㊪≁␏Ⓩ࡝ࢪࢹࣝࢪๆ⃥᫤࡞࣭ࣝ࣎ࢰ࣭ࡡὩᛮ࠿
୕᪴ࡌࡾࡆ࡛ࢅᣞᵾ࡛ࡊࡒࠊು⿭㐿ఎᏄࡡࣈ࣓࣭ࣞࢰ࣭㡷ᇡࢅ༟㞫ࡊࠉGUS 㐿ఎᏄ࡞㏻⤎ࡊࡒࡵࡡࢅ A.
nidulans ࡞ᑙථࡊࡒࠊ㧏ᾈ㏩ᅸๆ⃥ࡷ⤊㊪Ὡᛮ໩๠ฌ⌦࡞ࡻࡖ࡙࣭ࣝ࣎ࢰ࣭ࡡὩᛮ࠿୕᪴ࡊࠉAnHOG ⤊㊪
ࢅ㐵᩷ࡌࡾࡻ࠹࡝ን␏మࢅᐙ୹࡞ࡊࡒሔྙ࡞ࡢὩᛮࡢධࡂ୕᪴ࡊ࡝࠾ࡖࡒࠊ௧୕ࡡ⤎ᯕ࠾ࡼࠉᮇ࣭ࣝ࣎ࢰ࣭
Ⓠ⌟⣌ࡢ AnHOG ⤊㊪ࡡὩᛮິឺࢅ࣓ࢼࢰ࣭ࡌࡾ⣌࡛ࡊ࡙ MAP kinase HogA ࡡࣛࣤ㓗໩᳠ฝ࡛୩ࢆ࡚᭯ຝ
࡚࠵ࡾࡆ࡛࠿☔ヾࡈࡿࡒࠊ
Construction of a reporter gene expression system for the Aspergillus nidulans HOG pathway
Kentaro Furukawa, Takako Furukawa, Yukiko Hoshi, Natsuko Sato, Akira Yoshimi*, Tomonori Fujioka, Osamu
Mizutani and Keietsu Abe (Tohoku Univ., Grad. Sch. Agri. Sci., *Tohoku Univ., NICHe)
O-8
Aspergillus oryzae ࡞࠽ࡄࡾ steA ࣓࣌ࣞࢡࡡᶭ⬗よᯊ
᲻⏛ᐰெ,➁හ㐠㞕㸝᮶ா㎨ᕝኬ㝌࣬⏍∸ᕝᏕ㸞
A.nidulans ࡡ steA ◒ቪᰬ࡚ࡢ᭯ᛮ⬂Ꮔࡡᙟᠺ࠿ᢒโࡈࡿࡾࡆ࡛࠾ࡼࠉsteA ࡢ᭯ᛮ⬂Ꮔࡡᙟᠺࢅโᚒࡌࡾ㌹෕
ㄢ⟿ᅄᏄࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ࡛ࡊ࡙ሒ࿈ࡈࡿ࡙࠷ࡾࠊA. oryzae ࡡࢣࢿ࣑よᯊࡡ⤎ᯕࠉ᭯ᛮୠ௥ࢅᣚࡒ࡝࠷ A.
oryzae ࠾ࡼࡵ steA ࠿ぜ࠷ฝࡈࡿࡒࠊࡐࡆ࡚ࠉA. oryzae ࡞࠽ࡄࡾ steA ࡡᶭ⬗ࢅㄢ࡬ࡾࡒࡴࠉsteA 㐛๨Ⓠ⌟ᰬ࡛
◒ቪᰬࢅష⿿ࡊࠉ⾪⌟ᆵࢅふᐳࡊࡒࠊࡐࡡ⤎ᯕࠉsteA 㐛๨Ⓠ⌟ᰬࡢ㔕⏍ᰬ࡞Ẓ࡬࡙Ⳟ⣊ࡡ⏍⫩ᢒโ࡛ฦ⏍Ꮔ
ᙟᠺࡡᢒโ࠿ヾࡴࡼࡿࠉᾦమᇰ㣬᫤ࡡⳞ⣊࡞ࡢࠉࣁ࣭ࣜࣤ≟࡞⫟ኬࡊࡒ⣵⬂࠿ふᐳࡈࡿࡒࠊࡱࡒࠉsteA 㐛๨
Ⓠ⌟ᰬࡢ Yatalase ➴ࡡ⣵⬂ባ⁈よモⷾ࡞ᑊࡌࡾវུᛮ࠿୕᪴ࡊ࡙࠷ࡒࠊࡈࡼ࡞ࠉᾦమᇰ㣬᫤࡞࠽ࡄࡾฦἢࢰ
ࣤࣂࢠ㈻࡞ࡗ࠷࡙࣋ࣈࢲࢺ࣏ࢪࣆ࢔࣭ࣤ࢝ࣈࣛࣤࢹἪ࡚よᯊࡊࡒ࡛ࡆࢀࠉș-1,3-glucanase ➴ࡡ⣵⬂ባ⁈よ㓕
⣪ࡡฦἢ࠿୕᪴࠷࡙࠷ࡒࠊࡆࡡࡒࡴࠉࡆࡿࡼࡡ⣵⬂ባ⁈よ㓕⣪ࡡᙫ㡢࡚ steA 㐛๨Ⓠ⌟ᰬࡡ⣵⬂ባ࡞␏ᖏ࠿ᘤ
ࡀ㉫ࡆࡈࡿ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡒࠊ㓕ẍ࡚ࡢș-1,3-glucanase ࠿ࢪࢷ࣭ࢩ≁␏Ⓩ࡞⣵⬂ባࡡྙᠺ࡞㛭୙ࡊ࡙࠷ࡾࠊ
A. oryzae ࡡ steA ࡢᾦమᇰ㣬᫤࡞㌹෕ࡈࡿ࡙࠷ࡒࡆ࡛࠾ࡼࠉⳞ⣊ࡡఘ㛏࡞ఔ࠹⣵⬂ባࡡྙᠺࡱࡒࡢ්ᵋ⠇࡞㛭
୙ࡊ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊୌ᪁ࠉRT-PCR ࡞ࡻࡽ steA ࠿㌹෕ࡈࡿ࡙࠷࡝࠷ࡆ࡛ࢅ☔ヾࡊࡒ◒ቪᰬ࡚ࡢࠉ
⾪⌟ᆵࠉฦἢࢰࣤࣂࢠ㈻ࣂࢰ࣭ࣤ࡞㔕⏍ᰬ࡛ࡡᕣ␏ࡢヾࡴࡼࡿ࡝࠾ࡖࡒࠊ
Function analysis of steA in Aspergillus oryzae
Hiroto Morita, Michio Takeuchi
(Dept. of Biochemistry and Biotechnology, Tokyo Univ. of Agriculture and Technology)
30
O-9
Aspergillus nidulans ࡡᙟ㈻㌹ᥦ᫤࡞ᚪさ࡛ࡈࡿࡾฦᏄ࠽ࡻࡦ᧧ష࡞ࡗ࠷࡙ࡡ᳠ ゞ
᱋㔕ူ▦ 1, ౪⏛ᗛᏏ 2, ௿⸠㟱ኰ 2࣬3 㸝1 ಘኬ࣬⌦ࠉ2 ಘኬ㝌࣬ᕝࠉ3 ಘኬ࣬ධᏕᩅ⫩ᶭᵋ㸞
⣊≟Ⳟࡡࣈࣞࢹࣈࣚࢪࢹࢅ⏕࠷ࡒᙟ㈻㌹ᥦ࡞࠽࠷࡙ࠉ῟ຊࡌࡾ DNA ฦᏄ࠿᯹Ⰵమ࡞⤄ࡲ㎲ࡱࡿࡾࡱ࡚ࡡ
㐛⛤ࡢࠉ(1)ཋᙟ㈻⭯ࡡ㏳㐛ࠉ(2)⣵⬂㈻ࡡ㏳㐛ࠉ(3)ᰶ⭯ࡡ㏳㐛ࠉ࠽ࡻࡦ(4)᯹Ⰵమ࡫ࡡ⤄㎲ࡲࡡ 4 ṹ㝭࡞ኬื࡚
ࡀࡾࠊᮇ◂✪࡚ࡢࠉࡆࡡ࠹ࡔࠉཋᙟ㈻⭯ࡡ㏳㐛࡞ࡗ࠷࡙᳠ゞࡊࡒࠊᮞᩩ࡛ࡊ࡙ Aspergillus nidulans ࢅ⏕࠷ࡒࠊ
pH ࡛ᾈ㏩ᅸࢅㄢᩒࡊࠉᐄῺ࡚ࣈࣞࢹࣈࣚࢪࢹᠩ⃦ᾦࢅᚋࡒࠊ࢙࣎ࣛࢲࣝࣤࢡࣛࢤ࣭ࣜ(PEG)࡛࢜ࣜࢨ࣑ࢗ࢕
࢛ࣤ(Ca2+)࡛࡛ࡵ࡞ DNA ฦᏄࢅຊ࠻ࡒṹ㝭࡚ࡡ᧘ᢶ࠽ࡻࡦᕵ㔐᧧షࡢࠉᙟ㈻㌹ᥦຝ⋙࡞ᙫ㡢ࡊ࡝࠾ࡖࡒࠊPEG
ࡢࣈࣞࢹࣈࣚࢪࢹࡡ⼝ྙࢅಀ㐅ࡌࡾࠊࡱࡒࠉᮇⳞࡡⳞ⣊⣵⬂ࡢኣᰶ࡚࠵ࡽࠉ⼝ྙᚃࡡ⣵⬂࡞࠽ࡄࡾᰶࡡ්ฦ
㒼࠿ၡ㢗࡛࡝ࡼ࡝࠷ࠊࡐࡆ࡚ࠉ⼝ྙ࡛ DNA ฦᏄࡡཱིࡽ㎲ࡲ࡞ࡗ࠷࡙᳠ゞࡊࡒ࠿ࠉࡆࡿࡼࡢ┞㛭ࡊ࡝࠾ࡖࡒࠊ
ḗ࡞ࠉDNA ฦᏄ࡛ࣈࣞࢹࣈࣚࢪࢹࡡ┞பష⏕࡞ᑊࡌࡾࠉPEG ࡡᙲ๪࡞ࡗ࠷࡙᳠ゞࡊࡒࠊCa2+ࢅຊ࠻࡝࠷ሔྙࠉ
ᅂ཭ࡊࡒࣈࣞࢹࣈࣚࢪࢹ⏤ฦ࠾ࡼࠉ῟ຊࡊࡒ DNA ฦᏄࡢ᳠ฝࡈࡿ࡝࠾ࡖࡒࠊPEG ࢅ࢙࣎ࣛࢲࣝࣤ࢕࣐ࣤ(PEI)
࡞௥࠻ࡒሔྙ࡞ࡢ᳠ฝࡈࡿࡒࡆ࡛࠾ࡼࠉ∸⌦Ⓩ࡝┞பష⏕࡛࠷࠹Ⅴ࡚ࡢࠉDNA ฦᏄ࡛ཋᙟ㈻⭯㛣ࡡ㞹ẴⓏᩲ
ງࡡ୯࿰ຝᯕࡡ㔔さᛮ࠿♟ြࡈࡿࡒࠊࡈࡼ࡞ࠉCa2+ࡢࠉ࣏ࢡࢾࢨ࣑ࢗࠉࢼࢴࢢࣜ࢕࢛ࣤ➴࡚௥᭨ྊ⬗࡚࠵ࡖ
ࡒࡆ࡛࠾ࡼࠉ≁␏Ⓩ࡝ࢨࢡࢻࣜฦᏄ࡛ࡊ࡙࢙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪࡡㄇᑙ➴ࢅ௒ࡊ࡙ DNA ฦᏄࡡཱིࡽ㎲ࡲ࡞㛭୙
ࡊ࡙࠷ࡾྊ⬗ᛮࡢ఩࠷࡛⩻࠻ࡼࡿࡒࠊࡳࡊࢀ PEG ࠽ࡻࡦ PEI ࡡ⣵⬂Ẑᛮ࡞ᑊࡌࡾಕ㆜ຝᯕࡡ᪁࠿㔔さ࠾ࡵ▩
ࡿ࡝࠷ࠊ
Extracellular DNA uptake during genetic transformation of Aspergillus nidulans
Tetsuya Kuwano1, Yasutaka Yoda2, Yasuo Itoh2࣬3 (1Fac. Science, 2Grad.Sch. Dev. Sci., 3Sch. Gen. Ed. / Shinshu Univ.)
O-10
⣊≟Ⳟ Aspergillus nidulans ࡞࠽ࡄࡾ࢞ࢲࢻ࣭ࢭ㐿ఎᏄ⩄ࡡⓆ⌟よᯊ
⏛୯ᙤ㸡ᒜᓧᬍ୓ࠉᇷහ⿩஄ࠉኯ⏛᪺ᚠ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
⣊≟Ⳟ࡞࠽࠷࡙ࠉࡐࡡ⣵⬂ባࡡ୹さᵋᠺᠺฦ࡚࠵ࡾ࢞ࢲࣤࢅฦよࡌࡾ㓕⣪࢞ࢲࢻ࣭ࢭࡢᙟឺᙟᠺ࣬⥌ᣚ࡞㔔
さ࡝ᙲ๪ࢅᯕࡒࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡾࠊA. nidulans ࡡࢣࢿ࣑࡞ࡢࠉ⢶㈻ฦよ㓕⣪ฦ㢦ࣆ࢒࣐࣭ࣛ18 ࡞ᒌࡌࡾ
࢞ࢲࢻ࣭ࢭࡡὩᛮ୯ᚨ࡛┞ྜྷᛮࡡ࠵ࡾ࢓࣐ࢿ㓗㒼าࢅࢤ࣭ࢺࡌࡾ㡷ᇡ࠿ 18 ࣧᡜᏋᅹࡌࡾࡆ࡛࠿᪺ࡼ࠾࡞࡝
ࡖ࡙࠷ࡾࠊࡆࡡ 18 ⛸ࡡ࢞ࢲࢻ࣭ࢭ㐿ఎᏄ㸝chiAࠤR㸞ࡢࡐࡿࡼ࠿ࢤ࣭ࢺࡌࡾ࢓࣐ࢿ㓗㒼า࠾ࡼࡈࡼ࡞㸧ࡗࡡ
ࢠࣚࢪ࡞ฦ㢦ࡈࡿࠉ chiAࠉH ࠉR ࡢ class IIIࠉṟࡽࡡ 15 ⛸ࡢ class V ࡡ࢞ࢲࢻ࣭ࢭࢅࢤ࣭ࢺࡌࡾ࡛⩻࠻ࡼࡿࡾࠊ
௑ᅂࠉࡆࡿࡼࡡ࢞ࢲࢻ࣭ࢭ㐿ఎᏄ࡞ࡗ࠷࡙ࣛ࢓ࣜࢰ࢕࣑ RT-PCR Ἢࢅ⏕࠷࡙Ⓠ⌟よᯊࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉ
chiD ࡛ chiH ࡡ㌹෕⏐∸ࡢฦ⏍Ꮔᙟᠺㄇᑙ᮪௲ୖ࡚ࡐࡡ㔖࠿᛬⃥࡞ቌຊࡊࡒࠊࡱࡒࠉࡆࡿࡼ㌹෕⏐∸ࡡ㔖ࡢฦ
⏍ᏄࡡⓆⰾࢅㄇᑙࡌࡾ࡛᫤㛣࡛භ࡞఩ୖࡊࡒࠊࡆࡿࡼࡡࡆ࡛࠾ࡼ chiD ࠉH ࡢฦ⏍Ꮔᙟᠺ࡞㛭ࢂࡾᶭ⬗ࢅᣚࡗ
ࡆ࡛࠿♟ြࡈࡿࡒࠊୌ᪁ࠉchiA ࡡ㌹෕⏐∸ࡡ㔖ࡢฦ⏍Ꮔ⮤మ࡚ࡢᑛ࡝࠷࠿Ⓠⰾㄇᑙ᮪௲࡞ࢨࣆࢹᚃ࡞᛬⃥࡝
ቌຊ࠿ぜࡼࡿࡒࠊࡆࡡࡆ࡛࠾ࡼ chiA ࡡฦ⏍ᏄࡡⓆⰾ࡞࠽ࡄࡾᶭ⬗࠿♟ြࡈࡿࡒࠊ
Expression of chitinase genes in Aspergillus nidulans
Aya Tanaka, Harutake Yamazaki, Hiroyuki Horiuchi and Akinori Ohta
(Dept.of Biotechnol., Univ. of Tokyo)
31
O-11
》Ⰵ⭁ᮑⳞ Coniophora puteana ࡞ࡻࡾ᳔∸⣵⬂ባ⏜ᮮኣ⢶ࡡฦよ࣒࢜ࢼࢫ࣑
ຊ἖ఫ ᖲ㸡஫༎ᔋᅽ᪝Ꮔ㸡㩢ᓞḿᾀ㸝᮶ኬ㝌࣬㎨⏍⛁࣬⏍ᮞ⛁㸞
ᮄᮞ⭁ᮑⳞࡢࠉୌ⯙Ⓩ࡞⭁ᮑᚃ࡞࠽ࡄࡾᮞࡡⰅ࠾ࡼⓉⰅ⭁ᮑⳞ࡛》Ⰵ⭁ᮑⳞ࡞ฦ㢦ࡈࡿࡾࠊⓉⰅ⭁ᮑⳞ࠿
᳔∸⣵⬂ባࡡ୹ᠺฦ࡚࠵ࡾࢬ࣭ࣜࣞࢪࠉࣉ࣐ࢬ࣭ࣜࣞࢪ࠽ࡻࡦࣛࢡࢼࣤࢅฦよ࡚ࡀࡾࡡ࡞ᑊࡊ࡙ࠉ》Ⰵ⭁ᮑ
Ⳟࡢࣉ࣐ࢬ࣭ࣜࣞࢪ࡛㟸ᬏᛮࢬ࣭ࣜࣞࢪࢅ≁␏Ⓩ࡞ฦよࡊࠉᮄᮞࡡᙁᗐࢅⴥࡊࡂ఩ୖࡈࡎࡾࡆ࡛࠿▩ࡼࡿ࡙
࠷ࡾࠊᢰᏄⳞ Coniophora puteana㸝࢕ࢺࢰࢢ㸞ࡢࠉᮄ㏸పᏩࢅ຋໩ࡈࡎࡾ》Ⰵ⭁ᮑⳞࡡୌ⛸࡚࠵ࡾ࠿ࠉୌ⯙Ⓩ
࡝》Ⰵ⭁ᮑⳞ࡛ࡢ␏࡝ࡽࠉ⢶㈻ຊỀฦよ㓕⣪㸝GH㸞ࣆ࢒࣐࣭ࣛ7 ࡞ᒌࡌࡾࢬࣞࣄ࢛ࣃࢺ࣭ࣞࣚࢭ㸝Cel7㸞ࢅ
Ⳟమአ࡞⏍⏐ࡌࡾࡆ࡛࠿ሒ࿈ࡈࡿ࡙࠷ࡾࠊᮇ㓕⣪ࡢࠉᏄᄙⳞࡷⓉⰅ⭁ᮑⳞ࡞࠽࠷࡙ࡢ⤎ᬏᛮࢬ࣭ࣜࣞࢪࡡฦ
よ࡞࠽࠷࡙㔔さ࡝ᙲ๪ࢅᢰࡖ࡙࠷ࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾ࠿ࠉ》Ⰵ⭁ᮑⳞ࡞࠽ࡄࡾᶭ⬗ࡢ୘᪺࡚࠵ࡾࠊࡐࡆ࡚
ᮇ◂✪࡚ࡢ C. puteana ࠿⏍⏐ࡌࡾ Cel7 ࡡ࢞ࣔࣚࢠࢰࣚ࢕ࢫࢅ⾔ࡖࡒࠊ
ࢬ࣭ࣜࣞࢪࢅ⅛⣪″࡛ࡊࡒ࡛ࡀ࡞ C. puteana ࠿⏍⏐ࡌࡾ 2 ⛸㢦ࡡ Cel7 ࢅ࣑࢜ࣚࢠ࣏ࣞࢹࢡࣚࣆ࢔࣭࡞ࡻࡖ
࡙⢥⿿ࡊὩᛮࢅῼᏽࡊࡒ࡛ࡆࢀࠉ୦㓕⣪࡛ࡵ⤎ᬏᛮࢬ࣭ࣜࣞࢪࢅฦよ࡚ࡀ࡝࠾ࡖࡒࠊࡱࡒࠉ୦㓕⣪ࢅࢤ࣭ࢺ
ࡌࡾ cDNA ࢅࢠ࣭ࣞࢼࣤࢡࡊࠉ᥆ᏽ࢓࣐ࢿ㓗㒼าࢅ௙ࡡ⣊≟Ⳟ⏜ᮮࡡ Cel7 ࡛Ẓ㍉ࡊࡒ࡛ࡆࢀࠉC. puteana ⏜
ᮮࡡ㓕⣪࡞ࡢ⢶㈻⤎ྙ࣓ࢩ࣭ࣖࣜ㸝CBM㸞࠿Ꮛᅹࡊ࡝࠾ࡖࡒࡆ࡛࠾ࡼࠉ୦㓕⣪ࡢ CBM ࠿Ḗዯࡊ࡙࠷ࡾࡒࡴ
࡞⤎ᬏᛮࢬ࣭ࣜࣞࢪࢅฦよ࡚ࡀ࡝࠷࡛⩻࠻ࡼࡿࡒࠊࡐࡆ࡚௙ࡡ᳔∸⣵⬂ባ⏜ᮮࡡኣ⢶ࢅᇱ㈻࡛ࡊ࡙Ὡᛮῼᏽ
ࢅ⾔ࡖࡒ࡛ࡆࢀࠉ࡜ࡔࡼࡡ㓕⣪ࡵࢡࣜࢤ࣏ࣤࢻࣤ࡞ᑊࡌࡾὩᛮ࠿㧏࠾ࡖࡒࠊࡆࡿࡱ࡚ GH ࣆ࢒࣐࣭ࣛ7 ࡞ᒌࡌ
ࡾࢡࣜࢤ࣏ࣤࢻࢻ࣭ࢭࡢሒ࿈ౚ࠿࡝࠷ࡆ࡛࠾ࡼࠉᮇ㓕⣪ࡢ᩺ぞࢡࣜࢤ࣏ࣤࢻࢻ࣭ࢭ࡚࠵ࡾ࡛⩻࠻ࡼࡿࠉC.
puteana ࡞ࡻࡾ᳔∸⣵⬂ባ⏜ᮮኣ⢶ࡡฦよࢅ≁ᚡࡘࡄࡾ㓕⣪࡚࠵ࡾ࡛⩻࠻ࡼࡿࡒࠊ
Degradation of plant cell wall polysaccharide by brown-rot fungus Coniophora puteana
Taira Kajisa, Kiyohiko Igarashi,and Masahiro Samejima (Dept. Biomat. Sci., Univ. Tokyo)
O-12
Fusarium oxysporum f. sp. melonis ⏜ᮮࡡβ-1,6-࢝ࣚࢠࢰࢻ࣭ࢭࡡ⢥⿿࡛ࢠ࣭ࣞࢼࣤࢡ
ㆺཾ㞕သ㸡㜨ᮇ㱗ྒྷ,ᕖᓧ᮶ᙢ㸝ኬ㜨ᗋ❟ኬ࣬⏍࿤⎌ሾ㸞
㎾ᖳࠉII ᆵ࢓ࣚࣄࢿ࢝ࣚࢠࢰࣤࡢ NK ⣵⬂Ὡᛮ໩ࡷࢤࣝࢪࢷ࣭ࣞࣜྺ཭㜴ᐐ࡝࡜⛸ࠍࡡ⏍⌦Ὡᛮࢅᣚࡗࡆ
࡛࠾ࡼࠉ༈ⷾဗࡷࢦࣈ࣒ࣛࣤࢹ࡛ࡊ࡙ࡡฺ⏕࠿᭿ᙽࡈࡿ࡙࠷ࡾࠊࡊ࠾ࡊࠉII ᆵ࢓ࣚࣄࢿ࢝ࣚࢠࢰࣤฦよ㓕⣪
࡞㛭ࡌࡾ◂✪ࡢᑛ࡝ࡂࠉࡐࡡࡻ࠹࡝㓕⣪ࡢ II ᆵ࢓ࣚࣄࢿ࢝ࣚࢠࢰࣤࡡᵋ㏸よᯊࡷ⏍⌦ὩᛮⓆ⌟࡞࠾࠾ࢂࡾ⢶
㙈ᵋ㏸ࡡỬᏽ࡞࠽࠷࡙ᙁງ࡝ᡥṹ࡛࡝ࡾࠊࡐࡆ࡚ᮇ◂✪࡚ࡢࠉ࣏࢜ࣚࢵ⏜ᮮ II ᆵ࢓ࣚࣄࢿ࢝ࣚࢠࢰࣤ㸝LWAG㸞
ฦよ㓕⣪⏍⏐Ⳟࡡࢪࢠ࣭ࣛࢼࣤࢡࢅ⾔࠷ࠉ㓕⣪ࡡ⢥⿿࠽ࡻࡦࡐࡡ㐿ఎᏄࡡࢠ࣭ࣞࢼࣤࢡࢅモࡲࡒࠊ
ᅰቫࡻࡽ༟㞫ࡊࡒࢦࣤࣈࣜࢅᑊ㇗࡞ࠉࡐࡡ୕⃀ࢅ⢊㓕⣪ᾦ࡛ࡊ࡙⏕࠷ LWAG ࠾ࡼࡡฦよ⏐∸ࢅᣞᵾ࡞ࢪ
ࢠ࣭ࣛࢼࣤࢡࢅ⾔ࡖࡒࠊฦよ⏐∸ࡡฦᯊࡢࠉ㧏ᛮ⬗㝔࢕࢛ࣤஹᥦࢠ࣏ࣞࢹࢡࣚࣆ࢔࣭࡞ࡻࡽ⾔ࡖࡒࠊࡐࡡ⤎
ᯕࠉ୹さฦよ⏐∸࡛ࡊ࡙࢓ࣚࣄࢿ࣭ࢪࠉ࢝ࣚࢠࢹ࣭ࢪ࡛ኣ㔖ࡡ࢛ࣛࢥ⢶ࢅ⏍ᠺࡊࡒ Fusarium oxysporum f. sp.
Melonis 12S ᰬࢅ┘ⓏࡡⳞ࡛ࡊ࡙㐽ᢝࡊࡒࠊ
ࡱࡍࠉᮅ▩࢛ࣛࢥ⢶㐗㞫㓕⣪(FoGal1)ࡡ⢥⿿ࢅ⾔ࡖࡒࠊᇰ㣬୕⃀ࢅ࡞ࡻࡽ⢥⿿ࡊࠉSDS-PAGE ࡞࡙㞹Ẵὃ
ິⓏ࡞ᆍୌ࡝ᵾဗࢅᚋࡒࠊḗ࡞ࠉᮇ㓕⣪ࡡష⏕࡞ࡻࡽ LAG ࠾ࡼ㐗㞫ࡌࡾ࢛ࣛࢥ⢶ࢅࢣࣜࢀ㐛ࢅ⏕࠷࡙⢥⿿ࡊࠉ
ᵋᠺ⢶ฦᯊࠉTOF-MSࠉH/C-NMR ࡞ࡻࡽβ-1,6-࢝ࣚࢠࢹࣄ࢛࣭ࢪ࡚࠵ࡾ࡛Ửᏽࡊࡒࠊࡆࡡࡆ࡛࠾ࡼ FoGal1 ࡢ
LWAG ࡻࡽ≁␏Ⓩ࡞ഁ㙈ࢅวࡽฝࡌࡆ࡛࠿࡚ࡀࡾ㓕⣪࡚࠵ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊ
ḗ࡞ࠉFoGal1 ࡡ N ᮆ❻࢓࣐ࢿ㓗㒼า᝗ሒ࠾ࡼష⿿ࡊࡒࣈࣚ࢕࣏࣭ࢅ⏕࠷࡙ࠉfogal1 㐿ఎᏄࡡධ㛏 cDNA ࢅ
⋋ᚋࡊࡒࠊ⋋ᚋࡊࡒ fogal1 㐿ఎᏄࢅኬ⭘Ⳟ࡞ᙟ㈻㌹ᥦࡊࠉ⤄ᥦ࠻㓕⣪ࢅ⋋ᚋࡊࡒࠊ
Purification and gene cloning of β-1,6-galactanase of Fusarium oxysporum f. sp. melonis
Yuya Taniguchi, Tatsuji Sakamoto, Haruhiko Kawasaki
(Graduate School of Life and Environmental Sciences, Osaka Prefecture Univ.)
32
O-13
ࣛ࣍ࣆࣚࣄࣤ⏍⏐⣊≟Ⳟ Ashbya gossypii ࡡ௥ㅨよᯊ
㔘ᨳ ┷ 1㸡⏛ᓞ ㅅ 1㸡ᮌ 㱗ὒ㸦㸡2㸝1 㟴ᒱኬ࣬㎨࣬ᚺ⏍໩㸡2 㟴ᒱኬ࣬๭⛁ᢇ㝌࣬⤣ྙࣁ࢕࢛㸞
A. gossypii ࡢ⢶㈻ࡓࡄ࡚࡝ࡂ㸡᳔∸ἔࡷἔ⬙⣌ᗣᲘ∸ࢅ㈠໩ࡊ࡙ࣛ࣍ࣆࣚࣄࣤ㸝ࣄࢰ࣐ࣤ B2㸞࡞ንᥦ࡚ࡀ
ࡾࡒࡴ㸡ἔ⬙⣌ᗣᲘ∸ฺ⏕ࡡふⅤ࠾ࡼ㔔さ࡝Ⳟ࡚࠵ࡽ㸡ୠ⏲ࡡࣛ࣍ࣆࣚࣄࣤࡡ㸨๪௧୕࠿ᮇⳞ࡞ࡻࡖ࡙⏍⏐
ࡈࡿ࡙࠷ࡾࠊࡆࡿࡱ࡚࡞㸡ᙔ◂✪ᐄ࡞࠽࠷࡙ን␏ฌ⌦࡞ࡻࡽࣛ࣍ࣆࣚࣄࣤ㧏⏍⏐ᰬࢅཱིᚋࡊ࡙࠷ࡾࠊ௑ᅂ㸡
ን␏ᰬࡡࣛ࣍ࣆࣚࣄࣤྙᠺ㓕⣪㐿ఎᏄࡡ㌹෕㔖ࢅㄢ࡬ࡒ࡛ࡆࢀ㸡࢕ࢮࢠ࢙ࣤ㓗ࣛ࢓࣭ࢭ㐿ఎᏄ㸝ICL1㸞ཀྵࡦ
࢕ࢮࢠ࢙ࣤ㓗⬲Ề⣪㓕⣪㐿ఎᏄ㸝ICDH1㸞ࡡ㌹෕㔖࠿ቌኬࡊ࡙࠷ࡒࠊࡐࡆ࡚㸡ᮇⳞ࡞࠽࠷࡙୦㐿ఎᏄࢅࡐࡿ
ࡑࡿ◒ቪࡊ㸡ࣛ࣍ࣆࣚࣄࣤ⏍⏐ᛮ࡞୙࠻ࡾᙫ㡢ࢅㄢ࡬ࡒࠊ࢜ࢻ࣏࢕ࢨࣤ⪇ᛮ㐿ఎᏄࢅ࣏࣭࣭࡛࢜ࡊࠉ୦❻࡞
㐿ఎᏄ⨠ᥦࡡࡒࡴࡡ┞ྜྷ㒼าࢅᣚࡗ㐿ఎᏄ◒ቪ⏕࢜ࢬࢴࢹࢅ PCR ࡞࡙ᵋ⠇ࡊ㸡⣵⬂࡞ᑙථࡊࡒࠊ┘ᵾ㡷ᇡ࡚
ࡡ⨠ᥦ࠿☔ヾ࡚ࡀࡒᰬࢅ㐿ఎᏄ◒ቪᰬ࡛ࡊࡒࠊ୦◒ቪᰬࢅ⳧⛸ἔࢅ⅛⣪″࡞ᇰ㣬ࡊ࡙ฦᯊࡊࡒ⤎ᯕ㸡ࣛ࣍ࣆ
ࣚࣄࣤ⏍⏐㔖ࡢ㔕ᛮᰬ࡛Ẓࡊ࡙ ICDH1 ◒ቪᰬ࡚ࡢ 4.7㸚㸡ICL1 ◒ቪᰬ࡚ࡢ 27.7㸚࡛㢟ⴥ࡞఩ୖࡊࡒࠊࡆࡡࡆ
࡛࠾ࡼ㸡୦㓕⣪ࡢࣛ࣍ࣆࣚࣄࣤ⏍ྙᠺ࡞࠽࠷࡙㔔さ࡚࠵ࡾࡆ࡛࠿ࢂ࠾ࡖࡒࠊ
Functional analyses of metabolic enzymes for riboflavin synthesis in Ashbya gossypii
Shin Kanamasa1, Satoshi Tajima1, Enoch Y. Park1,2
(1Dept. Appl. Biol. Chem., Fac. Agric., Shizuoka Univ., 2Integ. Biosci., Grad. Sch. Sci. Technol., Shizuoka
Univ.)
O-14
》Ⰵ⭁ᮑⳞ࢛࢛ࢗࢫࣚࢰࢢ࡞࠽࠷࡙ࢨࣖࢗ㓗⪇ᛮ࡞ᐞ୙ࡌࡾࢰࣤࣂࢠ㈻ࡡᶭ⬗よ ᯊ
ῳ㑋▩ᶖ 1,ኃཬఘ࿰ 1,ᱭ⃕ಆ᪺ 1,▦ᓧୌྍ 1,ᓞ⏛ᖷኰ 2,᭱㒂Ṃᩝ 1 (1 ாኬ࣬⏍Ꮛ◂ࠉ2 ⚗஬ᕝኬ࣬ᕝ)
》Ⰵ⭁ᮑⳞ࠿ฦἢࡌࡾࢨࣖࢗ㓗ࡢࠉᮄ㈻ᠺฦࡡຊỀฦよࢅり፳ࡌࡾୌ᪁ࠉ㖙ࢅྱࡳᮄᮞ㜭⭁๠ࢅ↋Ẑ໩ࡌ
ࡾࠊ㖙⪇ᛮⳞ࢛࢛ࢗࢫࣚࢰࢢࡢࢨࣖࢗ㓗Ⓠ㓕࡞౪Ꮛࡊ࡙⏍⫩ࡊࠉࢨࣖࢗ㓗ࢅᇰᆀ୯࡞ኬ㔖࡞⵫✒ࡌࡾࠊୌ⯙
࡞ࢨࣖࢗ㓗ࡢ⏍∸࡞Ẑᛮࢅ♟ࡌࡒࡴࠉᮇ◂✪࡚ࡢ࢛࢛ࢗࢫࣚࢰࢢࡡࢨࣖࢗ㓗Ẑᛮᅂ㑂ᶭᵋࢅよ᪺ࡌࡾࡆ࡛ࢅ
┘Ⓩ࡛ࡊࡒࠊᚋࡼࡿࡾ▩ぜࡢࠉⳞࡡୌḗ⅛⣪௥ㅨᶭᵋࡡよ᪺ࠉ᩺ぞᮄᮞ㜭⭁๠㛜Ⓠ࡞㈠ࡌࡾࠊ࢛࢛ࢗࢫࣚࢰ
ࢢ cDNA ࣚ࢕ࣇ࣭ࣚࣛࢅ⏕࠷ࡒᙟ㈻㌹ᥦ࡞ࡻࡽࢨࣖࢗ㓗⪇ᛮࢅ⋋ᚋࡊࡒ㓕ẍ࠾ࡼࠉࡆࡿࡱ࡚ᶭ⬗ᮅ▩࡛ࡈࡿ
ࡒࢲ࢛ࣝࢺ࢞ࢨࣤᵕࢰࣤࣂࢠ㈻(TRP26)ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄࡡ࣓࣌ࣞࢡ cDNA FpTRP26 ࢅᚋࡒࠊFpTRP26 ᙟ
㈻㌹ᥦ㓕ẍ࡚ࡢࠉࢤࣤࢹ࣭࡛ࣞࣜẒ࡬⣵⬂හ࡚ࡡࢨࣖࢗ㓗㔖࠿΅ᑛࡊࡒࠊFpTRP26 ࡢ࢛࢛ࢗࢫࣚࢰࢢ࡞࠽࠷
࡙ࡵⓆ⌟࠿ヾࡴࡼࡿࠉᮇⳞ࡞࠽࠷࡙ࢨࣖࢗ㓗⪇ᛮ࡞ᐞ୙ࡌࡾ࡛♟ြࡈࡿࡒࠊ⌟ᅹ FpTRP26 ࡡᶭ⬗ࢅよᯊ୯࡚
࠵ࡾࠊ
The possible role of FpTRP26 involved in oxalic acid resistance of brown-rot fungus Fomitopsis palustris
Tomoki Watanabe1), Nobukazu Shitan1), Toshiaki Umezawa1), Kazufumi Yazaki1),Mikio Shimada2) , and
Hattori1) (1RISH, Kyoto Univ.) (2 Tech., Fukui Univ. of Technol.)
33
Takefumi
O-15
TPP ⤎ྙᆵࣛ࣍ࢪ࢕ࢴࢲࡡᶭ⬗ᨭን
ᒜහ㝧ᐰ 1,2㸡୔ይኬ㍔ 2㸡❉ᑈ㝧ᩝ 1㸡ఔක༡ 1㸡こᮟ㢟 1㸡ᮙᮇ├ᕤ 2,3 㸝1 Ⓣ㭧㒿㏸㸝ᰬ㸞࣬◂✪㛜Ⓠᐄ㸡2 ⏝
༞ኬ࣬FIBER㸡3 ⏝༞ኬ࣬⌦ᕝ㸞
࠘┘Ⓩ࠙ᠻࠍࡢࡆࡿࡱ࡚࡞㯔Ⳟ࠾ࡼࢲ࢓࣐ࣤࣅࣞࣛࣤ㓗(TPP)⤎ྙᆵࣛ࣍ࢪ࢕ࢴࢲࢅぜฝࡊࠉࡐࡡᶭ⬗ࢅ┷ᰶ
⏍∸࡚ิࡴ࡙᪺ࡼ࠾࡞ࡊࡒࠊᮇࣛ࣍ࢪ࢕ࢴࢲࡢ A. oryzae thiA ࡡ 5'㟸⩳ゼ㡷ᇡ(5'UTR)࡞Ꮛᅹࡌࡾ࢕ࣤࢹࣞࣤ୕
࡞ᅹࡽࠉTPP ࡛⤎ྙࡌࡾࡆ࡛࡚ࢪࣈࣚ࢕ࢨࣤࢡᵕᘟࢅን໩ࡈࡎࡾࠊࡆࡿ࡞ࡻࡽ࢕ࣤࢹࣞࣤࡡୌ㒂࠿ṟᏋࡊࡒ
ᮅᠺ⇅ mRNA ࠿㌹෕ࡈࡿࠉโᚒୖ࡞࠵ࡾ㐿ఎᏄࡡⓆ⌟ࢅᢒโࡌࡾࠊ᭯⏕ࢰࣤࣂࢠ㈻ࡡ⏍⏐ࢅ┘Ⓩ࡛ࡊࡒሔྙ
ࡢ㐿ఎᏄⓆ⌟ࢅㄇᑙࡌࡾࣛ࣍ࢪ࢕ࢴࢲ࠿ይࡱࡊ࠷࠿ࠉ┷ᰶ⏍∸࡚ࡢࡱࡓሒ࿈ࡈࡿ࡙࠷࡝࠷ࠊࡐࡆ࡚ TPP ⤎ྙ
ᆵࣛ࣍ࢪ࢕ࢴࢲࡡ࿔㎰㡷ᇡ࡞ን␏ࢅᑙථࡊࠉᮇᮮ࡛ࡢ㏣ࡡโᚒᵕᘟ࡞ᶭ⬗ᨭንࡌࡾࡆ࡛ࢅモࡲࡒࠊ
࠘᪁Ἢ࡛⤎ᯕ࠙ࣛ࣍ࢪ࢕ࢴࢲ࿔㎰㡷ᇡ࡫ࡡን␏ᑙථ࡞ࡻࡾโᚒᵕᘟࡡን໩ࢅ☔ヾࡌࡾࡒࡴࠉA. oryzae thiA ࡡ
5'UTR ୖὮ࡞ GUS ࣭ࣝ࣎ࢰ࣭㐿ఎᏄࢅ᥃⤾ࡊࡒⓆ⌟࢜ࢬࢴࢹࢅషᠺࡊࡒࠊᐙ୹࡛ࡊ࡙ A. oryzae niaD300ࠉᇰ
ᆀ࡛ࡊ࡙ CD ᭩ᑛᇰᆀࢅ⏕࠷ࡒࠊࡱࡍⓆ⌟࢜ࢬࢴࢹ࠾ࡼ TPP Ꮛᅹୖ࡚ṟᏋࡌࡾ࢕ࣤࢹࣞࣤ㒂ฦ㒼าࢅ๎㝎ࡌ
ࡾࡆ࡛࡚Ⓠ⌟≁ᛮ࠿㏣㌹ࡊࠉᇰᆀ࡞ࢲ࢓࣐ࣤࢅ 10ȣM ῟ຊࡌࡾࡆ࡛࡚ GUS Ὡᛮ࠿ࢲ࢓࣐ࣤ㟸Ꮛᅹୖࡡ 1.5
ಶ࡞ቌຊࡊࡒࠊࡈࡼ࡞୕オን␏㐿ఎᏄࡡ 5'ࢪࣈࣚ࢕ࢨࣤࢡ㒂న࡛ TPP ⤎ྙᆵࣛ࣍ࢪ࢕ࢴࢲ㛣ࡡ㡷ᇡࢅ 5 ࡱࡒ
ࡢ 9 ሲᇱ๎㝎ࡊࡒ⤎ᯕࠉࢲ࢓࣐ࣤᏋᅹୖ࡚ࡡὩᛮ࠿ࡐࡿࡑࡿ⣑ 5ࠉ4 ಶ࡞ቌຊࡊࡒࠊ௧୕࡞ࡻࡽࠉࢲ࢓࣐ࣤ῟
ຊ࡞ࡻࡽ㐿ఎᏄⓆ⌟ࢅㄇᑙࡌࡾࣛ࣍ࢪ࢕ࢴࢲࡡ㛜Ⓠ࡞ᠺຉࡊࡒࠊ
Functional changes of TPP-dependent riboswitch
Takahiro Yamauchi1,2, Daisuke Miyoshi2, Takafumi Kubodera1, Mitsuhiro Ban1, Akira Nishimura1, Naoki Sugimoto2,3
(1Hakutsuru Sake Brewing Co. Ltd., 2FIBER, Konan Univ., 3Fac. Sci. Eng., Konan Univ.)
O-16
㯔Ⳟ Aspergillus oryzae ࡡࢲ࢓࣐ࣤ㧏⏍⏐ᰬࡡฦᏄ⫩⛸
ᚠ஬⨶㔓࣬❉ᑈ㝧ᩝ࣬஫࿝ົஒ 1࣬ᒜୖఘ㞕࣬ఔ ක༡࣬こᮟ 㢟 㸝Ⓣ㭧㒿㏸◂✪㛜Ⓠࠉ1 ᮶໪ኬ㝌㎨࣬⏍
∸⏐ᴏ๭ᠺ㸞
ࢲ࢓࣐ࣤ 2 ࣛࣤ㓗(TPP)ࡢ࢙ࢾ࣭ࣜ࢟௥ㅨ࡞࠽ࡄࡾ㔔さ࡝⿭㓕⣪࡚࠵ࡽࠉ㓕ẍࢅ୯ᚨ࡞ࡐࡡ⏍ྙᠺ⤊㊪࠽ࡻ
ࡦโᚒᶭᵋ࠿㐿ఎᏄ࡚ࣝ࣊ࣜ࡮࡯よ᪺ࡈࡿ࡙࠷ࡾࠊୌ⯙Ⓩ࡞ࢲ࢓࣐ࣤྙᠺ࡞㛭୙ࡌࡾ㐿ఎᏄ⩄ࡢ᭩⤂⏍⏐∸
࡚࠵ࡾ TPP ࡞ࡻࡽࣆ࢔࣭ࢺࣁࢴࢠᢒโࢅུࡄࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾ࠿ࠉࡐࡡᢒโよ㝎࡞ࡻࡽࢲ࢓࣐ࣤࡡ⏍⏐
㔖࠿㢟ⴥ࡞ቌኬࡊࡒሒ࿈ౚࡢ࡝࠷ࠊᠻࠍࡢࠉࡆࡿࡱ࡚࡞㯔Ⳟ A. oryzae ࡡࢲ࢓࣐ࣤ㧏⏍⏐ᰬཱིᚋࢅ┘ᣞࡊࠉࢲ
࢓࣐ࣤࢅᵋᠺࡌࡾࢲ࢓ࢯ࣭ࣜ⎌ྙᠺ㐿ఎᏄ thiA ࡡ⬲ᢒโᰬࢅཱིᚋࡊࡒ࠿ࠉࢲ࢓࣐ࣤࡡ⏍ྙᠺ࡞㢟ⴥ࡝ቌኬࡢ
☔ヾࡈࡿ࡝࠾ࡖࡒࠊ
௑ᅂࡢ thiA ௧አ࡚ࠉࢲ࢓࣐ࣤ⏍ྙᠺ⣌࡞࠽ࡄࡾ࣭࢞࣎࢕ࣤࢹ࡛⩻࠻ࡼࡿࡾ᥆ᏽࣅ࣐ࣛࢩࣤ⎌ྙᠺ㓕⣪㐿ఎ
Ꮔ nmtA ࠽ࡻࡦࢲ࢓࣐ࣤ 2 ࣛࣤ㓗໩㓕⣪㐿ఎᏄ thiP ࡞ࡵ╌┘ࡊࡒࠊࡐࡡ⤎ᯕࠉthiAࠉnmtA ࠽ࡻࡦ thiP ࢅྜྷ᫤
࡞⬲ᢒโ໩ࡊࡒ᫤࡞ TPP ⏍ྙᠺࡡ㢟ⴥ࡝ቌኬ࠿☔ヾࡈࡿࡒࠊ
Molecular breeding of the thiamin-high-producing strain from Aspergillus oryzae
Misato Tokui, Takafumi Kubodera, Katsuya Gomi1, Nobuo Yamashita, Mitsuhiro Ban, Akira Nishimura
(Hakutsuru Sake Brewing CO., LTD.
1
Grad. Sch. Agri. Sci., Tohoku Univ.,)
34
O-17
Ềฦྱ㔖ࡡ␏࡝ࡾᅖమᇰ㣬᮪௲ୖ࡞࠽ࡄࡾ㯔Ⳟ㐿ఎᏄⓆ⌟ࡡよᯊ
ᑚᯐ ல⣎Ꮔ㸦ࠉఫ㔕 ඔ᫓㸦ࠉ஁⏛ ༡ඔ㸧ࠉ⛑ Ὂ஦㸧ࠉኬ⟺ ಘୌ㸦 㸝㸦㔘Ἁᕝᴏኬ࣬ࢣࢿ࣑◂ࠉ㸧᭮᰿
෗࣬⥪◂㸞
࠘┘Ⓩ࠙㯔Ⳟ Aspergillus oryzae ࡢࠉ㒿షࡽࡡ㝷ࠉ⡷࡝࡜ࡡᅖమ୕࡚ᇰ㣬ࢅ⾔࠹ᅖమᇰ㣬ࡡ᪁Ἢ࠿᤿ࡼࡿ࡙࠷
ࡾࠊࡐࡡ㝷ࠉᅖమᇰᆀࡡྱỀ⋙࡞ࡻࡽ⏍⏐ࡈࡿࡾ㓕⣪⩄࠿ንࢂࡾࡆ࡛࠿⤊㥺Ⓩ࡞▩ࡼࡿ࡙࠷ࡾ࠿ࠉ㐿ఎᏄࣝ
࡚࣊ࣜࡡよᯊࡢ࡝ࡈࡿ࡙࠷࡝࠷ࠊࡐࡆ࡚ࠉᠻࠍࡢỀฦ࠿㔔さ࡝ࣆ࢒ࢠࢰ࣭࡚࠵ࡾ࡛⩻࠻ࠉỀฦྱ㔖ࡡ␏࡝ࡾ
ᅖమᇰ㣬᮪௲࡞࠽ࡄࡾ㯔Ⳟࡡ㐿ఎᏄよᯊࢅ⾔ࡖࡒࠊỀฦྱ㔖࡛㐿ఎᏄⓆ⌟࡛ࡡ㛭㏻ᛮࢅ♟ࡌࡆ࡛࡞ࡻࡽࠉ㧏
ຝ⋙㓕⣪⏍⏐ᇰ㣬᮪௲ࡡᥞ♟࠿᭿ᙽࡈࡿࡾࠊ
࠘᪁Ἢ࠽ࡻࡦ⤎ᯕ࠙Ềฦྱ㔖ࡡ␏࡝ࡾࢤ࣑࢟ࣆࢪ࣏࡞࠽࠷࡙ A. oryzae RIB40 ᰬࢅ 30Υ48 ᫤㛣ᇰ㣬ࡊࡒᚃࠉ
RNA ࢅ᢫ฝࡊࡐࡿࡑࡿࡡ㐿ఎᏄࡡⓆ⌟ࢅࣛ࢓ࣜࢰ࢕࣑ PCR ࡞ࡻࡽよᯊࡊࡒࠊࡱࡒࠉ㯔Ⳟࡡࢣࢿ࣑よᯊ࡚᪺
ࡼ࠾࡛࡝ࡖࡒ 11,000 㐿ఎᏄࢅࢪ࣎ࢴࢹࡊࡒ㸹㹃㸶࢛ࣛࢥ࣏࢕ࢠࣞ࢓ࣝ࢕ࢅ⏕࠷ࠉ㐿ఎᏄࡡⓆ⌟ࣈࣞࣆ࢒࢕ࣜ
ࡡよᯊࢅ⾔ࡖࡒࠊࡆࡿ࡞ࡻࡽࠉỀฦྱ㔖౪ᏋⓏ࡞Ⓠ⌟㔖࠿␏࡝ࡾ㐿ఎᏄ⩄ࢅぜฝࡊࡒࠊ
Analysis of Aspergillus oryzae gene expression with different water content under solid-state culture condition
Akiko Kobayashi㸦, Motoaki Sano㸦, Hiromoto Hisada㸧, Yoji Hata㸧, Shinichi Ohashi㸦
(㸦KIT,
㸧
Res. Inst., Gekkeikan Sake Co.)
35
P-1
㓕ẍ Bni1 ࡡ┞ྜྷࢰࣤࣂࢠ㈻ AoBni1 ࢅ⏕࠷ࡒ㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ Spitzenkörper ࡡ
ྊっ໩
▴ᕖ⤦⌦㸡ḿ㊪ῗஒ, ᭯ᒱᏕ㸡໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
࠘┘Ⓩ࠙Spitzenkörper ࡢࠉ⣊≟ⳞࡡⳞ⣊඙❻࡞Ꮛᅹࡌࡾฦἢᑚ⬂࡞ᐣࡳ࢛ࣜ࢝ࢾ࡚ࣚࠉⳞ⣊ࡡఘ㛏୯ᚨ࡛
ࡊ഼࡙ࡂࠊSpitzenkörper ࡞ࡢࠉฦἢᑚ⬂࠿⵫✒ࡈࡿ࡙࠷ࡾ௙ࠉ࢙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪᑚ⬂ࡵᏋᅹࡌࡾࡆ࡛࠾ࡼࠉ
⣵⬂හࡡᑚ⬂㍲㏞ࡡධమാࢅ⌦よࡌࡾࡒࡴ࡞ࡵ㔔さ࡝࢛ࣜ࢝ࢾ࡚ࣚ࠵ࡾࠊ᭩㎾㓕ẍࡡ formin ࢰࣤࣂࢠ㈻࡚࠵
ࡾ Bni1 ࠿ Spitzenkörper ࡡᵋᠺᠺฦ࡚࠵ࡾࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊᮇ◂✪࡚ࡢ A. oryzae ࡞࠽ࡄࡾ Bni1 ┞ྜྷࢰ
ࣤࣂࢠ㈻ AoBni1 ࡞ EGFP ࢅ⼝ྙࡈࡎࡾࡆ࡛࡞ࡻࡽࠉA. oryzae ࡡ Spitzenkörper ࡡྊっ໩࡛ࠉࡐࡡふᐳࢅ⾔ࡖ
ࡒࠊ
࠘⤎ᯕ࠙A. oryzae ࢣࢿ࣑ࢸ࣭ࢰ࣭࣊ࢪ࠾ࡼ㓕ẍ BNI1 ࡛┞ྜྷᛮࢅ♟ࡌ㐿ఎᏄࢅぜ࠷ࡓࡊࠉAobni1 ࡛ྞࡘࡄ
ࡒࠊAoBni1 ࡢ A. nidulans ࡡ formin ࡚࠵ࡾ SepA ࡛ࡡ┞ྜྷᛮ࠿㧏ࡂࠉformin ࡞≁᭯ࡡࢺ࣒࢕࡚ࣤ࠵ࡾ FH2 ࡛
FH3 ࢅ᭯ࡊ࡙࠷ࡾࡆ࡛࠿ࢰࣤࣂࢠ㈻࣓ࢲ࣭ࣆ᳠⣬࡚᪺ࡼ࠾࡞࡝ࡖࡒࠊࡱࡒࠉFH1 ࢺ࣒࢕ࣤࡡ≁ᚡ࡚࠵ࡾࣈࣞ
ࣛࣤ࡞ᐣࢆࡓ㒼าࡵ᭯ࡊ࡙࠷ࡒࠊḗ࠷࡚ Aobni1 ࢅ PCR ࡞ࡻࡖ࡙༟㞫ࡊࠉegfp ࡛ࡡ⼝ྙ㐿ఎᏄࢅⓆ⌟ࡌࡾ A.
oryzae ᙟ㈻㌹ᥦమࢅཱིᚋࡊࡒࠊࡆࡡᙟ㈻㌹ᥦమࡡ㢟᚜㙶ふᐳࢅ⾔ࡖࡒ࡛ࡆࢀࠉⳞ⣊ࡡ඙❻ࡡ Spitzenkörper ᵕ
ᵋ㏸࡞ EGFP ⺧ක࠿ぜࡼࡿࡒࠊ⌟ᅹࡆࡡ Spitzenkörper ᵕᵋ㏸ࡡᛮ㈻ࡡよᯊࢅ㐅ࡴ࡙࠷ࡾࠊ
Observation of Spitzenkörper using a fusion protein of EGFP with AoBni1 in A. oryzae.
Eri ISHIKAWA, Jun-ya SHOJI, Manabu ARIOKA, Katsuhiko KITAMOTO (Dept. of Biotech., Univ. of Tokyo)
P-2
⺧කࢰࣤࣂࢠᵾㆉ࡞ࡻࡾ Aspergillus nidulans ඙❻ᠺ㛏ᶭᵋࡡよᯊ㸯᚜ᑚ⟮࡛࢓ࢠࢲࣤ
ࡡ㛭౿ࢅ୯ᚨ࡞
1
ᇷᑹူஒ㸡2 ⣒ ᭰ᾀ㸡3Naimeh Taheri-Talesh㸡2 ⳝờ㡝ᙢ㸡 3 Berl R. Oakley㸝1 ᚠᓞኬ㝌࣬ࣉࣜࢪࣁ࢕࢛㸡2 ྞ
ཿᒁኬ㝌༈⣌࣬ฦᏄᵾⓏ㸡3 ࢛ࣀ࢕࢛ᕗ❟ኬ࣬ฦᏄ㐿ఎ㸞
┷Ⳟࡡ඙❻ᠺ㛏ࡢࠉⳞ⣊඙❻࡞ᠺ㛏Ⅴ࠿㝀ᏽࡈࡿࡾ≁ᚡⓏ࡝ᠺ㛏ᵕᘟ࡚ࠉᠺ㛏࡞㛭୙ࡌࡾᵕࠍ࡝ฦᏄ⩄
ࡡᑻᅹ໩࡛༝ㄢࡡඔ࡞ᠺࡽ❟ࡖ࡙࠷ࡾࠊ⣵⬂㦭᰹ࡢࠉᠺ㛏Ⅴࡡ⥌ᣚࡷఘ㛏ཬᚺ࡞ᚪ㡪ࡡᙲ๪ࢅᯕࡒࡊ࡙࠷ࡾ
࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊᠻࠍࡢ GFP ᵾㆉࢲ࣭ࣖࣇࣛࣤࢅ⏕࠷ࡒよᯊ࡞ࡻࡽࠉA. nidulans ࡡ㏷࠷඙❻ᠺ㛏࡞࠽࠷࡙ࠉ
᚜ᑚ⟮࠿ᚪ㡪࡚࠵ࡾ஥ࠉᠺ㛏❻㎾എࡡ⣵⬂㈻᚜ᑚ⟮࠿ฦ⿛᭿࡞ࡵಕᣚࡈࡿࡾ஥ࢅ᪺ࡼ࠾࡞ࡊࡒࠊ࢓ࢠࢲࣤࡢ
ࡌ࡬࡙ࡡ⣵⬂ᠺ㛏࡞ᚪ㡪࡚࠵ࡾ஥࠿▩ࡼࡿ࡙࠷ࡾࠊࡐࡆ࡚ GFP ᵾㆉ࢓ࢠࢲࣤࢅ⏕࠷࡙᚜ᑚ⟮ᶭ⬗࡛ࡡ㛭౿ࢅ
ㄢ࡬ࡒࠊ࢓ࢠࢲࣤࡢࠉኣᩐࡡࣂࢴࢲ≟ࡡ㞗ྙమ࡛ࡊ࡙ᠺ㛏❻࠾ࡼᩐȣm හഁ࡞㞗୯ࡊ࡙ᑻᅹࡊ࡙࠷ࡒࠊ࢓ࢠ
ࢲࣤࡡᑻᅹࡢ᚜ᑚ⟮࡞౪Ꮛࡊ࡙࠽ࡽࠉ⣵⬂ᠺ㛏ࡢ࢓ࢠࢲࣤࡡᑻᅹ࡞౪Ꮛࡊ࡙࠷ࡒࠊᠺ㛏ࡡ㐔࠷ฝⰾ├ᚃࡡⳞ
⣊හࡡ࢓ࢠࢲࣤᑻᅹࡢࠉ㏷ࡂᠺ㛏ࡌࡾᠺ⇅ࡊࡒⳞ⣊඙❻࡛ࡐࡿ࡛ࡢ␏࡝ࡖ࡙࠷ࡒࠊA. nidulans ࡚ࡢࠉⳞ⣊ࡡ
ఘ㛏࡛ࡐࡿ࡞ఔ࠹ᠺ㛏㏷ᗐࡡቌຊ࡞ᚉࡖ࡙᚜ᑚ⟮࡛࢓ࢠࢲࣤ࠿༝ㄢࡊ࡙ᶭ⬗ࡌࡾᶭᵋ࠿☔❟ࡊࠉ⤽⤾Ⓩ࡚㏷
࠷඙❻ᠺ㛏ࢅྊ⬗࡞ࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡾࠊ⌟ᅹࠉ࢓ࢠࢲࣤ⤎ྙࢰࣤࣂࢠ㈻ࡷ SNARE ࡡᑻᅹࠉ࠽ࡻࡦᑻᅹ
โᚒ࡞࠽ࡄࡾ࢞ࢾࢨࣤࡡ㛭୙࡞ࡗ࠷࡙᳠ゞ୯࡚࠵ࡾࠊ
Essential role of microtubule and actin in hyphal tip growth of Aspergillus nidulans: analysis with live cell
imaging of proteins tagged with fluorescence proteins.
1
Tetsuya Horio, 2 Tomohiro Akashi, 3Naimeh Taheri-Talesh, 2Akihiko Kikuchi, 3Berl R. Oakley
(1 Health Bio.,
Tokushima Univ. Grad. Sch., 2 Mol. Mycol. Med., Nagoya Univ. Grad. Sch., 3Mol. Genet., Ohio State Univ.)
36
P-3
A. oryzae ࡡ Woronin body ᙟᠺ࡞㛭୙ࡌࡾ Aohex1 㐿ఎᏄࡡ◒ቪᰬ࡞࠽ࡄࡾ⾪⌟ᆵよᯊ
୷ᒜ₮ୌࠉ໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
࠘┘Ⓩ࠙Woronin body ࡢᏄᄙⳞ㢦࡛ୌ㒂ࡡ୘ᏰධⳞ㢦⣊≟Ⳟ࡞Ꮛᅹࡌࡾ࢛ࣜ࢝ࢾ࡚ࣚ࠵ࡽࠉ㝰ባᏇࢅራ࠷࡚
⁈Ⳟࡡఎ᧓ࢅ㜭ࡃ഼ࡀࢅ᭯ࡌࡾ 1)ࠊWoronin body ࡡᑻᅹࡢ㝰ባ㎾എࡡ࡮࠾ࠉⳞ⣊඙❻࡝࡜ืࡡⳞ⣊㒂న࡞ࡵ
ふᐳࡈࡿࠉ㝰ባᏇࢅራࡃ௧አࡡᶭ⬗ࢅࡵࡗࡆ࡛࠿⩻࠻ࡼࡿࡾ࠿ࠉࡆࡿࡼࡡᶭ⬗࡞ࡗ࠷࡙ࡢ࠵ࡱࡽࢂ࠾ࡖ࡙࠷
࡝࠷ࠊᠻࠍࡢࡆࡿࡱ࡚࡞ Woronin body ᙟᠺ࡞ᚪさ࡝ Aohex1 㐿ఎᏄࡡ ORF ධ㛏ࢅ◒ቪࡊࡒᰬࢅష⿿ࡊࡒ࠿ࠉ
㏳ᖏࡡᇰ㣬᮪௲࡚ࡢࢤࣤࢹ࣭ࣞࣜᰬ࡛ྜྷᵕࡡ⏍⫩ࢅ♟ࡊࡒ 2)ࠊᮇ◂✪࡚ࡢࠉAoHex1 ࡡ᩺ࡒ࡝ᶭ⬗ࢅ᪺ࡼ࠾࡞
ࡌࡾࡒࡴ࡞ Aohex1 㐿ఎᏄ◒ቪᰬࡡᵕࠍ࡝⏍⫩᮪௲࡚ࡡ⾪⌟ᆵよᯊࢅ⾔ࡖࡒࠊ
࠘᪁Ἢ࡛⤎ᯕ࠙Aohex1 㐿ఎᏄ◒ቪᰬࢅ྘⛸᮪௲㸝㧏Ὼࠉ㧏ᾈ㏩ᅸࠉᰜ㣬㣒㣱ࠉ྘⛸ⷾ๠῟ຊ࡝࡜㸞࡚ᇰ㣬ࡊࠉ
⏍⫩≁ᛮࢅレ⣵࡞ㄢ࡬ࡒࠊⷾ๠῟ຊ᮪௲࡚ᇰ㣬ࢅ⾔ࡖࡒ࡛ࡆࢀࠉβ-1,3-ࢡࣜ࢜ࣤྙᠺ㓕⣪㜴ᐐ๠ Micafungin
࠽ࡻࡦ β ⤎ྙᆵ⢶㙈⤎ྙᛮⰅ⣪ Congo red ࡡᏋᅹୖ࡚ Aohex1 㐿ఎᏄ◒ቪᰬࡡ⏍⫩఩ୖ࠿ふᐳࡈࡿࡒࠊࡆࡿࡼ
ࡢࠉAoHex1 ࠿⣵⬂ባࡡ࣓ࣛࢸࣛࣤࢡ࡞㛭୙ࡊ࡙࠷ࡾࡆ࡛ࢅ♟ြࡌࡾࡵࡡ࡚࠵ࡾࠊ⌟ᅹࠉAohex1 㐿ఎᏄ◒ቪ
ᰬ࠿ Micafungin ࠽ࡻࡦ Congo red ࡞វུᛮࢅ♟ࡌᶭᵋࡡよᯊࢅ⾔ࡖ࡙࠷ࡾࠊ
1) Maruyama et al., (2005) Biochem. Biophys. Res. Commun., 331, 1081-8.
2) හ⏛ࡼࠉ᪝ᮇ㎨ⰹ໩Ꮥఌ 2006 ᖳᗐኬఌさ᪠㞗ࠉp. 98
Phenotypic analysis of the disruptant of the Aohex1 gene required for Woronin body formation in A. oryzae
Juni-ichi Maruyama, Katsuhiko Kitamoto
(Dept. of Biotechnol., Univ. of Tokyo)
P-4
A. oryzae ࡡ Woronin body ᙟᠺ࡞ᚪさ࡝ Aohex1 㐿ఎᏄ࡞࠽ࡄࡾ㐽ᢝⓏࢪࣈࣚ࢕ࢨࣤࢡ
ࡡណ⩇࡞㛭ࡌࡾよᯊ
ᒷᓧ೸ኯ㑳㸡හ⏛༡ᩅ, Praveen Rao Juvvadi, ୷ᒜ₮ୌ, ໪ᮇົࡥࡆ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
࠘┘Ⓩ࠙Woronin body ࡢࠉᏄᄙⳞ㢦࠽ࡻࡦ㢦⦍ࡡ୘ᏰධⳞ㢦⣊≟Ⳟ࡞Ꮛᅹࡌࡾ࢛ࣜ࢝ࢾ࡚ࣚ࠵ࡽࠉⳞ⣊᥾ഭ
᫤࡞⁈Ⳟࡡఎ᧓ࢅ㜭ࡃࡒࡴ㝰ባᏇࢅራࡃᙲ๪࠿▩ࡼࡿ࡙࠷ࡾࠊA. oryzae ࡡ Woronin body ᙟᠺ࡞ᚪさ࡝ Aohex1
㐿ఎᏄࡡ㌹෕⏐∸ࡢ㐽ᢝⓏࢪࣈࣚ࢕ࢨࣤࢡࢅུࡄࠉࢪࣈࣚ࢕ࢨࣤࢡࡈࡿࡾሔྙ AoHex1 ࢰࣤࣂࢠ㈻ࡢ 50 ࢓࣐
ࢿ㓗▯ࡂ࡝ࡾ 1)ࠊୌ᪁࡚ࠉ࢓࢜ࣂࣤ࢜ࣄ Neurospora crassa ࡚ࡢ hex1 㐿ఎᏄࡡ㐽ᢝⓏࢪࣈࣚ࢕ࢨࣤࢡࡢぜࡼࡿ
ࡍࠉA. oryzae ࡛␏࡝ࡾ⌦⏜ࡢ୘᪺࡚࠵ࡾࠊᮇ◂✪࡚ࡢࠉAohex1 㐿ఎᏄ◒ቪᰬࡡ┞⿭⬗ࢅᣞᵾ࡛ࡊ࡙㐽ᢝⓏࢪ
ࣈࣚ࢕ࢨࣤࢡࡡណ⩇ࢅ᳠ゞࡊࡒࠊ
࠘᪁Ἢ࣬⤎ᯕ࠙఩ᾈ㏩ᅸࢨࣘࢴࢠ࡞ࡻࡽ඙❻⣵⬂ࢅ⁈Ⳟࡌࡾ࡛ࠉAohex1 㐿ఎᏄ◒ቪᰬ࡚ࡢ㝼᥃ࡌࡾ⣵⬂࡞⁈
Ⳟ࠿ఎ᧓ࡌࡾ 1)ࡡ࡚ࠉࡆࡡ⾪⌟ᆵࢅᣞᵾ࡞┞⿭よᯊࢅ⾔ࡖࡒࠊAohex1 㐿ఎᏄࡻࡽ࢕ࣤࢹࣞࣤࡡ㒂ฦࢅ㝎࠷ࡒ
ࢪࣈࣚ࢕ࢨࣤࢡᆵࢅⓆ⌟ࡌࡾࣈࣚࢪ࣐ࢺࠉཀྵࡦࢪࣈࣚ࢕ࢨࣤࢡ㒂న࡞ን␏ࢅຊ࠻㟸ࢪࣈࣚ࢕ࢨࣤࢡᆵࡡࡲࢅ
Ⓠ⌟ࡌࡾࣈࣚࢪ࣐ࢺࢅࠉAohex1 㐿ఎᏄ◒ቪᰬ࡞ᙟ㈻㌹ᥦࡊࡒࠊཱིᚋࡊࡒᰬ࡞ࡗ࠷࡙ࠉ఩ᾈ㏩ᅸࢨࣘࢴࢠ࡞ࡻ
ࡽ⁈Ⳟࡡఎ᧓ࢅふᐳࡊࡒࠊࡐࡡ⤎ᯕࠉࢪࣈࣚ࢕ࢨࣤࢡᆵ AoHex1 Ⓠ⌟ᰬࡢ⁈Ⳟࡡఎ᧓ࢅ㜭࠷ࡓࡆ࡛࠾ࡼࠉ┞
⿭࠿☔ヾࡈࡿࡒࠊୌ᪁࡚ࠉ㟸ࢪࣈࣚ࢕ࢨࣤࢡᆵࢅⓆ⌟ࡊࡒሔྙࠉ┞⿭ࡢ☔ヾࡈࡿ࡝࠾ࡖࡒࠊࡆࡡࡆ࡛࠾ࡼࠉ
Aohex1 㐿ఎᏄࡡ㌹෕⏐∸࠿ࢪࣈࣚ࢕ࢨࣤࢡࡈࡿࡾࡆ࡛࡞ࡻࡖ࡙ࠉAoHex1 ࠿㝰ባᏇࢅራࡃᶭ⬗ࢅࡵࡗࡆ࡛࠿
♟ြࡈࡿࡒࠊ⌟ᅹࠉࢪࣈࣚ࢕ࢨࣤࢡᆵ࡛㟸ࢪࣈࣚ࢕ࢨࣤࢡᆵࡡ AoHex1 ࢰࣤࣂࢠ㈻ࡡᛮ㈻ࢅよᯊࡌࡾ஢ᏽ࡚
࠵ࡾࠊ
1) Maruyama et al., (2005) Biochem. Biophys. Res. Commun. 33, 1081-8.
Functional analysis of two forms of the Woronin body protein AoHex1 expressed by alternative splicing.
Kentaro Iwasaki, Hirotaka Uchida, Praveen Rao Juvvadi, Jun-ichi Maruyama, Katsuhiko Kitamoto
(Dept. of Biotechnol., Univ. of Tokyo)
37
P-5
࢓࢜ࣂࣤ࢜ࣄ࡞࠽ࡄࡾ⺧කࢰࣤࣂࢠ࡚ᵾㆉࡊࡒ NC-RAS-2 㐿ఎᏄࡡྊっ໩
Ⱪἠᖶኯ㸡ᮟᒜ⫍Ꮔ 㸝㛭᮶Ꮥ㝌ኬ㝌࣬ᕝᏕ◂✪⛁㸞
⣊≟ⳞࡡⳞ⣊ᠺ㛏ࡢ඙❻ࡡࡲ࡚⾔ࢂࡿࡾ࠿ࠉ඙❻࡞࠽ࡄࡾᠺ㛏࡞ᚪさ࡝⣵⬂ባࡡ๑㥉మ࠽ࡻࡦ⣵⬂ባྙᠺ
㓕⣪ࠉࡱࡒⳞమአ࡞Ꮛᅹࡌࡾᰜ㣬ฦࢅฦよࡊฺ࡙⏕ࡌࡾࡒࡴࡡฦよ㓕⣪࡝࡜ࡢ඙❻ᑚ⬂㸝apical vesicles㸞࡞
ໜࡱࡿࡒᙟ࡚Ⳟ⣊ࡡ඙❻࡞㐘ࡣࡿࠉࡐࡆ࡚඙❻ᑚ⬂ࡡහᐖ∸࠿ exocytosis ࡞ࡻࡽ⣵⬂⭯አ࡞ฦἢࡈࡿࠉࡆࡿ
ࡼࡡฦἢࡈࡿࡒ๑㥉మࡷ㓕⣪ࢅ⏕࠷࡙⣵⬂ባ࠿ྙᠺࡈࡿࠉࡱࡒⳞమአ࡞ฦἢࡈࡿࡒฦよ㓕⣪࡞ࡻࡽⳞమአ࡞
Ꮛᅹࡌࡾ㧏ฦᏄᰜ㣬ฦࡢฦよࡈࡿࠉᑚࡈ࡝ฦᏄ࡛࡝ࡖ࡙Ⳟమහ࡞ྺ཭ࡈࡿฺ⏕ࡈࡿࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊ
⣊≟Ⳟࡡ࣓ࢸࣜ⏍∸࡛ࡊ࡙◂✪࡞⏕࠷ࡼࡿ࡙࠷ࡾ࢓࢜ࣂࣤ࢜ࣄࡡᙟឺⓏ✲↓ን␏మ smco7 ࡢཋ࢝ࣤ㐿ఎᏄ
ࡡୌࡗ࡚࠵ࡾ NC-RAS-2 ࡡ null mutant ࡚࠵ࡾࠊsmco7 ࡢ㔕⏍ᆵ࡛Ẓ㍉ࡌࡾ࡛Ⳟ⣊࠿⣵ࡂࠉⳞ⣊ᠺ㛏㏷ᗐ࠿ⴥࡊ
ࡂ㐔ࡂࠉࡱࡒ⣵⬂ባ࠿⷟ࡂࠉⳞమአฦよ㓕⣪Ὡᛮ࠿఩࠷ࠊࡆࡿࡼࡡ⤎ᯕࡢ NC-RAS-2 ࡡ௒ᅹࡌࡾಘྒఎ㐡⣌
࠿඙❻ᑚ⬂ࡡ㐘ᦑࡷ඙❻ᑚ⬂ࡡ exocytosis ࡡㄢ⟿࡞㛭୙ࡊ࡙࠷ࡾࡆ࡛ࢅ♟ြࡊ࡙࠷ࡾࠊ
ࡐࡆ࡚ࠉᮇ◂✪࡚ࡢ NC-RAS-2 㐿ఎᏄ࡞࣭ࣝ࣎ࢰ࣭࡛ࡊ࡙⥫Ⰵ⺧කࢰࣤࣂࢠ㈻ࢅࢤ࣭ࢺࡌࡾ GFP 㐿ఎᏄࢅ
㏻⤎ࡊ࡙⼝ྙ㐿ఎᏄࢅషᠺࡊࠉࡐࡡ⼝ྙ㐿ఎᏄࢅ NC-RAS-2 ࢰࣤࣂࢠ㈻࠿Ḗ᥾ࡊ࡙࠷ࡾ smco7 ࡞ᑙථࡊ࡙⼝
ྙࢰࣤࣂࢠ㈻ࢅྙᠺࡈࡎࠉsmco7 Ⳟ⣊හ࡚ࡡ NC-RAS-2 ࢰࣤࣂࢠ㈻ࡡᑻᅹᛮཀྵࡦⓆ⌟᫤᭿࡝࡜ࢅྊっ໩ࡊ࡙よ
ᯊࡊࠉ඙❻ᑚ⬂ࡡ㐘ᦑࡷ඙❻ᑚ⬂ࡡ exocytosis ࡛ࡡ㛭౿࡞㛭ࡌࡾ▩ぜࢅᚋࡻ࠹࡛ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙⾔ࢂ
ࡿࡒࠊsmco7 ⣵⬂හ࡞࠽ࡄࡾ⼝ྙ㐿ఎᏄࡡⓆ⌟ࡡよᯊ⤎ᯕ࡞ࡗ࠷࡙ሒ࿈ࡊࠉ඙❻ᑚ⬂ࡡ㐘ᦑࡷ඙❻ᑚ⬂ࡡ
exocytosis ࡛ࡡ㛭౿࡞ࡗ࠷࡙᳠ゞࡌࡾࠊ
Visualization of NC-RAS-2 protein tagged with fluorescence protein in Neurospora crassa
Kouta Moizumi,Tadako Murayama
(Grad.Sch. of Engineering., Kanto Gakuin Univ.)
P-6
N-⤎ྙᆵ⢶㙈ࣄ࣭ࢫࢅ⏕࠷ࡒ A. oryzae ᑚ⬂మහࣝࢠࢲࣤࡡཱིᚋ࡛ᑻᅹよᯊ
ῳ㑋ὀ♰ 1㸡ᮿᑹୌ㑳 1,2㸡୷ᒜ₮ୌ 3㸡໪ᮇົࡥࡆ 3㸡௿⸠ᖶᠺ 1,2㸝⌦◂ 1㸡CREST2㸡᮶ኬ㝌㎨⏍⛁࣬ᚺ⏍ᕝ 3 㸞
࠘┘Ⓩ࠙㯔Ⳟ Aspergillus oryzae ࡢඁࡿࡒࢰࣤࣂࢠ㈻ฦἢ⬗ࢅ᭯ࡊ࡙࠽ࡽࠉ␏⛸ࢰࣤࣂࢠ㈻ࢅ⏍⏐ࡌࡾ࣌ࢪࢹ
࡛ࡊ࡙ࡡฺ⏕࠿᭿ᙽࡈࡿ࡙࠷ࡾࠊᠻࠍࡢ A. oryzae ࡞࠽ࡄࡾࢰࣤࣂࢠ㈻⏍⏐⬗ງࢅ㧏ࡴࡾࡒࡴ࡞ࠉN-⤎ྙᆵ⢶
㙈ࢅ௒ࡊࡒࢰࣤࣂࢠ㈻ဗ㈻⟮⌦ᶭᵋ࡞㛭ࡌࡾ⌦よ࠿ᚪさ୘ྊḖ࡚࠵ࡾ࡛⩻࠻࡙࠷ࡾࠊᫎᖳᗐࠉN-⤎ྙᆵ⢶㙈
⤎ྙࣄ࣭ࢫࢅ⏕࠷ࡒ A. oryzae ⭯⏤ฦࡡよᯊࢅ⾔࠷ࠉ⢶㙈ヾㆉࢰࣤࣂࢠ㈻ࡡཱིᚋ࡞ᠺຉࡊ࡙࠷ࡾ 1)ࠊᮇⓆ⾪࡚
ࡢ Glc1Man9GlcNAc2 ⢶㙈⤎ྙࣄ࣭ࢫ࡞⤎ྙࡊࡒ⢶㙈ヾㆉࢰࣤࣂࢠ㈻ࡡྜྷᏽࠉ྘⛸⢶㙈࡞ᑊࡌࡾ⤎ྙ≁␏ᛮよ
ᯊࠉ࠽ࡻࡦ A. oryzae ⣵⬂හ࡞࠽ࡄࡾᑻᅹよᯊࢅ⾔ࡖࡒ⤎ᯕࢅሒ࿈ࡌࡾࠊ
࠘᪁Ἢ࡛⤎ᯕ࠙ᑚ⬂మࢅྱࡳ⭯⏤ฦࡢࠉA. oryzae Ⳟమࢅᾦమ✽⣪࡞࡙◒◃ᚃࠉ㐪ᚨฦ㞫Ἢ࡞౩ࡌࡾࡆ࡛࡞ࡻ
ࡽㄢ⿿ࡊࡒࠊᚋࡼࡿࡒᑚ⬂మ⏤ฦࢅ⏕࠷࡙ࠉGlc1Man9GlcNAc2 ࣄ࣭ࢫ࡞⤎ྙࡌࡾࢰࣤࣂࢠ㈻ࢅཱིᚋࡊࡒࠊࡆ
ࡡ⢶㙈⤎ྙࢰࣤࣂࢠ㈻ࡢࠉ㯔Ⳟࢣࢿ࣑ࢸ࣭ࢰ࣭࣊ࢪࢅฺ⏕ࡊࡒ LC/MS/MS よᯊࡻࡽ A. oryzae ࡞࠽ࡄࡾ calnexin
(CNX)࡚࠵ࡾ࡛ྜྷᏽࡈࡿࡒࠊࡈࡼ࡞ࠉ⛸ࠍࡡ⢶㙈⤎ྙࣄ࣭ࢫ࡛ࡡ⤎ྙᐁ㥺ࢅ⾔࠷ࠉA. oryzae ࡞࠽ࡄࡾ CNX ࠿
Glc1Man9 ⢶㙈≁␏Ⓩ࡞⤎ྙࡌࡾࡆ࡛ࢅ☔ヾࡊࡒࠊA. oryzae ⣵⬂හ࡞࠽ࡄࡾ CNX ࡡᑻᅹࢅㄢ࡬ࡾࡒࡴ࡞
CNX-EGFP ⼝ྙࢰࣤࣂࢠ㈻ࢅⓆ⌟ࡌࡾᙟ㈻㌹ᥦᰬࢅష⿿ࡊࠉྊっ໩ࢅモࡲࡒࠊEGFP ⺧ක࡛⣵⬂᯹Ⰵࡡふᐳ
ࡻࡽࠉCNX ࠿ᑚ⬂మ࡞ᑻᅹࡌࡾࡆ࡛࠿ᙁࡂ♟ြࡈࡿࡒࠊ⌟ᅹࠉ⛸ࠍࡡᑚ⬂మᆵ⢶㙈ࣄ࣭ࢫࢅ⏕࠷࡙ࠉ⢶㙈ヾ
ㆉࢰࣤࣂࢠ㈻ࡡよᯊࢅ⾔ࡖ࡙࠷ࡾࠊ
1) ୷ᒜࡼࠉ➠㸪ᅂ⣊≟ⳞฦᏄ⏍∸Ꮥࢤࣤࣆ࢒ࣝࣤࢪさ᪠㞗 p. 71
Identification and localization of intracellular lectins using N-glycan-conjugated beads in Aspergillus oryzae
Taisuke Watanabe1, Ichiro Matsuo1,2, Jun-ichi Maruyama3, Katsuhiko Kitamoto3, Yukishige Ito1,2
(RIKEN1, CREST2, Dept. of Biotech., Univ. of Tokyo3)
38
P-7
㯔Ⳟ A. oryzae ࡞࠽ࡄࡾᑚ⬂మࢪࢹࣝࢪࡡྊっ໩
ኬ㔕⤚Ꮔ㸡୷ᒜ₮ୌ㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
࠘┘Ⓩ࠙⣵⬂හ࡚⏍⏐ࡈࡿࡒฦἢࢰࣤࣂࢠ㈻ࡢࠉࡱࡍᑚ⬂మ࡚ဗ㈻⟮⌦ࢅུࡄࡾࠊࡆࡡ࡛ࡀࠉࣆ࢚࣭ࣜࢸ࢔
ࣤࢡ࡞኶ᩃࡊࡒንᛮࢰࣤࣂࢠ㈻࠿ᑚ⬂మහ࡞⵫✒ࡊࠉᑚ⬂మᶭ⬗࡞ㇿᢰ࠿࠾࠾ࡾࡆ࡛ࢅࠔᑚ⬂మࢪࢹࣝࢪࠕ
࡛࿣ࡩࠊᑚ⬂మࢪࢹࣝࢪ࡞ᚺ➽ࡊ࡙ࠉᑚ⬂మࢨࣔ࣋ࣞࣤ㐿ఎᏄࡡⓆ⌟ㄇᑙࠉࢰࣤࣂࢠ㈻ࡡ⩳ゼᢒโࠉንᛮࢰ
ࣤࣂࢠ㈻ࡡฦよ࣬㝎ཡࠉ࢓࣎ࢹ࣭ࢨࢪ࡝࡜࠿㉫ࡆࡾࠊ㯔Ⳟࡢ㧏࠷ࢰࣤࣂࢠ㈻⏍⏐⬗ງࢅᣚࡗ࠿ࠉິ∸ࡷ᳔∸
⏜ᮮࡡࢰࣤࣂࢠ㈻ࢅ⏍⏐ࡈࡎࡒሔྙ࡞ࡢୌ⯙࡞⏍⏐㔖࠿㧏ࡂ࡝࠷ࠊࡆࡡཋᅄ࡛ࡊ࡙ᑚ⬂మࢪࢹࣝࢪ࡞ࡻࡾฦ
よ࠿⩻࠻ࡼࡿ࡙࠷ࡾࠊࡐࡆ࡚ࠉᑚ⬂మࢪࢹࣝࢪᚺ➽⤊㊪ࡡ୕Ὦ࡞న⨠ࡌࡾ㐿ఎᏄ࡚࠵ࡾ㓕ẍ HAC1 ࣓࣌ࣞࢡ
AohacA ࢅ⏕࠷࡙ࠉᑚ⬂మࢪࢹࣝࢪࢅྊっ໩ࡌࡾ⣌ࡡᵋ⠇ࢅモࡲࡒࠊ
࠘᪁Ἢཀྵࡦ⤎ᯕ࠙A. niger ࡡ▩ぜ࠾ࡼࠉAohacA ࡢ 20 bp ࡡ࢕ࣤࢹࣞࣤࢅᣚࡔࠉᑚ⬂మࢪࢹࣝࢪ࡞ᚺ➽ࡊ࡙ࡆࡡ
࢕ࣤࢹࣞࣤ࠿ࢪࣈࣚ࢕ࢨࣤࢡࡈࡿࡾࡆ࡛࡚⩳ゼࡡ㝷࡞ࣆ࣭࣑ࣝࢨࣆࢹ࠿㉫ࡀࠉὩᛮᆵࡡ AohacA ࠿⩳ゼࡈࡿࡾ
࡛⩻࠻ࡼࡿࡒࠊࡐࡆ࡚ࠉAoHacA ࡡ㸸ᮆ❻ࡡ㌹෕Ὡᛮ໩ࢺ࣒࢕ࣤࢅ๎㝎ࡊࡒ㐿ఎᏄ AohacAΔC ࡞ egfp 㐿ఎᏄ
ࢅ⼝ྙࡈࡎࡒ AohacAΔC-egfp 㐿ఎᏄࢅ A. oryzae ࡞ᑙථࡊࡒࠊᚋࡼࡿࡒᰬ࡞ᑚ⬂మࢪࢹࣝࢪㄇᑙ๠࡚࠵ࡾࢩࢲ
࢛ࢪࣝ࢕ࢹ࣭ࣜ(DTT)ࢅ῟ຊࡊࡒ࡛ࡆࢀࠉDTT ࡞ᚺ➽ࡊ࡙⣵⬂හ࡞ EGFP ⺧ක࠿ふᐳࡈࡿࡒࠊࡱࡒࠉRT-PCR
࡞ࡻࡽ AohacA ࡡ 20 bp ࢕ࣤࢹࣞࣤࡡࢪࣈࣚ࢕ࢨࣤࢡ࠿ቌຊࡊ࡙࠷ࡾࡆ࡛࠿☔ヾࡈࡿࡒࠊ⌟ᅹࠉࡆࡡᰬ࡞␏⛸
ࢰࣤࣂࢠ㈻Ⓠ⌟ࣈࣚࢪ࣐ࢺࢅᑙථࡊࠉ␏⛸ࢰࣤࣂࢠ㈻⏍⏐᮪௲ୖ࡚ࡡᑚ⬂మࢪࢹࣝࢪᚺ➽ࢅ⺧ක㢟᚜㙶࡞ࡻ
ࡽふᐳࡊ࡙࠷ࡾࠊ
Visualization of ER stress in Aspergillus oryzae
Ayako Ohno, Jun-ichi Maruyama, Manabu Arioka, Katsuhiko Kitamoto
(Dept. of Biotechnol., Univ. of Tokyo)
P-8
㯔Ⳟ Aspergillus oryzae Ⳟ⣊ᇱ㒂࡞࠽ࡄࡾ࢛ࣜ࢝ࢾࣚᵋᠺᠺฦࡡ࢛࣭ࢹࣆ࢒ࢩ࣭࡞ࡻ
ࡾᾦ⬂࡫ࡡཱི㎲ࡲ
ḿ㊪ῗஒ㸡ⳝ㛣㝧ᚷ, ᭯ᒱᏕ㸡໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
࠘┘Ⓩ࠙⣊≟Ⳟ࡞࠽࠷࡙ᾦ⬂ࡢࠉⳞ⣊ࡡ㒂న࡞ᚺࡋࡒኣᵕ࡝ᙟឺࢅ♟ࡌ㸣ࡆࡿࡱ࡚ᠻࠍࡢ㯔Ⳟ A. oryzae ࡡ
ᾦ⬂ᙟឺࡡよᯊ࠾ࡼ㸡Ⳟ⣊ᇱᗇ㒂࡞࠽࠷࡙ᾦ⬂࠿⣵⬂ࡡኬ༖ࢅ༥ࡴࡾ࡮࡜࡞Ⓠ㐡ࡊ࡙࠷ࡾࡆ࡛㸡ࡈࡼ࡞ᾦ⬂
⭯࠿ᾦ⬂හ⭅࡫ཱིࡽ㎲ࡱࡿࡒ⑖㊟࠿࠵ࡾࡆ࡛ࢅぜ࠷ࡓࡊ࡙࠷ࡾ
1)
ࠊࡆࡡ⤎ᯕ࠾ࡼࠉⳞ⣊ᇱᗇ㒂࡞࠽࠷࡙ࡢ⣵
⬂㈻࠽ࡻࡦ࢛ࣜ࢝ࢾࣚᠺฦࡡໜᣋⓏ࡝ฦよ࠿㉫ࡆࡖ࡙࠷ࡾྊ⬗ᛮ࠿࠵ࡾ࡛⩻࠻㸡࢛ࣜ࢝ࢾࣚࡡິឺࢅふᐳࡌ
ࡾࡆ࡛࡞ࡻࡖ࡙᳠ッࢅ⾔ࡖࡒ㸣
࠘⤎ᯕ࠙྘࢛ࣜ࢝ࢾࣚࡡふᐳࡡࡒࡴࠉᙔ◂✪ᐄ࡚షᠺࡈࡿࡒࠉࣃࢪࢹࣤ H2Bࠉ࢛࣋ࣜ࢞ࢨࢮ࣭࣑ᑻᅹࢨࢡ
ࢻࣜࠉࡱࡒࡢࢠ࢙ࣤ㓗ྙᠺ㓕⣪࡞ࡐࡿࡑࡿ EGFP ࢅ௛ຊࡊࡒ⼝ྙࢰࣤࣂࢠ㈻ࢅⓆ⌟ࡌࡾᰬࢅ౐⏕ࡊࡒ㸣ࡆࡿ
ࡼࡡᰬࢅ 24 ᫤㛣ᇰ㣬ᚃふᐳࡊࡒ࡛ࡆࢀࠉ᪜࡞ሒ࿈ࡈࡿ࡙࠷ࡾࡻ࠹࡞ࡐࡿࡑࡿᰶ㸡࢛࣋ࣜ࢞ࢨࢮ࣭࣑ࠉ࣐ࢹࢤ
ࣤࢺࣛ࢓࡞⺧ක࠿ふᐳࡈࡿࡒ㸣ࡆࡿ࡞ᑊࡊ 48 ᫤㛣ᇰ㣬ᚃࡡⳞ⣊మ࡚ࡢධ࡙ࡡᰬ࡞࠽࠷࡙ࠉᾦ⬂ࡡⓆ㐡ࡊࡒᇱ
ᗇ㒂ࡡⳞ⣊࡚ୌ㒂ࡡ EGFP ⺧ක࠿ᾦ⬂හ⭅࡞ࡵふᐳࡈࡿࡒ㸣ୌ᪁࡚࢛࣭ࢹࣆ࢒ࢩ࣭࡞ᚪ㡪࡝ Aoatg8 ࢅḖ᥾ࡊ
ࡒᰬ࡞࠽࠷࡙྘⼝ྙࢰࣤࣂࢠ㈻ࢅⓆ⌟ࡈࡎふᐳࡊࡒ࡛ࡆࢀࠉධ࡙ࡡᰬ࡞࠽࠷࡙ 60 ᫤㛣௧୕ᇰ㣬ࢅ⾔ࡖ࡙ࡵ
EGFP ⺧කࡢ࡮࡛ࢆ࡜ᾦ⬂හ⭅࡞ふᐳࡈࡿ࡝࠾ࡖࡒࠊ௧୕ࡡ⤎ᯕ࠾ࡼࠉᇱᗇ㒂ࡡⳞ⣊࡞࠽࠷࡙ࡢ࢛࣭ࢹࣆ࢒
ࢩ࣭࡞ࡻࡽࠉᰶࡷ࢛࣋ࣜ࢞ࢨࢮ࣭࣑ࠉ࣐ࢹࢤࣤࢺࣛ࢓࡛࠷ࡖࡒ࢛ࣜ࢝ࢾࣚࡡᵋᠺᠺฦ࠿ᾦ⬂࡫ཱི࡛ࡽ㎲ࡱࡿࠉ
ฦよࡈࡿ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ
1) ḿ㊪ࡼࠉ➠஫ᅂ⣊≟ⳞฦᏄ⏍∸Ꮥࢤࣤࣆ࢒ࣝࣤࢪㅦⁿさ᪠㞗ࠉp41
Autophagic uptake of organelle components by vacuoles in mature hyphae of A. oryzae.
Jun-ya SHOJI, Takashi KIKUMA, Manabu ARIOKA, Katsuhiko KITAMOTO (Dept. Biotech., Univ. Tokyo)
39
P-9
ᢰᏄⳞ Coprinus cinereus ⏜ᮮ࢛࣭ࢹࣆ࢒ࢩ࣭㛭㏻ࢰࣤࣂࢠ㈻ Atg8 ࣓࣌ࣞࢡࡡᶭ⬗よ
ᯊ
ྚᮟ᪸ᚠ㸡ῳ㑋 ᙪ, 㯖⏛᜜ᙢ㸝㤮ᕖኬ࣬㎨㸞
࢛࣭ࢹࣆ࢒ࢩ࣭࡛ࡢࠉ┷ᰶ⣵⬂࡞࠽ࡄࡾࢰࣤࣂࢠ㈻ฦよᶭᵋࡡୌࡗ࡚࠵ࡽࠉ㟸≁␏Ⓩ࡞⣵⬂හ࡞Ꮛᅹࡌࡾ
࢛ࣜ࢝ࢾࣚࡷࢰࣤࣂࢠ㈻ࢅฦよࡌࡾ⣌࡚࠵ࡾࠊ㎾ᖳࠉ྘⛸⏍∸࡞࠽ࡄࡾ࢛࣭ࢹࣆ࢒ࢩ࣭㛭㏻㐿ఎᏄࡡ◒ቪᰬ
➴ࡡよᯊ࠾ࡼࠉ࢛࣭ࢹࣆ࢒ࢩ࣭࠿⣵⬂ฦ໩㐛⛤࡞࠽࠷࡙ࡵ㔔さ࡝ᙲ๪ࢅᣚࡗࡆ࡛࠿♟ြࡈࡿ࡙ࡀ࡙࠷ࡾࠊᮇ
◂✪࡚ࡢࠉᢰᏄⳞ Coprinus cinereus ࡞࠽ࡄࡾ࢛࣭ࢹࣆ࢒ࢩ࣭࡞ࡗ࠷࡙᪺ࡼ࠾࡛ࡌࡾࡒࡴࠉࡱࡍࠉྜྷᶭᵋࡡ࣏
࣭࣭࡛࢜⩻࠻ࡼࡿ࡙࠷ࡾ Atg8 ࣓࣌ࣞࢡࡡᶭ⬗よᯊࢅ⾔ࡖࡒࡡ࡚ࡆࡆ࡞ሒ࿈ࡌࡾࠊ
C. cinereus ࡡࢣࢿ࣑よㄖ᝗ሒ࡞ᇱࡘࡀ PCR Ἢ࠽ࡻࡦ RACE Ἢࢅ⏕࠷࡙ C. cinereus ࡻࡽ Atg8(CcAtg8)࣓࣌ࣞ
ࢡ cDNA ࢅཱིᚋࡊࡒࠊよᯊࡡ⤎ᯕࠉྜྷ cDNA ࡢ 126 ࢓࣐ࢿ㓗ṟᇱࢅࢤ࣭ࢺࡊ࡙࠷ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊ
Saccharomyces cerevisiae ࡞࠽࠷࡙ Atg8 ࡡḖ᥾ᰬࡢࠉ⬂Ꮔᙟᠺ⬗ࢅḖ኶ࡊ࡙࠷ࡾࡣ࠾ࡽ࡚࡝ࡂࠉ࢛࣭ࢹࣆ࢒
ࢩ࣭ㄇᑙ᮪௲࡞࠽࠷࡙ふᐳࡈࡿࡾ Atg8 ࡡᾦ⬂࡫ࡡᑻᅹࡵ⾔ࢂࡿ࡝ࡂ࡝ࡾࠊࡐࡆ࡚ࠉᚋࡼࡿࡒ CcAtg8 ࣓࣌ࣞ
ࢡ cDNA ࢅ S. cerevisiae ATG8 Ḗ᥾ᰬ࡞ᑙථࡊࠉࡐࡡ┞⿭ᛮ࡞ࡗ࠷࡙᳠ゞࡊࡒࠊࡐࡡ⤎ᯕࠉCcAtg8 ࣓࣌ࣞࢡ
cDNA ࢅᑙථࡊࡒᰬ࡚ࡢࠉ㔕⏍ᰬ࡛ྜྷᵕࡡ⬂Ꮔᙟᠺ⬗ࢅᅂᚗࡌࡾࡆ࡛࠿☔ヾࡈࡿࠉࡱࡒ⥫Ⰵ⺧කࢰࣤࣂࢠ㈻
ࢅ࣏࣭࣭࡛࢜ࡊࡒᑻᅹよᯊ࡚ࡢࠉCcAtg8 ࣓࣌ࣞࢡ࠿ᾦ⬂࡞ᑻᅹࡌࡾࡆ࡛࠿☔ヾࡈࡿࡒࠊ
௧୕ࡡ⤎ᯕࡢࠉC.cinereus ࡻࡽཱིᚋࡊࡒ CcAtg8 ࣓࣌ࣞࢡ cDNA ࠿ࠉS. cerevisiae හ࡞࠽࠷࡙࢛࣭ࢹࣆ࢒ࢩ
࣭㛭㏻㐿ఎᏄ࡛ࡊ࡙ᶭ⬗ࡊ࡙࠷ࡾࡆ࡛ࢅ♟ြࡌࡾࡵࡡ࡚࠵ࡾࠊ
Functional analysis of Atg8 homolog from Coprinus cinereus.
Masanori Yoshimura, Akira Watanabe, Yasuhiko Asada
(Dept. of Agriculture, Univ. of Kagawa)
P-10
㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ࢙ࣤࢺࢮ࣭࣑ᵕᵋ㏸మࡡよᯊ
ᵵཾ⿩ḗ㑳ࠉḿ㊪ῗஒࠉ᭯ᒱ Ꮥࠉ໪ᮇົࡥࡆ (᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ)
࠘┘Ⓩ࠙⣊≟Ⳟ࡞࠽࠷࡙࢙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪࡢࡱࡓࡐࡡᏋᅹ࠿᪺ࡼ࠾࡞࡝ࡖࡒṹ㝭࡚࠵ࡽࠉࡐࡡレ⣵࡝ᶭᵋ
ࡢ࡮࡛ࢆ࡜ࢂ࠾ࡖ࡙࠷࡝࠷ࠊᠻࠍࡢࡆࡿࡱ࡚࡞⣵⬂⭯ࢰࣤࣂࢠ㈻࡛ EGFP ࡛ࡡ⼝ྙࢰࣤࣂࢠ㈻࡚࠵ࡾ
AoUapC-EGFP ࢅ⏕࠷࡙⣊≟Ⳟ࡞࠽ࡄࡾ࢙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪࡡᏋᅹࢅิࡴ࡙♟ࡌ࡛࡛ࡵ࡞ࠉ࢙ࣤࢺࢦ࢕ࢹ࣭ࢨ
ࢪㄇᑙ᫤࡞ኣࡂふᐳࡈࡿࡾ࢙ࣤࢺࢮ࣭࣑ᵕᵋ㏸మࢅぜฝࡊ࡙࠷ࡾ 1) 2)ࠊࡱࡒ࢙ࣤࢺࢮ࣭࣑ᵕᵋ㏸మࡢ᚜ᑚ⟮౪
ᏋⓏ࡞⣵⬂හࢅⳞ⣊ࡡ㍂࡞Ἒࡖࡒິࡀࢅࡊ࡙࠷ࡾࡆ࡛ࢅ♟ࡊࡒࠊࡐࡆ࡚௑ᅂࡢ࢙ࣤࢺࢮ࣭࣑ᵕᵋ㏸మ࡞㛭ࡌ
ࡾࡻࡽレ⣵࡝よᯊࢅ⾔ࡖࡒࠊ
࠘᪁Ἢ࡛⤎ᯕ࢙࠙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪࡡㄇᑙᚃ࡞࢓ࢠࢲࣤ⬲㔔ྙ๠ࢅష⏕ࡈࡎࡾ࡛Ⳟ⣊හ࡞࢙ࣤࢺࢮ࣭࣑ᵕᵋ
㏸మࡢふᐳࡈࡿ࡝ࡂ࡝ࡖࡒࠊࡆࡡࡆ࡛࠾ࡼ࢓ࢠࢲࣤࡢ࢙ࣤࢺࢮ࣭࣑ᵕᵋ㏸మࡡᾦ⬂࡫ࡡິឺ࡞ᙫ㡢ࡊ࡙࠷࡝
࠷ࡆ࡛࠿♟ြࡈࡿࡒࠊࡈࡼ࡞ࠉ࢙ࣤࢺࢮ࣭࣑ᵕᵋ㏸మࡡ࣓࣭ࢰ࣭ࢰࣤࣂࢠ㈻࡛ࡡ㛭ࢂࡽࢅㄢ࡬ࡾࡒࡴ࡞ࢱ࢕
ࢼࣤ㔔㙈ཀྵࡦࢱ࢕ࢻࢠࢲࣤ々ྙమࡡḖ᥾ᰬࢅ⏕࠷࡙ふᐳࡊࡒ࡛ࡆࢀࠉ࡮࡛ࢆ࡜ࡡ࢙ࣤࢺࢮ࣭࣑ᵕᵋ㏸మࡢⳞ
⣊࡞Ἒࡖࡒ㐘ິࢅೳḾࡊࡒࠊࡆࡡࡆ࡛࠾ࡼࢱ࢕ࢼ࣓࣭ࣤࢰ࣭ࢰࣤࣂࢠ㈻ࡢ࢙ࣤࢺࢮ࣭࣑ᵕᵋ㏸మࡡິឺ࡞ୌ
㒂㛭୙ࡊ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ⌟ᅹ࢙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪࡡ᮪௲Ḗ᥾ᰬࢅష⿿ࡊࠉࡐࡿࢅ⏕࠷࡙࢙ࣤࢺࢮ
࣭࣑ᵕᵋ㏸మ࡞㛭ࡊ࡙᭞࡝ࡾよᯊࢅ⾔ࡖ࡙࠷ࡾࠊ
1) ᵵཾࡼࠉ➠ 5 ᅂ⣊≟ⳞฦᏄ⏍∸Ꮥࢤࣤࣆ࢒ࣝࣤࢪさ᪠㞗 p.42
2) Higuchi Y. et al. (2006) Biochem. Biophys. Res. Commun. 340, 784-791.
Analysis of the endosomal-like structures in Aspergillus oryzae
Yujiro Higuchi, Jun-ya Shoji, Manabu Arioka, Katsuhiko Kitamoto (Dept. of Biotech., Univ. of Tokyo)
40
P-11
㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ AoVps24 ࡡᾦ⬂ᙟᠺ࡞㛭ࡌࡾᶭ⬗ࡡよᯊ
㎦ᕦᬏ⣎㸡ⳝ㛣㝧ᚷ㸡ḿ㊪ῗஒ㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ (᮶ኬ㝌㹺㎨⏍⛁㹺ᚺ⏍ᕝ)
<┘Ⓩ>VPS (vacuolar protein sorting)㐿ఎᏄࡢฝⰾ㓕ẍ S. cerevisiae ࡞࠽࠷࡙ᾦ⬂㓕⣪࢜ࣜ࣍࢞ࢨ࣋ࣈࢲࢱ࣭ࢭ
Y (CPY)ࢅ⣵⬂አ࡞ฦἢࡌࡾን␏࡞㛭ࡌࡾ㐿ఎᏄ࡚࠵ࡽࠉࢰࣤࣂࢠ㈻㍲㏞ᶭᵋࡷᾦ⬂ᶭ⬗࡝࡜࡞㛭୙ࡌࡾࡆ࡛
࠿▩ࡼࡿ࡙࠷ࡾࠊᠻࠍࡢࡆࡿࡱ࡚࡞࢙ࣤࢺࢮ࣭࣑ࢅ௒ࡊࡒᾦ⬂࡫ࡡࢰࣤࣂࢠ㈻㍲㏞࡞㛭ࢂࡾ VPS24 ࡡ A.
oryzae ࡞࠽ࡄࡾ࣓࣌ࣞࢡ Aovps24 ࢅ༟㞫ࡊࠉEGFP ⼝ྙࢰࣤࣂࢠ㈻Ⓠ⌟ᰬࢅཱིᚋࡊࡒࠊࡱࡒྜྷ㐿ఎᏄ◒ቪᰬ࡚
ࡢⴥࡊ࠷⏍⫩㜴ᐐࡷⳞ⣊ࡡᙟឺ␏ᖏࠉฦ⏍Ꮔᙟᠺᶭ⬗ࡡḖ᥾࠿ふᐳࡈࡿࡒ
1)
ࠊᮇ◂✪࡚ࡢࡆࡿࡼࡡᰬࢅ⏕࠷
࡙ A. oryzae ࡞࠽ࡄࡾᾦ⬂ᶭ⬗ཀྵࡦᾦ⬂ᙟᠺࡡよᯊࢅ⾔ࡖࡒࠊ
<᪁Ἢཀྵࡦ⤎ᯕ>Aovps24 ◒ቪᰬ࡞ CPY-EGFP ཀྵࡦ EGFP-AoVam3 ࢅࡐࡿࡑࡿⓆ⌟ࡈࡎࠉᾦ⬂ཀྵࡦᾦ⬂⭯ࡡふ
ᐳࢅ⾔ࡖࡒࠊ ◒ቪᰬ࡞࠽࠷࡙ᾦ⬂ࡢᮅⓆ㐡࡚࠵ࡽࠉ㏳ᖏࡡ୷࠷ኬࡀ࡝ᾦ⬂ࡢふᐳࡈࡿ࡝࠾ࡖࡒࠊS. cerevisiae
ࡡ vps24 ን␏ᰬ࡚ࡢᾦ⬂ࡡ␏ᖏࡢふᐳࡈࡿࡍࠉⴥࡊ࠷⏍⫩㜴ᐐࡷᙟឺ␏ᖏࡢぜࡼࡿ࡝࠷ࡆ࡛࠾ࡼࠉAovps24
◒ቪᰬࡡ⾪⌟ᆵࡢᾦ⬂ᙟᠺࡡḖ᥾࡞ࡻࡾࡵࡡ࠿♟ြࡈࡿࡒࠊࡊࡒ࠿ࡖ࡙⣊≟Ⳟ࡚࠵ࡾ A. oryzae ࡞࠽࠷࡙ࡢᾦ
⬂ᶭ⬗࠿Ⳟ⣊ࡡᙟឺᙟᠺࡷฦ໩࡞ࡵ㛭୙ࡌࡾ࡛⩻࠻ࡼࡿࡾࠊࡱࡒ EGFP-AoVps24 ࢅ⏕࠷࡙ AoVps24 ࡡᾦ⬂Ⓠ
㐡᮪௲ୖ࡚ࡡᣪິࡡふᐳࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉEGFP-AoVps24 ࡚ྊっ໩ࡈࡿࡾᾦ⬂㎾എࡡᚃ᭿࢙ࣤࢺࢮ࣭࣑
ᵕᵋ㏸మ࠿ N ″ᯜῤ᮪௲ୖ࡞࠽࠷࡙ฆ㞗ࡌࡾᵕᏄ࠿ふᐳࡈࡿࡒࠊࡆࡡฆ㞗ࡊ࡙ぜࡼࡿࡾᵋ㏸మࡢ FM4-64 ࡞
ࡻࡽ᯹Ⰵࡈࡿࡾࡆ࡛࠾ࡼࠉᚃ᭿࢙ࣤࢺࢮ࣭࣑ᵕᵋ㏸మ࠿ฆ㞗ࡊࡒࡵࡡ࡚࠵ࡾ࡛⩻࠻ࡼࡿࠉ⌟ᅹࡆࡡᵋ㏸మࡡ
レ⣵ࢅよᯊ୯࡚࠵ࡾࠊ
1) Tatsumi et al.,(2006) Biochem. Biophys. Res. Commun. 347, 970-978
Functional analysis of AoVps24 for vacuolar formation in A. oryzae
Akinori Tatsumi, Takashi Kikuma, Jun-ya Shoji, Manabu Arioka, Katsuhiko Kitamoto
(Dept. of Biotechnology, Univ. of Tokyo)
P-12
࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ௛╌ჹᙟᠺ࡞ᑊࡌࡾ྘⛸ᢘ⏍∸㈻ࡡᙫ㡢
㛏ᓞ⏜ኚ㸡Ṃᕖ἖㸡᩟⸠ᙤ⳱㸡㙂ಲ㧏ᚷ 㸝᮶⌦ኬ⌦ᕝ㸞
࣏ࢠࣞࣚ࢕ࢺ⣌ᢘ⏍∸㈻ࡢཋᰶ⏍∸࡞ᑊࡊ࡙ࢰࣤࣂࢠྙᠺ㜴ᐐὩᛮࢅࡵࡗࠊ┷ᰶ⏍∸࡞ᑊࡊ࡙ࡵ㧏⃨ᗐ࡚
࣐ࢹࢤࣤࢺࣛ࢓࡝࡜ࡡཋᰶ⏍∸ᆵࣛ࣍ࢮ࣭࣑ࢅᣚࡗ࢛ࣜ࢝ࢾࣚ࡞ష⏕ࡊ࿣ྺ㜴ᐐ➴࠿㉫ࡆࡾࠊࡊ࠾ࡊ㎾ᖳ 14
ဤ⎌࣏ࢠࣞࣚ࢕ࢺ࠿┷ᰶ⏍∸࡞ᑊࡊ࡙ᚉᮮࡡష⏕ᶭష࡚ࡢㄕ᪺࡚ࡀ࡝࠷ష⏕ࢅ♟ࡌࡆ࡛࠿ሒ࿈ࡈࡿ࡙࠷ࡾࠊ
࢕ࢾ࠷ࡵࡔ⑋Ⳟ࡚ࡵ࿣ྺ㜴ᐐࢅ♟ࡈ࡝࠷⃨ᗐ࡚௛╌ჹᙟᠺ㜴ᐐ࠿ふᐳࡈࡿࡒࠊྜྷᵕࡡ௛╌ჹᙟᠺ㜴ᐐࡢࢠࣞ
࣑ࣚࣆ࢘ࢼࢤ࣭࡚ࣜ᳠ᏽࢅ⾔ࡖࡒሔྙ࡞ࡵふᐳࡈࡿࠉ௙࡞ࡵ௛╌ჹฦ໩࡞ᑊࡊྜྷᵕࡡᙫ㡢ࢅ୙࠻ࡾᢘ⏍∸㈻
࠿࠵ࡾࡡ࡚ࡢ࡝࠷࠾࡛⩻࠻ࡼࡿࡾࠊࡐࡆ࡚ࠉࢰࣤࣂࢠ㈻ྙᠺ㜴ᐐຝᯕࡡ࠵ࡾᢘ⏍∸㈻࡞㝀ࡼࡍᵕࠍ࡝ᢘ⏍∸
㈻ࢅ⏕࠷࡙࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ௛╌ჹฦ໩࡞ᑊࡌࡾᙫ㡢ࢅ᳠ᏽࡊࡒ࡛ࡆࢀࠉ┷ᰶ⣵⬂ࡡࢰࣤࣂࢠ㈻ྙᠺࢅ㜴ᐐ
ࡌࡾ glutarimide ⣌ᢘ⏍∸㈻ 9-Me streptimidoneࠉࢽࢠ࢛ࣝࢲࢺ⏍ྙᠺ⣌㜴ᐐ๠ Hadacidin ➴࡚ฦ⏍ᏄࡡⓆⰾ࠿
ྊ⬗࡝⃨ᗐ࡞࠽࠷࡙௛╌ჹฦ໩㜴ᐐ࠿ふᐳࡈࡿࡒࠊ
The effect of antibiotics on appressorial development of Magnaporthe oryzae.
Yuka Nagashima, Osamu Takekawa, Ayaka Saitoh, Takashi Kamakura.
(Tokyo Univ.of Sciense)
41
P-13
࢕ࢾ࠷ࡵࡔ⑋Ⳟ࡞࠽ࡄࡾᶭ᲌ๆ⃥ུᐖ࢜ࣜࢨ࣑ࢗ࢕࢛ࣤࢲࢾࣜ MID1 ࣓࣌ࣞࢡࡡ ᶭ
⬗よᯊ
௿⃕┷⃀㸡᯵ᮄ▩Ꮔ㸡㙂ಲ㧏ᚷ 㸝᮶⌦ኬ⌦ᕝ㸞
࢜ࣜࢨ࣑ࢗࡢ⣊≟Ⳟࡡ඙❻ᠺ㛏࡞࠽࠷࡙ࡐࡡ⃨ᗐ໖㒼࠿⏕࠷ࡼࡿࡾࡓࡄ࡚࡝ࡂࠉ஦ḗ࣒ࢴࢬࣤࢩ࣭࡛ࣔࡊ
࡙ᵕࠍ࡝ࢨࢡࢻࣜఎ㐡࡞㛭ࢂࡖ࡙࠷ࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊ࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ௛╌ჹฦ໩࡞࠽࠷࡙ࡵ࢜ࣜࢨࢗ
࣑ࢨࢡࢻࣜ࠿㔔さ࡝ᙲ๪ࢅᯕࡒࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊMID1 ࡢ S.cerevisiae ࡚Ⓠぜࡈࡿࡒᶭ᲌ๆ⃥ུᐖ
ᆵ࢜ࣜࢨ࣑ࢗ࢕࢛ࣤࢲࣔࢾ࡚ࣜ࠵ࡾ࠿ࠉࡆࡡ㐿ఎᏄࡢ⣊≟Ⳟ࡞࠽࠷࡙ᗀࡂಕᏋࡈࡿ࡙࠷ࡒࠊ᪝ᮇ⏐࢕ࢾ࠷ࡵ
ࡔ⑋Ⳟ P2 ᰬࢅࡵࡔ࠷࡙ MID1 ࣓࣌ࣞࢡ㐿ఎᏄࡡⓆ⌟よᯊࢅ RT-PCR ࡞ࡻࡽ⾔ࡖࡒ࡛ࡆࢀࠉᰜ㣬ᠺ㛏᫤ཀྵࡦ௛
╌ჹᙟᠺ᮪௲ୖࡡ୦᮪௲ୖ࡚Ⓠ⌟࠿ぜࡼࡿࡒࠊࡱࡒࠉࢣࢿ࣑ࢸ࣭ࢰࡻࡽ஢᝷ࡈࡿࡾ MID1 ࣓࣌ࣞࢡ㐿ఎᏄࡡ
ORF ࢅ┞ྜྷⓏ 2 ᅂ⤄ᥦ࠻࡞ࡻࡽ◒ቪࡊࡒ࡛ࡆࢀࠉ◒ቪᰬࡡ⏍⫩㏷ᗐ࣬௛╌ჹᙟᠺ⬗࡞㔕ᛮᰬ࡛ࡡ᭯ណ࡝ᕣࡢ
ぜࡼࡿ࡝࠾ࡖࡒࠊ⌟ᅹࠉ࿔ᅑࡡ࢜ࣜࢨ࣑ࢗ⎌ሾࡷࢪࢹࣝࢪ࡞ᑊࡌࡾᚺ➽࡞ࡗ࠷࡙レ⣵࡝᳠ゞࢅ⾔ࡖ࡙࠷ࡾࠊ
Functional analyses of mechanosensitive ion channel MID1 homolog in Magnaporthe oryzae.
Masumi Izawa, Tomoko Kashiwagi, Takashi Kamakura
㸝Tokyo Univ. of Sciense㸞
P-14
࢕ࢾ⛁᳔∸࠷ࡵࡔ⑋Ⳟ(Pyricularia spp.)࡞࠽ࡄࡾ㟸⑋ཋງ㐿ఎᏄ AVR-Pita ࡡᗑ஋᯹Ⰵ
మንິᶭᵋ
୯㤷࠷ࡘࡲ㸡☶㒂༐ᑖ㸡୯ᒁᩔᆍ㸡┶ᒜ⁘ᚷ㸡ᅰఫᖶ㞕 㸝♼ᡖኬ࣬㎨㸞
࠷ࡵࡔ⑋Ⳟ࡞࠽ࡄࡾᐞ⏍ᛮฦ໩ᶭᵋࢅࢣࢿ࣑ᵋ㏸࠾ࡼよ᪺ࡌࡾࡆ࡛ࢅ┘Ⓩ࡞㸡々ᩐ⛸ࡡ Pyricularia ᒌⳞ㞗
ᅆࡡẒ㍉ᰶᆵよᯊࢅ⾔ࡖࡒ㸣ࡐࡡ⤎ᯕ㸡ᮇⳞ㞗ᅆ࡞ࡢ㸡᯹Ⰵమᵋ㏸ࡡኣᵕ໩࡛࡛ࡵ࡞㟸⑋ཋງ㐿ఎᏄᗑࢅን
ິ࣬ฦᩋࡈࡎࡾᶭᵋ࠿Ꮛᅹࡌࡾࡆ࡛࠿♟ြࡈࡿࡒ㸣ࡆࡡ࠹ࡔ㸡ࡌ࡚࡞ࢠ࣭ࣞࢼࣤࢡࡈࡿ࡙࠷ࡾ㟸⑋ཋງ
㐿ఎᏄࡡࡥ࡛ࡗ AVR-Pita ࡡᗑ஋᯹Ⰵమ࡛࿔㎰ᵋ㏸ࢅレ⣵࡞᳠ゞࡊࡒ㸣ᮇ㐿ఎᏄࡡᗑ஋᯹Ⰵమ࡝ࡼࡦ࡞᯹Ⰵమ
୕ࡡᗑ஋న⨠ࡢ㸡ᖏ᯹Ⰵమ࠵ࡾ࠷ࡢ㐛๨᯹Ⰵమ㸡ࢷ࣒ࣞ࢓㎾എ࠵ࡾ࠷ࡢ㟸ࢷ࣒ࣞ࢓㡷ᇡ࡛࠷࠹ࡻ࠹࡞ᵕࠍ࡚㸡
ᮇ㐿ఎᏄ࠿ᐞ⏍ᛮฦ໩ࡡ㐛⛤࡚ࢣࢿ࣑୕ࢅࢱ࢕ࢻ࣐ࢴࢠ࡞ንິ࣬ฦᩋࡊࡒࡆ࡛࠿ึ᪺ࡊࡒ㸣ࡆࡡᶭᵋࢅよ᪺
ࡌࡾࡒࡴ㸡ᮇ㐿ఎᏄ࿔㎰㡷ᇡࢅࢠ࣭ࣞࢼࣤࢡࡊ㸡ᵋ㏸ࢅẒ㍉ࡊࡒ࡛ࡆࢀ㸡ᮇ㐿ఎᏄ୕Ὦ࠽ࡻࡦୖὮ࡞ᵕࠍ࡝
⤄ࡲྙࢂࡎ࡚ࣝࢹࣞࢹࣚࣤࢪ࣎ࢯࣤࡡ᩷∞➴࠿Ꮛᅹࡊࡒ㸣ࡆࡡࡆ࡛࠾ࡼ㸡࿔㎰࡞Ꮛᅹࡌࡾ㌹⛛ᅄᏄ࠿␏ᡜⓏ
⤄ࡲᥦ࠻ࡡ፳మ࡞࡝ࡽ㸡ᮇ㐿ఎᏄࢅ⛛ິࡈࡎࡒ࡛⩻࠻ࡼࡿࡒ㸣≁࡞࢕ࢾⳞ⩄࡚ࡢ㸡Ⳟ⩄හ࡚ࡵᗑ஋᯹Ⰵమࡡ
ንິ࣬ฦᩋ࠿⃥ࡊࡂ㉫ࡆࡖ࡙࠽ࡽ㸡࿔㎰࡞㞗✒ࡊࡒ㌹⛛ᅄᏄ㒼า࠿௙ࡡⳞ⩄࡛Ẓ࡬࡙ࡻࡽኣᵕ࡚࠵ࡖࡒ㸣ࡆ
ࡡࡻ࠹࡝࿔㎰ᵋ㏸ࡡኣᵕ໩࠿␏ᡜⓏ⤄ࡲᥦ࠻ࡡᶭఌࢅቌኬࡈࡎ㸡ࢤࣅ࣭ᩐࡷᗑ஋᯹Ⰵమࡡࡻࡽ⃥ࡊ࠷ንິᛮ
ࢅ࢕ࢾⳞ࡞௛୙ࡊࡒ࡛ᛦࢂࡿࡒ㸣
Dynamics of an avirulence gene locus, AVR-Pita, in Pyricularia spp.
Izumi Chuma, Chihiro Isobe, Hiotshi Nakayashiki, Shigeyuki Mayama, Yukio Tosa
(Fac. of Agriculture, Kobe Univ.)
42
P-15
࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ㟸⑋ཋᛮ㐿ఎᏄ Avr-pia ࡡࢠ࣭ࣞࢼࣤࢡ
᭧᰷㍜㞕 1㸡୔ᮄួ௒ 1㸡ᮿ஬ᖶኯ㑳 1㸡Ⱙ⃕Ṃெ 2 㸡㨛㢄ⱝᶖ 3 㸡ᖲඳ㔔ୌ஄ 1㸡୯ᓞᨼᙢ 4 㸡ෞ⏛ᡛ⏠ 5 㸝1 ໪ኬ㝌㎨࣬ᚺ⏕ⳞᏕ㸡2 ୯ኳ㎨◂㸡3 ㎨ᴏ⏍∸㈠″◂㸡4 ᮶໪㎨◂㸡5 ᨲ㏞ኬ㸞
ᠻࠍࡢ㸡࢕ࢾ㸲࠷ࡵࡔ⑋Ⳟࡡᐙ୹≁␏ᛮ┞பష⏕ࡡよᯊ࡛㸡ᐙ୹≁␏ᛮን␏ࡡᶭᵋࢅよᯊࡌࡾࡒࡴ㸡࢕ࢾ
ဗ⛸យ▩᪣ࡡࡵࡗ┷ᛮᢤᢘᛮ㐿ఎᏄ Pi-a ࡞ᑊࡌࡾ㟸⑋ཋᛮ㐿ఎᏄ࡚࠵ࡾ Avr-Pia ࡡࢠ࣭ࣞࢼࣤࢡࢅモࡲ࡙࠷
ࡾ㸣⌟ᅹࡱ࡚࡞࢕ࢾ࠷ࡵࡔ⑋Ⳟ Ina168 ᰬࢤࢪ࣐ࢺࢣࢿ࣑ࣚ࢕ࣇ࣭ࣚࣛ࠾ࡼ Avr-Pia 㡷ᇡࢅྱࡳ⣑ 40kb ࡡ࢕ࣤ
ࢦ࣭ࢹࢅྱࡳࢤࢪ࣐ࢺࢠ࣭ࣞࣤࢅᚋ࡙࠷ࡾ㸣ࡐࡆ࡚㸡࢕ࣤࢦ࣭ࢹ⣑ 40kb ࡡ㒼า࠾ࡼࢹࣚࣤࢪ࣎ࢯࣤ➴ࡡ᪜▩
ࡡཬᚗ㒼าࢅ㝎ࡂ㒂ฦࢅ PCR Ἢ࡞ࡻࡖ࡙ IࠤVI ࡡ 6 ࡗࡡ᩷∞࡞ฦࡄ㸡ืࠍ࡞ࢠ࣭ࣞࢼࣤࢡࡊࡒ㸣ࡆࡿࡼࢠࣞ
࣭ࣤࢅយ▩᪣࡞ᑊࡊ࡙⑋ཋᛮࢅࡵࡗ㸝avr-Pia㸞ን␏ᰬ Ina168m95-1 ࡞ᙟ㈻㌹ᥦࡊ㸡ࡆࡿࡼࡡᙟ㈻㌹ᥦమࢅយ
▩᪣࡞᥃⛸ࡊࡒ㸣ࡐࡡ⤎ᯕ㸡᩷∞ V ࢅᙟ㈻㌹ᥦࡊࡒࡵࡡࡓࡄ࠿㸡យ▩᪣࡞ᑊࡊ࡙㟸⑋ཋᛮࢅ♟ࡊࡒ㸣ࡱࡒ㸡
ྜྷᵕ࡞យ▩᪣࡞⑋ཋᛮࡡᅜሔฦ㞫ᰬ Ina86-137 ᰬ࡞᩷∞ V ࢅᙟ㈻㌹ᥦࡊࡒ࡛ࡆࢀ㸡យ▩᪣࡫ࡡ⑋ཋᛮࢅ኶ࡖ
ࡒ㸣᩷∞ V ࡡ㡷ᇡࢅቌᖕࡌࡾࣈࣚ࢕࣏࣭ࢅ⏕࠷࡙ᅜሔฦ㞫ᰬࡡࢣࢿ࣑ DNA ࡡ PCR ཬᚺࢅ⾔ࡖࡒ࡛ࡆࢀ㸡យ
▩᪣࡞㟸⑋ཋᛮࡡⳞᰬࡡࡲ࠾ࡼྜྷ᩷∞ࡡቌᖕ࠿ぜࡼࡿࡒ㸣௧୕ࡻࡽ㸡᩷∞ V ࡞ Avr-Pia ࢅྱࡳࡆ࡛࠿♟ြࡈ
ࡿࡒ㸣᩷∞ V ࡢධ㛏 3,531bp ࡚㸡හᅹࡌࡾ᭩ࡵ㛏࠷ 255bp ࡡ ORF ࡞⣵Ⳟ⏜ᮮ cytochrome ᵕࢰࣤࣂࢠ㈻࡫ࡡᘽ
࠷┞ྜྷᛮ࠿ࡲࡼࡿࡒ㸣
Molecular cloning of Avr-Pia, the avirulence gene in Magnaporthe oryzae toward the rice blast resistance gene
Pi-a㸣
Teruo Sone1, Shinsuke Miki1, Kotaro Matsui1, Taketo
4
5 1
Ashizawa2, Hideki Kito3, Kazuyuki Hirayae2, Toshihiko
2
Nakajima , and Fusao Tomita .( Hokkaido Univ., NARC, 3NIAS, 4NARCT, 5Univ. of Air)
P-16
Biotinylated NeoechinulinA ໩Ꮥྙᠺ⤊㊪ࡡᨭၻ࡛ Aspergillus ᒌࡡ NeoechinulinA ⏍ྙ ᠺ
㐿ఎᏄ⩄ࡡ᥀⣬
➁㔕ḿὊ㸡ኬ▴㯖⏜, ᇷහ᱀Ꮔ, 㙂ಲ㧏ᚷ 㸝᮶⌦ኬ⌦ᕝ㸞
Neoechinulin A ࡢ 1㸞ᢘ㓗໩ష⏕ࠉ2㸞⣵⬂ࡡ NADH㸢ࢸࣃࢺࣞࢣࢻ࣭ࢭὩᛮࡡ୕᪴ష⏕ࠉཀྵࡦ 3㸞⣵⬂ࡡ
Caspase-3 ࡡὩᛮᢒโష⏕ࠉࡡᑛ࡝ࡂ࡛ࡵ㸨ࡗࡡᶭ⬗ࢅ᭯ࡊ࡙࠽ࡽࠉSIN-㸦㸝3-(4-morpholinyl)sydnonimine
hydrochlorid㸞࡞ࡻࡽㄇᑙࡈࡿࡾ♼⤊⣵⬂Ṓ࡞ᑊࡊ࡙ࠉ㧏࠷ᢒโష⏕ࢅ♟ࡌࠊNeoechinulin A ࡢୌ㓗໩✽⣪ࡷ
ࢪ࣭ࣂ࣭࢛࢞ࢨࢺ࡛࠷ࡖࡒ⏍⌦Ὡᛮ∸㈻࡞ࡢᙫ㡢ࢅ୙࠻ࡍ࡞ᢘ㓗໩ష⏕ࢅ♟ࡌ໩ྙ∸࡚࠵ࡽࠉ⏍మහ࡫ࡡᙫ
㡢ࢅ⩻࠻ࡒሔྙ㟸ᖏ࡞᭯⏕࡚࠵ࡾࠊNeoechinulin A ࡢࠉ≁࡞ฦ໩ PC12 ⣵⬂ཀྵࡦิ௥ᇰ㣬♼⤊⣵⬂࡞࠽࠷࡙⣵
⬂Ṓࢅᢒโࡊࡒࡆ࡛࠾ࡼࠉ♼⤊⣵⬂࡞ᑊࡊ࡙≁␏Ⓩ࡞ష⏕ࡌࡾྊ⬗ᛮ࠿㧏࠷࡛⩻࠻ࡼࡿࡾࠊ
ࡆࡿࡼ⮾࿝῕࠷Ὡᛮࢅᣚࡖ࡙࠷ࡾ Neoechinulin A ࡓ࠿ࠉ⏍ྙᠺ࡞㛭ࡾ㐿ఎᏄࡢᮅࡓ༟㞫࣬よᯊࡈࡿ࡙࠷࡝
࠷ࠊࡻࡖ࡙ᠻࠍࡢ Neoechinulin A ࡡ⏍ྙᠺ࡞㛭ࡾ㐿ఎᏄࡡ༟㞫࡛よᯊࢅ┘ᣞࡊࡒࠊNeoechinulin A ⏍ྙᠺ⤊㊪
࡚ᚪさ࡝㓕⣪ࡢ᭩⤂⏐∸࡚࠵ࡾ Neoechinulin A ࡞ば࿰ᛮ࠿࠵ࡾ࡛஢ῼࡈࡿࡾࡡ࡚ࠉT7 ࣆ࢒࣭ࢩࢸ࢔ࢪࣈࣝ࢕
Ἢ࡞ࡻࡖ࡙ྙᠺ⤊㊪࡞㛭ࢂࡾ㐿ఎᏄࡡ༟㞫࡛よᯊࢅモࡲࡒࠊࡱࡒࠉT7 ࣆ࢒࣭ࢩࢸ࢔ࢪࣈࣝ࢕Ἢࢅ⾔࠹㝷࡞ᚪ
さ࡚࠵ࡖࡒ Neoechinulin A ࡡࣄ࢛ࢲࣤ໩ㄇᑙమࡡ໩Ꮥྙᠺࢅ⾔࠷ࠉ᪜▩ࡡྙᠺ᪁Ἢࡡᨭၻࢅモࡲࡒࠊ
Improvement of chemical synthesis of biotinylated Neoechinulin A and search for Aspergillus spp. genes involved
in the neoechinulin A biosynthetic pathway.
Masahiro Takeno, Mayu Oishi, Momoko horiuchi, and Takashi Kamakura.
(Tokyo Univ.of Science)
43
P-17
Aspergillus fumigatus ࠿⏍⏐ࡌࡾࢹࣛࣈࣞࢪࢰࢲࣤ㢦ࡡ⏍ྙᠺ㐿ఎᏄࢠࣚࢪࢰ࣭
ຊ⸠├ᶖ㸡㧏ᮄᾇࠉὰぜ⾔ᘧࠉ᤻ㆺ⚵᫓㸡㛏⏛⿩஄ 㸝⌦◂࣬୯ኳ◂࣬ᢘ⏍∸㈻㸞
Aspergillus fumigatus BM939 ᰬ࠿⏍⏐ࡌࡾࢹࣛࣈࣞࢪࢰࢲࣤ㢦ࡢ⣵⬂࿔᭿ࢅโᚒࡌࡾ⏍∸Ὡᛮࢅ᭯ࡌࡾࢩ
ࢢࢹࣅ࣋ࣚࢩࣤ໩ྙ∸࡚࠵ࡾࠊ᚜ᑚ⟮࠿ฦᏄᵾⓏ࡚࠵ࡽࠉࡐࡡ㔔ྙࢅ㜴ᐐࡌࡾࡆ࡛࡞ࡻࡽ⣵⬂࿔᭿ࢅฦ⿛᭿
࡞ೳḾࡈࡎࡾࠊᵕࠍ࡝㢦⦍మࡡ୯࡞ࡢ᚜ᑚ⟮㔔ྙ࡞ᑊࡊ࡙ಀ㐅Ὡᛮࢅ♟ࡌࡵࡡࡵ࠵ࡽࠉࢹࣛࣈࣞࢪࢰࢲࣤࡡ
ࣆ࢒࣭࣏ࢤ࢓ࡢᢘ⭐⑾๠ࡡ࣭ࣛࢺ໩ྙ∸࡛ࡊ࡙᭯᭻࡚࠵ࡾࠊ
ࢹࣛࣈࣞࢪࢰࢲࣤ㢦ࡡ᥆ᏽ⏍ྙᠺ⤊㊪ࢅᇱ࡞ࠉࢹࣛࣈࢹࣆ࢒࡛ࣤࣈࣞࣛࣤࡡ⦨ྙཬᚺࢅᢰ࠹㟸ࣛ࣍ࢮ࣭࣑
ᆵ࣋ࣈࢲࢺྙᠺ㓕⣪㸝NRPS㸞࡞ຊ࠻ࠉࣈࣝࢼࣜᇱ㌹⛛㓕⣪㸝DMATSࠉࢩ࣒ࢲࣜ࢓࣭ࣛࣜࢹࣛࣈࢹࣆ࢒ࣤྙ
ᠺ㓕⣪㸞
ࠉP450 㓗໩㓕⣪ࠉ࣒ࢲࣜᇱ㌹⛛㓕⣪ࡡ㛭୙ࢅ஢᝷ࡊࡒࠊA. fumigatus ࢣࢿ࣑୯࡞ぜฝࡈࡿࡾ㸫⛸㢦ࡡ
dimodular NRPS ࡡ࠹ࡔࠉ⏍ྙᠺ࡫ࡡ㛭୙࠿஢᝷ࡈࡿࡒ㓕⣪㐿ఎᏄࡌ࡬࡙࠿㎾എ࡞Ꮛᅹࡌࡾࡡࡢ㸦⛸㢦ࡓࡄ࡚
࠵ࡖࡒࠊ᭩㎾ࠉMaiya ࡼࡢᮇ NRPS 㐿ఎᏄࢅ A. nidulans ࡞࠽࠷࡙㐛๨Ⓠ⌟ࡈࡎࡾࡆ࡛࡞ࡻࡽࠉࢹࣛࣈࣞࢪࢰ
ࢲࣤ㢦⏍ྙᠺࡡิṹ㝭࡚࠵ࡾ brevianamide F㸝cyclo-L-Trp-L-Pro㸞ࡡ⏍ྙᠺ࡞㛭୙ࡊ࡙࠷ࡾࡆ࡛ࢅሒ࿈ࡊ࡙࠷
ࡾ㸝Maiya et al., ChemBioChem 2006, 7, 1062-1069㸞ࠊࡊ࠾ࡊ࡝࠿ࡼࠉࢣࢿ࣑よㄖᰬ㸝Af293㸞࡚ࡢࢹࣛࣈࣞࢪࢰ
ࢲࣤ㢦࠿᳠ฝࡈࡿ࡙࠽ࡼࡍࠉࢿࢴࢠ࢓ࢗࢹ࡞ࡻࡾ⏍ྙᠺ㐿ఎᏄ⩄ࡡᶭ⬗ྜྷᏽ࡞ࡢ⮫ࡖ࡙࠷࡝࠷ࠊ⌟ᅹࠉࢹࣛ
ࣈࣞࢪࢰࢲࣤ㢦⏍⏐ᰬࢅᑊ㇗࡞ು⿭ NRPSࠉDMATS 㐿ఎᏄࡡࢿࢴࢠ࢓ࢗࢹᰬࢅష⿿୯࡚࠵ࡾࠊࡱࡒࠉࢣࢿ࣑
よㄖᰬ࡛ࢹࣛࣈࣞࢪࢰࢲࣤ⏍⏐ᰬࡡࢹࣛࣈࣞࢪࢰࢲࣤ㢦ࡡ⏍⏐⬗ࡡᕣ␏ࡡཋᅄࢅ᪺ࡼ࠾࡞ࡌࡾࡒࡴࠉ⏍⏐ᰬ
࠾ࡼࡡ⏍ྙᠺ㐿ఎᏄࢠࣚࢪࢰ࣭ࡡࢠ࣭ࣞࣤ໩࡛ࢣࢿ࣑㒼าࡡẒ㍉ࢅ⾔ࡖ࡙࠷ࡾࠊ
Studies on tryprostatin biosynthesis gene cluster of Aspergillus fumigatus BM939.
Naoki Kato, Hiroshi Takagi, Yukihiro Asami, Hideaki Kakeya, Hiroyuki Osada
(Antibiotics Lab., Discovery Res. Institute, RIKEN)
P-18
㯔Ⳟ A. oryzae ࡡ III ᆵ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕 ⣪㐿ఎᏄࡡᶭ⬗よᯊ
ເࠎᗛ௥ 1㸡⸠஬ ໌ 1㸡Praveen Rao Juvvadi2㸡໪ᮇົࡥࡆ 2㸡ᾇ⩹ሪ㇇ 1
㸝1 ᮶ኬ㝌࣬ⷾ, 2 ᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
࠘┘Ⓩ࠙
ࡆࡿࡱ࡚࡞⣊≟Ⳟࡡ III ᆵ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪
㸝PKS㸞
࡛ࡊ࡙ิࡴ࡙ࡡౚ࡛࡝ࡾ 4 ࡗࡡ㐿ఎᏄ㸝csyAࠉ
csyBࠉcsyC ࠽ࡻࡦ csyD㸞ࢅ A. oryzae ࡡࢣࢿ࣑୯࡞ぜฝࡊࠉcsyA㸡B㸡D ࡡ 3 㐿ఎᏄ࠿ A. oryzae ࡚Ⓠ⌟ࡊ࡙࠷
ࡾࡆ࡛ࢅ☔ヾࡊࡒࠊIII ᆵ PKS 㐿ఎᏄࡢࠉࡐࡡり፳ࡌࡾཬᚺࡡኣᵕᛮ࠾ࡼࠉ⌟ᅹࠉ㟸ᖏ࡞Ἰ┘ࡈࡿ࡙࠷ࡾ࠿ࠉ
csy 㐿ఎᏄࢅྱࡳ⣊≟Ⳟ III ᆵ PKS 㐿ఎᏄࡢࠉ⣌⤣ᶖよᯊ࡞ࡻࡽ᳔∸ࡷࣁࢠࢷࣛ࢓ࡡ III ᆵ PKS 㐿ఎᏄ࡛ࡢ␏
࡝ࡾࢡ࣭ࣜࣈࢅషࡾࡆ࡛࠾ࡼࠉࡐࡡᶭ⬗࡞⮾࿝࠿ᣚࡒࡿ࡙࠷ࡾࠊᮇ◂✪࡚ࡢࠉA. oryzae ࡡ csy ྘㐿ఎᏄࡡᶭ
⬗ࢅよ᪺ࡌࡾࡒࡴࠉ㐿ఎᏄ◒ቪᰬࡡష⿿ࠉ࠽ࡻࡦࠉㄇᑙⓆ⌟࡞ࡻࡾ PKS ⏐∸ࡡྜྷᏽࢅモࡲࡒࠊ
࠘᪁Ἢ࠽ࡻࡦ⤎ᯕ࠙A. oryzae RIB40 ⏜ᮮࡡ NSR13 ᰬࢅᐙ୹࡛ࡊࠉ࣏࣭࣭࢜㐿ఎᏄ adeA ࡛ࡡ┞ྜྷ⤄ᥦ࠻࡞ࡻ
ࡽࠉcsyA 㐿ఎᏄࡡ◒ቪᰬࢅཱིᚋࡊࡒࠊᮇ◒ቪᰬࡢࠉ⏍⫩ࡷⳞ⣊࣬ฦ⏍ᏄⰅ⣪࡝࡜ࡡ⾪⌟ᆵ࡞㐢࠷ࡢヾࡴࡼࡿ
࡝࠾ࡖࡒࠊḗ࡞ࠉcsyA 㐿ఎᏄࢅ amyB ࣈ࣓࣭ࣞࢰ࣭ୖ࡞⤄ࡲ㎲ࢆࡓⓆ⌟ࣈࣚࢪ࣐ࢺࢅᵋ⠇ࡊࠉA. oryzae M-2-3
ᰬ࡞ᙟ㈻㌹ᥦ࣬ᑙථࡊࡒࠊ࣏ࣜࢹ࣭ࢪᏋᅹୖ࡞ csyA 㐿ఎᏄࢅㄇᑙⓆ⌟ࡈࡎࠉcsyA ≁␏Ⓩ࡝໩ྙ∸ࡡ᳠ฝࢅモ
ࡲࡒࠊㄇᑙᇰᆀ୕ΰ࡛Ⳟమࠉࡐࡿࡑࡿࢅ㓉㓗࢙ࢲ࡚ࣜ᢫ฝࡊࠉTLCࠉHPLC ࡞ࡻࡽฦᯊࡊࡒࠊࡐࡡ⤎ᯕࠉᇰ
ᆀࠉⳞమ࡛ࡵ࡞ࢤࣤࢹ࣭ࣞࣜࡡᙟ㈻㌹ᥦమ࡞ࡢぜࡼࡿ࡝࠷々ᩐࡡ໩ྙ∸࠿ヾࡴࡼࡿࡒࠊࡐࡆ࡚ࠉኬ㔖ᇰ㣬ࡊ
ࡒㄇᑙᇰᆀ୕ΰࡻࡽ୹⏍ᠺ∸࡛⩻࠻ࡼࡿࡾ໩ྙ∸ࢅ᢫ฝࡊࠉࡆࡿࢅ࣑࢜ࣚࢠ࣏ࣞࢹ࡞ࡻࡽ༟㞫ࠉ⢥⿿ࡊࡒࠊ
⌟ᅹࠉNMR ࡝࡜࡞ࡻࡽࡐࡡᵋ㏸よᯊࢅ㐅ࡴ࡙࠷ࡾࠊ
Functional analysis of type III polyketide synthase genes from Aspergillus oryzae
Yasuyo Seshime1, Isao Fujii1, Praveen Rao Juvvadi2, Katsuhiko Kitamoto2, Yutaka Ebizuka1
(1Grad. School of Pharm. Sciences, Univ. of Tokyo, 2Dept. of Biotechnology, Univ. of Tokyo)
44
P-19
⧖㏁ࡊᆵࢰ࢕ࣈϨ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪ࡡⓆ⌟࡛ᶭ⬗よᯊ
᲻ཾ ᬓ⨶ࠉ⸠஬ ໌ࠉᾇ⩹ሪ ㇇㸝᮶ኬ㝌࣬ⷾ㸞
6-࣒ࢲࣜࢦࣛࢲࣜ㓗ྙᠺ㓕⣪㸝MSAS㸞ࡢࠉ⣊≟Ⳟࡡࢰ࢕ࣈ I ⧖㏁ࡊᆵ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪㸝PKS㸞࡛ࡊ
࡙᭩ᑚ࡚࠵ࡽࠉࡐࡡᵋ㏸࡛ཬᚺᶭᵋࡡよ᪺ࡡࡻ࠷ࢰ࣭ࢣࢴࢹ࡚࠵ࡾ࡛⩻࠻ࡼࡿࡾࠊᠻࠍࡢࠉAspergillus terreus
⏜ᮮࡡ MSAS ࡚࠵ࡾ ATX ࡞ࡗ࠷࡙ࠉ㓕ẍⓆ⌟⣌ࢅ⏕࠷࡙༟≺࡚ࡢ୘Ὡᛮ࡝ N ᮆḖ኶మ࡛ C ᮆḖ኶మࢅභⓆ
⌟ࡈࡎࡾࡆ࡛࡞ࡻࡽࠉATX ࡡཬᚺ୯ᚨࡢᑛ࡝ࡂ࡛ࡵ㸧ࡗࡡ࣎ࣛ࣋ࣈࢲࢺ㙈ࡡ┞பష⏕࡞ࡻࡽᵋᠺࡈࡿ࡙࠷ࡾ
ࡆ࡛ࢅ♟ࡊ࡙ࡀࡒࠊ
௑ᅂࠉḖ኶㡷ᇡࢅࡻࡽ㛏ࡂࡊࡒን␏మࡡභⓆ⌟࡞ࡻࡽࠉཬᚺ୯ᚨࡡ්ᵋᠺ࡞᭩ᑚ㝀ᚪさ࡝㡷ᇡࢅ᪺ࡼ࠾࡞
ࡊࡒࠊMSAS ࡢ⦨ྙ㓕⣪ KSࠉ࢓ࢨࣜᇱ㌹⛛㓕⣪ ATࠉ⬲Ề㓕⣪ DHࠉࢢࢹ㑇ඔ㓕⣪ KRࠉ࢓ࢨ࣭ࣜ࢞ࣔࣛࣕࣈ
ࣞࢷ࢕ࣤ ACP ࠾ࡼ࡝ࡾ࣎ࣛ࣋ࣈࢲࢺ㙈࠿ப࠷࡞ head-to-tail ᵋ㏸ࢅ࡛ࡖ࡙Ὡᛮ୯ᚨࢅᵋᠺࡊࠉᶭ⬗ࡊ࡙࠷ࡾ࡛
⩻࠻ࡼࡿ࡙࠷ࡒࡆ࡛࠾ࡼࠉATX ࡡ྘り፳ࢺ࣒࢕ࣤὩᛮ୯ᚨን␏మࢅ⏕࠷ࡒ᳠ゞࡵ⾔ࡖࡒࠊ྘ን␏మࢅⓆ⌟ࡈ
ࡎࡾ࡛ࠉKR ን␏మ࠿ࢹࣛࢢࢰ࢕ࢺ୯㛣మࡡ⎌໩మ࡚࠵ࡾ triacetic acid lactone㸝TAL㸞ࢅྙᠺࡊࡒ࡞ࡵ㛭ࢂࡼ
ࡍࠉDH-KR භን␏మ࠿ TAL ࢅྙᠺࡊ࡝࠾ࡖࡒࡆ࡛࠾ࡼࠉDH ࡢ୯㛣మࡡ⬲Ềཬᚺ௧አࡡཬᚺࡵり፳ࡌࡾྊ⬗
ᛮ࠿♟ြࡈࡿࡒࠊ␏⛸ን␏మࡡභⓆ⌟࡞࠽࠷࡙ࡢࠉ␏࡝ࡾ࣎ࣛ࣋ࣈࢲࢺ୕ࡡ KS ࡛ ATࠉKS ࡛ DHࠉAT ࡛ DHࠉ
KR ࡛ ACP ࠿ཬᚺ୯ᚨࡡᵋᠺ࡞㛭ࢂࡾࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡽࠉࡆࡿࡱ࡚ᥞ၌ࡈࡿ࡙࠷ࡒ༟⣟࡝ head-to-tail ࣓ࢸ
࡚ࣜࡢㄕ᪺࡚ࡀ࡝࠷ࢺ࣒࢕ࣤ㛣┞பష⏕ࡡᏋᅹ࠿ぜฝࡈࡿࡒࠊATX ࡢ㸩㔖మ࡚ᶭ⬗ࡌࡾࡆ࡛࠾ࡼࠉ㸧ࡗࡡ࣎
ࣛ࣋ࣈࢲࢺ㙈࠿ head-to-head ࡚㎾᥃ࡊࠉࡆࡿ࠿ࡈࡼ࡞㸩㔖మࢅᙟᠺࡌࡾ᩺ࡒ࡝࣓ࢸࣜࢅᥞ᱄ࡊࡒࠊ
Expression and functional analysis of iterative type I polyketide synthase
Tomomi Moriguchi, Isao Fujii, Yutaka Ebizuka
(Graduate School of Pharmaceutical Sciences, The Univ. of Tokyo)
P-20
㯔Ⳟ A. oryzae ࡡ⧖㏁ࡊᆵࢰ࢕ࣈϨ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪㐿ఎᏄࡡⓆ⌟࡛ᶭ⬗よᯊ
Ꮼᒁᬓ༡ 1㸡ເࠎᗛ௥ 1, ໪ᮇົࡥࡆ 2, ⸠஬໌ 1, ᾇ⩹ሪ㇇ 1 㸝᮶ኬ㝌࣬ⷾ 1, ᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ 2㸞
㯔Ⳟ A.oryzae RIB40 ࡡࢣࢿ࣑よᯊ࡞ࡻࡽᮇⳞ࡞ࡢኣᩐࡡ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪㸝PKS㸞㐿ఎᏄ࠿Ꮛᅹࡌࡾ
ࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊᏰධ㛏ࡡ⧖㏁ࡊᆵࢰ࢕ࣈϨPKS 㐿ఎᏄࡢ 28 ಴Ꮛᅹࡊࠉࡐࡡࢺ࣒࢕ࣤᵋᠺ࠾ࡼࠉⰶ㤮
᪐໩ྙ∸ࢅ⏍ᠺࡌࡾ࡛⩻࠻ࡼࡿࡾⰶ㤮᪐ᆵ PKS ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄࡢ 13 ಴㸝Ao1-1~13㸞ࠉ⬙⫣᪐໩ྙ∸ࢅ⏍
ᠺࡌࡾ㑇ඔᆵ PKS ࡡ㐿ఎᏄࡢ 12 ಴㸝Ao2-1~12㸞ࠉࡱࡒࠉ㟸ࣛ࣍ࢮ࣭࣑࣋ࣈࢲࢺྙᠺ㓕⣪࡛ࡡࣀ࢕ࣇࣛࢴࢺᆵ
PKS ࡡ㐿ఎᏄࡢ 3 ಴㸝Ao3-1~3㸞࡚࠵ࡖࡒࠊࡊ࠾ࡊࠉࡆࡿࡱ࡚࡞ A. oryzae ࡻࡽࡆࡿࡼ PKS ࡡ⏐∸࡛⩻࠻ࡼࡿ
ࡾ໩ྙ∸ࡡ༟㞫ࡢሒ࿈ࡈࡿ࡙࠽ࡼࡍࠉࡐࡡᶭ⬗ࢅ᪺ࡼ࠾࡞ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙ࠉࡆࡿࡼ PKS ࢅ㡨ḗㄇᑙⓆ
⌟ࡈࡎࠉࡐࡡ⏍ᠺ⏐∸ࢅྜྷᏽࡌࡾࡆ࡛࡛ࡊࡒࠊ
ࡱࡍࠉⰶ㤮᪐ᆵ PKS 㐿ఎᏄ Ao1-1 ࢅ⣊≟Ⳟ⏕Ⓠ⌟࣊ࢠࢰ࣭pTAex3R ࡞ Gateway Ἢ࡞ࡻࡽᑙථࡊࠉⓆ⌟ࣈࣚ
ࢪ࣐ࢺࢅᵋ⠇ࠉA.oryzae M-2-3 ᰬ࡞ᙟ㈻㌹ᥦ࣬ᑙථࡊࡒࠊᚋࡼࡿࡒᙟ㈻㌹ᥦమࢅㄇᑙᇰ㣬ࡊࠉ⏍⏐໩ྙ∸ࢅ
HPLC ࡚ฦᯊࡊࡒ࡛ࡆࢀࠉᇰ㣬ᾦ࣬Ⳟమ᢫ฝᾦ࡛ࡵ࡞ࢤࣤࢹ࣭ࣞࣜࡡᙟ㈻㌹ᥦమ࡞ࡢᏋᅹࡊ࡝࠷≁␏Ⓩ࡝㸧
ࡗࡡ໩ྙ∸࠿᳠ฝࡈࡿࠉ⌟ᅹࠉࡐࡡ༟㞫ࠉྜྷᏽ࡞ࡗ࠷࡙᳠ゞ୯࡚࠵ࡾࠊࡱࡒࠉ௙ࡡ PKS 㐿ఎᏄ࡞ࡗ࠷࡙ࡵྜྷ
ᵕ࡞Ⓠ⌟⣌ࡡᵋ⠇࡛よᯊࢅ㐅ࡴ࡙࠷ࡾࠊ
Expression and functional analysis of typeϨpolyketide synthase genes from A. oryzae
Tomohiro Moriya1, Yasuyo Seshime1, Katsuhiko Kitamoto2, Isao Fujii1, Yutaka Ebizuka1
(1Grad. School of Pharm. Sciences, Univ. of Tokyo, 2Dept. of Biotechnology, Univ. of Tokyo)
45
P-21
Penicillium luteo-aurantium ࡡ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪㐿ఎᏄࡡᶭ⬗よᯊ
ఫ㔕 ⿩ᇱ㸡⸠஬ ໌㸡ᾇ⩹ሪ ㇇ 㸝᮶ኬ㝌࣬ⷾ㸞
⣵⬂࿔᭿ࡡೳḾ࡝࡜ᵕࠍ࡝Ὡᛮࢅࡵࡗࣚࢸ࢔ࢨࢤ࣭ࣜࡢࠉࡐࡡ࣏ࢠࣞࣚࢠࢹࣤᵋ㏸࠾ࡼ࣎ࣛࢢࢰ࢕ࢺ࡛⩻
࠻ࡼࡿࡾ࠿ࠉࡐࡡ⏍ྙᠺ㐿ఎᏄࡢᮅࡓྜྷᏽࡈࡿ࡙࠷࡝࠷ࠊࡐࡆ࡚ࠉࣚࢸ࢔ࢨࢤ࣭ࣜ⏍⏐Ⳟ࡚࠵ࡾ Penicillium
luteo-aurantium ࡻࡽ᩺ぞ PKS 㐿ఎᏄࡡࢠ࣭ࣞࢼࣤࢡࢅモࡲࠉࡆࡿࡱ࡚࡞㑇ඔᆵ PKS ࢅࢤ࣭ࢺࡌࡾ pkHS1 ࠉ
PKS/NRPS ࡡࣀ࢕ࣇࣛࢴࢺ㐿ఎᏄ࡚࠵ࡾ pkHS5ࠉⰶ㤮᪐ᆵ PKS ࢅࢤ࣭ࢺࡌࡾ pkHS12 ࡡධ㛏㒼าࢅỬᏽࡊࡒࠊ
pkHS1 ࡞㛭ࡊ࡙ࡢࠉGateway cloning Ἢࢅ⏕࠷࡙⣊≟ⳞⓆ⌟⏕࣊ࢠࢰ࣭pTAex3R ࡞ᑙථࡊࡒࣈࣚࢪ࣐ࢺ
pTA-PkHS1 ࡚ Aspergillus oryzae ࢅᙟ㈻㌹ᥦࡊࠉ⏍⏐∸ࡡฦᯊࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉᇰ㣬ᾦࡡ HPLC ฦᯊ࡚ࠉ
ࢤࣤࢹ࣭ࣞࣜ࡞ࡢぜࡼࡿ࡝࠷≁␏Ⓩ࡝ࣅ࣭ࢠࢅ᳠ฝࡊࡒࠊᵾဗ࡛ࡡẒ㍉࠾ࡼࠉࡆࡿࢅ࢛ࣜࢬࣛࣤ㓗࡚࠵ࡾ࡛
ྜྷᏽࡊࡒࠊPkHS1 ࡢࢢࢹ㑇ඔ㓕⣪ KR ࢺ࣒࢕ࣤࠉ⬲Ề㓕⣪ DH ࢺ࣒࢕ࣤࢅ᭯ࡌࡾࡆ࡛࠾ࡼ㑇ඔᆵ PKS ࡛⩻࠻
ࡼࡿࡒ࠿ࠉࡐࡡࢺ࣒࢕ࣤ㒼⨠ࡢ 6-࣒ࢲࣜࢦࣛࢲࣜ㓗ྙᠺ㓕⣪ࡡ C ᮆ࡞࢓ࢨࣜᇱ㌹⛛㓕⣪ AT ࢺ࣒࢕ࣤࢅ௛ຊ
ࡊࡒࡵࡡ࡚࠵ࡽࠉ࢓ࢬࢲࣜᇱࢅࢪࢰ࣭ࢰ࣭࡛ࡊ࡙࢛ࣜࢬࣛࣤ㓗ྙᠺ㓕⣪ᵕ࡞㛚⎌ࡊࠉ࢛ࣜࢬࣛࣤ㓗ࢅ⏍ᠺࡊ
ࡒࡵࡡ࡛⩻࠻ࡼࡿࡾࠊࣚࢸ࢔ࢨࢤ࣭ࣜࡢ࢛ࣜࢬࣛࣤ㓗ࡡ㒂ฦᵋ㏸ࢅࡵࡗࡆ࡛࠾ࡼࠉืࡡ PKS/FAS ࠿⏍ᠺࡊࡒ
࢓ࢨࣜ㙈ࢅࢪࢰ࣭ࢰ࣭࡛ࡊ࡙ PkHS1 ࡡ AT ࠿ུࡄཱིࡽࠉ㸨ฦᏄࡡ㓉㓗༟నࡡ⦨ྙࠉ㛚⎌ཬᚺ࡞ࡻࡽࠉࣚࢸ࢔
ࢨࢤ࣭ࣜࡡ⅛⣪㦭᰹ࢅ⏍ᠺࡌࡾྊ⬗ᛮ࠿⩻࠻ࡼࡿࡾࠊPkHS5 ࡞㛭ࡊ࡙ࡵࠉGateway cloning Ἢࢅ⏕࠷࡙ష⿿ࡊ
ࡒࣈࣚࢪ࣐ࢺ pTA-PkHS5 ࢅ A. oryzae ࡞ᑙථࡊࠉᚋࡼࡿࡒᙟ㈻㌹ᥦమࡡ⏍⏐∸࡞ࡗ࠷࡙ฦᯊࢅ㐅ࡴ࡙࠷ࡾࠊ
Functional analysis of polyketides synthase genes from Penicillium luteo-aurantium
Hiroki Sano, Isao Fujii, Yutaka Ebizuka
(Graduate School of Pharmaceutical Sciences, Univ. of Tokyo)
P-22
⣊≟Ⳟⰶ㤮᪐࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪ࡡࢠࣚ࢕ࢭࣤࢦ࢕ࢠ࣭ࣚࢭࢺ࣒࢕ࣤ
ᐋᓧ ಆࠉ⸠஬ ໌ࠉᾇ⩹ሪ ㇇ 㸝᮶ኬ㝌࣬ⷾ㸞
Aspergillius fumigatus ࡡ࣒ࣚࢼࣤ⏍ྙᠺ࡞㛭ࢂࡾ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪㸝PKS㸞࡚࠵ࡾ Alb1p ࡢࣉࣈࢰࢢࢰ
࢕ࢺ࡚࠵ࡾࢻࣆࢹࣅࣞࣤ໩ྙ∸ YWA1 ࢅ⏍ᠺࡌࡾࠊࡐࡡ㸸ᮆ㡷ᇡࡢࢠࣚ࢕ࢭࣤᆵࡡཬᚺ࡞ࡻࡽ YWA1 ࡡ㸧
⎌┘ࡡ㛚⎌ࢅり፳ࡌࡾࢠࣚ࢕ࢭࣤࢦ࢕ࢠ࣭ࣚࢭࢺ࣒࢕ࣤ CYC ࡚࠵ࡾࠊAlb1p ࡡ CYC Ḗ኶మ࡚࠵ࡾ Cd-3 ࢅⓆ
⌟ࡈࡎࡾ࡛࢕ࢮࢠ࣏ࣛࣤᆵࡡࣉࣈࢰࢢࢰ࢕ࢺ໩ྙ∸࡚࠵ࡾ DHCI ࠿⏍⏐ࡈࡿࡾ࠿ࠉCd-3 ࡛භ࡞ CYC ࢺ࣒࢕
ࣤࢅ≺❟ࡡࢰࣤࣂࢠ࡛ࡊ࡙Ⓠ⌟ࡈࡎࡾ࡛ YWA1 ࠿⏍⏐ࡈࡿࡾࡆ࡛࠾ࡼࠉ
CYC ࢺ࣒࢕ࣤ࠿≺❟ࡡ㓕⣪ࢰࣤࣂࢠ
࡛ࡊ࡙ Cd-3 ୕ࡡࣉࣈࢰࢢࢰ࢕ࢺ୯㛣మ࡞഼ࡀࠉࢠࣚ࢕ࢭࣤ⎌໩ཬᚺࢅり፳࡚ࡀࡾࡆ࡛ࢅሒ࿈ࡊࡒࠊ
ࡐࡆ࡚ࠉ௑ᅂࠉCYC ࢺ࣒࢕ࣤࡡᶭ⬗ࢅࡈࡼ࡞よᯊࡌࡾࡒࡴࠉCYC ࢰࣤࣂࢠࡡኬ⭘Ⳟ࡚ࡡⓆ⌟࡛⢥⿿ࢅモ
ࡲ ࡒ ࠊ Alb1p ࡡ CYC ࢺ ࣒ ࢕ ࣤ 㐿 ఎ Ꮔ ࢅ ኬ ⭘ Ⳟ Ⓠ ⌟ ⏕ ࣊ ࢠ ࢰ ࣭ pET21a ࡞ ᑙ ථ ࡊ ࠉ Ⓠ ⌟ ࣈ ࣚ ࢪ ࣐ ࢺ
pET21a-alb1p-CYC ࢅᵋ⠇ࡊࡒࠊࡆࡿࢅኬ⭘Ⳟ BL21 Codon plus ࡞ᑙථࡊⓆ⌟ࢅモࡲࡒ࡛ࡆࢀࠉ25˚Cࠉ18 hr ㄇ
ᑙᇰ㣬࡚᭩ࡵࡻࡂྊ⁈ᛮ⏤ฦ࡞Ⓠ⌟ࡌࡾࡆ࡛࠿☔ヾࡈࡿࡒࠊHis tag ࢅ௛ຊࡊࡒ CYC ࢰࣤࣂࢠࡢࠉNi affinity
column ࡚࡮࡯༟ୌࡡࢰࣤࣂࢠ࡛ࡊ࡙⢥⿿ࡈࡿࠉenterokinase ฌ⌦࡞ࡻࡽ His tag ࢅ㝎ཡࡊࡒࠊ⌟ᅹࠉࡆࡡ CYC
ࢰࣤࣂࢠ࡛ Cd-3 ࡞ࡻࡾ YWA1 ࡡ in vitro ྙᠺ⣌ࡡ්ᵋ⠇ࢅ᳠ゞ୯࡚࠵ࡾࠊ
Studies on the Claisen cyclase domains of fungal aromatic polyketide synthases
Shun Miyazaki, Isao Fujii㸡Yutaka Ebizuka
(Graduate School of Pharmaceutical Sciences, The Univ. of Tokyo)
46
P-23
㓕ẍࢅ࣌ࢪࢹ࡛ࡊࡒ Aspergillus fumigatus DHN-࣒ࣚࢼࣤ⏍ྙᠺ⣌ࡡ්ᵋᠺ
༞㒂 ኚᕵ㸡⸠஬ ໌ࠉᾇ⩹ሪ ㇇ 㸝᮶ኬ㝌࣬ⷾ㸞
ࣃࢹ࢓ࢪ࣋ࣜ࢟ࣜࢪ⑍ࡡ୹ࡒࡾ⑋ᅄⳞ࡚࠵ࡾ Aspergillus fumigatus ࡡ⬂ᏄⰅ⣪⏍ྙᠺ㐿ఎᏄࢠࣚࢪࢰ࣭࡞ࡢࠉ
ⰶ㤮᪐⧖㏁ࡊᆵࢰ࢕ࣈϨ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪㸝PKS㸞ࢅࢤ࣭ࢺࡌࡾ alb1 ࢅྱࡳ 6 ࡗࡡ㐿ఎᏄ࠿Ꮛᅹࡌࡾࠊ
ࣉࣈࢰࢢࢰ࢕ࢺྙᠺ㓕⣪࡚࠵ࡾ Alb1p ࡞ࡻࡽࢻࣆࢹࣅࣞࣤ YWA1 ࠿⏍ᠺࡊࠉࡆࡿ࠿ࢠࣚࢪࢰ࣭ࢅᵋᠺࡌࡾഁ
㙈ว᩷㓕⣪ࠉ㑇ඔ㓕⣪ࠉ⬲Ề㓕⣪࡝࡜࡞ࡻࡽࠉ㡨ḗࠉ1,8-dihyroxynaphthalene㸝DHN㸞࡫࡛ንᥦࡈࡿࠉ᭩ᚃ࡞
DHN ࠿ࣆ࢘ࢿ࣭ࣜ㓗໩㓕⣪࡞ࡻࡽ㔔ྙࡊ࡙ DHN-࣒ࣚࢼࣤ࠿⏍ᠺࡌࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊࡊ࠾ࡊࠉࢠࣚࢪࢰ
࣭හ࡞ࢤ࣭ࢺࡈࡿࡾ㸧ࡗࡡ㓗໩㓕⣪ Abr1pࠉAbr2p ࡡ࡜ࡔࡼ࠿ DHN ࡡ㔔ྙࢅり፳ࡌࡾ࠾࡝࡜୘᪺࡝Ⅴࡵኣ࠷ࠊ
ᮇ◂✪࡞࠽࠷࡙ࡢࠉA. fumigatus ࡡ DHN-࣒ࣚࢼࣤ⏍ྙᠺ࡞㛭ࢂࡾ྘㓕⣪ࡡᶭ⬗ࢅྜྷᏽࡌࡾ࡛࡛ࡵ࡞ࠉ㓕ẍࢅ
࣌ࢪࢹ࡛ࡊࡒ⣊≟Ⳟ஦ḗ௥ㅨ⏍ྙᠺ⣌ࡡ්ᵋᠺ࣓ࢸ࡛ࣜࡊ࡙ DHN-࣒ࣚࢼࣤ⏍ྙᠺ⣌ࡡᵋ⠇ࢅ┘Ⓩ࡛ࡊࡒࠊ
ࡱࡍࠉ㓕ẍහ࡚Ⓠ⌟ࡊࡒ PKS ࢅὩᛮᆵ࡛ࡌࡾࡒࡴࠉAspergillus nidulans ࡡ࣌ࢪ࣌ࣂࣤࢷࢷ࢕ࣤ㌹⛛㓕⣪㐿
ఎᏄ npgA ࢅ᯹Ⰵమ࡞⤄ࡲ㎲ࢆࡓ㓕ẍࢅష⿿ࡊࡒࠊࡆࡿ࡞ alb1 ࡡⓆ⌟ࣈࣚࢪ࣐ࢺ pYES-alb1 ࢅᑙථࡊࠉࡆࡡ
ᙟ㈻㌹ᥦ㓕ẍ࠿ YWA1 ࢅ⏍⏐ࡌࡾࡆ࡛ࢅ☔ヾࡊࡒࠊḗ࡞ࠉYWA1 ࡡഁ㙈ว᩷㓕⣪㐿ఎᏄ ayg1 ࢅභⓆ⌟ࡈࡎࠉ
1,3,6,8-tetrahydroxynaphthalene㸝T4HN㸞ࡡ⏍⏐ࢅ☔ヾࡊࡒࠊ⤾࠷࡙ࠉT4HN 㑇ඔ㓕⣪ࢅࢤ࣭ࢺࡌࡾ࡛஢᝷ࡈࡿ
ࡒ arp2 㐿ఎᏄࢅභⓆ⌟ࡈࡎࡾࡆ࡛࡞ࡻࡽ syctalone ࠿⏍ᠺࡌࡾࡆ࡛ࢅ☔ヾࡊࡒࠊ⌟ᅹࠉṟࡽࡡ㸨ࡗࡡ㐿ఎᏄ
arp1ࠉabr1ࠉabr2 ࡞ࡗ࠷࡙ࡵ྘Ⓠ⌟ࣈࣚࢪ࣐ࢺࢅᵋ⠇ࠉ㡨ḗࠉ㓕ẍ࡞ᑙථࡊࠉよᯊࢅ㐅ࡴ࡙࠷ࡾࠊ
Reconstitution of DHN-melanin biosynthetic pathway of Aspergillus fumigatus in yeast
Natsuki Nambu, Isao Fujii, Yutaka Ebizuka
(Graduate School of Pharmaceutical Sciences, The Univ. of Tokyo)
P-24
GFP ⼝ྙ⺦Ⓣ㈻ࢅ⏕࠷ࡒ㯔Ⳟࡡࢨࢸࣞࣆ ࢚࢓⏍⏐ㄢ⟿ᅄᏄ(SreAo)ࡡᑻᅹよᯊ
ῳ㎰஁ᩏࠉఫ⸠ฺḗ㸝ᒷᡥ⏍ᕝ◂㸞
㯔Ⳟ(A.oryzae)ࡢΰ㒿ࡡ₧ᅹⓏ࡝╌Ⰵཋᅄ࡛࡝ࡖ࡙࠷ࡾࣆ࢘ࣛࢠࣛࢨࣤࢅฦἢࡌࡾࠊࣆ࢘ࣛࢠࣛࢨࣤࡢࢨࢸ
ࣞࣆ࢚࢓ࡡୌ⛸࡚࠵ࡽ㕪Ḗஇ᫤࡞ㄇᑙⓏ࡞ฦἢ⏍⏐ࡈࡿࡾࠊSreAo ࡢࣆ࢘ࣛࢠࣛࢨࣤ⏍⏐โᚒᅄᏄ㐿ఎᏄ࡚
࠵ࡽࠉ㕪ఴ๨⎌ሾୖ࡚ࣆ࢘ࣛࢠࣛࢨࣤྙᠺࡡ➠ୌṹ㝭࡚࠵ࡾ࢛ࣜࢼࢲࣤ-N5-࢛࢞ࢨࢣࢻ࣭ࢭ㐿ఎᏄ㸝dffA㸞ࢅ
㌹෕࡚ࣝ࣊ࣜᢒโࡌࡾ࠿ࠉSreAo ࡞ࡻࡾ dffA ㌹෕ࡡ ON/OFF ࡡโᚒᶭᵋࡢ༎ฦ࡞ࡢ᪺ࡼ࠾࡚࡝࠷ࠊ
௑ᅂᠻࠍࡢࠉSreAo ࡛ GFP ࡛ࡡ⼝ྙ⺦Ⓣ㈻ࢅ㯔Ⳟ࡚Ⓠ⌟ࢅࡈࡎࠉࡐࡡ⣵⬂හ࡚ࡡᑻᅹᛮࢅㄢ࡬ࡒࠊ㧏Ⓠ⌟ࣈ
࣓࣭ࣞࢰ࡚࠵ࡾ amyB ࣈ࣓࣭ࣞࢰࢅ⏕࠷࡙ GFP ⼝ྙ⺦Ⓣ㈻ࢅⓆ⌟ࡈࡎࡒ࡛ࡆࢀࠉGFP ⺧කࡡ⣵⬂ᰶ࡫ࡡᑻᅹ
࠿ふᐳࡈࡿࡒࠊࡱࡒࠉGFP ⼝ྙ⺦Ⓣ㈻ࡢ sreAo Ḗ᥾ᰬࡡ⾪⌟ᆵࢅ㔕ᛮᆵ࡞ᚗᖉࡈࡎࡒࡆ࡛࠾ࡼࠉSreAo ࠿ᰶ
ࢰࣤࣂࢠ㈻࡛ࡊ࡙ᶭ⬗ࡊ࡙࠷ࡾࡆ࡛࠿☔ヾࡈࡿࡒࠊࡱࡒ⌟ᅹࠉᇰᆀ୯ࡡ㕪ࡡᏋᅹࡡ᭯↋࡞ࡻࡖ࡙ GFP ⼝ྙ⺦
Ⓣ㈻ࡡᑻᅹᛮ࠿ን໩ࡌࡾ࠾࡜࠹࠾ࢅ᳠ゞࡊ࡙࠽ࡽࠉࡐࡡ⤎ᯕࡵెࡎ࡙Ⓠ⾪ࡌࡾ஢ᏽ࡚࠵ࡾࠊ
Analysis of localization of SreAo, a siderophore biosynthesis regulator of Aspergillus oryzae, as GFP fusion
protein
Hisayuki Watanabe, Toshitsugu Sato (IBRC)
47
P-25
⣒㯔Ⳟ㸝Monascus pilosus㸞࡞ࡻࡾ஦ḗ௥ㅨ⏐∸ࡡ⏍⏐
୔Ꮹ๙ྍ 1㸡ᘿ᪺Ễ 1,2㸡හᐣ஁⨶Ꮔ 3㸡ἑ㔕ຩெ 1 㸡㔕ᓧಘ⾔ 1 㸡⏐ᮇᘧ஄ 1㸡⛼ᇄ㈶஦ 3
㸝1 ᒱᒜ┬ᕝᢇࢬ㸡2JSPS㸡3 ᒱᒜኬ㝌࣬⏍∸ᶭ⬗໩Ꮥ㸞
⣒㯔Ⳟ㸝Monascus pilosus㸞ࡢࣞࣁࢪࢰࢲࣤ㸝ࢤࣝࢪࢷ࣭ࣞࣜ఩ୖష⏕㸞ࡷ㉝ⰅⰅ⣪㸝ኮ↓Ⰵ⣪㸞࡝࡜ࡡ᭯⏕
࡝஦ḗ௥ㅨ⏐∸ࢅ⏍⏐ࡌࡾ࠿ࡐࡡㄢ⟿࡞ࡗ࠷࡙ࡢ࡮࡛ࢆ࡜᪺ࡼ࠾࡞ࡈࡿ࡙࠷࡝࠷ࠊ⣒㯔Ⳟ࡞ࡻࡾ஦ḗ௥ㅨ⏐∸
ࡡ㧏⏍⏐ࢅ┘Ⓩ࡞⅛⣪″ࡷ✽⣪″࡝࡜ࡡᙫ㡢ࢅㄢ࡬ࠉ⣒㯔Ⳟ࡞ࡻࡾ஦ḗ௥ㅨ⏐∸ࡡ⏍⏐࠿ cAMP ⤊㊪ࢅ⤊ࡾᙁ
࠷ࢡࣜࢤ࣭ࢪᢒโࢅུࡄࡾࡆ࡛ࢅ᪺ࡼ࠾࡞ࡊࡒࠊࡐࡊ࡙㐲ว࡝ᇰ㣬ᇱ㈻ࡡㄢ⿿࡞ࡻࡽࣁ࢕࢛࣏ࢪᙟᠺࢅ⥌ᣚࡊ
࡝࠿ࡼࢡࣜࢤ࣭ࢪᢒโࢅᅂ㑂ࡌࡾࡆ࡛࡚஦ḗ௥ㅨ⏐∸ࡡ㧏⏍⏐ࢅᐁ⌟࡚ࡀࡾࡆ࡛ࢅ♟ࡊࡒࠊྜྷ᫤࡞ࢡࣜࢤ࣭ࢪ
ᢒโ࠿よ㝎ࡈࡿࡒን␏ᰬࢅฦ㞫ཱིᚋࡊࡆࡿࡱ࡚࡞࡝࠷㧏⏍⏐ࢅ㐡ᠺࡊࡒࠊࡱࡒ⣊≟Ⳟࡡ஦ḗ௥ㅨࢅโᚒࡌࡾᅄ
Ꮔ LaeA ࡡ⣒㯔Ⳟ࣓࣌ࣞࢡ MpLaeA ࢅྜྷᏽཱིᚋࡊࡐࡡโᚒᶭᵋ࡛஦ḗ௥ㅨ࡫ࡡᐞ୙ࢅㄢ࡬ࡒࠊࡐࡡ⤎ᯕࠉMpLaeA
ࡡ mRNA ࡢ 5’ᮆ❻㟸⩳ゼ㡷ᇡ࡚㐽ᢝⓏࢪࣈࣚ࢕ࢨࣤࢡࢅུࡄ㛏㙈ᆵ࡛▯㙈ᆵ࡛࡝ࡾࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊ
ࡆࡡ㐽ᢝⓏࢪࣈࣚ࢕ࢨࣤࢡࡢᇰ㣬ᇱ㈻࡝࡜࡞ࡻࡾโᚒࢅུࡄࠉ▯㙈ᆵ࡫ࡢᵋᠺⓏ࡞ࢪࣈࣚ࢕ࢨࣤࢡࡈࡿ࡙࠷ࡒ
ࡡ࡞ᑊࡊࠉ㛏㙈ᆵ࡫ࡡࢪࣈࣚ࢕ࢨࣤࢡࡢ࣋ࣈࢹࣤ࡝࡜ࡡ᭯ᶭ✽⣪″ࡡᏋᅹ࡞ࡻࡽᢒโࡈࡿࡒࠊࡐࡊ࡙⮾࿝῕࠷
ࡆ࡛࡞㛏㙈ᆵ࡫ࡡࢪࣈࣚ࢕ࢨࣤࢡ࠿ᢒ࠻ࡼࡿࡾ᮪௲ୖ࡚ࣞࣁࢪࢰࢲࣤ࠿⏍⏐ࡈࡿ࡙࠷ࡾࡆ࡛ࢅぜ࠷ࡓࡊࡒࠊ㛏
㙈ᆵࡡࡵࡗ㛏࠷ 5’㟸⩳ゼ㡷ᇡ࡞㟸⩳ゼⓏ࡝ຝᯕࡵ♟ြࡈࡿࡾࡆ࡛࠾ࡼࠉ㛏㙈ᆵ࡫ࡡᢒโ࠿▯㙈ᆵ࡞ࡻࡾ⩳ゼࢅ
ಀ㐅ࡈࡎࠉࡐࡡ⤎ᯕࣞࣁࢪࢰࢲࣤ࠿⏍⏐ࡈࡿࡒ࡛⩻࠻࡙࠷ࡾࠊࡈࡼ࡞࢓ࣤࢲࢬࣤࢪ㙈ᑙථ࡞ࡻࡾ MpLaeA ࡡ㌹
෕ᢒโ࡞ࡻࡖ࡙ࣞࣁࢪࢰࢲࣤࡡ⏍⏐࠿ᢒ࠻ࡼࡿࡒࡆ࡛࠾ࡼࠉ⣒㯔Ⳟ࡞࠽࠷࡙ࡵᮇ㐿ఎᏄ࠿஦ḗ௥ㅨࡡโᚒ࡞ᐞ
୙ࡊ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ
Production of secondary metabolites by Monascus pilosus
Tsuyoshi Miyake1, Ming-Yong Zhang1,2, Kumiko Uchitomi3, Isato Kono1, Nobuyuki Nozaki1, Hiroyuki Sammoto1,
Kenji Inagaki3 (1Ind Tech Ctr, Okayama Pref, 2JSPS, 3Dept Biofunct Chem, Grad Sch, Okayama Univ)
P-26
࢞ࣔ࣊ࢵⴆ㯜⑋Ⳟ Cong:1-1 ࡛ࡐࡡ⑋ཋᛮḖ᥾ᰬ Cong:1-2 ࡞࠽ࡄࡾࢰࣤࣂࢠ㈻Ⓠ⌟
ࣂࢰ࣭ࣤࡡよᯊ
ᒱ㒂᪺Ꮔ, ྚ⏛㝧ᘇ*࣬᭯ᮇ ⿩**࣬ᇸ๑ ├**࣬᲻ ⩟྿***࣬ᑈᒱ ᚥ࣬᭯Ờ ງ㸝㎨ᕝኬ࣬*㎨⎌◂࣬**⌦◂࣬
***㎨ኬ㸞
ᅰቫ⑋ཋᛮᏄࡡ࠹Ⳟ࡚࠵ࡾ࢞ࣔ࣊ࢵⴆ㯜⑋Ⳟ Fusarium oxysporum f. sp. conglutinans Cong:1-1 ࡡ⤽௥ᇰ㣬
୯࡞⑋ཋᛮࢅḖ᥾ࡊࡒᰬ Cong:1-2 ࠿ฝ⌟ࡊࡒࠊCong:1-2 ࢅ࠵ࡼ࠾ࡋࡴ࢞ࣔ࣊ࢵ࡞ฌ⌦ࡌࡾ࡛ࠉCong:1-1 ࡞ࡻ
ࡾ⑋ᐐ࠿ᢒโࡈࡿࡾࡆ࡛࠾ࡼࠉCong:1-2 ࡢ࢞ࣔ࣊ࢵⴆ㯜⑋࡞ᑊࡌࡾ⏍∸㜭㝎ὩᛮࢅᣚࡗࠊCong:1-1 ࡛ Cong:1-2
㛣࡚ࡡⓆ⌟ࢰࣤࣂࢠ㈻ࡡᕣ␏ࢅㄢᰕࡌࡾࡆ࡛࡚⑋ཋᛮ㛭㏻ᅄᏄࠉ⏍∸㜭㝎Ὡᛮ㛭㏻ᅄᏄࡡ᥀⣬ࢅモࡲ࡙࠷ࡾࠊ
10 ᪝㛣ᣲ࡛࠹ᇰ㣬ࡊࡒⳞమ࠾ࡼࢰࣤࣂࢠ㈻ࢅ᢫ฝࡊࠉ஦ḗඔ㞹ẴὃິἪ࡞ࡻࡖ࡙ฦ㞫ᒈ㛜ࡊࠉࢰࣤࣂࢠ㈻ࢪ
࣎ࢴࢹࡡᕣ␏ࢅㄢᰕࡊࡒࠊࡐࡡ⤎ᯕࠉ⣑ 300 ࡡࢪ࣎ࢴࢹࢅࡐࡿࡑࡿ᳠ฝࡊࠉᕣࡡ࠵ࡾࢰࣤࣂࢠ㈻ࢅ々ᩐぜฝ
ࡊࡒࠊࡆࡡ࠹ࡔࠉCong:1-2 ࡚ᅸ಻Ⓩ࡞ኣࡂⓆ⌟ࡊ࡙࠷ࡾ᥆ᏽ࢛࢞ࢨࢺࣛࢱࢠࢰ࣭ࢭࢰࣤࣂࢠ㈻ (ODX1) ࡞Ἰ
┘ࡊࠉよᯊࢅ⾔ࡖࡒ࠿ࠉ⑋ཋᛮ࡞ࡢ㛭୙ࡊ࡙࠷࡝࠾ࡖࡒࠊ
Comparison of protein expression patterns between cabbage yellows fungus Cong:1-1 and its virulenceloss-mutant Cong:1-2.
Akiko Okabe, Takanobu Yoshida, Naoshi Domae, Shougo Mori, Thoru Teraoka, Tutomu Arie (Tokyo Univ. of Agric.
& Tech., Natn. Inst. for Agro-Env. Sci., RIKEN, Tokyo Univ. of Agric.)
48
P-27
Colletotrichum acutatum ࡡ࣊ࢿ࣐ࣜ⪇ᛮ㐿ఎᏄ CaBEN1 ࡢș-ࢲ࣭ࣖࣇࣛࣤ㐿ఎᏄࡡ㌹
෕࡞㛭ࢂࡖ࡙࠷ࡾ
୯⏿Ⰳ஦࣬୯㔕ḿ᪺ 㸝㎨◂ᶭᵋ ᯕᶖ◂㸞
ᯕᶖࢅࡢࡋࡴ㎨ష∸ࡡ⅛⑃⑋ࢅᘤࡀ㉫ࡆࡌ Colletotrichum acuatum ࡢࠉ࣊ࢿ࣐ࣜ➴ࡡ࣊ࣤࢫ࢕࣐ࢱࢯ࣭ࣜ⣌
ṽⳞ๠࡞఩វུᛮࢅ♟ࡌࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࠊୌ⯙Ⓩ࡞ࠉⳞ㢦ࡡ࣊ࢿ࣐ࣜ⪇ᛮࡢࠉⷾ๠ࡡᵾⓏ࡚࠵ࡾș-ࢲࣖ
࣭ࣇࣛࣤࡡ࢓࣐ࢿ㓗⨠ᥦ࡞㉫ᅄࡌࡾ࠿ࠉC.acutatum ࡚ࡢืࡡᶭᵋࡡ㛭୙࠿♟ြࡈࡿ࡙࠽ࡽࠉࡐࡡレ⣵ࡢ୘᪺
࡚࠵ࡾࠊᠻࠍࡢࠉࣀ࢕ࢡ࣏ࣞ࢕ࢨࣤ⪇ᛮ㐿ఎᏄ࡚ᙟ㈻㌹ᥦࡊࡒ⣑ 3,600 ᰬࡡ୯࠾ࡼ࣊ࢿ࣐ࣜវུᛮን␏ᰬ
CAT7-150 ࢅ㐽ᢜࡊࠉ࣊ࢿ࣐ࣜ⪇ᛮ㛭㏻㐿ఎᏄ CaBEN1 ࢅ༟㞫ࡊ࡙࠷ࡾࠊCaBEN1 ࡢࠉGibberella zeae ࡷ
Neurospora crassa ࡡ leucin zipper motif ࢅ᭯ࡌࡾᶭ⬗ᮅ▩ࢰࣤࣂࢠ㈻࡛㧏࠷┞ྜྷᛮࢅ♟ࡌ 818 ࢓࣐ࢿ㓗ࢅࢤ࣭
ࢺࡌࡾ࡛᥆ᏽࡈࡿࡒࠊୌ᪁ࠉCAT7-150 ࡛㔕⏍ᰬ CAB03 ࠾ࡼ㸧⛸ࡡș-ࢲ࣭ࣖࣇࣛࣤ㐿ఎᏄ㸝CaTUB1 ࠽ࡻࡦ
CaTUB2㸞ࢅࡐࡿࡑࡿ༟㞫࣬よᯊࡊࡒ࠿ࠉ୦ᰬࡡ㛣࡞㒼าࡡ㐢࠷ࡢヾࡴࡼࡿ࡝࠾ࡖࡒࠊCAB03 ࠽ࡻࡦ CaBEN1
㐿ఎᏄ◒ቪᰬ࡞࠽ࡄࡾ CaTUB1 ࠽ࡻࡦ CaTUB2 ࡡⓆ⌟≁ᛮࢅࢿ࣭ࢧࣤよᯊࡊࡒ࡛ࡆࢀࠉCAB03 ࡚ࡢ࣊ࢿ࣐ࣜ
ฌ⌦࡞ࡻࡖ࡙ CaTUB1 ࡡ㌹෕࠿㢟ⴥ࡞Ὡᛮ໩ࡈࡿࡾࡡ࡞ᑊࡊࠉCaBEN1 㐿ఎᏄ◒ቪᰬ࡚ࡢ㌹෕ࡡὩᛮ໩ࡢධ
ࡂぜࡼࡿ࡝࠾ࡖࡒࠊࡆࡿࡼࡡ⤎ᯕ࠾ࡼࠉC.acutatum ࡡ࣊ࢿ࣐ࣜ⪇ᛮ࡞㛭୙ࡌࡾ CaBEN1 ࡢ CaTUB1 ࡡ㌹෕ᅄ
Ꮔ࡚࠵ࡾྊ⬗ᛮ࠿♟ြࡈࡿࡒࠊ
CaBEN1 required for benomyl resistance regulates the transcription of beta-tubulin gene in Colletotrichum
acutatum
Ryoji Nakaune, Masaaki Nakano
(NIFTS, NARO)
P-28
ࣛࣤࢥᩤⅤⴘⴝ⑋Ⳟࡡ AM Ẑ⣪⏍ ྙᠺ㐿ఎᏄࢠࣚࢪࢰ࣭࡞ࢤ࣭ࢺࡈࡿࡾ Zn(II)2Cys6
ࢰ࢕ࣈ㌹෕โᚒᅄᏄࡡᶭ⬗
᧓ᮇ౅᪺㸡ඡ⋚ᇱୌ᭹ 1㸡ᒜᮇᖷ༡ 2㸡ᑹㆺ ᾀ 1 㸡᯼᳔ᑠᚷ (ྞኬ㝌⏍㎨࣬1 㫵ཱིኬ㎨࣬2 ᒱᒜኬ㎨)
ࣛࣤࢥᩤⅤⴘⴝ⑋Ⳟࡢ㸡ᐙ୹≁␏ⓏẐ⣪㸝AM Ẑ⣪㸞ࢅ⏍⏐ࡊ㸡Ẑ⣪វུᛮࡡࣛࣤࢥဗ⛸࡞ࡡࡲᩤⅤᛮ⑋ᐐࢅᘤ
ࡀ㉫ࡆࡌ㸣඙࡞㸡ᮇⳞ࠾ࡼ 2 ಴ࡡ AM Ẑ⣪⏍ྙᠺ㐿ఎᏄ㸝AMT ࠽ࡻࡦ AMT2㸞ࢅྜྷᏽࡊࡒ㸣ࡈࡼ࡞㸡IFO8984
ᰬࡡ AMT 㐿ఎᏄࢅྱࡳ BAC ࢠ࣭ࣞࣤࢅ㐽ᢜࡊ㸡ࡐࡡሲᇱ㒼า(118 kb)ࢅỬᏽࡊࡒ࡛ࡆࢀ㸡AMT㸡AMT2 ࡞ຊ
࠻ 22 ಴ࡡ᥆ᏽㄖࡲᯗ㸝ORF㸞࠿ぜฝࡈࡿࡒ㸣Ẑ⣪⏍⏐᫤࡛㟸⏍⏐᫤࡞࠽ࡄࡾࡆࡿࡼ ORF ࡡⓆ⌟ࣝ࣊ࣜࢅẒ
㍉ࡊࡒ࡛ࡆࢀ㸡AMT㸡AMT2 ࠽ࡻࡦ 10 ಴ࡡ ORF ࠿Ẑ⣪⏍⏐᫤࡞㧏Ⓠ⌟ࡌࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡽ㸡AMT 㐿ఎ
Ꮔ࡛ࡆࡿࡼ ORF ࢅྱࡳ⣑ 62 kb ࡡ㡷ᇡ࠿Ẑ⣪⏍ྙᠺ㐿ఎᏄࢠࣚࢪࢰ࣭࡚࠵ࡾࡆ࡛࠿♟ြࡈࡿࡒ㸣ࡆࡡࢠࣚࢪ
ࢰ࣭࡞ࡢ㸡2 ḗ௥ㅨ㛭㏻㓕⣪࡛┞ྜྷ࡝ ORF ࡞ຊ࠻㸡Zn(II)2Cys6 ࢰ࢕ࣈࡡ㌹෕โᚒᅄᏄࢅࢤ࣭ࢺࡌࡾ ORF13
࠿Ꮛᅹࡌࡾ㸣ࡐࡆ࡚㸡ORF13 ࡡ㐿ఎᏄ◒ቪᰬࢅషฝࡊ㸡ࡐࡡẐ⣪⏍ྙᠺ࡞࠽ࡄࡾᶭ⬗ࢅよᯊࡊࡒ㸣㔕⏍ᰬ࡛
◒ቪᰬࡡẐ⣪⏍⏐㔖ࢅẒ㍉ࡊࡒ࡛ࡆࢀ㸡◒ቪᰬ࡚ࡢࡐࡡ⏍⏐㔖࠿ⴥࡊࡂቌຊࡌࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒ㸣ࡈ
ࡼ࡞㸡Ẑ⣪⏍ྙᠺ㐿ఎᏄࢠࣚࢪࢰ࣭ࡡ ORF ࡡⓆ⌟ࣝ࣊ࣜ࠿㔕⏍ᰬ࡞Ẓ࡬◒ቪᰬ࡚୕᪴ࡌࡾࡆ࡛ࢅぜฝࡊࡒ㸣
ࡱࡒ㸡ORF13 㧏Ⓠ⌟ᰬࢅషฝࡊࡒ࡛ࡆࢀ㸡㧏Ⓠ⌟ᰬ࡚ࡢ㐿ఎᏄࢠࣚࢪࢰ࣭ࡡ ORF ࡡⓆ⌟ࣝ࣊ࣜ࠿఩ୖࡊ㸡Ẑ
⣪⏍⏐㔖࠿΅ᑛࡌࡾࡆ࡛ࢅ☔ヾࡊࡒ㸣௧୕ࡡ⤎ᯕࡢ㸡ORF13㸝AMTR1 ࡛࿤ྞ㸞࠿ AM Ẑ⣪⏍ྙᠺ㐿ఎᏄ⩄ࡡ
ㇿࡡ㌹෕โᚒᅄᏄࢅࢤ࣭ࢺࡌࡾࡆ࡛ࢅ♟ࡊࡒ㸣
Function of a Zn(II)2Cys6-type transcription regulator embedded in the AM-toxin biosynthesis gene cluster of
the apple pathotype of Alternaria alternata
Yoshiaki Harimoto, Motoichiro Kodama1, Mikihiro Yamamoto2, Hiroshi Otani1, Takashi Tsuge
(Grad. Sch. Bioagric. Sci., Nagoya Univ., 1Fac. Agric., Tottori Univ., 2Fac. Agric., Okayama Univ.)
49
P-29
ࢻࢨ㯦ᩤ⑋Ⳟࡡ㈇✰Ⳟ⣊࡞࠽ࡄࡾὩᛮ㓗⣪⛸(ROS)ࡡᶭ⬗よᯊ
⋖ᗛὒࠉờ⏛೸ୌࠉ୯ᒁᩔᆍࠉᮌᮠඒ㸝♼ᡖኬ࣬⮤↓⛁Ꮥ㸞
ᐙ୹౴ථ᫤࡞⏍ᠺࡌࡾࢻࢨ㯦ᩤ⑋Ⳟࡡ ROS ࡡᙲ๪ࢅ᪺ࡼ࠾࡛ࡌࡾࡒࡴ࡞ࠉ⣵⬂Ꮥ࣬ฦᏄ⏍∸ᏕⓏᡥἪࢅ⏕
࠷࡙よᯊࡊࡒࠊ⑋ཋⳞࡡ⬂Ꮔᠩ⃦ᾦࢅࢻࢨⴝ࡞᥃⛸ࡌࡾ࡛ࠉࢻࢨⴝ୕࡚⬂ᏄⓆⰾࡊࡒᚃࠉⓆⰾ⟮඙❻࠿⭶ࡼ
ࡲ௛╌ჹࢅᙟᠺࡌࡾࠊࡐࡡ௛╌ჹ࠾ࡼ㈇✰Ⳟ⣊࠿⏍ࡋࢻࢨⴝࡡࢠࢲࢠࣚ࡞౴ථࡌࡾ᫤ࠉវུᛮࢻࢨⴝࡡ㈇✰
Ⳟ⣊࡞ኬぞᶅ࡝ H2O2 ⏍ᠺ࠿ヾࡴࡼࡿࠉࡐࡿ࡞ఔࡖ࡙Ⳟࡢ㧏㢎ᗐ࡞ᐙ୹࡫౴ථࡊࡒࠊᢤᢘᛮࢻࢨⴝࡡ㈇✰Ⳟ⣊
࡚ࡢᑚぞᶅ࡝ H2O2 ⏍ᠺ࠿ヾࡴࡼࡿࠉᐙ୹࡫ࡡⳞࡡ౴ථ⋙ࡢ఩࠾ࡖࡒࠊࡊ࠾ࡊࠉH2O2 ࡡᾐཡ๠࡛ࡊ࡙࢓ࢪࢤࣜ
ࣄࣤ㓗ࢅ῟ຊࡊࡒ⬂Ꮔᠩ⃦ᾦࢅ᥃⛸ࡌࡾ࡛ࠉ୦ဗ⛸ⴝ࡞࠽ࡄࡾ㈇✰Ⳟ⣊࡚ H2O2 ⏍ᠺ㔖࠿΅ᑛࡊࠉࡐࡿ࡞┞㛭
ࡊ࡙វུᛮࢻࢨⴝ࡚ᐙ୹࡫ࡡ౴ථ⋙࠿఩ୖࡊࡒࠊ௧୕ࡡ⤎ᯕࡢࠉᮇⳞࡡ㈇✰Ⳟ⣊࡞࠽ࡄࡾ H2O2 ࡡ⏍ᠺࡢ౴␆
ງ࡞㛭ࢂࡾ஥ࢅ♟ြࡌࡾࠊḗ࡞ࠉ⑋ཋᛮ࡫ࡡ㛭୙ࢅッ᪺ࡌࡾࡒࡴ࡞ ROS ⏍ᠺ࡞㛭ࢂࡾ NADPH oxidase (NOX)
ࡡ༟㞫ࢅモࡲࡒࠊࡆࡿࡱ࡚࡞Ⳟ㢦࡚ሒ࿈ࡈࡿ࡙࠷ࡾ NOX 㐿ఎᏄࢅẒ㍉ࡊࠉ⦨ྙࣈࣚ࢕࣏࣭ࢅスゝᚃࠉᮇⳞࡡ
ࢣࢿ࣑ DNA ࢅ㗢ᆵ࡞ࡊ࡙ PCR ࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉ᪜ሒࡡ NOXA 㐿ఎᏄ࡛㧏࠷┞ྜྷᛮࢅ♟ࡌ PCR ᩷∞࠿
ᚋࡼࡿࠉࡐࡿࢅࣈ࣭ࣞࣇ࡛ࡊ࡙⏕࠷ᮇⳞࢣࢿ࣑ࡡࢤࢪ࣐ࢺࣚ࢕ࣇ࣭ࣚࣛ࠾ࡼ㝟ᛮࢠ࣭ࣞࣤࢅ㐽ᢜࡊࠉᏰධ㛏
㐿ఎᏄࢅ༟㞫ࡊࡒࠊ
Role of reactive oxygen species (ROS) on penetration peg of Alternaria alternata Japanese pear pathotype
Gang-Su HYON, Kenichi Ikeda, Hitoshi Nakayashiki, Yukio Tosa, Pyoyun Park
(Grad.Sch.of Science and Technology,Kobe Univ.)
P-30
Fusarium oxysporum ࡡ⬂Ꮔᙟᠺ㛭㏻㌹෕ᅄᏄ Ren1 ࡢல◢㓗㑇ඔ㓕⣪㐿ఎᏄࢅโᚒࡌ
ࡾ
㣜⏛♰ୌ㑳㸡ᑚཋᨼ᪺㸡᯼᳔ᑠᚷ 㸝ྞኬ㝌⏍㎨㸞
F. oxysporum ࡢ㸡ᑚᆵ⬂Ꮔ㸡ኬᆵ⬂Ꮔ࠽ࡻࡦཉ⭯⬂Ꮔࡡ 3 ⛸㢦ࡡ↋ᛮ⬂Ꮔࢅᙟᠺࡌࡾ㸣ᮇⳞࡢ㸡ᰜ㣬㇇ᐣ
࡝Ᏸධᇰᆀ࡚ࡢ⬂Ꮔࢅᙟᠺࡊ࡝࠷࠿㸡࢜ࣜ࣍࢞ࢨ࣒ࢲࣜࢬ࣭ࣜࣞࢪࢅ୹࡝⅛⣪″࡛ࡌࡾ CMC ᇰᆀ࡚ࡢ࡮࡛
ࢆ࡜Ⳟ⣊ᠺ⫩ࡎࡍ㸡ᑚᆵ⬂Ꮔ࡛ኬᆵ⬂Ꮔࢅኬ㔖࡞ᙟᠺࡌࡾ㸣඙࡞㸡EST㸝Expressed Sequence Tag㸞よᯊ࡞ࡻ
ࡖ࡙୦ᇰᆀ࡞࠽ࡄࡾⓆ⌟㐿ఎᏄ⩄ࢅẒ㍉よᯊࡊࡒ㸣Ᏸධᇰᆀᇰ㣬᫤࡛ CMC ᇰᆀᇰ㣬᫤ࡡ cDNA ࣚ࢕ࣇࣚࣛ
࣭ࢅష⿿ࡊ㸡ࡐࡿࡑࡿ⣑ 1,300 ࢠ࣭ࣞࣤࡡ㒂ฦሲᇱ㒼าࢅỬᏽࡊࡒ㸣Ửᏽ㒼าࡡẒ㍉࡞ࡻࡖ࡙㸡୦ࣚ࢕ࣇࣚ
࣭ࣛ࠾ࡼࡐࡿࡑࡿ⣑ 640 ಴ࡡ␏࡝ࡾ㐿ఎᏄ⏜ᮮࡡ≺❟ࢠ࣭ࣞࣤࢅྜྷᏽࡊࡒ㸣ࡱࡒ㸡ᮇⳞࡡ⬂Ꮔᙟᠺ࡞୘ྊḖ
࡝㌹෕โᚒᅄᏄࢅࢤ࣭ࢺࡌࡾ REN1 㐿ఎᏄࢅྜྷᏽࡊࡒ㸣ࡐࡆ࡚㸡⬂Ꮔᙟᠺ᫤ࡡⓆ⌟㐿ఎᏄ࡞ࡗ࠷࡙㸡㔕⏍ᰬ
࡛ ΔREN1 ን␏ᰬ࡞࠽ࡄࡾⓆ⌟ࣝ࣊ࣜࢅࣛ࢓ࣜࢰ࢕࣑ PCR Ἢ࡞ࡻࡖ࡙Ẓ㍉ࡊ㸡Ren1 ࡞ࡻࡖ࡙ḿ࡞โᚒࡈࡿࡾ
23 ಴ࡡು⿭㐿ఎᏄࢅ㐽ᢜࡊࡒ㸣௑ᅂ㸡ࡆࡿࡼ㐿ఎᏄࡡ࠹ࡔ◢㓗ሲฺ⏕ᛮ࡞㛭୙ࡌࡾல◢㓗㑇ඔ㓕⣪㐿ఎᏄ
㸝FoNIIA ࡛࿤ྞ㸞࡞ࡗ࠷࡙㸡ࡐࡡ⬂Ꮔᙟᠺ࡞࠽ࡄࡾᶭ⬗ࢅよᯊࡊࡒ㸣ΔREN1 ን␏ᰬ࡞࠽ࡄࡾ FoNIIA ࡡⓆ⌟
ࣝ࣊ࣜࡢ㔕ᛮᰬࡡ 1/100 ⛤ᗐ࡚࠵ࡽ㸡ல◢㓗ᇰᆀ࡚ࡡ ΔREN1 ን␏ᰬࡡᠺ⫩ࡢ㔕⏍ᰬ࡞Ẓ࡬᭯ណ࡞఩ୖࡊࡒ㸣
ୌ᪁ࠉ◢㓗㑇ඔ㓕⣪㐿ఎᏄࡡⓆ⌟ࣝ࣊ࣜࡢ୦Ⳟᰬ࡚ྜྷᵕ࡚࠵ࡖࡒ㸣ΔFoNIIA ን␏ᰬࢅషฝࡊ㸡ࡐࡡᑚᆵ⬂Ꮔ㸡
ኬᆵ⬂Ꮔ࠽ࡻࡦཉ⭯⬂Ꮔࡡᙟᠺ⬗ࢅ᳠ᏽࡊࡒ࡛ࡆࢀ㸡ኬᆵ⬂Ꮔࡡᙟᠺᩐ࠿᭯ណ࡞΅ᑛࡌࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝
ࡖࡒ㸣௧୕ࡡ⤎ᯕࡢ㸡Ren1 ࡞ࡻࡖ࡙โᚒࡈࡿࡾல◢㓗㑇ඔ㓕⣪࠿ኬᆵ⬂Ꮔᙟᠺᩐ࡞㛭୙ࡌࡾࡆ࡛ࢅ♟ࡊࡒ㸣
Conidiation-Related Transcription Factor Ren1 Regulates the Nitrite Reductase Gene in Fusarium oxysporum
Yuichiro Iida, Toshiaki Ohara, Takashi Tsuge
(Grad. Sch. Bioagric. Sci., Nagoya Univ.)
50
P-31
Fusarium oxysporum ࡡࣅࣜࣄࣤ㓗ࢸ࢜ࣜ࣍࢞ࢨ࣭ࣚࢭ㐿ఎᏄࡡྜྷᏽ
ᮇᒜ យ㸡ᑈᒱ ᚥࠉ᭯Ờ ງ 㸝᮶ா㎨ᕝኬ࣬㝌㎨⏍∸โᚒ㸞
ஹ㒼୘ධᛮᏄࡡ࠹Ⳟ Fusarium oxysporum ࡢࠉᅰቫఎ᯹ᛮࡡ᳔∸⑋ཋⳞ࡚ࠉ᳔∸᰷㒂࠾ࡼ౴ථࡊࠉᑙ⟮හࢅ
ⶕᘇࡊࠉධ㌗ࢅⴆ෺࣬ᯜṒࡈࡎࡾࠊᮇⳞࡢࠉ㓗⣪ฦᅸ࠿఩࠷࡛᝷ᏽࡈࡿࡾ᳔∸ࡡᑙ⟮හ࡚⏍⫩ࡌࡾࡆ࡛࠾ࡼࠉ
ᑙ⟮හ࡚ࡢ᎒Ẵ࿣ྺࢅ⾔࠹ࡆ࡛࠿᥆ᐳࡈࡿࡾࠊࡱࡒࠉᮇⳞࡢ᎒Ẵ࿣ྺࡡ㸦ࡗ࡚࠵ࡾ࢓ࣜࢤ࣭ࣜⓆ㓕⬗ࢅᣚࡗ
ࡆ࡛ࡵ▩ࡼࡿ࡙࠷ࡾࠊࡐࡆ࡚ࠉᮇ◂✪࡚ࡢ࢓ࣜࢤ࣭ࣜⓆ㓕࡛⑋ཋᛮ࡛ࡡ㛭㏻ᛮ࡞ࡗ࠷࡙ㄢᰕࡌࡾࡒࡴࠉይẴ
࿣ྺ࡛࢓ࣜࢤ࣭ࣜⓆ㓕ࡡฦᒪⅤ௛㎾࡚ᶭ⬗ࡌࡾࣅࣜࣄࣤ㓗ࢸ࢜ࣜ࣍࢞ࢨ࣭ࣚࢭ㸝PDC㸞࡞╌┘ࡊࡒࠊ F.
oxysporum ࡚ࡢ PDC 㐿ఎᏄࡢᮅྜྷᏽ࡚࠵ࡖࡒࡒࡴࠉ㎾⦍⛸࡚࠵ࡾ F. graminearum ࡡࢸ࣭ࢰ࣭࣊ࢪ୕ࡡࠉ௙Ⳟ
ࣅࣜࣄࣤ㓗ࢸ࢜ࣜ࣍࢞ࢨ࣭ࣚࢭ࡛┞ྜྷᛮࡡ㧏࠷ᶭ⬗ᮅ▩ࢰࣤࣂࢠ㈻㸝#XM_390010.1㸞ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ᝗
ሒࢅᇱ࡞ࠉ⦨㔔 PCRࠉTAIL-PCR ࡞ࡻࡖ࡙ࠉF. oxysporum ࠾ࡼ 3554bp ࡡ᩷∞ࢅࢠ࣭ࣞࢼࣤࢡࡊࡒࠊࡆࡡ DNA
᩷∞୯࡞ࡢࠉ3 ࡗࡡ࢙ࢠࢮࣤ࠾ࡼ࡝ࡾ᥆ᏽ ORF㸝1710 bp㸞࠿ᏋᅹࡊࠉPDC ࡛᝷ᏽࡈࡿࡾ 570 ࢓࣐ࢿ㓗࠾ࡼ࡝
ࡾࢰࣤࣂࢠ㈻ࢅࢤ࣭ࢺࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡒࠊࡆࡡ㐿ఎᏄࢅ pdc1 ࡛ࡊࡒࠊPDC1 ࡢࠉ࢓࣐ࢿ㓗㒼า࡚ F.
graminearum ࡡ᥆ᏽࢰࣤࣂࢠ㈻࡛ 96.1%ࠉAsperugillus fumigatus ࡡ PDC㸝#XM_74419.1㸞࡛ 63.5㸚ࠉA.oryzae
ࡡ PDC㸝#AF098293.1㸞࡛ 62㸚ࠉSaccharomyces cerevisiae ࡡ PDC1㸝#NC_001144.4㸞࡛ 47%ࡡ┞ྜྷᛮࢅ♟ࡊࡒࠊ
ࡱࡒࠉPDC ࡞㧏ࡂಕᏋࡈࡿࡾ࢓࣐ࢿ㓗࣓ࢲ࣭ࣆ㸝-GDG-࠾ࡼ-NN-㸞࠿Ꮛᅹࡊࡒ㸝aa. 448-476㸞
ࠊࢣࢿ࣐ࢴࢠࢦ
ࢧࣤよᯊࡡ⤎ᯕࠉpdc1 ࡢ F. oxysporum ࢣࢿ࣑୕࡞ 1 ࢤࣅ࣭Ꮛᅹࡊࡒࠊ A putative pyruvate decarboxylase gene in Fusarium oxysporum
Ai Motoyama, Tohru Teraka, Tsutomu Arie
(Tokyo Univ. of Agric. & Tech.)
P-32
ᾈ㏩ᅸ࡞ࡻࡽⓆ⌟࠿ንິࡌࡾ㯔Ⳟࡡ㐿ఎᏄ⩄࡛ࡐࡡࣈ࣓࣭ࣞࢰ࣭ࡡฺ⏕
⧂⏛ ೸ࠉ1 ᆊᮇ࿰ಆࠉ2 ᭯㤷ᑋⱝࠉ1 ኬ໪⏜౅ࠉ1 ᰘⷭࢱࣚࣚࢴࢹࠉ1 ᒜ⏛ ಞࠉ1,3 ᒷୖ࿰⿩ࠉ4 ⚽⏛ ಞ
㸝㔘Ἁᕝኬࢣࢿ࣑◂ࠉ1 㒿⥪◂ࠉ2 ┬❟ᗀᓞኬࠉ3 ᗀኬ඙❻◂ࠉ4 ᐁ㊰ዥᏄኬ㸞
ᾈ㏩ᅸ࡞ᑊࡌࡾᚺ➽ࡢࠉ㓕ẍ࠾ࡼဳ஘㢦࡞⮫ࡾ┷ᰶ⏍∸࡞࠽࠷࡙ᬉ㐚Ⓩ࡞Ꮛᅹࡊࠉレ⣵࡝◂✪࠿࡝ࡈࡿ࡙
࠷ࡾࠊ≁࡞㯔Ⳟ Aspergillus oryzae ࡚ࡢࠉ㯔㏸ࡽ(ᅖమᇰ㣬)ࡡᕝ⛤࡞࠽࠷࡙⎌ሾヾㆉࡡ㔔さ࡝ᅄᏄ࡛⩻࠻ࡼࡿࠉ
ᐁ㝷࡞ AtfB ࡷ HogA ࡝࡜ᾈ㏩ᅸโᚒ⣌࠿㔔さ࡚࠵ࡾࡆ࡛࠿♟ࡈࡿ࡙࠷ࡾࠊࡐࡆ࡚ࠉᾈ㏩ᅸࡡን໩࡞ᚺࡋ࡙Ⓠ
⌟࠿ㄇᑙࡈࡿࡾ㐿ఎᏄ⩄ࢅよᯊࡊࠉࡐࡡࣈ࣓࣭ࣞࢰ࣭ࢅฺ⏕ࡊࡒⓆ⌟࣊ࢠࢰ࣭ࡡ㛜Ⓠࢅ⾔ࡖࡒࠊ
A. oryzae RIB40 ᰬࢅ DP ᇰᆀ࡞࡙ 24 ᫤㛣๑ᇰ㣬ࡊࡒᚃࠉ0.8M NaCl ࠽ࡻࡦ 1.2M Sorbitol ࢅ῟ຊࡊࡒᇰᆀ
࡞⛛ࡊࠉ⤊᫤Ⓩ࡝࣏࢕ࢠࣞ࢓ࣝ࢕よᯊࢅ⾔࡝࠷Ⓠ⌟࠿ንິࡌࡾ㐿ఎᏄࡡ᥀⣬ࢅ⾔࡝ࡖࡒࠊࡐࡡ⤎ᯕࠉ5 ಶ௧
୕Ⓠ⌟࠿୕᪴ࡌࡾ㐿ఎᏄ⩄࡛ࡊ࡙ 19 ⛸㢦ࡡ㐿ఎᏄࢅぜฝࡊࡒࠊ࠹ࡔ 5 ࡗࡢᾈ㏩ᅸ࡞㗞ᨼ࡞ࠉኬࡀࡂᚺ➽ࡊࡒ
ࡆ࡛࠾ࡼࠉGUS ࣭ࣝ࣎ࢰ࣭よᯊࢅ⾔࡝ࡖࡒࠊ⤎ᯕࠉ➴ᘿ᮪௲࡚ㄇᑙྊ⬗࡚ࠉ࠾ࡗཚᐠ࡝โᚒ࠿ྊ⬗࡝ 2 ࡗࡡ
ࣈ࣓࣭ࣞࢰ࣭ࢅཱིᚋࡊࡒࠊࡱࡒࠉᮇࣈ࣓࣭ࣞࢰ࣭ࢅฺ⏕ࡊࠉᾈ㏩ᅸ࡚ㄇᑙࡈࡿࡾⓆ⌟⣌ࢅᵋ⠇ࡊࡒࠊ
Isolation of Isoosmotic Up-regulated Gene (IUG) promoters in A. oryzae
Ken Oda, 1Kazutoshi Sakamoto, 2 Toshihide Arima, 1 Yuka Okita, 1Dararat Kakizono, 1Osamu Yamada, 1,3Kazuhiro
Iwashita, 4Osamu Akita
(KIT, 1NRIB, 2Prefect. Univ. of Hiroshima, 3Hiroshima Univ., 4Jissen Women’s Univ.)
51
P-33
㯑Ⳟ Aspergillus oryzae ࡡ bZIP ᆵ㌹෕โᚒᅄᏄ㐿ఎᏄ atfA ࡡよᯊ
ᆊᮇ࿰ಆ 1㸡ᒜ⏛ಞ 1㸡ኬ໪⏜౅ 1㸡ᒷୖ࿰⿩ 1㸡⚽⏛ಞ 2㸡஫࿝ົஒ 3㸡୔୕㔔᪺ 1 㸝1 㒿⥪◂㸡2 ᐁ㊰ዥᏄኬ㸡3
᮶໪ኬ㝌࣬㎨㸞
⡷㯔ࠉࡨࡌࡱ㯑࡝࡜ࡡᅖమᇰ㣬࡚ࡢࠉᾦమᇰ㣬ࡷᐨኮᇰᆀࡻࡽࡵỀฦὩᛮ࠿఩ࡂࠉࡆࡿ࠿㯑Ⳟࡡᅖమᇰ㣬
࡞࠽ࡄࡾ≁ᚡⓏ࡝㐿ఎᏄⓆ⌟ࢅ⏍ࡲฝࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊࡆࡿࡱ࡚ᠻࠍࡢᅖమᇰ㣬᫤ࡡ≁ᚡⓏ࡝㐿
ఎᏄⓆ⌟ࢅโᚒࡌࡾ㌹෕โᚒᅄᏄࡡ≁ᏽࢅモࡲࠉATF/CREB ࣆ࢒࣐࣭ࣛ࡞ᒌࡌࡾ atfB 㐿ఎᏄ࠿ᅖమᇰ㣬ᚃ᭿
ࡡ㐿ఎᏄⓆ⌟ࢅโᚒࡌࡾࡆ࡛ࢅ᪺ࡼ࠾࡞ࡊ࡙ࡀࡒࠊ㯑Ⳟ࡞ࡢ AtfB ࢰࣤࣂࢠ㈻ࡡ᥆ᏽ DNA ⤎ྙࢺ࣒࢕࡛ࣤ࡮
࡯ྜྷୌࡡ࢓࣐ࢿ㓗㒼าࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ atfA ࠿Ꮛᅹࡊ࡙࠷ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖ࡙࠷ࡾࠊ
௑ᅂᠻࠍࡢ atfA 㐿ఎᏄࡡᶭ⬗ࢅよᯊࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙ atfA ◒ቪᰬࢅషᠺࡊよᯊࡊࡒ࡛ࡆࢀࠉฦ⏍Ꮔࡡ
Ⓠⰾ⋙࠿ኬࡀࡂ఩ୖࡊ࡙࠷ࡒࠊᾦమᇰ㣬୯࡚ࡡⳞమ㔖ࡡ᥆⛛ࢅふᐳࡊࡒ⤎ᯕ atfA ◒ቪᰬࡡⳞమ㔖ࡢ㔕⏍ᰬࡻ
ࡽ఩ࡂ᥆⛛ࡊࡒ࠿ࠉࡆࡡⳞమ㔖ࡡ఩ୖࡢⓆⰾ⋙࡞౪Ꮛࡊ࡙࠽ࡽࠉᾦమᇰ㣬୯࡚ࡡⳞ⣊ᠺ㛏ࡢ㔕⏍ᰬ࡛ྜྷ➴࡚
࠵ࡾࡆ࡛࠿☔ヾࡈࡿࡒࠊࡱࡒ atfA ◒ቪᰬࡢࡨࡌࡱᅖమᇰ㣬㸝‭ᗐ 25%㸞࡞࠽࠷࡙⏍⫩࠿ᴗ❻࡞㐔ࡿࠉᇰ㣬⎌
ሾ୯ࡡ‭ᗐࢅ 95%࡞㧏ࡴࡾࡆ࡛࡞ࡻࡽ⏍⫩࠿ᅂᚗࡌࡾ࡛࠷࠹⌟㇗࠿ぜࡼࡿࡒࠊࡈࡼ࡞ᾈ㏩ᅸࢅ㧏ࡴࡒᐨኮᇰ
ᆀ୕࡚ࡡ⏍⫩ࡡ㐔ࡿࡵふᐳࡈࡿࡒࡆ࡛࠾ࡼ atfA ࡢ఩ỀฦὩᛮ㸝㧏ᾈ㏩ᅸ㸞⎌ሾ࡫ࡡᚺ➽࡞ࡵ㔔さ࡚࠵ࡾࡆ࡛
࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊ
Analysis of atfA, a transcription-factor gene of Aspergillus oryzae
Kazutoshi Sakamoto1, Osamu Yamada1, Yuka Okita1, Kazuhiro Iwashita1, Osamu Akita1, Katsuya Gomi2, Shigeaki
Mikami1 (1National Research Institute of Brewing, 2Jissen Women’s Univ., 3 Tohoku Univ., Grad. Sch. Agri. Sci.)
P-34
࢓࢜ࣂࣤ࢜ࣄࡡᾈ㏩ᅸࢪࢹࣝࢪ࡚ㄇᑙࡈࡿࡾࢡࣛࢬ࣭ࣞࣜྙᠺ࡞࠽ࡄࡾ glycerol
dehydrogenase ࡡᙲ๪
ᒜୖ࿰Ᏻ 1ࠉᆊ㔕┷ᖲ 2ࠉሲ⃕࠵ࡍࡈ 1ࠉ⸠ᮟ┷ 1 㸝1 ᮶Ὂኬ࣬⏍࿤ࠉ2 ᮶Ὂኬ᳔࣬ᶭ◂ࢬ㸞
⏍∸ࡢᾈ㏩ᅸࢪࢹࣝࢪ࡞ᚺ➽ࡊ࡙⣵⬂හ࡞ࢡࣛࢬ࣭ࣞࣜࢅ⵫✒ࡌࡾ࠿ࠉࢡࣛࢬ࣭ࣞࣜྙᠺ㓕⣪࡛ࡊ࡙ฝⰾ
㓕ẍࡢ Gpd1(glycerol-3-phosphate dehydrogenase)ࢅࠉ⣊≟Ⳟࡢ glycerol dehydrogenase㸝GLD㸞ࢅ౐⏕ࡊ࡙࠷ࡾ
࡛ሒ࿈ࡈࡿ࡙࠷ࡾࠊ࢓࢜ࣂࣤ࢜ࣄ࡞ࡢࠉ
GLD Ὡᛮࢅࡵࡗ࡛᥆ᏽࡈࡿࡾ㐿ఎᏄ࠿ᩐ⛸Ꮛᅹࡌࡾ࠿ࠉࡐࡡ࠹ࡔ gcy-1
࡛ gcy-3 㐿ఎᏄࡡⓆ⌟࠿ᾈ㏩ᅸฌ⌦࡞ࡻࡽ OS-2 MAP ࢞ࢻ࣭ࢭ౪ᏋⓏ࡞ㄇᑙࡈࡿࡾࠊࡐࡆ࡚ࠉgcy-1 ◒ቪᰬࢅ
షᠺࡊࠉᾈ㏩ᅸវུᛮ࠽ࡻࡦ GLD 㓕⣪Ὡᛮࢅㄢ࡬ࡒࠊࡐࡡ⤎ᯕࠉgcy-1 ◒ቪᰬࡢࠉᾈ㏩ᅸវུᛮࢅ♟ࡌ࡛ྜྷ
᫤࡞ࠉ
GLD Ὡᛮ࠿ᾐ኶ࡊ࡙࠽ࡽࠉࣆࣜࢩ࢛࢞ࢮࢼࣜࡷᾈ㏩ᅸฌ⌦ࡊࡒሔྙ࡞ࡵ㓕⣪Ὡᛮࡢ᳠ฝࡈࡿ࡝࠾ࡖࡒࠊ
ࡊ࠾ࡊࠉᾈ㏩ᅸฌ⌦࡞ࡻࡽ⵫✒ࡌࡾࢡࣛࢬ࣭ࣞࣜࡢࠉ㔕⏍ᰬࡻࡽࡢ఩࠷ࡵࡡࡡࠉgcy-1 ◒ቪᰬ࠽ࡻࡦ os-2 ን␏
ᰬ࡚㢟ⴥ࡞ヾࡴࡼࡿࡒࠊୌ᪁ࠉᾈ㏩ᅸវུᛮ cut ን␏ᰬࡢࠉGLD 㓕⣪Ὡᛮࡢ㔕⏍ᰬྜྷᵕ࡞ᾈ㏩ᅸฌ⌦࡚ㄇᑙ
ࡈࡿࡒ࠿ࠉࢡࣛࢬ࣭ࣞࣜࡡ⵫✒ࡢධࡂ࠽ࡆࡼ࡝࠾ࡖࡒࠊࡆࡿࡼࡡ㐿ఎᏄ㛣ࡡ஦㔔ን␏ᰬࢅషᠺࡊᾈ㏩ᅸវུ
ᛮࢅẒ㍉ࡊࡒࠊࡐࡡ⤎ᯕࠉᾈ㏩ᅸវུᛮࡢ(cut os-2)㸳(os-2ࠉgcy-1 os-2)㸳(cutࠉgcy-1ࠉcut gcy-1)㸳(㔕⏍ᰬ)࡚
࠵ࡖࡒࠊgcy-1 ን␏ࡢࠉos-2 ࠽ࡻࡦ cut ን␏ᰬࡡᾈ㏩ᅸវུᛮ࡞࡮࡛ࢆ࡜ᙫ㡢ࢅ୙࠻࡝࠷ࡆ࡛࠾ࡼࠉgcy-1 ࠿
OS-2 ࡞ࡻࡽㄢ⟿ࡈࡿ࡙࠷ࡾ୹さ࡝ GLD 㐿ఎᏄ࡚࠵ࡾ࡛⩻࠻ࡼࡿࡒࠊୌ᪁ࠉcut os-2 ᰬࡢࡐࡿࡑࡿࡡ༟≺ᰬࡻ
ࡽࡵ㧏࠷ᾈ㏩ᅸវུᛮࢅ♟ࡊࡒࡆ࡛࠾ࡼࠉᾈ㏩ᅸᚺ➽࡞ࡻࡾࢡࣛࢬ࣭ࣞࣜྙᠺ࡞ࡢ OS-2 MAP ࢞ࢻ࣭ࢭ࠽ࡻ
ࡦ GLD ࡞౪Ꮛࡊ࡝࠷ྙᠺ⤊㊪࠿࠵ࡾ࡛⩻࠻ࡼࡿࡒࠊ
Role of glycerol dehydrogenase on glycerol synthesis in response to osmotic stress in Neurospora crassa
Kazuhiro Yamashita1, Shinpei Banno2, Azusa Shiozawa1, and Makoto Fujimura1 (1Life Sci., Toyo Univ., 2PRPC.,Toyo
Univ.)
52
P-35
࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ Hik1 ࡡ᝗ሒఎ㐡⣌ࡡよᯊ
᲻⏛┷⣟ 1,2㸡Ꮻఫ⨶ㄵ 2㸡ᕝ⸠ಆ❮ 1㸡ᮇᒜ㧏ᖶ 1 㸝1 ⌦◂࣬୯ኳ◂ࠉ2 ᮶Ὂኬ࣬ᕝ㸞
࠘┘Ⓩ࠙࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡࣀ࢕ࣇࣛࢴࢺᆵࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ Hik1 ࡡ null ን␏మ㸝Δhik1 ᰬ㸞ࡢ㧏⃨ᗐࡡ⢶
࡞ᑊࡌࡾ㧏ᾈ㏩ᅸវུᛮࢅ♟ࡌࡡ࡞ᑊࡊࠉⅤን␏మ㸝hik1-M16 ᆵን␏ᰬ㸞ࡢ㧏ᾈ㏩ᅸ㉰វུᛮཀྵࡦ⑋ཋᛮࡡ
఩ୖࢅ♟ࡌࠊ㧏ᾈ㏩ᅸࢪࢹࣝࢪᚺ➽࡞㛭୙ࡌࡾ MAP ࢞ࢻ࣭ࢭ Osm1 ࡡὩᛮ໩ࣝ࣊ࣜࡡよᯊ࠾ࡼࠉhik1-M16
ᰬ࡚ࡢ㔕⏍ᰬࡷΔhik1 ᰬ࡛Ẓ㍉ࡊ࡙ Osm1 ࡡὩᛮ໩⬗࠿఩ୖࡊ࡙࠷ࡾࡆ࡛ࢅ᪺ࡼ࠾࡞ࡊ࡙࠷ࡾࠊ௑ᅂࠉࡻࡽレ
⣵࡝よᯊࢅ⾔ࡖࡒࠊ
࠘᪁Ἢ࣬⤎ᯕ࠙hik1-M16 ᆵን␏మࡡ㧏ᾈ㏩ᅸ㉰វུᛮࡡཋᅄࢅㄢ࡬ࡾࡒࡴࠉOsm1 ௧አࡡ MAP ࢞ࢻ࣭ࢭ࡚⑋
ཋᛮ࡞ᚪ㡪ࡡ Mps1 ࡛ Pmk1 ࡡὩᛮ໩ࢅὩᛮᆵ p44/42 MAP ࢞ࢻ࣭ࢭᢘమࢅ⏕࠷࡙よᯊࡊࡒࠊࡐࡡ⤎ᯕࠉ㔕⏍
ᰬ࡚ࡢᮅฌ⌦࡛㧏ᾈ㏩ᅸฌ⌦࡞࠽࠷࡙ Mps1 ࡡὩᛮ໩ࡢ࡮࡛ࢆ࡜ぜࡼࡿ࡝࠾ࡖࡒࠊୌ᪁ࠉhik1-M16 ᰬ࡚ࡢ㧏
ᾈ㏩ᅸฌ⌦࡚ࡢࠉ㔕⏍ᰬ࡛ྜྷᵕ࡞Ὡᛮ໩ࡢ࡮࡛ࢆ࡜ヾࡴࡼࡿ࡝࠾ࡖࡒ࠿ࠉᮅฌ⌦࡞࠽࠷࡙ࡢ Mps1 ࠿Ὡᛮ໩
ࡈࡿࡒࠊࡆࡡࡆ࡛࠾ࡼࠉhik1-M16 ᰬ࡚ࡢ Mps1 ࡡ᝗ሒఎ㐡⣌࠿ᨡ஗ࡈࡿ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ
Analysis of the signal transduction pathway of a histidine kinase Hik1 of the rice blast fungus
Masumi Morita1,2, Ron Usami2, Toshiaki Kudo1, Takayuki Motoyama1
(1Discovery Res. Institute, RIKEN, 2Fac. Technol., Univ. Toyo)
P-36
⣊≟Ⳟ Aspergillus nidulans ࡞࠽ࡄࡾኣᵕ࡝㓗໩ࢪࢹࣝࢪᚺ➽ᶭᵋࡡよᯊ
῞㔕♰ᗀࠉⴏཋኬ♰ࠉ㔘୷ாᏄࠉຊ⸠㞖ኃࠉᑚᯐူኰࠉỀ㔕⊓㸝ྞኬ㝌࣬⏍࿤㎨Ꮥ㸞
His-Asp ࣛࣤ㓗࣭ࣛࣝ⣌ࡢࣁࢠࢷࣛ࢓࠾ࡼ㧏➴᳔∸࡞ࡱ࡚ᬉ㐚Ⓩ࡞Ꮛᅹࡌࡾ⎌ሾᚺ➽᝗ሒఎ㐡ᶭᵋ࡚࠵ࡾࠊ
ᠻࠍࡢ⣊≟Ⳟࡡᬉ㐚Ⓩ࡝⎌ሾᚺ➽࣒࢜ࢼࢫ࣑ࡡよ᪺ࢅ┘ᣞࡊࠉA. nidulans ࡞࠽ࡄࡾ His-Asp ࣛࣤ㓗࣭ࣛࣝ⣌ࡡ
మ⣌Ⓩ࡝ᶭ⬗よᯊࢅ⾔ࡖ࡙࠷ࡾࠊࡆࡿࡱ࡚࡞ His-Asp ࣛࣤ㓗࣭ࣛࣝ⣌ࡡᵋᠺᅄᏄ࡚࠵ࡾ Histidine kinase (HK)ࠉ
HPt ᅄᏄࠉResponse regulator (RR)ࢅࡐࡿࡑࡿ㸦㸪⛸㢦ࠉ㸦⛸㢦ࠉ㸩⛸㢦ྜྷᏽࡊࠉࡆࡡ࠹ࡔࡡ㸧ࡗࡡ RR㸝SskAࠉ
SrrA㸞࠿㓗໩ࢪࢹࣝࢪᚺ➽࡞㛭୙ࡊ࡙࠷ࡾࡆ࡛ࢅ᪺ࡼ࠾࡞ࡊࡒࠊࡱࡒࠉ㓕ẍ࡞࠽࠷࡙ࡢ AP-1 type ㌹෕ᅄᏄࡵ
୹さ࡝㓗໩ࢪࢹࣝࢪᚺ➽ᅄᏄ࡚࠵ࡾࡆ࡛࠾ࡼࠉA. nidulans ࡡ┞ྜྷᅄᏄࡵెࡎ࡙ᶭ⬗よᯊࢅモࡲࡒࠊAP-1 type
㌹෕ᅄᏄ࡞≁ᚡⓏ࡝ N ᮆ❻ࡡ bZIP ࢺ࣒࢕ࣤࠉC ᮆ❻ࡡ CRD ࣓ࢲ࣭ࣆࢅ࡛ࡵ࡞᭯ࡌࡾᅄᏄࡢࠉA. nidulans ࡡ
ࢣࢿ࣑୯࡞ୌࡗ᥆ᏽࡈࡿࡒࠊࡐࡡᅄᏄࢅ NapA (A. nidulans AP-1)࡛ࡊ࡙◒ቪᰬࢅష⿿ࡊࠉ㓗໩ࢪࢹࣝࢪᚺ➽ࡡ
よᯊࢅ⾔ࡖࡒ࡛ࡆࢀࠉNapA ◒ቪᰬࡢ SskA ࡷ SrrA ࡡ◒ቪᰬ࡛ྜྷᵕ࡞ H2O2ࠉt-BOOH ࡞វུᛮࢅ♟ࡊࠉࡈࡼ
࡞ࡢ Menadione ࡞ᑊࡊ࡙ࡵវུᛮࢅ♟ࡌࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊࡐࡡୌ᪁࡚ NapA ࡡ◒ቪࡢࠉSskA ࡷ SrrA
ࡡ◒ቪ࡛ࡢ␏࡝ࡽࠉฦ⏍Ꮔᙟᠺ⬗ࡷᙟឺ࡞ࡢධࡂᙫ㡢ࢅཀྵ࡯ࡈ࡝࠾ࡖࡒࠊࡈࡼ࡞ࠉ㓕ẍ࡞࠽࠷࡙ AP-1 type
㌹෕ᅄᏄࡷ RR㸝SskAࠉSrrA ࡡ┞ྜྷᅄᏄ㸞ࡢࠉ㔔㔘ᒌࡷ Methylglyoxal (MG) ࡡࢪࢹࣝࢪᚺ➽࡞ࡵ㛭୙ࡊ࡙࠷
ࡾࡆ࡛࠿▩ࡼࡿ࡙࠽ࡽࠉA. nidulans ࡞࠽ࡄࡾࡆࡿࡼࡡࢪࢹࣝࢪᚺ➽ᶭᵋ࡞ࡗ࠷࡙ࡵよᯊࡊࡒ࠷࡛⩻࠻࡙࠷ࡾࠊ
ࡆࡡࡻ࠹࡞ࠉ㓕ẍ࡛⣊≟Ⳟࡡ㛣࡚ಕᏋࡈࡿ࡙࠷ࡾࢪࢹࣝࢪᚺ➽ᅄᏄ࡞↌Ⅴࢅᙔ࡙ࡒよᯊ࠾ࡼࠉ⣊≟Ⳟࡡ⎌ሾ
ᚺ➽ᶭᵋࡡධమാࡡ⌦よࢅ┘ᣞࡊ࡙࠷ࡾࠊ
Analysis of the components involved in oxidative stress responses in Aspergillus nidulans
Yoshihiro Asano, Daisuke Hagiwara, Kyoko Kanamaru, Masashi Kato, Tetsuo Kobayashi, Takeshi Mizuno
(Grad. Sch. Bioagricultural Sci. Nagoya Univ.)
53
P-37
⣊≟Ⳟ Aspergillus nidulans ࡞࠽ࡄࡾ஦ᠺฦᛮ᝗ሒఎ㐡⣌ࣛࣤ㓗ᇱ௯௒ᅄᏄ YpdA
ࡡよᯊ
ఫ⸠ዄ὘Ꮔ, ཿᕖ೸ኯ㑳, 㜷㒂ᩏᝃ (᮶໪ኬ㝌࣬㎨࣬ᚺ⏍⛁)
┷ᰶ⏍∸ࡡᾈ㏩ᅸᚺ➽◂✪ࡢฝⰾ㓕ẍ(Saccharomyces cerevisiae)࡞࠽࠷࡙᭩ࡵ㐅ᒈࡊ࡙࠽ࡽࠉ㧏ᾈ㏩ᅸᚺ➽
HOG (High-Osmolarity Glycerol response) ⤊㊪ࡡ࡮࡯ධᐖ࠿᪺ࡼ࠾࡞ࡈࡿ࡙࠷ࡾࠊSln1p-Ypd1p-Ssk1p ࡢࠉ
His-Asp ࣛࣤ㓗ᇱ࣭ࣛࣝࢅᙟᠺࡌࡾᾈ㏩ᅸࢬࣤࢦ࣭஦ᠺฦᛮ᝗ሒఎ㐡⣌࡚࠵ࡽࠉୖὮࡡ Hog1p MAPK
(mitogen-activated protein kinase) cascade ࢅㇿ࡞โᚒࡊ࡙࠷ࡾࠊ⣊≟Ⳟࡡ࣓ࢸࣜ⏍∸࡚࠵ࡾ Aspergillus nidulans
ࡢࠉ㓕ẍ HOG ⤊㊪ᵋᠺ㐿ఎᏄࡡ࢛ࣜࢮࣞࢡࢅࡌ࡬࡙᭯ࡌࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖ࡙࠷ࡾࠊ㓕ẍࡡࣛࣤ㓗ᇱ௯௒
ᅄᏄ Ypd1p ࡡḖ᥾ࡢ Hog1pMAPK ࢅᵋᠺⓏ࡞Ὡᛮ໩ࡌࡾࡒࡴ࡞⮬Ṓࢅ♟ࡌࡆ࡛࠿ሒ࿈ࡈࡿ࡙࠷ࡾ࠿ࠉᠻࠍࡢ
௧๑ࠉA. nidulans ࡡ Ypd1p ࢛ࣜࢮࣞࢡ࡚࠵ࡾ YpdA ࠿ࡆࡡ⮬Ṓᛮࢅᢒโ࡚ࡀࡾࡆ࡛ࢅ᪺ࡼ࠾࡞ࡊࡒࠊࡱࡒࠉ
㓕ẍ࡚ࡢ Ypd1p ࡛┞பష⏕ࡌࡾ histidine kinase ࢅ Sln1p ࡊ࠾ᣚࡒ࡝࠷࠿ࠉA. nidulans ࡢ YpdA ࠿々ᩐࡡ histidine
kinase ࠾ࡼࣛࣤ㓗ᇱ㌹⛛ࢅུࡄࡾྊ⬗ᛮ࠿࠵ࡽࠉ஦ᠺฦᛮ᝗ሒఎ㐡⣌࠿々㞟໩ࡊ࡙࠷ࡾྊ⬗ᛮ࠿♟ြࡈࡿࡾࠊ
ᠻࠍࡢ alcA ࣈ࣓࣭ࣞࢰ࣭ࢅ⏕࠷ࡒࢤࣤࢸ࢔ࢨࣘࢻࣜ ypdA 㐿ఎᏄⓆ⌟ᢒโᰬ (ypdAΔ) ࢅ㏸ᠺࡊࡒࠊࡆࡡᰬ
ࡢ ypdA Ⓠ⌟ࢅᢒโࡊࡒሔྙ࡞ࡡࡲ㢟ⴥ࡝⏍⫩㜴ᐐࢅ♟ࡊࡒ஥࠾ࡼࠉA. nidulans ࡵฝⰾ㓕ẍ࡛ྜྷᵕ࡞ ypdA 㐿
ఎᏄࢅḖ᥾ࡌࡾ࡛⮬Ṓࢅ♟ࡌྊ⬗ᛮ࠿♟ြࡈࡿࡒࠊࡈࡼ࡞ࠉA. nidulans HogA MAPK (㓕ẍ Hog1pMAPK ࣓࣌
ࣞࢡ) ࠿ ypdAΔ ᰬ࡚ࡢࠉᾈ㏩ᅸๆ⃥ࡡ᭯↋࡞㛭ࢂࡼࡍᖏ࡞Ὡᛮ໩≟ឺ࡞࠵ࡾ஥࠿ふᐳࡈࡿࡒࠊࡱࡒࠉypdAΔ ᰬ
࡞࠽࠷࡙ HogA ࡡ୕ὮᅄᏄ࡚࠵ࡾ PbsB MAPKK (㓕ẍ Pbs2pMAPKK ࣓࣌ࣞࢡ) ࢅ◒ቪࡊࡒ࡛ࡆࢀ฽Ṓᛮࡡᅂ
㑂࠿ぜࡼࡿࡒࠊ௧୕ࡡ஥࠾ࡼࠉYpdA ࡢ Aspergillus nidulans HOG ⤊㊪ (AnHOG ⤊㊪) ࡡᵋᠺᅄᏄ࡚࠵ࡽࠉYpdA
Ḗ᥾࡞ࡻࡾ⏍⫩㜴ᐐࡡཋᅄࡡୌࡗ࡛ࡊ࡙ AnHOG ⤊㊪ࡡ᧘஗࠿㉫ࡆࡖ࡙࠷ࡾ஥࠿⩻࠻ࡼࡿࡒࠊ
Analysis of ypdA gene encoding a two-component histidine-containing phosphotransfer YpdA in the filamentous
fungus Aspergillus nidulans
Natsuko Sato, Kentaro Furukawa, Keietsu Abe (Tohoku Univ., Grad. Sch. Agri. Sci.)
P-38
Aspergillus nidulans ࣝࢪ࣎ࣤࢪ࣭ࣝ࢟ࣖࣝࢰ࣭SskA, SrrA ࡡ᫤㛣ⓏⓆ⌟ንິ࡛㎨ⷾ
វུᛮ࡛ࡡ㛭㏻
ᮿᯐⰃ༡ࠉ㔘୷ாᏄࠉⴏཋኬ♰ࠉ୷஬ῗୌ㑳ࠉỀ㔕⊓ࠉຊ⸠㞖ኃࠉᑚᯐူኰ 㸝ྞኬ㝌⏍࿤㎨࣬⏍∸ᶭᵋ㸞
[┘Ⓩ] ⣊≟Ⳟ Aspergillus nidulans ࡢࠉ௙ࡡኣࡂࡡ⣊≟Ⳟྜྷᵕࠉࣆࣜࢩ࢛࢞ࢮࢼࣜࠉ࢕ࣈࣞࢩ࢛ࣤ➴ࡡ㎨ⷾ࡞ᑊࡊ࡙ࠉ
Nik-1 ᆵ histidine kinase (nikA) ౪ᏋⓏ࡝វུᛮࢅ♟ࡌࠊA. nidulans ࡞ࡢࠉHis-Asp ࣛࣤ㓗࣭ࣛࣝᆵ᝗ሒఎ㐡⣌࡞࠽ࡄࡾ
ୖὮᅄᏄ࡚࠵ࡾ Response Regulator (RR) ࠿ 3 ⛸(SskA, SrrA, SrrC) ᏋᅹࡊࠉSskA, SrrA 㢦జ㐿ఎᏄࡢ⣊≟Ⳟࠉ㓕ẍ࡞㧏
ᗐ࡞ಕᏋࡈࡿ࡙࠷ࡾࠊᠻࠍࡢࠉSskA, SrrA ࡛㎨ⷾវུᛮ࡛ࡡ㛭㏻ࢅ᪺ࡼ࠾࡞ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙ࠉNikA, SskA, SrrA 㐿
ఎᏄࡡ᫤㛣ⓏⓆ⌟ንິ࡛྘㐿ఎᏄ◒ቪᰬࡡฦ⏍ᏄⓆⰾ᫤࡞࠽ࡄࡾ㎨ⷾࡡᙫ㡢ࢅよᯊࡊࡒࠊ
[⤎ᯕ] ࣆࣜࢩ࢛࢞ࢮࢼࣜࠉ࢕ࣈࣞࢩ࢛ࣤᏋᅹୖ࡚ࡢࠉA. nidulans ࡡ⏍⫩ࡢ㜴ᐐࡈࡿࡾࠊSskA, SrrA ༟≺◒ቪᰬࡡ⏍⫩
ࡵ୕オ㎨ⷾ࡞ᑊࡊ࡙ࠉវུᛮࢅ♟ࡌ࠿ࠉ஦㔔◒ቪᰬࡢ NikA ◒ቪᰬ࡛ྜྷ➴ࡡ⪇ᛮ࡛࡝ࡾ(ⴏཋࡼࠉᮇࢤࣤࣆ࢒ࣝࣤࢪཾ
㢄Ⓠ⾪)ࠊࣛ࢓ࣜࢰ࢕࣑ PCR ࡞ࡻࡾ⤊᫤Ⓩ࡝Ⓠ⌟ࣈࣞࣆ࢒࢕ࣜよᯊࡡ⤎ᯕࠉNikA mRNA ࡢᖏ࡞㧏ࣝ࣊ࣜᏋᅹࡊࠉฦ
⏍Ꮔᙟᠺ᭿࡞ࡢࡈࡼ࡞Ⓠ⌟ࣝ࣊ࣜ࠿୕᪴ࡊࡒࠊࡱࡒࠉSskA ࡢ࡮࡯ᵋᠺⓏ࡞Ⓠ⌟ࡊ࡙࠷ࡾࡡ࡞ᑊࡊࠉSrrA ࡢฦ⏍Ꮔᙟᠺ
ᚃ᭿࡞㧏Ⓠ⌟ࡊ࡙࠷ࡒࠊࡆࡿࡢࠉSskA ࡛ SrrA ࡡᶭ⬗ࡌࡾ᫤᭿࠿␏࡝ࡾࡆ࡛ࢅ♟ࡊ࡙࠽ࡽࠉᚉࡖ࡙ࠉSskAࠉSrrA ◒ቪ
ࡡᙫ㡢࠿ᇰ㣬᫤᭿࡞ࡻࡽ␏࡝ࡾྊ⬗ᛮ࠿⩻࠻ࡼࡿࡒࠊࡐࡆ࡚ࠉᮇሒ࿈࡚ࡢฦ⏍Ꮔ࠾ࡼࡡⓆⰾ࡛ࡐࡡᚃࡡⳞ⣊ᠺ㛏࡞୙
࠻ࡾ SskAࠉSrrA ◒ቪࡡᙫ㡢ࢅ㢟᚜㙶ୖ࡚ふᐳࡊࡒࠊSskAࠉSrrA ࠷ࡍࡿࡡ◒ቪᰬ࡞࠽࠷࡙ࡵࠉ୕オ㎨ⷾ࡞ࡻࡾⓆⰾ㜴
ᐐࡢぜࡼࡿ࡝࠾ࡖࡒ࠿ࠉSskA ◒ቪᰬ࡚ࡢᩐ᫤㛣࡚Ⳟ⣊ᠺ㛏࠿ೳḾࡊࠉⳞ⣊࠿⭶₮ࡊ࡙ᩐ⌌≟࡛࡝ࡖࡒࠊSrrA ◒ቪᰬ࡚
ࡢ⏍⫩㏷ᗐ࠿ⴥࡊࡂ఩ୖࡊࠉ㧏ᗐ࡞ࣇࣚࣤࢲࣤࢡࡊࡒም࠿ᙟᠺࡈࡿࡒࠊ
Differential roles of response regulators, SskA and SrrA, in fungicide sensitivity in Aspergillus nidulans
Yoshihiro Matsubayashi, Kyoko Kanamaru, Daisuke Hagiwara, Junichiro Marui, Takeshi Mizuno, Masashi Kato, Tetsuo
Kobayashi (Grad. Sch. of Bioagricultural Sciences, Nagoya Univ.)
54
P-39
In vitro ࣛࣤ㓗࣭ࣛࣝ⣌ࢅ⏕࠷ࡒ Aspergillus nidulans His-Asp ࣛࣤ㓗࣭ࣛࣝࢾࢴࢹ࣠
࣭ࢠࡡよᯊ
᮶ಘᏳࠉ㔘୷ாᏄࠉᮿ㮭᫓์ࠉᒜ⠓㈏ྍࠉỀ㔕⊓ࠉຊ⸠㞖ኃࠉᑚᯐူኰ 㸝ྞኬ㝌࣬⏍࿤㎨㸞
His-Asp ࣛࣤ㓗࣭ࣛࣝ᝗ሒఎ㐡⣌ࡢ⏍∸࡞࠽ࡄࡾ⎌ሾវ▩࣬⣵⬂හ᝗ሒఎ㐡ᶭᵋ࡚࠵ࡽࠉཋᰶ⏍∸ࡓࡄ࡚࡝
ࡂࠉ㓕ẍࡷ㧏➴᳔∸࡞࠽࠷࡙ࡵࡐࡡᏋᅹ࠿᪺ࡼ࠾࡛࡝ࡖ࡙࠷ࡾࠊ࣓ࢸࣜ⣊≟Ⳟ࡚࠵ࡾ A. nidulans ࡞ࡢ
Histidine Kinase㸝HK㸞㸦㸪⛸㢦ࠉHPt㸦⛸㢦ࠉResponse Regulator㸝RR㸞㸩⛸㢦ࡡᏋᅹ࠿ࢣࢿ࣑᝗ሒࡻࡽ᥆ᏽ
ࡈࡿ࡙࠷ࡾࠊᮇ◂✪࡚ࡢࡆࡿࡼࡌ࡬࡙ࡡᅄᏄࡡ cDNA ࢅཱིᚋࡊࠉࣛࢤࣤࣄࢻࣤࢹࢰࣤࣂࢠ㈻ࢅ⢥⿿ࡌࡾࡆ࡛
࡞ࡻࡽࠉࣛࣤ㓗࣭ࣛࣝࢾࢴࢹ࣭࣠ࢠࢅᡋ␆Ⓩ࡞ in vitro よᯊࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊࡒࠊ௑ᅂᠻࠍࡢ HPt ࡻࡽୖὮ
ࡡࢨࢡࢻࣜఎ㐡⤊㊪࡞↌Ⅴࢅࡊ࡯ࡽࠉin vitro ࣛࣤ㓗ᇱ㌹⛛ᐁ㥺ࢅ⾔ࡖࡒࠊኬ⭘Ⳟࡡ௥⾪Ⓩ࡝ HK ࡚࠵ࡾ ArcB
ࢅ㐛๨Ⓠ⌟ࡊࡒ⣵⬂㈻⭯ࢅࣛࣤ㓗ᇱࡡࢺࢻ࣭࡛ࡊ⣊≟Ⳟ HPt ࠽ࡻࡦ RR ࡫ࡡࣛࣤ㓗ᇱ㌹⛛ࢅよᯊࡊࡒ࡛ࡆࢀࠉ
HPt(YpdA)࡫ࡡࣛࣤ㓗ᇱ㌹⛛࡛ࠉࡈࡼ࡞ YpdA ࠾ࡼ RR㸝SrrA㸞࡫ࡡࣛࣤ㓗ᇱ㌹⛛ࢅ☔ヾࡊࡒࠊࡈࡼ࡞ࠉ᥆ᏽ
ࣛࣤ㓗໩⟘ᡜࢅ⨠ᥦࡊࡒን␏ࢰࣤࣂࢠ㈻㸝YpdAH85QࠉSrrAD385E㸞ࢅㄢ⿿ࡊ in vitro よᯊࡌࡾࡆ࡛࡞ࡻࡽࠉᐁ㝷
࡞ࡆࡿࡼࡡ࢓࣐ࢿ㓗ṟᇱ࠿ࣛࣤ㓗ᇱ㌹⛛࡞㛭୙ࡌࡾࡆ࡛ࢅッ᪺ࡊࡒࠊ
In vitro analysis of His-Asp phosphorelay in Aspergillus nidulans
Nobuhiro Azuma, Kyouko Kanamaru, Akinori Matushika, Takashi Yamashino, Takeshi Mizuno, Masashi Kato, Tetuo
Kobayashi
(Graduate School of Bioagricultural Sciences, Nagoya Univ.)
P-40
Aspergillus nidulans Histidine Kinase ࡡ㒂న≁␏ⓏⓆ⌟
㕝ᮄ㯖Ꮔ㸡㔘୷ாᏄ, ຊ⸠㞖ኃ, ᑚᯐူኰ 㸝ྞኬ㝌⏍࿤㎨࣬⏍∸ᶭᵋ㸞
࠘┘Ⓩ࠙⣊≟Ⳟ Aspergillus nidulans ࡡ His-Asp ࣛࣤ㓗࣭ࣛࣝ⣌ᵋᠺᅄᏄ࡛ࡊ࡙ࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ (HK)
15 ⛸ࠉࣝࢪ࣎ࣤࢪ࣭ࣝ࢟ࣖࣝࢰ࣭ (RR) 4 ⛸ࠉHPt ᅄᏄ 1 ⛸ࡡᏋᅹ࠿᥆ᏽࡈࡿ࡙࠷ࡾ࠿ࠉࡆࡿࡼࡡᅄᏄ࠿
វ▩ࡌࡾ⎌ሾࢨࢡࢻࣜࡡ⛸㢦ࡷࡐࡡᚺ➽ᶭᵋࡡኬ㒂ฦࡢ࠷ࡱࡓよ᪺ࡈࡿ࡙࠷࡝࠷ࠊࡱࡒࠉ15 ⛸㢦ࡡ HK ࠾
ࡼఎ㐡ࡈࡿࡾ࡛⩻࠻ࡼࡿࡾ々ᩐࡡࢨࢡࢻࣜࢅࠉ࠷࠾࡞ࡊ࡙ࡒࡖࡒ 1 ⛸㢦ࡡ HPt ࢅ௒ࡊ࡙ࡐࡡୖὮࡡ RR ࡫
࡛ఎ㐡ࡌࡾࡡ࠾ࠉࡐࡡレ⣵࡞ࡗ࠷࡙ࡵ᪺ࡼ࠾࡞ࡈࡿ࡙࠷࡝࠷ࠊࡐࡡ࣒࢜ࢼࢫ࣑ࡡୌࡗ࡛ࡊ࡙ࠉࡐࡿࡑࡿࡡ HK
࠿Ⳟమහࡡ␏࡝ࡾሔᡜࠉ␏࡝ࡾ᫤᭿࡞Ⓠ⌟ࡌࡾࡆ࡛࡚ࠉࡐࡿࡑࡿ␏࡝ࡾࢨࢡࢻࣜࢅୖὮ࡫࡛ఎ㐡ࡊ࡙࠷ࡾྊ
⬗ᛮࢅ⩻࠻ࠉ྘ HK ࡡⓆ⌟㒂నࡷⓆ⌟᮪௲ࡡよᯊࢅ⾔࠹ࡆ࡛࡞ࡊࡒࠊ
࠘᪁Ἢ࡛⤎ᯕ࠙PCR ࡚ቌᖕࡊࡒ 15 ⛸㢦ࡡ྘ HK ࡡࣈ࣓࣭ࣞࢰ࣭㡷ᇡࡡୖὮ࡞࣭ࣝ࣎ࢰ࣭㐿ఎᏄ࡛ࡊ࡙
GFP 㐿ఎᏄࢅ㏻⤎ࡊࠉA.nidulans ࡞ᑙථࡊࡒࠊᚋࡼࡿࡒᰬࢅᵕࠍ࡝ᇰ㣬᮪௲࡚ᇰ㣬ࡊࠉ⺧ක㢟᚜㙶ふᐳࢅ⾔
࠹ࡆ࡛࡚ࠉࡐࡿࡑࡿࡡ HK ࡡⓆ⌟㒂నࡡよᯊࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉ㸝㸦㸞Ⳟ⣊࠽ࡻࡦ㡤ᄙ࠾ࡼ⬂Ꮔ࡞࠾ࡄ࡙
ࡡⳞమධమ࡚⺧ක࠿ࡲࡼࡿࡒࡵࡡࠉ㸝㸧㸞㡤ᄙ࠾ࡼ⬂Ꮔ࡞࠾ࡄ࡙ࡡ㒂ฦ࡞ᙁ࠷⺧ක࠿ふᐳࡈࡿࡒࡵࡡࠉ㸝㸨㸞
௑ᅂࡡふᐳ᮪௲࡚ࡢⓆ⌟࠿ぜࡼࡿ࡝࠾ࡖࡒࡵࡡࡡ 3 ⛸㢦࡞ኬืࡈࡿࡒࠊHK ࡡ㒂న≁␏ⓏⓆ⌟࡛ᶭ⬗ฦᢰࡡ
㛭౿࡞ࡗ࠷࡙⩻ᐳࡌࡾࠊ
Site-specific expression of the Histidine kinases in Aspergillus nidulans.
Asako Suzuki, Kyoko Kanamaru, Masashi Kato, Tetsuo Kobayashi
(Dept. of Biological Mechanisms and Functions, Graduate School of Bioagricultural Science, Nagoya Univ.)
55
P-41
࢕ࣤࢣ᰷ࣤ⭁⑋Ⳟ࡞࠽ࡄࡾ MAPK 㐿ఎᏄ◒ቪᰬࡡషฝ࡛ࡐࡡ⑋ཋᛮ
໪ᔪຩ඼㸡㡪㈙ᬍ஁, Ⓤ⏣‫‮‬᭹ 㸝ᒪ㜟ኬ㝌࣬㎨㸞
⣵⬂ࡢኣᵕ࡝⎌ሾࢨࢡࢻࣜࢅུࡄ࡙⏍⌦≟ឺࢅን໩ࡈࡎࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࠊ⣵⬂හࡡࢨࢡࢻࣜఎ㐡⣌
࡞ࡢ mitogen-activated protein kinase (MAPK)ࡷ cAMP-dependent protein kinase ࡝࡜࠿࠵ࡽࠉ࢕ࢾ࠷ࡵࡔ⑋Ⳟ
(Magnaporthe grisea)࡝࡜ࡡ᳔∸⑋ཋᛮ⣊≟Ⳟ࡞࠽࠷࡙ࡢࢨࢡࢻࣜఎ㐡⣌ࡡ⑋ཋᛮ࡫ࡡ㛭୙࠿ヾࡴࡼࡿ࡙࠷ࡾࠊ
ᮇ◂✪࡚ࡢ࢕ࣤࢣ᰷ࣤ⭁⑋Ⳟ Fusarium solani f. sp. phaseoli ࡞࠽ࡄࡾ MAPK ࡡ⑋ཋᛮ࡫ࡡ㛭୙ࢅㄢ࡬ࡒࠊࡱ
ࡍ㎾⦍Ⳟ࡚࠵ࡾ࢙ࣤࢺ᰷ࢗ⭁⑋Ⳟ F. solani f. sp. pisi ࡚᪺ࡼ࠾࡞ࡈࡿ࡙࠷ࡾ FUS3/KSS1 ࢰ࢕ࣈࡡ MAPK 㐿ఎᏄ
ࡡሲᇱ㒼าࢅࡵ࡛࡞ PCR ⏕ࣈࣚ࢕࣏࣭ࢅスゝࡊࠉF. solani f. sp. phaseoli ࠾ࡼ MAPK ࣓࣌ࣞࢡ(FspMAPK1)ࢅ
ࢠ࣭ࣞࢼࣤࢡࡊࡒࠊFspMAPK1 ࠿ࢤ࣭ࢺࡌࡾ࢓࣐ࢿ㓗㒼า࡞ࡢ᥆ᏽࣛࣤ㓗໩㒂న࡚࠵ࡾ TEY 㒼า࠿Ꮛᅹࡊࡒࠊ
ࢠ࣭ࣞࢼࣤࢡࡊࡒ FspMAPK1 ࡞ࣀ࢕ࢡ࣏ࣞ࢕ࢨࣤ⪇ᛮ㐿ఎᏄࢅ᣼ථࡊ࡙㐿ఎᏄ◒ቪ⏕ࣈࣚࢪ࣐ࢺ࣊ࢠࢰ࣭ࢅ
ᵋ⠇ࡊࠉ࢙࣎ࣛࢲࣝࣤࢡࣛࢤ࣭ࣜἪ࡞ࡻࡽ F. solani f. sp. phaseoli ࢅᙟ㈻㌹ᥦࡊࡒࠊᚋࡼࡿࡒᙟ㈻㌹ᥦమࡡ୯
࠾ࡼࠉPCR ཀྵࡦ FspMAPK1 ࢅࣈ࣭ࣞࣇ࡛ࡊࡒࢦࢧࣤࣀ࢕ࣇࣛࢱ࢕ࢭ࣭ࢨࣘࣤ࡞ࡻࡽ FspMAPK1 ◒ቪᰬࢅ㐽ᢜ
ࡊࡒࠊ࢕ࣤࢣࣤࡡࡵࡷࡊୖ⫿㍂ࢅฺ⏕ࡌࡾ᪁Ἢ࡚ FspMAPK1 ◒ቪᰬࡡ⑋ཋᛮࢅㄢ࡬ࡒ⤎ᯕࠉFspMAPK1 ◒ቪ
ᰬࡡ⑋ཋᛮࡢኬࡀࡂ఩ୖࡊ࡙࠷ࡒࠊᚉࡖ࡙ࠉ௙ࡡ᳔∸⑋ཋᛮ⣊≟Ⳟ࡛ྜྷᵕ F. solani f. sp. phaseoli ࡞࠽࠷࡙ࡵ
MAPK ࠿⑋ཋᛮ࡞㛭୙ࡊ࡙࠷ࡾࡆ࡛࠿♟ࡈࡿࡒࠊ
MAPK gene desruption and it’s affection of pathogenicity of Fusarium solani f. sp. phaseoli
Yusuke Kitajima, Haruhisa Suga, Mituro Hyakumachi (Gifu Univ.)
P-42
Neurospora crassa ࡡ grg-1 (glucose-repressible gene)ࡢ OS-2 MAP ࢞ࢻ࣭ࢭ࡞ࡻࡽⓆ⌟ㄢ
⟿ࢅུࡄࡾ
ῳ㑌⟿Ꮔ㸦ࠉᆊ㔕┷ᖲ 2 ࠉሲ⃕࠵ࡍࡈ 1ࠉⴘྙ์ᖶ㸨ࠉᮄᮟ┷㸨ࠉ⸠ᮟ┷ 1 (㸦᮶Ὂኬ㹺⏍࿤㸡2 ᮶Ὂኬ㹺᳔ᶭ◂ࢬ,㸨⌦◂㹺୯ኳ◂)
࢓࢜ࣂࣤ࢜ࣄࡡ OS-2(MAP ࢞ࢻ࣭ࢭ)ࡢࠉᾈ㏩ᅸࢬࣤࢦ࣭(ࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ)࠾ࡼࡡࢨࢡࢻࣜࢅུࡄᾈ㏩
ᅸᚺ➽࡞㔔さ࡝ᙲ๪ࢅᢰࡖ࡙࠷ࡾࠊୌ᪁ࠉPKAC-1(A ࢞ࢻ࣭ࢭ)ࡢࠉࢡࣜࢤ࣭ࢪࢬࣤࢦ࣭(G ࢰࣤࣂࢠུᐖమ?)
ࡡୖὮ࡚⬂Ꮔᙟᠺ࡛Ⳟ⣊ᙟᠺࡡࢪ࢕ࢴࢲࣤࢡ࡞㛭୙ࡊ࡙࠷ࡾࠊos-2;pkac-1 ஦㔔ን␏ᰬ࡝࡜ࡡᾈ㏩ᅸវུᛮよ
ᯊ࠾ࡼ୦⤊㊪ࡡࢠࣞࢪࢹ࣭ࢠ࠿♟ြࡈࡿ࡙࠷ࡾ࠿ࠉレ⣵ࡢ୘᪺࡚࠵ࡾࠊ௑ᅂࠉgrg-1(glucose-repressible gene)
㐿ఎᏄࡡⓆ⌟࠿ࠉOS-2 ࢅὩᛮ໩ࡌࡾ fludioxonil ࠽ࡻࡦᾈ㏩ᅸฌ⌦࡚㢟ⴥ࡞ㄇᑙࡈࡿࡾࡆ࡛࠿ࠉࣛ࢓ࣜࢰ࢕࣑
RT-PCR よᯊ࠾ࡼ᪺ࡼ࠾࡞࡝ࡖࡒࠊgrg-1 ࡡࣈ࣓࣭ࣞࢰ࣭㡷ᇡࡡ࢙࣒ࣝࣤࢹ㒼าࢅ᳠⣬ࡊࡒ࡛ࡆࢀࠉ
CRE(cAMP-Responsive Element)ࡷ STRE(Stress Responsive Element)࠿Ꮛᅹࡊࡒࠊgrg-1 㐿ఎᏄࡡⓆ⌟ᵕᘟࢅභ↌
Ⅴ࣭ࣝࢧ࣭㢟᚜㙶ࢅ⏕࠷࡙よᯊࡌࡾࡒࡴ࡞ࠉgrg-1 ࡡࣈ࣓࣭ࣞࢰ࣭࡞ EGFP 㐿ఎᏄࢅ㏻⤎ࡊࡒࣈࣚࢪ࣐ࢺ
(pGRGFP)ࢅషᠺࡊࡒࠊpGRGFP ᑙථᰬࡢࠉfludioxonil ࡷᾈ㏩ᅸฌ⌦࡞ࡻࡽᙁ࠷⺧කࢅ♟ࡌࡆ࡛࠿☔ヾࡈࡿࡒࠊ
⌟ᅹࠉpGRGFP ࢅฺ⏕ࡊ࡙ࠉⓆ⌟ㄢ⟿࡛࢙࣒ࣝࣤࢹ࡛ࡡ㛭㏻࡞ࡗ࠷࡙よᯊࢅ⾔ࡖ࡙࠷ࡾࠊgrg-1 ࡢ⣊≟Ⳟ࡞≁
᭯ࡡ㐿ఎᏄ࡚࠵ࡽࡐࡡᶭ⬗ࡢ୘᪺࡚࠵ࡾ࠿ࠉOS ⤊㊪ࡡୖὮ࡚⢶᩺⏍ Key 㓕⣪ࡡ㐿ఎᏄ(fbp-1 ࡷ pck-1)ࡷ⬂Ꮔ
≁␏Ⓩ࢜ࢰ࣭ࣚࢭ cat-1 ࠿ㄢ⟿ࡈࡿ࡙࠷ࡾࡆ࡛࡛ྙࢂࡎࡾ࡛ࠉOS ⤊㊪࠿⢶″ࡡᙫ㡢ࢅུࡄ࡙࠷ࡾྊ⬗ᛮ࠿♟
ြࡈࡿࡾࠊ
Expression of glucose-repressible gene, grg-1, is regulated by OS-2 MAP kinase
Setsuko Watanabe1,Shinpei Banno㸧,Azusa Shiozawa1,Noriyuki Ochiai㸨,Makoto Kimura㸨,Makoto Fujimura1 (1Life
Sci.,Toyo Univ.,2PRRC.,Toyo Univ.,㸨Env.Mol.Biol.,RIKEN.)
56
P-43
Neurospora crassa ࡡ OS-2 MAP ࢞ࢻ࣭ࢭࡡୖὮࡡ㌹෕ㄢ⟿ᅄᏄࡡྜྷᏽ
ሲ⃕࠵ࡍࡈ 1㸡ᆊ㔕┷ᖲ 2㸡ᒜୖ࿰Ᏻ 1㸡ῳ㑌⟿Ꮔ 1 㸡⚗᲻ᩝᗛ 1 㸡⸠ᮟ┷ 1
㸝1 ᮶Ὂኬ࣬⏍࿤, 2 ᮶Ὂኬ᳔࣬ᶭ◂ࢬ㸞
N.crassa ࡡ OS-2 MAP ࢞ࢻ࣭ࢭࡢᾈ㏩ᅸࢪࢹࣝࢪ࡝࡜࡞ࡻࡽὩᛮ໩ࡈࡿࠉࢡࣛࢬ࣭ࣞࣜྙᠺ㓕⣪(gcy-1, gcy-3,
dak-1)ࠉ࢜ࢰ࣭ࣚࢭ(ctt-1, cat-1)ࡷ⢶᩺⏍ key 㓕⣪(fbp-1, pck-1)ࡡ㐿ఎᏄⓆ⌟ࢅㄇᑙࡌࡾࠊࡊ࠾ࡊࠉࡆࡿࡼࡡ OS-2
౪Ꮛᛮ㐿ఎᏄࡡ㌹෕ㄢ⟿㐿ఎᏄࡢ≁ᏽࡈࡿ࡙࠷࡝࠷ࠊࡐࡆ࡚ࠉ㓕ẍࡡ Hog1 MAP ࢞ࢻ࣭ࢭୖὮࡡ㌹෕ㄢ⟿㐿
ఎᏄ sko1㸡msn2/4 ࠽ࡻࡦ yap1 ࡡ࣓࣌ࣞࢡ࡚࠵ࡾ atf-1 (CREB-type), msn-1 (STRE-type), nap-1 (AP-1-type)ࡡ◒ቪ
ᰬࢅ┞ྜྷ⤄ࡲᥦ࠻Ἢ࡞ࡻࡽ༟㞫ࡊࠉࡐࡡよᯊࢅ⾔ࡖࡒࠊos-2 ን␏ᰬࡢࣆࣜࢩ࢛࢞ࢮࢼࣜ࡞⪇ᛮ࠽ࡻࡦᾈ㏩ᅸ
࡞វུᛮࢅ♟ࡌ࠿ࠉ࠷ࡍࡿࡡ◒ቪᰬࡵ㔕⏍ᰬ࡛ྜྷᵕࡡវུᛮࢅ♟ࡊࡒࠊࡊ࠾ࡊࠉnap-1 ◒ቪᰬࡢฝⰾ㓕ẍࡡ
yap1 ን␏ᰬྜྷᵕ࡞࣒ࢻࢩ࢛ࣤ࡝࡜ࡡ㓗໩Ⓩࢪࢹࣝࢪ࡞វུᛮࢅ♟ࡊࠉmsn-1 ◒ቪᰬࡢᙁ࠷࣒ࢻࢩ࢛ࣤ⪇ᛮࢅ
♟ࡊࡒࠊࣛ࢓ࣜࢰ࢕࣑ PCR ࢅ⏕࠷࡙྘◒ቪᰬࡡ OS-2 ౪Ꮛᛮࡡ㐿ఎᏄ⩄ࡡⓆ⌟ㄇᑙࢅㄢ࡬ࡒ࡛ࡆࢀࠉmsn-1
࠽ࡻࡦ nap-1 ◒ቪᰬ࡚ࡢ㔕⏍ᰬ࡛㢟ⴥ࡝ᕣ࠿ヾࡴࡼࡿ࡝࠾ࡖࡒ࠿ࠉatf-1 ◒ቪᰬ࡚ࡢ gcy-1 ࢅࡢࡋࡴ 7 ⛸㢦ࡡ
OS-2 ౪Ꮛᛮ㐿ఎᏄࡡⓆ⌟ㄇᑙ࠿㢟ⴥ࡞఩ୖࡊ࡙࠷ࡾࡆ࡛࠿ึ᪺ࡊࡒࠊࡆࡡࡆ࡛࠾ࡼࠉATF-1 ࠿ OS-2 MAP ࢞
ࢻ࣭ࢭࡡୖὮࡡ㌹෕ㄢ⟿ᅄᏄ࡚࠵ࡾ࡛᥆ᏽࡈࡿࡒࠊࡊ࠾ࡊࠉatf-1 ◒ቪᰬࢅ㔕⏍ᰬࡷ os-2 ን␏ᰬ࡛ஹ㒼ࡊࡒ࡛
ࡆࢀࠉᏄᄙ⬂Ꮔࡢḿᖏ࡞ᙟᠺࡈࡿࡾࡵࡡࡡࠉࡐࡡⓆⰾ⋙ࡢ 50%⛤ᗐ࡚࠵ࡽࠉatf-1 ◒ቪࢅࡵࡗᏄᏘ࠿ධࡂᚋࡼ
ࡿ࡝࠷ࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊ⌟ᅹࠉࡆࡡཋᅄࢅㄢᰕࡌࡾ࡛࡛ࡵ࡞ࠉࣈ࣓࣭ࣞࢰ࣭よᯊࢅ⾔ࡖ࡙࠷ࡾࠊ
Identification of transcriptional factor regulated by OS-2 MAP kinase in Neurospora crassa
Azusa Shiozawa1, Shinpei Banno2, Kazuhiro Yamashita1, Setsuko Watanabe1, Fumiyasu Fukumori1, and Makoto
Fujimura1
(1Life Sci.,Toyo Univ., 2PRRC.,Toyo Univ.)
P-44
The nsdC gene encoding a novel positive regulator of sexual development of Aspergillus
nidulans is regulated by complicated transcriptional and post-transcriptional control.
Hye-Ryoun Kim and Dong-Min Han (Division of Life Science, Wonkwang University, Korea )
The nsdC gene which is predicted to encode a putative transcription factor carrying a novel type of zinc finger DNA
binding domain consisting of two C2H2 and a C2HC motifs acts as a positive regulator of sexual development in A.
nidulans. Two distinct transcripts, 3.2 and 2.8 kb, are synthesized and the smaller one differentially accumulates in
various stages of growth and development as well as under the varying nutritional conditions. 5’ RACE and Northern
blot analysis revealed that there were at least two alternative transcription initiation sites which might be responsible for
two kinds of transcript in different size. Two alternative transcription termination sites were also found through 3’
RACE and Northern blot analysis of nsdC deletion mutant. The gene carries two relatively long introns in its 5’UTR,
one of which is spliced alternatively. The results suggest that the expression of nsdC is controlled by complicated
transcriptional and post transcriptional regulations.
57
P-45
Molecular analysis of the rapamycin-sensitive and cycloheximide-resistant ActA1 gene in
Aspergillus nidulans
Sun-Hee Noh and Suhn-Kee Chae (Dept. Biochemistry and Biomed RRC, Paichai Univ. Daejeon, 302-735 Korea)
The TOR (Target Of Rapamycin) pathway has been implicated in regulation of cellular responses to nutrients.
During the course of screening of rapamycing-sensitive and -resistant mutants in A. nidulans, we observed that
rapamycin sensitivities of FGSC stock strains were correlated to the presence of the ActA1 mutation in the strains. In
order to isolate the cognate gene responsible to rapamycin-sensitivity, we took an approach of complementation of an
ActA1 mutant with a genomic DNA library. Transformant was selected based on their wild-type sensitivity to
rapamycin. The yeast GCN20-like gene was identified from the complementing region in the transformant. Sequencing
analysis of its gDNA and cDNA predicted that the 2,596 bp-long ORF encoded a putative ATP-binding cassette protein
and was interrupted by 8 introns. This ActA1 complementing DNA region was mapped to chromosome III near ActA
indicating that the complementing clone was not a suppressor of ActA1. The ActA1 mutant allele was amplified and
sequenced. The mutant allele ActA1 contained point mutations at 393rd and 394th amino acid residues. ΔActA displayed
resistance to cycloheximide, but failed to show rapamycin sensitivity. Although ǼGCN20 of S. cerevisiae is sensitive
to 5'-MT, over-expression of ActA did not complement its 5'-MT sensitivity. [Supported by KRF]
P-46
N.crassa ࡞࠽ࡄࡾࢨࢡࢻࣜ㐿ఎᏄ ncSCD2, ncSHK1 ࡡよᯊ
ᕖᮟ┷ᚷ,ୌ▴᫓ᙢ 㸝᮶ὊኬᏕ࣬⏍࿤⛁Ꮥ◂✪⛁㸞
S.pombe ࡚ࡢࠉRAS1 ࡞ࡻࡾโᚒࢅུࡄࡾ SCD1 ࢨࢡࢻࣜఎ㐡⤊㊪࠿Ꮛᅹࡊࠉ⣵⬂ࡡᙟឺᙟᠺࡷ㞜㞕㛣ࡡஹ
㒼࡞㛭ࡌࡾㄢ⟿ࡊ࡙࠷ࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࠊࡆࡡ SCD1 ⤊㊪࡚ࡢࠉscaffolding ࢰࣤࣂࢠ㈻࡚࠵ࡾ SCD2ࠉ
small-GTPase ࡚࠵ࡾ CDC42ࠉࡐࡡ GDP/GTP ஹᥦᅄᏄ࡚࠵ࡾ SCD1ࠉࡐࡊ࡙ kinase Ὡᛮࢅᣚࡗ SHK1 ࠿々ྙ
మࢅᙟᠺࡊࠉୖὮ࡫ࡡࣛࣤ㓗໩ࢨࢡࢻࣜࡡఎ㐡ࢅ⾔࡝ࡖ࡙࠷ࡾࠊྜྷࡋࢨࢡࢻࣜఎ㐡⤊㊪୕࡚ࠉSCD2 ࡛ SHK1
ࡢ༝ㄢࡊ഼࡙࠷࡙࠷ࡾࠊ
N.crassa ࡡࢣࢿ࣑ࢸ࣭ࢰ࣭࣊ࢪ୕࡞࠽࠷࡙ࡵࠉࡆࡿࡼ S.pombe ࡡ SCD2 ࡛ SHK1 ࡞㧏࠷┞ྜྷᛮࢅᣚࡗ㡷ᇡ
࠿Ꮛᅹࡊࠉࡐࡿࡑࡿ ncSCD2ࠉncSHK1 ࡛ࡊࡒࠊࡐࡊ࡙ N.crassa ࡞࠽ࡄࡾࡐࡿࡼࡡᶭ⬗ࢅㄢ࡬ࡾࡒࡴࠉncSCD2ࠉ
ncSHK1 㐿ఎᏄࢅ◒ቪࡊ࡙ࡐࡡᶭ⬗ࡡよᯊࢅ⾔࡝ࡖࡒࠊ
୦㐿ఎᏄ◒ቪᰬࡢⳞ⣊ࡡఘ㛏ᠺ㛏࠿㐔ࡂࠉⳞ⣊඙❻࡚ࡡ␏ᖏ࡝ฦᒪࡷ㟸ᖏ࡞≻࠷㝰ባ㛣㝰ࡡᙟᠺ࡛࠷ࡖࡒ
ᙟឺ␏ᖏࠉஹ㒼⬗ࡡḖዯ࡛࠷ࡖࡒභ㏳ࡡ⾪⌟ᆵࢅ♟ࡊࡒࠊࡊ࠾ࡊ୦㐿ఎᏄ◒ቪᰬ㛣ࡡᙟ㈻࡞ࡢ࠷ࡂࡗ࠾ࡡ㐢
࠷࠿ぜࡼࡿࠉ≁࡞ ncSHK1 㐿ఎᏄ◒ቪᰬ࡞㛭ࡊ࡙ࡢᾈ㏩ᅸ⪇ᛮࢅ♟ࡌ࡛࠷ࡖࡒᙟ㈻ࢅ♟ࡊࡒࠊ୦㐿ఎᏄ◒ቪ
ᰬ࡞ࡢኣᑛࡡᙟ㈻ࡡ㐢࠷ࡢ࠵ࡾ࠿ࠉ
ࡷࡢࡽ N.crassa ࡞࠽࠷࡙ࡵ SCD1 ⤊㊪࠿⣵⬂ࡡᙟឺᙟᠺࡷஹ㒼ࢅโᚒࡊࠉ
ࡐࡡ⤊㊪୕࡚ ncSCD2 ࠉncSHK1 ࡢᶭ⬗ࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡾࠊࡱࡒ ncSHK1 㐿ఎᏄ◒ቪᰬ࠿ᾈ㏩ᅸ⪇ᛮࢅ♟
ࡊࡒࡆ࡛࠾ࡼࠉN.crassa ࡚ࡢ SCD1 ⤊㊪࠿ᾈ㏩ᅸㄢ⟿ࢅᢰ࠹ࢨࢡࢻࣜఎ㐡⤊㊪࡛ࢠࣞࢪࢹ࣭ࢠࡊ࡙࠷ࡾྊ⬗
ᛮ࠿࠵ࡾ࡛⩻࠻ࡼࡿࡾࠊ
Characterization of signaling gene ncSCD2, ncSHK1 of N.crassa
Masashi Kawamura, Akihiko Ichiishi
(Grad . Sch . of Life Sciences , Toyo Univ)
58
P-47
Neurospora crassa ࡡ NER 㐿ఎᏄ mus-43, mus-44 ࡡよᯊ
ఫ⸠ ḿொ㸡ୌ▴ ᫓ᙢ㸝᮶Ὂኬ࣬⏍࿤⛁Ꮥ㒂㸞
⣰አ⥲࠿ࡵࡒࡼࡌ DNA ᥾ഭࡢ୹࡞ TC(6-4)ක⏐∸ ((6-4)PP) ࡛ࢨࢠࣞࣇࢰࣤᆵࣅ࣐ࣛࢩࣤ஦㔖మ (CPD)
࡞௥⾪ࡈࡿࠉࢽࢠ࢛ࣝࢲࢺ㝎ཡಞᚗ (nucleotide excision repair : NER) ࡢࡆࡿࡼࡡ DNA ᥾ഭࢅヾㆉࡊಞᚗࢅ⾔
࠹ࠊ
࢓࢜ࣂࣤ࢜ࣄࡡ NER ࡞㛭୙ࡌࡾ mus-43 ࡛ mus-44 ን␏ᰬࡢࠉ⣰አ⥲ࠉ4NQO ࡡን␏ཋ࡞ᑊࡊ࡙វུᛮࢅ♟
ࡊࡒࡆ࡛࠾ࡼࠉಞᚗ⣌࡞㛭୙ࡊ࡙࠷ࡾࡆ࡛ࢅ☔ヾࡊࡒ㹶
ࡆࡿࡼࡡን␏ᰬ࡛࢓࢜ࣂࣤ࢜ࣄ࡚ࡆࡿࡱ࡚࡞༟㞫ࡈࡿ࡙࠷ࡾ௙ࡡಞᚗ⣌ን␏ᰬ࡛஦㔔ን␏ᰬࢅష⿿ࡊࠉ࢙ࣅ
ࢪࢰࢨࢪよᯊࢅ⾔ࡖࡒ⤎ᯕࠉ୦ን␏ᰬࡡ⣰አ⥲࡞ᑊࡌࡾ⏍Ꮛ⋙࠿ NER ን␏ᰬ࡚࠵ࡾ mus-38 ን␏ᰬ࡛㟸ᖏ࡞
జࡒ⤎ᯕࢅ♟ࡊࡒࡆ࡛࠾ࡼ NER ⣌࡞㛭୙ࡊ࡙࠷ࡾࡆ࡛ࢅ☔ヾࡊࡒࠊࡱࡒࠉmus-38 ን␏ᰬ࡚௧๑࡞ሒ࿈ࡈࡿ࡙
࠷ࡾකᅂᚗ⬗ࡡḖ᥾࠿ mus-44 ን␏ᰬ࡚ࡵྜྷᵕ࡞ࡲࡼࡿࠉmus-43 ን␏ᰬ࡞࠽࠷࡙ࡡකᅂᚗ⋙ࡢ㔕⏍ᰬࡻࡽࡢ఩
࠷ࡵࡡࡡࠉmus-38,mus-44 ን␏ᰬࡻࡽࡢ㧏࠷ࡆ࡛ࢅ☔ヾࡊࡒࠊ
ࣂࣤࢹࢷࣤ㓗さịᛮᰬࢅ⏕࠷࡙ࠉ୦ን␏ᰬࡡᚗᖉ✲↓ን␏⋙ࢅㄢ࡬ࡒ⤎ᯕࠉ୦ን␏ᰬ࡛ࡵ࡞㔕⏍ᰬࡻࡽࡵ
㧏࠷ᚗᖉ⋙ࢅ♟ࡊࡒࠊ
⣰አ⥲࡞ࡻࡽㄇ㉫ࡈࡿࡾ(6-4)PPࠉCPD ࡡ஦ࡗࡡ DNA ᥾ഭ࡞ᑊࡌࡾ㝎ཡ⬗࡞ࡗ࠷࡙ࠉ(6-4)PP ࠽ࡻࡦ CPD
࡞ᑊࡌࡾᢘమࢅ⏕࠷࡙ῼᏽࢅ⾔ࡖࡒࠊCPD ࡡ᥾ഭࡢ mus-43,mus-44 ୦ን␏ᰬ࡛ࡵ㔕⏍ᰬ࡛ྜྷ⛤ᗐࡡ㝎ཡ⬗ࢅ
᭯ࡊࡒࠊࡐࡿࡼࡡ⤎ᯕࡵྙࢂࡎ࡙ሒ࿈ࡌࡾࠊ
Functional analyses of the NER gene, mus-43 and mus-44 from Neurospora crassa
Masahito Sato, Akihiko Ichiishi
(Toyo, Univ. Fac. of Life Sciences)
P-48
㯔Ⳟࡡ DNA ligase (lig4) 㐿ఎᏄ◒ቪ࡞ࡻࡾ㧏㢎ᗐ┞ྜྷ⤄ᥦ࠻ᐙ୹ࡡ㏸ᠺ
Ềㆺ἖ 1 㸡ᕝ⸠Ὂᖲ 1, ᮿ὾▩ᕤ 1, ஬୕ᘧୌ 2, 㜷㒂ᩏᝃ 3, ஫࿝ົஒ 1 㸝1 ᮶໪ኬ㝌㎨࣬⏍∸⏐ᴏ๭ᠺࠉ 2 ᇳ
⋚ኬ⌦ࠉ 3 ᮶໪ኬ㝌㎨࣬ᚺ⏍໩㸞
㎾ᖳࠉ࢓࢜ࣂࣤ࢜ࣄ࡝࡜࡞࠽࠷࡙㟸┞ྜྷᮆ❻⤎ྙཬᚺ࡞㛭୙ࡌࡾ㐿ఎᏄ࡚࠵ࡾ ku70, ku80 ࡡ࣓࣌ࣞࢡ㐿ఎ
Ꮔࢅ◒ቪࡌࡾࡆ࡛࡚┞ྜྷ⤄ᥦ࠻ຝ⋙࠿๸Ⓩ࡞୕᪴ࡌࡾ࡛࠷࠹⤎ᯕ࠿ሒ࿈ࡈࡿࡒࠊࡆࡿࡼࡡ⤎ᯕ࠾ࡼ㟸┞ྜྷᮆ
❻⤎ྙࢅᢒโࡌࡾࡆ࡛࡚┞ྜྷ⤄ᥦ࠻㢎ᗐ࠿୕᪴ࡌࡾࡆ࡛࠿♟ࡈࡿࡒࠊ㯔Ⳟ࡞࠽࠷࡙ࡵ ku70, ku80 ࣓࣌ࣞࢡ㐿
ఎᏄ◒ቪᰬ࠿ᚋࡼࡿ࡙࠷ࡾ࠿ࠉ࢓࢜ࣂࣤ࢜ࣄ࡝࡜࡞Ẓ࡬ࡐࡡ┞ྜྷ⤄ᥦ࠻㢎ᗐࡢ఩࠷ࡆ࡛࠿ሒ࿈ࡈࡿ࡙࠷ࡾࠊ
ࡐࡆ࡚ᮇ◂✪࡚ࡢࠉ㯔Ⳟࡡ᭞࡝ࡾ㎷㏷࡝㐿ఎᏄ◒ቪᰬ㏸ᠺࢨࢪࢷ࣑ࡡ☔❟ࢅ┘Ⓩ࡛ࡊࠉ㟸┞ྜྷᮆ❻⤎ྙཬᚺ
࡞࠽࠷࡙ DNA ࡡ⤎ྙཬᚺ࡞㛭୙ࡊ࡙࠷ࡾ lig4 ࣓࣌ࣞࢡ (ligD) ࡡ㐿ఎᏄ◒ቪᰬࡡ㏸ᠺ࡛┞ྜྷ⤄ᥦ࠻ຝ⋙ࡡ
᳠ᏽࢅ⾔ࡖࡒࠊࣈࣞࢷ࢓࣭ࢭࡡⓆ⌟࡞㛭୙ࡌࡾ㌹෕ᅄᏄ࡚࠵ࡾ prtR 㐿ఎᏄࢅ⏕࠷࡙┞ྜྷ⤄ᥦ࠻ຝ⋙ࢅ᳠ᏽ
ࡊࡒ࡛ࡆࢀࠉሒ࿈ࡈࡿ࡙࠷ࡾ㯔Ⳟ ku70, ku80 㐿ఎᏄ◒ቪᰬࡻࡽࡵ┞ྜྷ⤄ᥦ࠻ຝ⋙࠿୕᪴ࡊ࡙࠷ࡒࠊ⌟ᅹࠉprtR
㐿ఎᏄ௧አࡡ㐿ఎᏄࢅ⏕࠷࡙┞ྜྷ⤄ᥦ࠻ຝ⋙ࢅ᳠ゞ୯࡚࠵ࡾࠊ
Highly efficient gene targeting of DNA ligase IV gene disruption in Aspergillus oryzae .
Osamu Mizutani1, Youhei Kudou1, Tomomi Matsuura1, Hirokazu Inoue2, Keietsu Abe1, Katsuya Gomi1 (1 Tohoku
Univ.,Grad.Sch.Agri.Sci, 2Saitama Univ.)
59
P-49
㯔Ⳟ㸝Aspergillus oryzae㸞࡞࠽ࡄࡾ ORF හ࡫ࡡ poly(A)௛ຊࢨࢡࢻࣜᑙථ࡞ࡻࡾ␏ᖏ
poly(A)௛ຊ
⏛୯⍖ᕤ 1, ᚠᒱ᪸ᩝ 1,2, 㧏ᮄᚰ 3, ᩺ㆺᑠᘧ 1, ஫࿝ົஒ 1㸝1 ᮶໪ኬ࣬㎨࣬⏍∸⏐ᴏ๭ᠺ⛁Ꮥ, 2 㔕⏛⏐ᴏ⛁Ꮥ◂
✪ᡜ, 3 ࢿ࣍ࢧ࢕࣑ࢩࣔࣂࣤ㸞
㯔Ⳟ㸝Aspergillus oryzae㸞ࢅᐙ୹࡛ࡊࡒ␏⛸ࢰࣤࣂࢠ㈻ࡡ⏍⏐㔖ࡢ⮤ᕤࡡࢰࣤࣂࢠ㈻࡛Ẓ㍉ࡌࡾ࡛ⴥࡊࡂᑛ
࡝ࡂࠉ⏍⏐㔖ࡡᨭၻ࠿᭿ᙽࡈࡿ࡙࠷ࡾࠊࡆࡿࡱ࡚࡞ᠻࠍࡢࠉࢱࢼ⏜ᮮࢰࣤࣂࢠ㈻ Der f 7 ࡡࢤࢺࣤࢅ㯔Ⳟ࡞᭩
㐲໩ࡌࡾࡆ࡛࡞ࡻࡽࠉmRNA 㔖ࡡቌຊ࡞ࡻࡾ Der f 7 ⏍⏐㔖ࡡᨭၻ࡞ᠺຉࡊ࡙࠷ࡾࠊࢾ࢕ࢷ࢔ࣇ࡝ Der f 7 (Der
ntv) ࡡ mRNA 㔖࠿΅ᑛࡌࡾཋᅄ࡛ࡊ࡙ࠉDer ntv ࡡ ORF හ࡞࠽࠷࡙ poly(A)࠿௛ຊࡊ࡙࠷ࡒࡆ࡛࠾ࡼࠉ␏ᖏ
poly(A)௛ຊ࡞ࡻࡾ nonstop mRNA ฦよ⤊㊪࡞ࡻࡾฦよࢅུࡄ࡙࠷ࡾྊ⬗ᛮ࠿⩻࠻ࡼࡿࡒࠊࡊ࠾ࡊࠉ┷ᰶ⏍∸
࡚ poly(A)௛ຊࢨࢡࢻ࡛ࣜࡊ࡙ᶭ⬗ࡌࡾ AAUAAA 㒼า࠿ Der ntv ORF හ࡞ࡢヾࡴࡼࡿ࡝࠾ࡖࡒࠊࡐࡆ࡚ࠉ㯔
Ⳟ࡞࠽࠷࡙ poly(A)௛ຊࢨࢡࢻ࡛ࣜࡊ࡙ᶭ⬗ࡌࡾ㒼าࢅ᪺ࡼ࠾࡞ࡌࡾࡒࡴࠉ㯔Ⳟ EST ࢠ࣭ࣞࣤ 3’ഁࢨ࣭ࢠ࢙
ࣤࢪࢅ⾔ࡖࡒ⤎ᯕࠉAAUAAA 㒼า࠿Ꮛᅹࡌࡾࢠ࣭ࣞࣤࡢ࡮࡛ࢆ࡜ᏋᅹࡎࡍࠉAUUUUࠉAUUUAࠉUAAAUࠉ
AAUAU ➴ࡡ㒼าࢅᣚࡗࢠ࣭ࣞࣤ࠿ኣᩐᏋᅹࡊࡒࠊDer ntv ࡷืࡡ␏⛸㐿ఎᏄ࡞࠽ࡄࡾ ORF හࡡ␏ᖏ poly(A)
௛ຊ㒂నࡡ୕Ὦ࡞ࡵࡆࡿࡼࡡ㒼า࠿Ꮛᅹࡊ࡙࠽ࡽࠉࡱࡒࠉ㯔Ⳟ⮤㌗ࡡ㐿ఎᏄࡡ ORF හ࡞ࡆࡿࡼࡡ㒼าࢅᑙථ
ࡌࡾࡆ࡛࡞ࡻࡽⓆ⌟㔖࠿఩ୖࡊࡒࡆ࡛࠾ࡼࠉ㯔Ⳟ࡞࠽࠷࡙ࡢࡆࡿࡼࡡ㒼า࠿ poly(A)௛ຊࢨࢡࢻ࡛ࣜࡊ࡙ᶭ⬗
ࡊ࡙࠷ࡾྊ⬗ᛮ࠿♟ြࡈࡿࡒࠊ
Premature polyadenylation by inserting putative polyadenylation signal sequences within ORF in Asepergillus
oryzae
Mizuki Tanaka1, Masafumi Tokuoka1,2, Shinobu Takagi3, Takahiro Shintani1, Katsuya Gomi1
(1Div. Biosci. Biotech. Future Bioind., Grad. Sch. Agric. Sci., Tohoku Univ., 2Noda Institute for Scientific Research,
3
Novozymes Japan Ltd.)
P-50
㯔Ⳟ(Aspergillus oryzae)ࡡ DNA ࢹࣚࣤࢪ࣎ࢯࣤ Crawler ࡡ㌹⛛Ὡᛮ࡛
transposase 㐿ఎᏄࡡ 3’-RACE よᯊ
ᑚ➗ཋ ༡ಘ㸦ࠉᑚ⏷ ᾀ㸧ࠉ⛑ Ὂ஦㸧ࠉ㧏ᶣ ▹⧂㸦ࠉ஫࿝ ົஒ㸨
㸝㸦⚽⏛┬⥪㣏◂࣬㓕⣪᚜⏍∸ࠉ㸧᭮᰿෗࣬⥪◂ࠉ㸨᮶໪ኬ㝌㎨࣬⏍∸⏐ᴏ๭ᠺ㸞
࠘ ┘Ⓩ࠙ᐁ⏕㯔Ⳟ Aspergillus oryzae OSI1013 ᰬ࠾ࡼぜฝࡈࡿࡒࢹࣚࣤࢪ࣎ࢯࣤ Crawler ࡢ Tc1/mariner
superfamily ࡞ᒌࡌࡾὩᛮᆵࡡ DNA ࢹࣚࣤࢪ࣎ࢯ࡚ࣤ࠵ࡾࠊฦ⏍Ꮔࡡ Cu ฌ⌦ࡷ㧏Ὼฌ⌦࡝࡜ࡡࢪࢹࣝࢪ᮪௲
ୖ࡞࠽ࡄࡾ niaD ࡷ crnA 㐿ఎᏄ࡫ࡡ㌹⛛᣼ථ࠽ࡻࡦ㌹⛛⬲㞫Ὡᛮ࠿㯔Ⳟ࡚ิࡴ࡙♟ࡈࡿࡒ㸦㸞ࠊୌ᪁ࠉࢿ࣭ࢧ
ࣤよᯊ࡚ࡢ㌹⛛Ὡᛮ࠿ヾࡴࡼࡿࡾ࠾ྫྷ࠾࡞࠾࠾ࢂࡼࡍࠉࢪ࣒࢓࡝ 2 ᮇࡡࢨࢡࢻࣜ࠿ヾࡴࡼࡿࡒࡆ࡛࠾ࡼࠉ
OSI1013 ᰬ࡚ࡢ transposase 㐿ఎᏄ࠾ࡼ 2 ⛸௧୕ࡡ㹢RNA ࠿⏍ᠺࡈࡿ࡙࠷ࡾࡆ࡛࠿᥆ᏽࡈࡿࡒࠊࡐࡆ࡚ࠉ㯔Ⳟ
࡞࠽ࡄࡾࢹࣚࣤࢪ࣎ࢯࣤ㌹⛛Ὡᛮ࡞ᑊࡌࡾโᚒᶭᵋࡡよᯊ࡛ࡐࡡฺ⏕ࢅ┘Ⓩ࡞ࠉCrawler ࡡ㌹෕ᵕᘟ࡞ࡗ࠷࡙
᳠ゞࢅ⾔ࡖࡒࠊ
࠘᪁Ἢ࡛⤎ᯕ࠙㛏ࡈࡡ␏࡝ࡾ Crawler ㌹෕⏐∸ࢅよᯊࡌࡾࡒࡴࠉCuSO㸩῟ຊᇰ㣬༇ࡡ total RNA ࢅ⏕࠷ࡒ
3’-RACE ࢅ⾔ࡖࡒࠊ3’-RACE ⏐∸ࡡ PCR ࡡ⤎ᯕࠉࢿ࣭ࢧࣤ࣬ࣇࣞࢴࢹ࡛ྜྷᵕࡡࢪ࣒࢓࡝ 2 ᮇࡡࣁࣤࢺ࠿ᚋࡼ
ࡿࠉሲᇱ㒼าỬᏽ࡞ࡻࡽ transposase ORF හ㒂࡚ࡵ poly(A)௛ຊ࠿㉫ࡀ࡙࠷ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊࡈࡼ࡞ࠉ
ࡐࡡ୕Ὦᇡ࡚ࡢ 118bp ࡡࢪࣈࣚ࢕ࢨࣤࢡࡵヾࡴࡼࡿࠉᵕࠍ࡝㛏ࡈࡡ RACE ࢠ࣭ࣞࣤࡵኣࡂᚋࡼࡿ࡙࠷ࡾࠊୌ
᪁ࠉ࢕ࣤࢰࢠࢹ࡝ධ㛏 RACE ࢠ࣭ࣞࣤࡢᑛ࡝࠾ࡖࡒࠊ ௧୕ࡡࡆ࡛࠾ࡼࠉ㌹⛛Ὡᛮࢅ♟ࡌࢹࣚࣤࢪ࣎ࢯࣤ
Crawler ࡚ࡢࠉORF හ࡚ࡡ poly(A)௛ຊࡷࢪࣈࣚ࢕ࢨࣤࢡ࡞ࡻࡽ୘Ᏸධ࡝㹢RNA ࠿⏍ᠺࡈࡿࠉnon-stop mRNA
decay ⤊㊪࡝࡜࡞ࡻࡾฦよࡷࣈࣞࢬࢴࢨࣤࢡ࡞ࡻࡖ࡙ Crawler ࡡ㌹⛛Ὡᛮ࠿โᚒࡈ࡙࠷ࡾྊ⬗ᛮ࠿♟ြࡈࡿࡒࠊ
1)ᑚ➗ཋࡼࠉ᪝ᮇ㎨ⰹ໩Ꮥఌ 2006 ᖳᗐኬఌさ᪠ࠉ㹥92
Transposition and expression of DNA transposon Crawler in Aspergillus oryzae
Hironobu Ogasawara1, Hiroshi Obata2, Yoji Hata2, Saori Takahashi1, Katsuya Gomi3
(1Akita Res. Inst. Food and Brewing, 㸨Res.Inst.,Gekkeikan Sake Co., 3Div. Biosci. Biotech. Future Bioind., Grad.
Sch. Agric. Sci., Tohoku Univ.㸡)
60
P-51
᰺ᇰ୘Ⰳ⑍≟ࢅ♟ࡌ࢙ࢿ࢞ࢰࢢ࡚᳠ฝࡈࡿࡒ dsRNA ࢗ࢕ࣜࢪ
ᶋ஬ᑋ㑳ࠉ▹ᕖᨳⱝࠉ㤷᭨⏜⨶ (᲻ᯐ⥪◂)
Ⳟ⣊࠿୘ᆍୌ࡝⏍⫩ࢅ♟ࡌࡻ࠹࡞࡝ࡖࡒ࢙ࢿ࢞ࢰࢢࡡ᰺ᇰⳞᰬ(㸸ᰬ)࡞ࡗ࠷࡙ࠉ⑋ཋᅄᏄࡡ᥀⣬ࢅ⾔ࡖࡒ
࡛ࡆࢀࠉฦᏄ㔖⣑ 15kb ࡡ dsRNA ࢗ࢕ࣜࢪ࡞វ᯹ࡊ࡙࠷ࡾࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊᮇࢗ࢕ࣜࢪࡢࠉ㸸ᰬࡡ⫩
ᠺ࡞౐⏕ࡈࡿࡒばⳞᰬ࡚ࡢ᳠ฝࡈࡿ࡝࠾ࡖࡒࡆ࡛࠾ࡼࠉ᰺ᇰ㐛⛤࡚វ᯹ࡊࡒࡵࡡ࡛᥆ᏽࡈࡿࡾࠊᮇࢗ࢕ࣜࢪ
ࡢ PEG ࡛ NaCl Ꮛᅹୖ࡚ỷẂࡌࡾ࠿ࠉ㞹Ꮔ㢟᚜㙶ふᐳ࡞ࡻࡖ࡙⢇Ꮔ࠿☔ヾ࡚ࡀ࡝࠾ࡖࡒࡆ࡛࠾ࡼࠉአ⓮ࢰࣤ
ࣂࢠ㈻࡞ࡻࡾ⢇Ꮔᵋ㏸ࢅᣚࡒ࡝࠷⛸㢦ࡡࢗ࢕ࣜࢪ࡚࠵ࡾ࡛᥆ᏽࡈࡿࡾࠊࡱࡒࠉⳞ⣊ࢅ◒◃ࡌࡾ㝷࡞ࢠࣞࣞࣆ
࢚࣑ࣜࢅ῟ຊࡊࡒ᫤ࡷࠉࢗ࢕ࣜࢪ⏤ฦࢅ࣒ࣤࣇ࡚ࣚࣤ℈㐛ࡌࡾ࡛ dsRNA ࠿᳠ฝࡈࡿ࡝ࡂ࡝ࡾࡆ࡛࠾ࡼࠉ⭯ᠺ
ฦ࡛㛭୙ࡊ࡙࠷ࡾྊ⬗ᛮࡵ♟ြࡈࡿ࡙࠷ࡾࠊ᢫ฝࡈࡿࡒᮇ dsRNA ࠾ࡼࣚࣤࢱ࣑ࣈࣚ࢕࣏࣭ࢅ⏕࠷ࡒ RT-PCR
࡞ࡻࡖ࡙ cDNA ࢠ࣭ࣞࣤࢅష⿿ࡊࠉሲᇱ㒼าࡡỬᏽࢅ⾔ࡖࡒࠊ
dsRNA virus found in a degenerate strain of the cultivated Flammulina velutipes.
Toshiro Yokoi, Masahide Sunagawa, Yumi Magae
(Forestry & Forest Products Res.Inst.)
P-52
ዃᙟ⑍≟ࢅ♟ࡌⳞᗃ᰺ᇰࢨ࢕ࢰࢢ࡚᳠ฝࡈࡿࡒ㸧ᮇ㙈 RNA ࢗ࢕ࣜࢪ
㤷᭨⏜⨶㸡▹ᕖᨳⱝࠉᶋ஬ᑋ㑳 㸝᲻ᯐ⥪◂㸞
Ⳟᗃࢨ࢕ࢰࢢ(Lentinula edodes)࡚ࠉᏄᐁమ࠿ዃᙟ࡞࡝ࡾ⿍ᐐ࠿྘ᆀ࡚⏍ࡋ࡙࠷ࡾࠊࡐࡡ⑋≟ࡢࠉⱴࡡⴥࡊ
࠷ᚈ㛏ࠉഘࡡᑚᆵ໩ࠉⱴࠉഘࡡ⾪㟻ࡡࡈࡈࡂࡿࠉ࡝࡜࡚࠵ࡾࠊࡐࡡཋᅄ࡛⩻࠻ࡼࡿࡾࢗ࢕ࣜࢪ࠿᳠ฝࡈࡿࡒࠊ
Ꮔᐁమ⤄⧂ࢅࣛࣤ㓗ࣁࢴࣆ࢒࣭୯࡚◒◃ࡊࡒᚃࠉ0.5M NaCl+ 0.7% PEG 6000 ࡞ࡻࡖ࡙ỷẂࡊ࡙ࡂࡾ⏤ฦࢅ㞹
Ꮔ㢟᚜㙶࡚ふᐳࡊࡒ⤎ᯕࠉዃᙟᏄᐁమ⤄⧂୯࡞ࣃ࣓≟⢇Ꮔ࠿᳠ฝࡈࡿࡒࠊᮇࢗ࢕ࣜࢪࡢࠉዃᙟᏄᐁమࢅ⏍ࡋ
ࡒᇰᆀ࠾ࡼྜྷ᫤࡞Ⓠ⏍ࡊ࡙࠷ࡾḿᖏᏄᐁమࡡ⤄⧂୯࡞ࡵᏋᅹࡊ࡙࠷ࡒࠊࡊ࠾ࡊࠉ⛸Ⳟ࡝ࡼࡦ࡞ᇰᆀࡡ୯ᚨ㒂
࠾ࡼฦ㞫ࡊࡒⳞ⣊୯࡞ࡢヾࡴࡼࡿ࡝࠾ࡖࡒࡆ࡛࠾ࡼࠉ᰺ᇰ㐛⛤࡚᩺ࡒ࡞វ᯹ࡊࡒࡵࡡ࡛᥆ᏽࡈࡿࡾࠊࢗ࢕ࣜ
ࢪࡡᰶ㓗ࢅ᢫ฝࡊࡒ࡛ࡆࢀࠉ4 ᮇࡡ dsRNA ࠿᳠ฝࡈࡿࡒࠊࡐࡡ࠹ࡔ᭩ࡵኬࡀ࡝ฦᏄ㔖 5.7Kb ࡡ dsRNA ࡢḿ
ᖏ,ዃᙟࡡ࡜ࡔࡼࡡᏄᐁమ࡞ࡵභ㏳࡞Ꮛᅹࡊ࡙࠷ࡒ࠿ࠉ௙ࡡ dsRNA ࡡ㞹Ẵὃິࣂࢰ࣭ࣤ࡞ࡢḿᖏ࡛ዃᙟᏄᐁ
మࡡ㛣࡞㐢࠷࠿ぜࡼࡿࡒࠊ≁࡞ࠉḿᖏᏄᐁమࡢ⣑ 500bp ࡡኬࡀࡈࡡ≁ᚡⓏ࡝ dsRNA ࢅኣࢤࣅ࣭ಕᣚࡊ࡙࠷ࡒࠊ
ࡆࡡ dsRNA ࡡධ㛏ࡡ㒼าࢅࢠ࣭ࣞࢼࣤࢡࠉࢨ࣭ࢠ࢙ࣤࢪࡊ࡙よᯊࢅ⾔ࡖࡒࠊᮇ㐿ఎᏄࡢࠉࢗ࢕ࣜࢪវ᯹ᚃࡡ
Ꮔᐁమࡡዃᙟ໩ࢅᢒโࡌࡾ satellite RNA ࡡྊ⬗ᛮࡵ࠵ࡾ࡛᥆ᏽࡈࡿࡾࠊ
dsRNA virus particles found in the cultivated Lentinula edodes that produces abnormal fruit bodies.
Yumi Magae, Masahide Sunagawa, Toshiro Yokoi
(Forestry & Forest Products Res. Inst.)
61
P-53
᪝ᮇ⏐ࠉ࣊ࢹࢻ࣑⏐ࡡல⛸㛣᥃ྙమࢠࣞ࢓࣠ࣄࢰࢢᰬ࡞࠽ࡄࡾ ITS 㒼าࡡዃዼ࡝ୌ
⮬
Truong Binh-Nguyen1,2, ᒱᓧᏏ᫆ 3㸡
Le Xuan-Tham4,㕝ᮄᙪ 1㸝1 ༐ⴝኬ࣬㝌⮤↓⛁Ꮥ, ᩅ⫩, 2 Tropical Biology Inst.
Vietnam, 3 ࠾ࡍࡈ DNA ◂, 4 Vietnam Atomic Energy Commission㸞
㣏⏕࢞ࢿࢤ࡚࠵ࡾࢠࣞ࢓࣠ࣄࢰࢢ Pleurotus cystidiosus subsp. abalonus ࡢࠉdikaryon ࡡ↋ᛮ⬂Ꮔࢅషࡾࠊᠻࠍࡢ
rRNA 㐿ఎᏄࡡ ITS 㒼า࠿ࠉ࣊ࢹࢻ࣑⏐࡛᪝ᮇ⏐ࡡࢠࣞ࢓࣠ࣄࢰࢢᰬ࡚ዃዼ࡞ୌ⮬ࡊ࡙࠷ࡾࡵࡡࢅぜࡗࡄࡒࠊ
࠷ࡍࡿࡡᰬࡵ ITS 㒼า࠿㸧ࡗࡡᰶ࡚ኬࡀࡂ␏࡝ࡖ࡙࠽ࡽࠉல⛸㛣᥃ྙమ࡛⩻࠻ࡼࡿࡾ࠿ࠉࡐࡡ ITS 㒼าࡡ⤄
ࡲྙࢂࡎ࠿࣊ࢹࢻ࣑⏐࡛᪝ᮇ⏐࡚࡮࡯ୌ⮬ࡊ࡙࠷ࡾࠊୠ⏲୯ࡡࢠࣞ࢓࣠ࣄࢰࢢࡡ௯㛣ࡡ ITS 㒼าࡢࡌ࡚࡞ኣ
ࡂሒ࿈ࡈࡿ࡙࠷ࡾ࠿ࠉࡆࡡࡻ࠹࡝㞟⛸⣌⤣ࡢ▩ࡼࡿ࡙࠷࡝࠷ࠊ୦ᰬ࠾ࡼฦ㞫ࡊࡒ monokaryon ࡢᠺ⫩ࡷᙟឺ࠿
␏ᖏ࡚࠵ࡽࠉ࠷ࡍࡿࡡ dikaryon ᰬࡵኣࡂࡡ຋ເን␏ࢅ⵫✒ࡊ࡙࠷ࡾࡻ࠹࡞ࡲ࠻ࡾࠊࡊࡒ࠿ࡖ࡙୦࢞ࢿࢤᰬࡢ
භ㏳ࡡ dikaryon ࢅ♵඙࡛ࡊ࡙㟸ᖏ࡞㛏࠷ୠ௥ࢅ↋ᛮ⏍Ṣ࡞ࡻࡖ࡙⤊࡙ࡀࡒྊ⬗ᛮ࠿♟ြࡈࡿࡾࠊࡐࡡࡻ࠹࡝
࢞ࢿࢤ࠿᪝ᮇ࡛࣊ࢹࢻ࣑࡞⏍᜝ࡊ࡙࠷ࡒ஥ᐁࢅࠉ㛏Ờᩝ᪺ࢅ⠇࠷ࡒཿ௥ெ࠿₆Ằ᪐ࡡ౴ථ࠾ࡼ࣊ࢹࢻ࣑ࡷ᪝
ᮇ࡞㏠ࡿࡒ࡛࠷࠹⩻ཿᏕⓏ▩ぜ࡛⤎ࡦࡗࡄࡾࡆ࡛ࡢྊ⬗ࡓࢀ࠹࠾ࠊ
Conservation of a combination of two ITS alleles in Japanese and Vietnamese Coremiopleurotus
Binh-Nguyen Truong1,2, Koei Okazaki3, Xuan-Tham Le4, Akira Suzuki1 (1Grad. Sci. Tech., Facul. of Edu., Chiba Univ.,
2
Trop. Biol. Inst. Vietnam, 3Kazusa DNA Res. Inst., 4 Vietnam Atomic Energy Comm.)
P-54
Agrobacterium ࢅ⏕࠷ࡒᢰᏄⳞ ࢙ࢿ࢞ࢰࢢ ࡡᙟ㈻㌹ᥦἪࡡ㛜Ⓠ
ᒱᮇᬓඖ,ᒜ⏛㞖ெ,⛼ෞ ⪵,ᒱᓧක㞕,ୖᆊ ㄌ 㸝ಘᕗኬ࣬⦼⥌࣬ᚺ⏍⛁,࣌ࢠࢹࡀࡡࡆ⥪ྙ◂✪ᡜ㸞
࢙ࢿ࢞ࢰࢢ Flammulina velutipes ࡡᏄᐁమᙟᠺᶭᵋࢅよ᪺ࡌࡾࡒࡴࠉᏄᐁమᙟᠺ᫤࡞≁␏Ⓩ࡞Ⓠ⌟ࡌࡾ㐿ఎ
Ꮔ⩄ࢅ differential display Ἢ࡞ࡻࡽ༟㞫ࡊࡒࠊ㸦㸞ࡆࡿࡼ㐿ఎᏄࡡᶭ⬗よᯊࡡࡒࡴ࡞ࡢ㸡⠾౼࡚ຝ⋙Ⓩ࡝ᙟ㈻㌹
ᥦἪ࠿ᚪさ࡚࠵ࡾࠊࡐࡆ࡚㸡Agrobacterium ࢅ⏕࠷ࡒ F. velutipes ࡞ᑊࡌࡾᙟ㈻㌹ᥦἪࢅ᳠ゞࡊࡒࠊF. velutipes
⏜ᮮࡡ glyceraldehyde-3-phosphate dehydrogenase 㐿ఎᏄ㸝gpd㸞ࣈ࣓࣭ࣞࢰ࣭โᚒୖ࡞ hygromycin B ⪇ᛮ㐿ఎ
Ꮔ㸝hph㸞ࢅ㒼⨠ࡊࡒࣁ࢕ࢻ࣭ࣛ࣊ࢠࢰ࣭ࢅᵋ⠇ࡊ㸡 A. tumefaciens LBA4404 ᰬ࡞ᑙථࡊࡒࠊᮇ࣊ࢠࢰ࣭ಕᣚ
ᰬ⣵⬂࡛ F. velutipes 々ᰶⳞ⣊∞ࢅ࢓ࢬࢹࢨࣛࣤࢥࣤᏋᅹୖ࡚භᇰ㣬ࡊࡒ࡛ࡆࢀ㸡㐽ᢝᇰᆀ୕࡞ hygromycin B
⪇ᛮࡡ F. velutipes ࢤࣞࢼ࣭࠿්⌟ᛮࡻࡂฝ⌟ࡊࡒࠊࡆࡡ࡛ࡀ㸡⪇ᛮࢤࣞࢼ࣭ࡡฝ⌟࡞ࡢ࢓ࢬࢹࢨࣛࣤࢥࣤࡡ
῟ຊ࠿ᚪ㡪࡚࠵ࡖࡒࠊࡈࡼ࡞භᇰ㣬᮪௲ࢅ᳠ゞࡊ㸡ࡻࡽኣࡂࡡ⪇ᛮࢤࣞࢼ࣭ࢅᚋࡾࡆ࡛࠿࡚ࡀࡾ᭩㐲᮪௲ࢅ
☔❟ࡊࡒࠊฝ⌟ࡊࡒ F. velutipes ࢤࣞࢼ࣭ࡢ hygromycin B ⪇ᛮࡡᙟ㈻ࢅᏭᏽ࡞ಕᣚࡊࡒࠊ≺❟࡞ᚋࡼࡿࡒ⪇ᛮ
ᰬⳞ⣊࠾ࡼࢣࢿ࣑ DNA ࢅㄢ⿿ࡊ㸡
hph 㐿ఎᏄࢅࣈ࣭ࣞࣇ࡛ࡊ࡙ࢦࢧࣤࣀ࢕ࣇࣛࢱ࢕ࢭ࣭ࢨࣘࣤよᯊࢅ⾔ࡖࡒࠊ
ࡐࡡ⤎ᯕ㸡ࡌ࡬࡙ࡡᰬ࡞࠽࠷࡙ hph 㐿ఎᏄࡡࢨࢡࢻࣜ࠿᳠ฝࡈࡿ㸡㸦ࢤࣅ࣭ࡡ T-DNA ࠿ࢣࢿ࣑୕ࡡࣚࣤࢱ࣑
࡝న⨠࡞᣼ථࡈࡿ࡙࠷ࡾࡆ࡛࠿ࢂ࠾ࡖࡒࠊ௑ᚃࠉࡆࡡᙟ㈻㌹ᥦἪࢅ⏕࠷࡙ RNAi Ἢࢅ⏕࠷ࡒ㐿ఎᏄⓆ⌟ᢒโ
⣌ࢅᵋ⠇ࡊ㸡Ꮔᐁమ≁␏Ⓩ࡞Ⓠ⌟ࡌࡾᮅ▩㐿ఎᏄࡡᶭ⬗ࢅよᯊࡌࡾ஢ᏽ࡚࠵ࡾࠊ
㸦㸞
M. Yamada, S. Sakuraba, K. Shibata, G. Taguchi, S. Inatomi, M. Okazaki, and M. Shimosaka, FEMS
Microbiol.Lett.254,165-172 (2006).
Agrobacterium-mediated transformation of the basidiomycete Flammulina velutipes
Tomomitsu Okamoto, Masato Yamada, Satoshi Inatomi, Mitsuo Okazaki,and Makoto Shimosaka
(Dept. Appl. Biol., Fac. Tex. Sci. Tech., Shinshu Univ., Mushroom Lab. Hokuto Co.)
62
P-55
㧏ὩᛮࣛࢡࢼࣤฦよⓉⰅ⭁ᮑⳞ Phanerochaete sordida YK-624 ᰬࡡᙟ㈻㌹ᥦ⣌ᵋ⠇
ᒜᓃ㈶἖㸡ᮄᮟಆ஄ࠉ⩹⏛Ⱪ㸝᮶໪㎨◂ࢬࣤࢰ࣭㸞ࠉᮙ὾❟ᶖࠉᖲ஬ᾀᩝ㸝㟴ᒱኬᏕ㎨Ꮥ㒂㸞
⮤↓⏲ࡻࡽ㐽ᢜࡈࡿࡒ㧏ὩᛮࣛࢡࢼࣤฦよⳞ Phanerochaete sordida㸝࿰ྞ㸰ࢗࢪ࢞࢕ࣞ࢜࣠ࢰࢢ㸞 YK-624
ᰬࡢࠉ㐽ᢝⓏࣛࢡࢼࣤฦよ⬗㸝ࢬ࣭ࣜࣞࢪฦよງ࡞ᑊࡌࡾࣛࢡࢼࣤࡡ┞ᑊⓏ࡝ฦよງ㸞࠿㟸ᖏ࡞㧏࠷ⓉⰅ⭁
ᮑⳞ࡚࠵ࡾࠊᮇ◂✪࡞࠽࠷࡙ࡢࠉP. sordida YK-624 ᰬ࠿ᣚࡗ㐽ᢝⓏࣛࢡࢼࣤฦよ⬗ງࡡฦᏄᶭᵋࢅよ᪺ࡊࠉ
ࡱࡒ⏐ᴏⓏ࡞ฺ⏕ྊ⬗࡝⛤ᗐࡱ࡚ࣛࢡࢼࣤฦよງࢅ᭞࡞ྡྷ୕ࡈࡎࡾࡆ࡛ࢅ┘ᵾ࡛ࡊ࡙࠷ࡾࠊࡐࡡࡒࡴ࡞ࡢ㐿
ఎᏄᑙථ⣌ࡡὩ⏕࠿Ḗ࠾ࡎ࡝࠷࠿ࠉ⌟ᅹࡡ࡛ࡆࢀ P. sordida ࡡᙟ㈻㌹ᥦ⣌ࡢሒ࿈ࡈࡿ࡙࠷࡝࠷ࡒࡴࠉP. sordida
YK-624 ᰬࡡᙟ㈻㌹ᥦ⣌ᵋ⠇࡞╌ᡥࡊࡒࠊ
P. sordida YK-624 ᰬࡢࠉᙟ㈻㌹ᥦ⣌࠿᪜࡞☔❟ࡊ࡙࠷ࡾⓉⰅ⭁ᮑⳞ P. chrysosporium ࡛␏࡝ࡽฦ⏍Ꮔࢅᙟ
ᠺࡎࡍࠉࡱࡒᏄᐁమࢅᙟᠺࡈࡎ࡙⬂Ꮔࢅᅂ཭ࡌࡾ᮪௲ࡵぜฝࡈࡿ࡙࠷࡝࠷ࠊࡐࡆ࡚ᮇ◂✪࡚ࡢࠉᰜ㣬Ⳟ⣊࠾
ࡼషᠺࡊࡒࣈࣞࢹࣈࣚࢪࢹ࡞⣰አ⥲ࢅ↯ᑏࡊ࡙ን␏ᑙථࢅ⾔࠷ࠉ5-FOA Ꮛᅹୖ࡚ࣈࣞࢹࣈࣚࢪࢹ්⏍ࢅ⾔ࢂ
ࡎࡒ⤎ᯕࠉኣᩐࡡࢗࣚࢨࣜさịᛮࢠ࣭ࣞࣤࢅᚋࡒࠊ྘ࢠ࣭ࣞࣤࡡࢗࣚࢨࣜྙᠺ㐿ఎᏄ URA3ࠉURA5 ࢅㄢ࡬ࡒ
⤎ᯕࠉ୦㐿ఎᏄ୕࡞ን␏ࢅᣚࡗኣࡂࡡࢠ࣭ࣞࣤࢅྜྷᏽࡊࡒ࠿ࠉࡐࡡ࠹ࡔ URA5 ࡡࢤ࣭ࢺ㡷ᇡ୯࡞ 43b.p.ࡡḖ኶
ࢅᣚࡗࢠ࣭ࣞࣤ UV2-#64 Ⳟᰬࢅ⏕࠷࡙ᙟ㈻㌹ᥦᐁ㥺ࢅ⾔ࡖࡒࠊᙟ㈻㌹ᥦ࣏࣭࣭࢜㐿ఎᏄ࡛ࡊ࡙ URA5 ධ㛏
5kb.p.ࢅᣚࡗࣈࣚࢪ࣐ࢺࢅ⏕࠷ࠉUV2-#64 Ⳟᰬࡻࡽషᠺࡊࡒࣈࣞࢹࣈࣚࢪࢹࢅ PEG Ἢ࡞ࡻࡖ࡙ᙟ㈻㌹ᥦࡊࡒ
⤎ᯕࠉኣᩐࡡࢗࣚࢨࣜ㟸さịᛮࢠ࣭ࣞࣤࢅᚋࡒࠊᙟ㈻㌹ᥦຝ⋙ࡢ 1 clone/105㹳106 ࣈࣞࢹࣈࣚࢪࢹ⛤ᗐ࡚࠵ࡖ
ࡒࠊ
Transformation by complementation of a uracil auxotroph of the hyper lignin-degrading basidiomycete
Phanerochaete sordida YK-624.
Kenji Yamagishi, Toshiyuki Kimura, Sigeru Oita (National Agricultural Research Center for Tohoku Region),
Tatsuki Sugiura, Hirofumi Hirai (Faculty of Agriculture, Shizuoka University)
P-56
࢓ࢡࣞࣁࢠࢷ࣑ࣛࢗࢅ⏕࠷ࡒᮄᮞ⭁ᮑᢰᏄⳞࡡᙟ㈻㌹ᥦ
ຊ἖ఫ ᖲ㸡஫༎ᔋᅽ᪝Ꮔ㸡㩢ᓞḿᾀ㸝᮶ኬ㝌࣬㎨⏍⛁㸞
ᮄᮞ⭁ᮑᢰᏄⳞࡢࠉⳞమአ࡞㓕⣪ࢅฦἢࡌࡾࡆ࡛࡚ᮄᮞࢅฦよࡌࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࠊࡐࡡᮄᮞฦよ⬗
ງࡡ㧏ࡈ࠾ࡼࠉࡆࡿࡱ࡚࡞ᮄᮞฦよ㓕⣪࡞ᑊࡊ࡙㓕⣪ᏕⓏࠉฦᏄ⏍∸ᏕⓏ࡝◂✪࠿レ⣵࡞⾔ࢂࡿ࡙ࡀ࡙࠷ࡾࠊ
ࡐࡡ⤎ᯕࠉ㓕⣪ࡡᮄᮞฦよ࡞࠽ࡄࡾ⏍⌦Ⓩ࡝ᶭ⬗よᯊࠉࡱࡒࡢ㓕⣪ࡡኬ㔖⏍⏐࡝࡜ࡡⅤ࡞࠽࠷࡙ࠉᮄᮞ⭁ᮑ
ᢰᏄⳞࡡᙟ㈻㌹ᥦἪࡡ☔❟࠿᭻ࡱࡿ࡙࠷ࡾࠊࡐࡆ࡚ࠉᮇ◂✪࡞࠽࠷࡙ࠉ࢓ࢡࣞࣁࢠࢷ࣑ࣛࢗࢅ⏕࠷ࡒᮄᮞ⭁
ᮑᢰᏄⳞ࡫ࡡᙟ㈻㌹ᥦἪࢅ☔❟ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊࡒࠊ
ᮄᮞ⭁ᮑᢰᏄⳞ Phanerochaete chrysosporium ࡡࢣࢿ࣑᝗ሒࢅฺ⏕ࡊ࡙ࠉࢡࣛࢬࣜ࢓ࣜࢸࣃࢺ-3 ࣛࣤ㓗ࢸࣃ
ࢺࣞࢣࢻ࣭ࢭ㸝GPD㸞㐿ఎᏄࠉࡐࡊ࡙࢓ࢠࢲࣤ㐿ఎᏄࡡࣈ࣓࣭ࣞࢰ࣭࡛ࢰ࣭࣐ࢾ࣭ࢰ࣭ࢅࢠ࣭ࣞࢼࣤࢡࡊࡒࠊ
ࡐࡿࡼࡡࣆࣚࢡ࣒ࣤࢹ࡛ࠉࣀ࢕ࢡ࣏ࣞ࢕ࢨࣤࣆ࢚ࢪࣆ࢚ࢹࣚࣤࢪࣆ࣭࢘ࣚࢭ㐿ఎᏄࢅ Double-Joint PCR Ἢ
1)
࡞ࡻࡽࣀ࢕ࢡ࣏ࣞ࢕ࢨࣤ⪇ᛮ㐿ఎᏄࣆࣚࢡ࣒ࣤࢹࢅష⿿ࡊࡒࠊࡐࡿࡑࡿࡡࣆࣚࢡ࣒ࣤࢹࢅࣁ࢕ࢻ࣭ࣛ࣊ࢠࢰ
࣭pCAMBIA0380 ࡞ᑙථࡌࡾࡆ࡛࡚ᢰᏄⳞ⤄ࡲᥦ࠻࣊ࢠࢰ࣭ࢅᵋ⠇ࡊࡒࠊᢰᏄⳞ⤄ࡲᥦ࠻࣊ࢠࢰ࣭ࢅᑙථࡊ
ࡒ Agrobacterium tumefaciens ࡛ࠉP. chrysosporium ࡡⳞ⣊ࢅභᇰ㣬ࡌࡾࡆ࡛࡚ᢰᏄⳞࡡᙟ㈻㌹ᥦࢅ⾔ࡖࡒࠊᙟ
㈻㌹ᥦᚃࠉࣀ࢕ࢡ࣏ࣞ࢕ࢨࣤࢅྱࡳᇰᆀ࡞࠽࠷࡙⏍㛏ࡊࠉ⤊௥ࡊࡒⳞ⣊ࡻࡽ᢫ฝࡊࡒࢣࢿ࣑ࢅ㗢ᆵ࡞ࠉࣀ࢕
ࢡ࣏ࣞ࢕ࢨࣤ㐿ఎᏄࡡࣈࣚ࢕࣏࣭ࢅ⏕࠷࡙ PCR ࢅ⾔ࡖࡒ⤎ᯕࠉGPD 㐿ఎᏄࠉ࢓ࢠࢲࣤ㐿ఎᏄࣈ࣓࣭ࣞࢰ࣭ࠉ
ࡐࡊ࡙ CaMV35S ࣈ࣓࣭ࣞࢰ࣭ࢅ౐⏕ࡊࡒ࣊ࢠࢰ࣭࡞࠽࠷࡙ᙟ㈻㌹ᥦమ࠿ᚋࡼࡿࡒࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊ
1): Yu, JH. et al. Fungal Genet. Biol. (2004)
Transformation of wood-rot basidiomycetes mediated Agrobacterium tumefaciens
Taira Kajisa, Kiyohiko Igarashi, Masahiro Samejima
(Univ. of Tokyo)
63
P-57
࢓ࢡࣞࣁࢠࢷ࣑ࣛࢗἪ࡞ࡻࡾࢨ࢕ࢰࢢ (Lentinula edodes) ᙟ㈻㌹ᥦ⣌ࡡ☔❟
ᐋᮇᖶᜠ*㸡ᖲᒱⰃ㍜*㸡ᒜᓃኬ㍔*㸡᫤ᮇᬊீ**㸡ᑹㆺ ᾀ*㸡๑ᕖ஦ኯ㑳*㸡ඡ⋚ᇱୌ᭹*
(*㫵ཱིኬ࣬㎨㸡**Ⳟ⶷◂✪ᡜ)
࢓ࢡࣞࣁࢠࢷ࣑ࣛࢗ (Agrobacterium tumefaciens) ࢅฺ⏕ࡊࡒ AtMT (A. tumefaciens-mediated transformation) Ἢ࡞
ࡻࡾ⣊≟Ⳟࡡᙟ㈻㌹ᥦࡢ㸡ࣈࣞࢹࣈࣚࢪࢹㄢ⿿࠿୘さ࡝࡜ኣࡂࡡฺⅤࢅ᭯ࡊ࡙࠷ࡾࠊᮇ◂✪࡚ࡢ㸡AtMT Ἢ
࡞ࡻࡾࢨ࢕ࢰࢢ㸝L. edodes㸞ᙟ㈻㌹ᥦ⣌ࡡ☔❟ࢅ┘ᣞࡊࡒࠊAgaricus ⏜ᮮ GPD 㐿ఎᏄࣈ࣓࣭ࣞࢰ࣭โᚒୖࡡ
ࣀ࢕ࢡ࣏ࣞ࢕ࢨࣤ B㸝HygB㸞ᢤᢘᛮ㐿ఎᏄ (hph) ࢅ㐽ᢝ࣏࣭࣭࡛࢜ࡌࡾࣁ࢕ࢻ࣭ࣛ࣊ࢠࢰ࣭ࢅᑙථࡊࡒ࢓ࢡ
ࣞࣁࢠࢷ࣑ࣛࢗᩐ⣌⤣࡛㸡ࢨ࢕ࢰࢢࡡୌᰶ࠽ࡻࡦ஦ᰶⳞ⣊మᰬᩐ⣌⤣ࡡ⤄ࡲྙࢂࡎࢅ⏕࠷ࡒࠊࢨ࢕ࢰࢢᇰ㣬
Ⳟ⣊మࣇࣞࢴࢠ࡛࢓ࢡࣞࣁࢠࢷ࣑ࣛࢗᠩ⃦ᾦࢅΊྙࡊ㸡΅ᅸᾈ㏩ฌ⌦ࡊࡒᚃ㸡㐽ᢝᇰᆀ࡚㐽ᢜࡊࡒࠊࡐࡡ⤎
ᯕ㸡HygB ྱ᭯ᇰᆀ୕࡚⏍⫩ࡌࡾኣᩐࡡࢤࣞࢼ࣭ࢅᚋࡒࠊᙟ㈻㌹ᥦຝ⋙ࡢ㸡౐⏕ࡊࡒ࢓ࢡࣞࣁࢠࢷ࣑ࣛࢗ⣌⤣
࡛ࢨ࢕ࢰࢢ⣌⤣ࡡ⤄ࡲྙࢂࡎ࡞ࡻࡽኬࡀࡂ␏࡝ࡖࡒࠊぜ࠾ࡄ୕ࡡᙟ㈻㌹ᥦຝ⋙㸝HygB ⪇ᛮࢤࣞࢼ࣭ᩐ㸤Ⳟ⣊
మࣇࣞࢴࢠᩐ㸞ࡢ㸡᭩ኬ 13%࡚࠵ࡖࡒࠊPCR Ἢ࡞ࡻࡽ㸡ᙟ㈻㌹ᥦᰬ࡞࠽࠷࡙ࡢ⤽௥ᚃࡵ hph ࠿Ꮽᏽࡊ࡙ಕᣚ
ࡈࡿ࡙࠷ࡾࡆ࡛࠿♟ࡈࡿࡒࠊࡱࡒ㸡ᙟ㈻㌹ᥦᰬࡻࡽᏄᐁమࢅㄇᑙࡊ㸡ᑙථ hph ࡡⓆ⌟ࢅ RT-PCR ࡞ࡻࡽ☔ヾ
ࡊࡒ࡛ࡆࢀ㸡Ꮔᐁమࡡഘ࠽ࡻࡦᯮ㒂ฦ࡞࠽࠷࡙ࡵⓆ⌟࠿ヾࡴࡼࡿࡒࠊ௧୕ࡡ⤎ᯕ㸡AtMT Ἢࢅฺ⏕ࡌࡾࡆ࡛
࡞ࡻࡽ㸡ࢨ࢕ࢰࢢ࡞࠽࠷࡙⠾౼࠾ࡗ㧏ຝ⋙࡚ᙟ㈻㌹ᥦమࢅషฝࡌࡾࡆ࡛࠿ྊ⬗࡛࡝ࡖࡒࠊ
Agrobacterium tumefaciens-mediated transformation of shiitake, Lentinula edodes
Yukie Miyamoto*, Yoshiki Hiraoka*, Daisuke Yamagishi*, Keisuke Tokimoto**, Hiroshi Otani*, Nitaro Maekawa*
and Motoichiro Kodama* (*Fac. Agric, Tottori Univ., **Tottori Mycol. Inst.)
P-58
Aspergillus oryzae ⏜ᮮ 㹨 C ࣏࣭࣭࢜ࢅฺ⏕ࡊࡒ Aspergillus oryzae ࡡᙟ㈻㌹ᥦ
こ ᬏᏄ㸡ᆎᇄ㝧஄㸡ᗀᖏḿெ 㸝ኬ㛭࣬⥪◂㸞
࠘┘Ⓩ࠙sC (ATP sulfurylase)ࡢ Aspergillus oryzae ࡡᙟ㈻㌹ᥦ࡞ࡻࡂฺ⏕ࡈࡿࡾ࣏࣭࣭࢜ࡡୌࡗ࡚࠵ࡾ࠿ࠉ
Aspergillus nidulans ⏜ᮮࡡ sC 㐿ఎᏄ࠿ᗀࡂ౐ࢂࡿ࡙࠽ࡽࠉࡆࡡሔྙ A. oryzae ࡚ࡢ᯹Ⰵమ୕࡞ࣚࣤࢱ࣑࡝⤄
ᥦ࠻࠿㉫ࡆࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࠊࣚࣤࢱ࣑࡝⤄ᥦ࠻ࡢ᫤࡞ይࡱࡊࡂ࡝࠷⤎ᯕࢅࡵࡒࡼࡌሔྙࡵ࠵ࡾࠊࡱࡒ
A. oryzae ⏜ᮮࡡ sC 㐿ఎᏄࢅ A. oryzae ࡡ┞ྜྷⓏ⤄ᥦ࠻࡞ฺ⏕ࡊࡒሒ࿈ࡢぜᙔࡒࡼ࡝࠷ࠊࡐࡆ࡚ᮇ◂✪࡚ࡢ
A. oryzae RIB40 ⏜ᮮࡡ sC 㐿ఎᏄࢅ࣏࣭࣭࡛࢜ࡌࡾࣈࣚࢪ࣐ࢺࢅᵋ⠇ࡊࠉA. oryzae RIB40 ⏜ᮮࡡ sC㸢ᰬ࡚࠵
ࡾ NS4 ᰬࡡᙟ㈻㌹ᥦࢅモࡲࡒࠊ
࠘᪁Ἢཀྵࡦ⤎ᯕ࣭࠙ࣝ࣎ࢰ࣭㐿ఎᏄ࡚࠵ࡾ uidA ࡡⓆ⌟ࣈࣚࢪ࣐ࢺࢅᵋ⠇ࡊࠉ
⎌≟ࡡࡱࡱࠉ࠵ࡾ࠷ࡢ sC ࣏࣭࣭࢜හ࡚ 1 ⟘ᡜว᩷ࡊ├㙈≟࡞ࡊ࡙ NS4 ᰬࢅᙟ㈻㌹ᥦࡊࡒ࡛ࡆࢀࠉ├㙈≟࡞
ࡊࡒࣈࣚࢪ࣐ࢺࢅ⏕࠷ࡒሔྙ࡞⎌≟ࡡࡱࡱ⏕࠷ࡒሔྙࡡ 10 ಶ௧୕ࡡຝ⋙࡚ᙟ㈻㌹ᥦమࢅཱིᚋࡊࡒࠊࡱࡒࢦࢧ
ࣤよᯊࡡ⤎ᯕࠉኬ༖࡚᯹Ⰵమ୕ࡡ sC 㐿ఎᏄᗑ࡫ࡡ┞ྜྷⓏ࡝⤄ᥦ࠻࠿㉫ࡆࡖ࡙࠷ࡾࡆ࡛ࢅ☔ヾࡊࠉ᣼ථࡈࡿࡒ
ࣈࣚࢪ࣐ࢺࡡࢤࣅ࣭ᩐࡢ 1-8 ⛤ᗐ࡛ᵕࠍ࡚࠵ࡖࡒࠊḗ࡞ A. oryzae ࡡ୹さ࡝ฦἢ㓕⣪࡚࠵ࡾࢰ࢜࢓࣐࣭ࣚࢭࡡ
Ⓠ⌟ࣈࣚࢪ࣐ࢺࢅᵋ⠇ࡊࠉNS4 ᰬࢅᙟ㈻㌹ᥦࡊࡒࠊཱིᚋࡊࡒು⿭ 48 ᰬࡡ࠹ࡔ SDS-PAGE ࡞ࡻࡖ࡙᭩ࡵ㧏࠷
⏍⏐ᛮࢅ☔ヾࡊࡒᰬࡢࠉࢦࢧࣤよᯊࡡ⤎ᯕࠉ┞ྜྷⓏ࡞ 8-10 ࢤࣅ࣭࠿᣼ථࡈࡿ࡙࠷ࡾࡆ࡛ࢅ☔ヾࡊࡒࠊᮇⳞᰬ
ࡡࢩ࣭ࣔᇰ㣬ࢅモࡲࡒ࡛ࡆࢀࠉᇰᆀ୯࡞⣑ 10g/l ࡡࢰ࢜࢓࣐࣭ࣚࢭࢅฦἢࡊࡒࠊ
Transformation of Aspergillus oryzae by the homologous sC gene.
Akiko Nishi, Takayuki Bogaki, Masato Hirotsune
(Gen. Res. Lab., Ozeki Corp.)
64
P-59
Aspergillus aculeatus ࡞࠽ࡄࡾ sC 㐽ᢝ࣏࣭࣭࢜ࢅ⏕࠷ࡒᙟ㈻㌹ᥦ⣌ࡡ㛜Ⓠ
㊂❟ ᾀ⨶, ㆺ ಞ἖, ⅛ㆺ 㡨ୌ, ᕖཾ ๙ྒྷ㸝㜨ᗋኬ㝌࣬⏍࿤⎌㸞, Ⲡ஬ ᇱኰ (୯㒂ኬ࣬ᚺ⏍),
㔘ᨳ ┷ (㟴ᒱኬ㝌࣬ᚺ⏍໩)
࠘┘Ⓩ࠙Aspergillus aculeatus ࠿⏍⏐ࡌࡾኣ⛸㢦ࡡࢬ࣭ࣜࣚࢭࡢࢬ࣭ࣜࣞࢪࡡ⢶໩ງ࡞ඁࡿࠉ྘⛸ࢬ࣭ࣜࣚࢭ
㐿ఎᏄࡡⓆ⌟ㄢ⟿ᶭᵋࢅฦᏄ࡚᪺ࣝ࣊ࣜࡼ࠾࡞ࡌࡾࡆ࡛ࡢᴗࡴ࡙⮾࿝῕࠷ࠊࡆࡿ࡞ࡢ A. aculeatus ⮤㌗ࢅᐙ
୹࡛ࡊࡒᙟ㈻㌹ᥦ⣌ࢅ㛜Ⓠࡌࡾࡆ࡛࠿ᚪ㡪࡚࠵ࡾࠊࡐࡆ࡚ࠉ◪㓗௥ㅨ࡞㛭ࢂࡾ ATP ࢪࣜࣆ࣭ࣛࣚࢭ㐿ఎᏄ
(sC) ࢅ㐽ᢝ࣏࣭࣭࡛࢜ࡊࡒ A. aculeatus ࡡᙟ㈻㌹ᥦ⣌ࢅ☔❟ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊࡒࠊ
࠘᪁Ἢཀྵࡦ⤎ᯕ࠙ࡱࡍࠉsC Ḗ᥾ን␏ᰬ (sC-) ࡡཱིᚋࢅモࡲࡒࠊ◪㓗ࡡ࢓ࢻࣞࢡ࡚࠵ࡾࢬࣝࣤ㓗ྱ᭯ᇰᆀ࡚⏍
⫩ࡊࡒࢬࣝࣤ㓗⪇ᛮᰬࡡ࠹ࡔࠉ◪㓗ሲࢅ༟ୌ◪㯜″࡛ࡊ࡙ྜྷ໩ฝᮮࡍࠉ୞ࡗࢠ࣑ࣞ㓗វུᛮࢅ♟ࡊࡒ㸫ᰬࢅ
sC- ು⿭ᰬ࡛ࡊ࡙༟㞫ࡊࡒࠊು⿭ᰬࡡࢦࢧࣤࣇࣞࢴࢹฦᯊ࡞ࡻࡽࠉsC ᗑన࡚ࡡኬぞᶅ࡝Ḗ᥾࠿㉫ࡆࡖ࡙࠷࡝
࠷ࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡽࠉ┞ྜྷ⤄ᥦ࠻ࡡᐙ୹࡛ࡊฺ࡙⏕ྊ⬗࡚࠵ࡾࡆ࡛࠿ࢂ࠾ࡖࡒࠊḗ࡞ࠉ᪜▩ࡡ Aspergillus ᒌ
sC ࡡಕᏋ㡷ᇡࢅᇱ࡞ష⿿ࡊࡒࣈࣚ࢕࣏࣭ࢅ⏕࠷ࡒ PCR ࡞ࡻࡽࠉA. aculeatus sC ࡡ㒂ฦ DNA ᩷∞ࢅཱིᚋࡊࡒࠊ
ᚋࡼࡿࡒ DNA ᩷∞ࢅࣈ࣭ࣞࣇ࡛ࡊ࡙ࢦࢧࣤࣇࣞࢴࢹฦᯊࢅ⾔࠷ࠉ⣑㸬kb ࡡ Pst I ᾐ໩᩷∞࠿ A. aculeatus
sC ධ㛏ࢅྱࡳ஥ࢅぜ࠷ࡓࡊࠉࢪࢠ࣭ࣛࢼࣤࢡ࡞ࡻࡽ sC ධ㛏ࢅཱིᚋࡊࡒࠊࢠ࣭ࣞࣤ໩ࡊࡒ sC ࢅ⏕࠷࡙ sCು⿭ᰬࢅᙟ㈻㌹ᥦࡊࠉ◪㓗ሲࢅ༟ୌ◪㯜″࡛ࡊ࡙⏍⫩࡚ࡀࡾᙟ㈻㌹ᥦమ࠿ᚋࡼࡿࡒࡒࡴࠉsC 㐽ᢝ࣏࣭࣭࢜࡞
ࡻࡾᙟ㈻㌹ᥦ⣌࠿☔❟࡚ࡀࡒࠊ௑ᚃࡢࡆࡡ⣌ࢅ⏕࠷ࠉ┞ྜྷ⤄ᥦ࠻ࢅモࡲࠉ⤄ᥦ࠻ຝ⋙࣬ࢤࣅ࣭ᩐ࡝࡜࡞ࡗ࠷
࡙᳠ゞࡊ࡙࠷ࡂࠊ
Development of a transformation system for Aspergillus aculeatus using the sC gene as selectable marker
Hiromi Adachi, Shuji Tani, Junichi Sumitani, Takashi Kawaguchi (Life & Env. Science, Osaka Pref. Univ.), Motoo
Arai (Biochem., Chubu Univ.), Shin Kanamasa (Biochem., Shizuoka Univ.)
P-60
Aspergillus aculeatus ࡞࠽ࡄࡾ trpC 㐿ఎᏄࢅ⏕࠷ࡒᙟ㈻㌹ᥦ⣌ࡡ㛜Ⓠ
๑⏛ඁ௒㸡㔘ᨳ┷ 1 , ㆺಞ἖ , ⅛ㆺ㡨ୌ , Ⲡ஬ᇱኰ 2 , ᕖཾ๙ྒྷ㸝㜨ᗋኬ㝌࣬ᚺ⏍⛁, 1 㟴ᒱኬ࣬ᚺ⏍໩, 2 ୯㒂
ኬ࣬ᚺ⏍㸞
࠘┘Ⓩ࠙trpC 㐿ఎᏄࡢࠉࢤࣛࢪ࣐㓗࠾ࡼࢹࣛࣈࢹࣆ࢒ࣤྙᠺ࡞⮫ࡾཬᚺࡡහࠉ୔⟘ᡜࢅり፳ࡌࡾኣᶭ⬗ࢰ
ࣤࣂࢠ㈻ࢅࢤ࣭ࢺࡊ࡙࠷ࡾࠊࡆࡡ㐿ఎᏄḖ᥾ን␏ᰬࡢࠉ୯㛣௥ㅨ⏐∸ࡡ㢦జమ࡚࠵ࡾࣆ࢛ࣜࣞ࢓ࣤࢪࣚࢼࣜ
㓗㸝5-FAA㸞࡞⪇ᛮ࡞࡝ࡾ஥ࠉࡱࡒ trpC ࡡᑙථ࡞ࡻࡽࢹࣛࣈࢹࣆ࢒ࣤさịᛮ࠿┞⿭ࡈࡿࡾ஥࠾ࡼࠉtrpC ࢅ
࣏࣭࣭࡛࢜ࡊࡒᙟ㈻㌹ᥦ⣌ࡢࠉ࣎ࢩࢷ࢔ࣇࢬࣝࢠࢨࣘࣤ࡞ࡻࡽຝ⋙ࡻࡂᵋ⠇࡚ࡀࡾ≁㛏ࢅ᭯ࡊ࡙࠷ࡾࠊࡐࡆ
࡚ࠉᮇ◂✪࡚ࡢࠉ A. aculeatus ࡡ㧏ࢰࣤࣂࢠ㈻ฦἢ⬗ࠉ⢶໩ງ࡞ඁࡿࡒࢬ࣭ࣜࣚࢭ๠ࡡ᭯ຝฺ⏕ࢅ᭩⤂┘ᵾ
࡛ࡊ࡙ࠉࡱࡍᮇⳞࡡ trpC ࢅ㐽ᢝ࣏࣭࣭࡛࢜ࡊࡒᙟ㈻㌹ᥦ⣌ࢅᵋ⠇ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊࡒࠊ
࠘᪁Ἢ࣬⤎ᯕ࠙௙ࡡ Aspergillus trpC 㐿ఎᏄࡡಕᏋ㡷ᇡࢅᇱ࡞スゝࡊࡒࣈࣚ࢕࣏࣭ࢅ⏕࠷࡙ PCR ࢅ⾔࠷ࠉ
⣑ 700bp ࡡ㒂ฦ trpC ᩷∞ࢅཱིᚋࡊࡒࠊࡆࡡ㒼าࢅࡵ࡛࡞షᠺࡊࡒ DNA ࣈ࣭ࣞࣇࢅ⏕࠷ࡒࠉࢦࢧࣤࣇࣞࢴ
ࢹฦᯊࡷࠉࢨࣘࢴࢹ࢝ࣤࢠ࣭ࣞࢼࣤࢡ࡞ࡻࡽࠉtrpC ධ㛏ࢅྱࡳ⣑ 9 kb ࡡ᩷∞ࢅཱིᚋࡊࡒࠊ⌟ᅹࠉࣆ࣭࣑ࣝ
ࢨࣆࢹን␏ࢅᑙථࡊࡒ trpC ᨭⰃ㐿ఎᏄࢅ⏕࠷࡙㔕⏍ᰬࢅᙟ㈻㌹ᥦࡊࠉ5-FAA ⪇ᛮࡡ⾪⌟ᆵࢅᣞᵾ࡞ࡊ࡙ࠉ
trpC 㐿ఎᏄን␏ᰬࡡ༟㞫ࢅ┘ᣞࡊ࡙࠷ࡾࠊ
Development of a transformation system for Aspergillus aculeatus using the trpC gene as a selectable marker
Yusuke Maeda , Shuji Tani , Junichi Sumitani , Takashi Kawaguchi (Life & Env. Science Osaka Pref . Univ.) , Motoo
Arai (Biochem., Chubu Univ.),Shin Kanamasa (Biochem., Shizuoka Univ.)
65
P-61
A. tamarii niaD 㐿ఎᏄࡡⓆ⌟よᯊ
ᮄᮟኣỜ 1 ᴃᮇ៿ୌ 1 ໪ᮇ์⾔ 2 㕝ᮄ⪵ 1 ᯵ᮄ㇇ 1 㸝1 ㎨◂ᶭᵋ࣬㣏⥪◂, 2 យ▩⏐ᢇ◂࣬㣏ဗᕝᢇ㸞
࠘┘Ⓩ࠙ሲฦྱ᭯㔖࠿㧏࠷㓲ἔ⢉୕࡚⏍⫩ࡌࡾ㯔Ⳟ࡛ࡊ࡙ࠉAspergillus tamarii ST-2 ࢅ㐽ᢜࡊࡒࠊᮇᰬ࠿㓲ἔ
⢉୕࡚⏍⫩࡚ࡀࡾ⌦⏜ࢅฦᏄ⏍∸ᏕⓏᡥἪ࡞ࡻࡽよ᪺ࡌࡾࡒࡴࠉᙟ㈻㌹ᥦ࣏࣭࣭࡛࢜ࡊ࡙◢㓗㑇ඔ㓕⣪㐿ఎ
Ꮔ niaD ࢅ PCR ࡞ࡻࡽฦ㞫ࡊࠉࡐࡡᵋ㏸ࢅよ᪺ࡊࡒ㸝ᖲᠺ 17 ᖳᗐᮇኬఌさ᪠㞗 p64㸞
ࠊA. oryzae niaD ࡛ࡡሲ
ᇱ㒼าẒ㍉ࡡ⤎ᯕࠉORF 㡷ᇡཀྵࡦ 3’ഁ㟸⩳ゼ㡷ᇡ࡚ࡢ┞ྜྷᛮ࠿㧏ࡂ 95%࡚࠵ࡖࡒ࠿ࠉࣈ࣓࣭ࣞࢰ࣭㡷ᇡ࡛᝷
ᏽࡈࡿࡾ㒼าࢅྱࡳ 5’ഁ㟸⩳ゼ㡷ᇡ࡚ࡢ⣑ 81%࡚࠵ࡖࡒࠊࡆࡡฦ㞫ࡊࡒ㒼าࢅࣈ࣭ࣞࣇ࡞ࠉA. tamarii ST-2
ᰬࡡࢣࢿ࣑ࢅ㗢ᆵ࡞ࢦࢧࣤࣇࣞࢴࢹよᯊࢅ⾔ࡖࡒ࡛ࡆࢀࠉ஢᝷ࡈࡿࡾኬࡀࡈࡡࣁࣤࢺ௧አ࡞࣏࢕ࢻ࣭ࣁࣤࢺ
࠿ 1 ᮇ☔ヾࡈࡿࡒࠊࡐࡆ࡚ࠉฦ㞫ࡊࡒ㒼า࠿ A. tamarii ࡡ niaD 㐿ఎᏄ࡚࠵ࡾࡆ࡛ࡡ☔ヾࡡୌࡗ࡛ࡊ࡙ࠉࢿࢧ
ࣤࣇࣞࢴࢹよᯊ࡞ࡻࡾⓆ⌟☔ヾࢅ⾔ࡖࡒࠊ
࠘᪁Ἢ࣬⤎ᯕ࠙◢㓗ሲࠉல◢㓗ሲࠉࢡࣜࢰ࣐ࣤࠉࢡࣜࢰ࣐ࣤ㓗ሲࢅ။ୌࡡ✽⣪″࡛ࡊࡒࢵ࢒࣋ࢴࢠࢺࢴࢠࢪ
ᾦమᇰᆀࢅష⿿ࡊࠉA. tamarii ST-㸧ࢅ᥃⛸ࡊ࡙ 2 ᪝㛣 30Υ࡚ᣲ┖ᇰ㣬ࢅ⾔ࡖࡒࠊᇰ㣬Ⳟమࡻࡽධ RNA ࢅ᢫ฝ
ࡊࠉA. tamarii niaD ࡡୌ㒂ࢅࣈ࣭ࣞࣇ࡛ࡊ࡙ࠉࢿࢧࣤࣇࣞࢴࢹよᯊࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉ✽⣪″ࢅ◢㓗ሲ࠵ࡾ
࠷ࡢல◢㓗ሲ࡛ࡊࡒ㝷ࡢ niaD ࡡⓆ⌟࠿᳠ฝࡈࡿࡒࠊୌ᪁ࠉ✽⣪″ࢅࢡࣜࢰ࣐ࣤ࠵ࡾ࠷ࡢࢡࣜࢰ࣐ࣤ㓗ሲ࡛ࡊ
ࡒ㝷ࡢ niaD ࡡⓆ⌟ࡢ᳠ฝࡈࡿ࡝࠾ࡖࡒࠊࡆࡡ⤎ᯕࡢࠉJohnstone ࡼ㸝Gene 90 (1990) 181-192㸞ࡡሒ࿈࡞ࡻࡾࠉ
Aspergillus nidulans niaD ࡡⓆ⌟โᚒ࡞㛭ࡌࡾ▩ぜ࡛□┢ࡊ࡝࠾ࡖࡒࠊࡆࡡࡆ࡛࠾ࡼࠉA. tamarii ࡡ niaD 㐿ఎ
Ꮔࡵ௙ࡡ Aspergillus ᒌࡡࡵࡡ࡛ྜྷᵕࡡⓆ⌟โᚒࢅུࡄࡾ࡛⩻࠻ࡼࡿࡾࠊ
Expression analysis of niaD gene isolated from A. tamarii
Tae Kimura1, Ken-Ichi Kusumoto1, Noriyuki Kitamoto2, Satoshi Suzuki1, Yutaka Kashiwagi1
(1Natl. Food Res. Inst., 2Food Res. Center, Aichi Ind. Technol. Inst.)
P-62
㯑Ⳟ DNAchip ࡞ࡻࡾ㯑Ⳟ Genotyping ࡡ᳠ゞ
ᑚᯐ㟱඼㸦,2ࠉᆊᮇ࿰ಆ㸧ࠉᒜ⏛ಞ㸧ࠉᒷୖ࿰⿩㸦㸡㸧ࠉୖ㣜ொ㸦㸡㸧ࠉ୔୕㔔᪺㸧 㸝㸦ᗀᓞኬ࣬඙❻◂ࠉ㸧㒿⥪◂㸞
㯑Ⳟࡢࠉΰ㒿ࡷ㓲ἔࠉ࿝ძ࡝࡜ࡡ㔂㏸࡞౐⏕ࡈࡿࠉ᪝ᮇࡡ㣏ᩝ໩࡛῕ࡂ㛭ࢂࡖ࡙࠽ࡽᅗⳞ࡛⛘ࡈࡿࡾ᚜⏍
∸࡚ࠉ2005 ᖳ࡞ࡢ㒿⥪◂ᡜⶮࡡ㯑Ⳟ RIB40 ᰬࡡධࢣࢿ࣑ࢨ࣭ࢠ࢙ࣤࢪ࠿よㄖࡈࡿࡒࠊᮇⳞᰬࡢࠉ㔕⏍࡞㎾࠷
Ⳟᰬ࡛ゕࢂࡿ࡙࠷ࡾࠊୌ᪁ࠉᐁ㝷ࡡ㔂㏸⌟ሔ࡚ࡢࠉΰ㒿ࡷ㓲ἔ࡝࡜ࡡ⏕㏭ࠉ࠵ࡾ࠷ࡢ⿿ဗࡡ㧏ဗ㈻໩࡞ࡗ࡝
࠿ࡾࡻ࠹࡝ኣᵕ࡝㯑Ⳟ࠿༟㞫ࠉ⫩⛸ࡈࡿฺ⏕ࡈࡿ࡙ᮮ࡙࠷ࡾࠊࡊ࠾ࡊࠉࡆࡿࡱ࡚㐿ఎⓏᏕⓏ࡝ཱི᡽࠿㞬ࡊ࠷
࡝࡜ࡡ⌦⏜࡚ࡆࡿࡼࡡⳞᰬࡡ≁ᚡ࡞ࡗ࠷࡙ࡢࠉ༎ฦ࡞⏍໩ᏕⓏ࣬ฦᏄ⏍∸ᏕⓏ࡝◂✪࠿࡝ࡈࡿ࡙࠽ࡼࡍࠉⳞ
ᰬࡡ⫩⛸㐽ᢜࡢࠉᏻ⬗フ౮ࡷ⤊㥺࡞ኣࡂ౪Ꮛࡊ࡙࠷ࡾࠊ㒿⥪◂࡚ࡢࠉ㯑Ⳟ RIB40 ᰬࡡࢣࢿ࣑᝗ሒ࡞ᇱࡘࡀ㯑
Ⳟ DNAchip㸝࢓ࣆ࢔࣒ࢹࣛࢠࢪ♣⿿㸞ࢅషᠺࡊࡒࠊ㯑Ⳟ DNAchip ࡢࠉධ㐿ఎᏄ㸝12,074 㐿ఎᏄ㸞ࡡⓆ⌟よᯊ
ࡓࡄ࡚࡝ࡂࠉཋ⌦୕ࢨ࣭ࢢࣤࢪࡡᕣࢅ᳠ッࡌࡾࡆ࡛࠿ྊ⬗࡚ࠉⳞᰬ㛣ࡡࢣࢿ࣑ࣝ࣊ࣜࡡ㐢࠷ࢅよᯊࡌࡾࡆ࡛
࠿ྊ⬗࡚࠵ࡾࠊࡐࡆ࡚྘⛸㯑Ⳟࡡ Genotyping ࠿ྊ⬗࠾࡜࠹࠾ࢅ᳠ゞࡌࡾࡒࡴ࡞ࠉ㒿⥪◂ᡜⶮࡡ RIB40ࠉRIB128
ᰬ࡝࡜ᩐ⛸ࡡᰬ࡞ࡗ࠷࡙㯑Ⳟ DNAchip よᯊࢅ⾔ࡖࡒࠊࡱࡍࠉRIB40 ᰬ࡚ࡢ㧏࠷ࢨࢡࢻࣜೋཀྵࡦ්⌟ᛮࢅ♟ࡊ
ࢣࢿ࣑࢓ࣝ࢕࡛ࡊ࡙౐⏕ฝᮮࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊࡗࡘ࠷࡙ࠉRIB128 ᰬ࡞ࡗ࠷࡙よᯊࡊࡒ࡛ࡆࢀࠉRIB40
ᰬ࡛␏࡝ࡽࠉ177 㐿ఎᏄ࡞ࡗ࠷࡙ࢨࢡࢻࣜ࠿ぜࡼࡿࡍࠉḖ᥾ࡊ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊࡐࡡ௙ࡡⳞᰬ࡞࠽
࠷࡙ࡵྜྷᵕ࡞ࢣࢿ࣑࡚ࣝ࣊ࣜࡡ᪺☔࡝㐢࠷࠿ぜࡼࡿࡒࠊࡱࡒ᪜࡞᪺ࡼ࠾࡞࡝ࡖ࡙࠷ࡾ࢓ࣆࣚࢹ࢞ࢨࣤྙᠺ㐿
ఎᏄ࡞ࡗ࠷࡙ࡵよᯊࢅ⾔ࡖࡒ⤎ᯕࠉࡆࡿࡱ࡚ࡡ PCR ࡞ࡻࡾฦ㢦࡛ྜྷࡋ⤎ᯕࢅᚋࡾࡆ࡛࠿࡚ࡀࡒࠊࡈࡼ࡞಴ࠍ
ࡡ㐿ఎᏄ࡞ࡗ࠷࡙ྜྷࡋ AFL ࢡ࣭ࣜࣈ࡞ฦ㢦ࡈࡿ࡙࠷࡙ࡵࠉAFL ྙᠺ㐿ఎᏄࡡୌ㒂ࡡ㒼า࠿ን໩ࡊ࡙࠷ࡾྊ⬗
ᛮ࠿࠵ࡖࡒࠊࡆࡡࡻ࠹࡞㯑Ⳟ DNAchip ࢅ⏕࠷࡙ࠉ㯑Ⳟࡡ Genotyping ࠿ྊ⬗࡚࠵ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ
Application of Aspergillus oryzae DNA chip for rapid genotyping of A. oryzae species.
Yasunori Kobayashi㸦,2, K. Sakamoto㸧, O. Yamada㸧, K. Iwashita㸦㸡㸧, H. Shimoi㸦㸡㸧, S. Mikami㸧
(㸦Hiroshima Univ.,
㸧
NRIB)
66
P-63
㈃ᰜ㣬᮪௲ୖ࡚Ⓠ⌟ࡌࡾ㯔ⳞⳞమහࢰࣤࣂࢠ㈻ࡡよᯊ
໪ᕖ἖ᜠ㸡ఫ㔕ඔ᫓㸡ᇸᮇකᏄ㸡⏣⏛㞖஄*㸡ኬ⟺ಘୌ㸝㔘Ἁᕝኬ࣬ࢣࢿ࣑◂ *⏐⥪◂㸞
㸱┘Ⓩ㸳㯔Ⳟ Aspergillus oryzae ࡢ᪝ᮇࡡఎ⤣Ⓩ㔂㏸ࡷ㓕⣪๠ࡡᕝᴏ⏍⏐࡚ࡡฺ⏕࡝࡜⏐ᴏⓏ࡞㟸ᖏ࡞
㔔さ࡝᚜⏍∸࡚࠵ࡽࠉ㎾ᖳ࡚ࡢ㐿ఎᏄ࡚ࣝ࣊ࣜࡡ◂✪ࡵ⢥ງⓏ࡞㐅ࡴࡼࡿ࡙࠷ࡾࠊᫎᖳⓆ⾪ࡈࡿࡒ㯔Ⳟධ
ࢣࢿ࣑よㄖࡡᏰ஡ࢅུࡄࠉ࣎ࢪࢹࢣࢿ࣑◂✪ࡡୌ❻࡛ࡊ࡙ࠉᮇ◂✪ᡜ࡚ࡢࢰࣤࣂࢠ㈻ࡡⓆ⌟ࢅ⥑⨮Ⓩ࡞よ
ᯊࡌࡾࡒࡴ᭯ຝ࡝ᡥṹ࡚࠵ࡾ஦ḗඔ㞹ẴὃິἪࢅ⏕࠷ࡒ㯔Ⳟࡡࣈࣞࢷ࢛࣭࣑よᯊࢅ⾔ࡖ࡙࠷ࡾࠊ
㯔Ⳟࡢᅖమᇰ㣬ࡷᾦమᇰ㣬࡝࡜ࡐࡡᇰ㣬᮪௲࡞ࡻࡽⓆ⌟ࡌࡾࢰࣤࣂࢠ㈻⩄࠿ኬࡀࡂን໩ࡌࡾࡒࡴࠉᵕࠍ
࡝ᇰ㣬᮪௲ୖ࡚ࡡよᯊ࠿ᚪさ࡛ࡈࡿࡾࠊ௑ᅂᐣᰜ㣬ᾦమᇰᆀ࡛㈃ᰜ㣬ᾦమᇰᆀ࡚྘ࠍᇰ㣬ࡊࡒ㯔Ⳟࢅ⏕࠷
࡙よᯊࢅ⾔࠷ࠉẒ㍉᳠ッࡊࡒࡡ࡚ሒ࿈ࡌࡾࠊ
㸱᪁Ἢ㸳ᐣᰜ㣬ᾦమᇰᆀ࡛ࡊ࡙ YPD ᾦమᇰᆀࠉ㈃ᰜ㣬ᾦమᇰᆀ࡛ࡊ࡙ Czapek-dox ᾦమᇰᆀࢅ⏕࠷࡙
A.oryzae RIB40 ᰬࢅᇰ㣬ࡊࠉࡐࡿࡑࡿࡡⳞమࡻࡽࢰࣤࣂࢠ㈻ࢅ᢫ฝ࣬⢥⿿ࡊࠉ஦ḗඔ㞹Ẵὃິࢅ⾔ࡖࡒࠊ
᳠ฝࡈࡿࡒࢪ࣎ࢴࢹࢅวࡽฝࡊࠉ㈻㔖ฦᯊჹ࡞ࡻࡾࢰࣤࣂࢠ㈻ࡡྜྷᏽࢅ⾔ࡖࡒࠊ
Proteome analysis of intracellular proteins from Aspergillus oryzae grown under the poor nutrient culture
conditions.
Harue Kitagawa, Motoaki Sano, Mitsuko Dohmoto, Masayuki Machida*, Shinichi Ohashi
(KIST, *AIST)
P-64
⡷ࢽ࢜ᇰᆀ࡚㧏Ⓠ⌟ࡌࡾ㯔Ⳟ㐿ఎᏄ⩄ࡡよᯊ
ᑹ㛭೸஦㸡ᒜ୯ᓤ*㸡ఫ㔕ඔ᫓㸡⠇㔕༜ኰ*㸡ኬ⟺ಘୌ 㸝㔘Ἁᕝኬ࣬ࢣࢿ࣑◂ *⠇㔕㣏ဗᕝᴏ㸞
㸱┘Ⓩ㸳⡷ࢽ࢜ࡢ㇇ᐣ࡝ᰜ㣬ࡷᶭ⬗ᛮᠺฦࢅྱࢆ࡚࠷ࡾ࠿ࠉ⡷ἔࠉ㣣ᩩࠉᶭ⬗ᛮᠺฦࡡ᢫ฝࠉᅰቫᨭⰃ๠࡝
࡜࡛ࡊ࡙ࡡฺ⏕࡞㝀ࡼࡿ࡙࠷ࡾࠊ⡷ࢽ࢜ࡡ྘⛸᭯⏕ᠺฦࢅྊ⁈໩࡚ࡀࡾ㣏ဗ⏕㓕⣪๠ࡡ⛸㢦࠿ᑛ࡝ࡂࠉࣁ࢕
࢛ࢤࣤࣁ࣭ࢩࣘࣤ⣪ᮞ࡛ࡊ࡙ࡡ᭞࡝ࡾ᭯ຝὩ⏕࠿᭻ࡱࡿ࡙࠷ࡾࠊ㯔Ⳟࡢ㓕⣪ࡡ᐀ᗔ࡚࠵ࡽࠉ12,000 㐿ఎᏄࡡ
࠹ࡔᶭ⬗ᮅ▩㐿ఎᏄ࠿ 8,000 㐿ఎᏄ⛤ᗐᏋᅹࡊࠉ⡷ࢽ࢜ࡡ᭯⏕ᠺฦࢅྊ⁈໩࡚ࡀࡾ㯔Ⳟࡡ㐿ఎᏄ⩄ࡡ⤄ྙࡎ
ࡡ㛜Ⓠࠉ᩺ぞຊỀฦよ㓕⣪㐿ఎᏄࡡཱིᚋࠉᶭ⬗ᛮࣁ࢕࢛ࢤࣤࣁ࣭ࢩࣘࣤ⣪ᮞ㛜Ⓠ࡫ࡡᒈ㛜ࢅ┘ᣞࡊ࡙࠷ࡾࠊ
㸱᪁Ἢ࠽ࡻࡦ⤎ᯕ㸳᪝ᮇ㒿㔂㏸᫤࡞ᤴฝࡌࡾ㉝ࢽ࢜(㣜⡷ࡡ⡷ࢽ࢜)࡛୯ࢽ࢜(හഁࡡ⡷ࢽ࢜)ࢅ㯔Ⳟࡡᾦమᇰ
㣬᫤ࡡ⅛⣪″ࠉ✽⣪″࡛ࡊ࡙῟ຊࡊࠉᇰ㣬Ⳟమࡻࡽ RNA ᢫ฝࠉDNA ࣏࢕ࢠࣞ࢓ࣝ࢕⏕ࣈ࣭ࣞࣇࡡㄢ⿿ࠉ㐿
ఎᏄࡡⓆ⌟ࣈࣞࣆ࢒࢕ࣜࢅよᯊࡊࡒࠊよᯊ࡞ࡢ 11,000DNA ࣏࢕ࢠࣞ࢓ࣝ࢕ࢅ౐⏕ࡊࡒࠊࡈࡼ࡞Ⓠ⌟㔖࠿ኣ࠷
㐿ఎᏄ࡞ࡗ࠷࡙ࡢࠉࣛ࢓ࣜࢰ࢕࣑ PCR Ἢ࡚ࡡⓆ⌟㔖ࡵ᳠ゞࡊࡒࠊよᯊࡊࡒࢸ࣭ࢰࡻࡽࠉಘ㢏࡚ࡀࡾࢸ࣭ࢰࢅ
ᚋࡾࡒࡴ࡞ࠉⓆ⌟ንິࡡᕣ࠿ 2 ಶ௧୕ࡡ㐿ఎᏄࢅࣅࢴࢠ࢓ࢴࣈࡊࡒ⤎ᯕࠉ㉝ࢽ࡚࢜ࡢ 64 ಴ࡡ㐿ఎᏄ࠿≁␏Ⓩ
࡞㧏Ⓠ⌟ࡊࠉࡐࡡ࠹ࡔᶭ⬗᪜▩ࡡ㓕⣪㐿ఎᏄ࠿ 34 ಴ࠉᶭ⬗ᮅ▩ࡡࢰࣤࣂࢠ㈻㐿ఎᏄ࠿ 13 ಴࠵ࡖࡒࠊࡆࡿ࡞
ᑊࡊ࡙୯ࢽ࡚࢜ࡢࠉ70 ಴ࡡ㐿ఎᏄ࠿㧏Ⓠ⌟ࡊࠉࡐࡡ࠹ࡔᶭ⬗᪜▩ࡡ㓕⣪㐿ఎᏄ࠿ 29 ಴ࠉᶭ⬗ᮅ▩ࡡࢰࣤࣂ
ࢠ㈻㐿ఎᏄ࠿ 16 ಴࠵ࡖࡒࠊ୦⡷ࢽ࢜ࡡ㧏⃨ᗐࡡᐨኮᇰᆀ࡚ࡡⓆ⌟Ẓ㍉࡞ࡗ࠷࡙ࡵሒ࿈ࡌࡾࠊ
Analysis of the specific gene cluster under a rice bran cultivation in Aspergillus oryzae
Kenji Ozeki, Takashi Yamanaka*, Motoaki Sano, Takuo Tsuno*, Shinichi Ohashi
(K.I.T., *Tsuno Food Industrial Co.)
67
P-65
㯑Ⳟ DNAchip ࡡ㛜Ⓠ࡛ࢣࢿ࣑よᯊࠉⓆ⌟よᯊ࡫ࡡฺ⏕
ᒷୖ࿰⿩㸦㸡㸧ࠉᆊᮇ࿰ಆ㸧ࠉᒜ⏛ಞ㸧ࠉ୔୕㔔᪺㸧 㸝㸦ᗀᓞኬ࣬඙❻◂ࠉ㸧㒿⥪◂㸞
㯜㯑Ⳟ㸝Aspergillus oryzae㸞ࡢࠉΰ㒿ࡷ㓲ἔࠉ࿝ძ࡝࡜ᠻ࠿ᅗࡡఎ⤣ⓏⓆ㓕⏐ᴏ࡞୘ྊḖ࡚࠵ࡾࡓࡄ࡚࡝ࡂࠉ
྘⛸ࢰࣤࣂࢠ㈻ࡡ౩⤝″ࠉ࠵ࡾ࠷ࡢኣ⛸ኣᵕ࡝ࢰࣤࣂࢠ㈻⏍⏐ࡡࡒࡴࡡ࣌ࢪࢹ࡛ࡊ࡙ᗀࡂฺ⏕ࡈࡿ࡙࠷ࡾ㔔
さ࡝᚜⏍∸࡚࠵ࡾࠊᫎᖳࠉ㒿⥪◂ᡜⶮࡡ㯑Ⳟ RIB40 ᰬࡡධࢣࢿ࣑ࢨ࣭ࢠ࢙ࣤࢪ࠿ප㛜ࡈࡿࠉࢣࢿ࣑᝗ሒࢅฺ
⏕ࡊࡒ◂✪ࡡⓆᒈ࠿᭿ᙽࡈࡿ࡙࠷ࡾࠊࡆࡿࡱ࡚࡞ࠉᙔ◂✪ᡜ࡚ࡢ࣎ࢪࢹࢣࢿ࣑◂✪ࡡᇱᮇⓏ࢕ࣤࣆ࡛ࣚ࡝ࡾ
cDNA ࣏࢕ࢠࣞ࢓ࣝ࢕㸝NRIB3000 ࢓ࣝ࢕㸞ࢅ㛜Ⓠࡊࡒࠊࡊ࠾ࡊࠉNRIB3000 ࢓ࣝ࢕࡚ࡢධ࡙ࡡ㐿ఎᏄࢅ⥑⨮
ࡊ࡙࠷࡝࠾ࡖࡒࡆ࡛࠾ࡼࠉ㯑Ⳟࡡධ㐿ఎᏄࡡⓆ⌟ࢅ㧏⢥ᗐ࡞්⌟ᛮⰃࡂよᯊࡌࡾࡆ࡛ࠉࡈࡼ࡞ࡢࠉChip on Chip
࡝࡜ࢣࢿ࣑㒼า࡞౪Ꮛࡊࡒよᯊ➴ࢅ⾔࠹ࡆ࡛ࢅ┘Ⓩ࡞ࠉ㯑Ⳟ DNAchip㸝Affymetryx ♣⿿㸞ࢅ㛜Ⓠࡊࡒࠊ
ᮇ㯑Ⳟ DNAchip ࡢࠉ㸦㐿ఎᏄ࠵ࡒࡽ 11 ⤄ࡡࣂ࣭ࣆ࢘ࢠࢹ࣏ࢴࢲࠉ࣐ࢪ࣏ࢴࢲࣈ࣭ࣞࣇࢬࢴࢹࢅ᭯ࡊࠉ࠾
ࡗࠉ࡮࡛ࢆ࡜᧧ష࠿⮤ິ໩ࡈࡿ࡙࠷ࡾࡒࡴࠉᐁ㥺⩽ࡡᢇ㔖࡞ࡻࡼࡍ㧏⢥ᗐ࠾ࡗ්⌟ᛮⰃࡂⓆ⌟よᯊࢅ⾔࠹ࡆ
࡛࠿ྊ⬗࡚࠵ࡾࠊᠻࠍࡢࠉ㯜㯑Ⳟ RIB40 ᰬࡡࢣࢿ࣑㒼าࢅ࣭࣊ࢪ࡞ධ㐿ఎᏄ㸝13,000 㐿ఎᏄ㸞ࡡࣈ࣭ࣞࣇࢬ
ࢴࢹࡡࢸࢧ࢕ࣤࢅ⾔࡝࠷ࠉ㯑Ⳟ DNAchip ࢅషᠺࡊࡒࠊࡗࡘ࠷࡙ࠉAspergillus oryzae RIB40 ᰬࢅ⏕࠷ࠉ᭩ᑛᇰ
ᆀ࡞ࡻࡽ≺❟࡞ㄢ⿿ࡊࡒモᩩ࡞ࡗ࠷්࡙⌟ᛮࢅ☔ヾࡊࡒ࡛ࡆࢀࠉ㧏࠷⢥ᗐ්࡛⌟ᛮࢅ☔ヾࡌࡾࡆ࡛࠿ฝᮮࡒࠊ
ᮇ㯑Ⳟ DNAchip ࡢࠉཋ⌦Ⓩ࡞ࡢࢣࢿ࣑ࢨ࣭ࢢࣤࢪ➴ࡡᕣ␏ࡷḖ᥾ࠉ㔔々➴ࢅ᳠ฝࡌࡾࡆ࡛ࡵྊ⬗࡚࠵ࡾࡆ࡛
࠾ࡼࠉ㯑Ⳟ DNAchip ࡡࢸࢧ࢕ࣤ࡞౐⏕ࡊࡒ RIB40 ᰬࡡධࢣࢿ࣑ DNA ࢅㄢ⿿ࡊࠉ᯹Ⰵమ DNA ࡡ᳠ฝࢅ⾔ࡖ
ࡒ࡛ࡆࢀࠉධ㐿ఎᏄ࡞ࡗ࠷්࡙⌟ᛮࡡⰃይ࡝ࢨࢡࢻࣜ࠿᳠ฝࡈࡿࡒࠊ⌟ᅹࠉᮇ㯑Ⳟ DNAchip ࢅฺ⏕ࡊ࡙ࠉᵕࠍ
࡝⎌ሾᅄᏄ࡞ᑊࡌࡾ㐿ఎᏄⓆ⌟ࡡᚺ➽࡞ࡗ࠷࡙᳠ゞࢅ⾔ࡖ࡙࠷ࡾࠊ
Development of Aspergillus oryzae DNAchip and its application.
Kazuhiro Iwashita㸦㸡㸧, K. Sakamoto㸧, O. Yamada㸧, , H. Shimoi㸦㸡㸧, S. Mikami㸧
(㸦Hiroshima Univ.,
㸧
NRIB)
P-66
㯔Ⳟ CCAAT-box ⤎ྙᅄᏄ࡞ࡻࡖ࡙โᚒࡈࡿࡾ㐿ఎᏄࡡ⥑⨮Ⓩよᯊ
㧏ᶣ᪺⌌㸡ᮙᒜ⣟ஒ㸡ྙ⏛⚵▦㸡ఫ㔕ඔ᫓㸟㸡ᑚᯐူኰ㸡ຊ⸠㞖ኃ㸝ྞኬ㝌⏍࿤㎨࣬⏍∸ᶭᵋࠉ㸟㔘Ἁᕝᴏኬ࣬
ࢣࢿ࣑◂㸞
⣊≟Ⳟ CCAAT ⤎ྙᅄᏄ࡚࠵ࡾ Hap 々ྙమࡢ HapB/C/E ࡡ 3 ࡗࡡࢦࣇࣗࢼࢴࢹ࠾ࡼᵋᠺࡈࡿ࡙࠽ࡽࠉኣࡂ
ࡡ⏐ᴏ୕㔔さ࡝㐿ఎᏄࡡ㌹෕ࢅಀ㐅ࡊ࡙࠷ࡾࠊࡆࡿࡱ࡚࡞ Hap 々ྙమࡡᛮ㈻ࡢレ⣵࡞よᯊࡈࡿ࡙ࡀࡒ࠿ࠉโ
ᚒࡈࡿࡾ㐿ఎᏄ⩄ࡢ౪↓࡛ࡊ࡙᪺ࡼ࠾࡚ࡢ࡝࠷ࠊࡆࡡࡒࡴࠉ㯔Ⳟඁᛮ㜴ᐐᆵ㸝ࢺ࣐ࢻࣤࢹࢾ࢝ࢷ࢔ࣇ㸞ን␏
ᰬࢅ⏕࠷࡙ DNA ࣏࢕ࢠࣞ࢓ࣝ࢕よᯊࢅモࡲࡒࠊ
ᠻࠍࡢࡌ࡚࡞ඁᛮ㜴ᐐᆵን␏㐿ఎᏄࡡ⋋ᚋ࡞ᠺຉࡊ࡙࠽ࡽࠉ࣓ࢸࣜ⣊≟Ⳟ Aspergillus nidulans ࡞࠽࠷࡙ࡐ
ࡡ᭯ຝᛮࢅ☔ヾࡊ࡙࠷ࡾࠊࡆࡡ㐿ఎᏄࢅ㯔Ⳟ࡞ᑙථࡊࠉ㯔Ⳟ DNA ࣏࢕ࢠࣞ࢓ࣝ࢕よᯊࢅ⾔ࡖࡒࠊよᯊࡡ⤎ᯕࠉ
Hap 々ྙమ࡞ࡻࡖ࡙㸧ಶ௧୕ಀ㐅ࡈࡿࡾ㐿ఎᏄ࠿ 50 ಴ぜ࠷ฝࡈࡿࡒࠊHap 々ྙమ࠿ኣࡂࡡ㐿ఎᏄࡡ㌹෕ಀ㐅
࡛ࡊ഼࡙࠷࡙࠷ࡾ࡛࠷࠹ࡆࡿࡱ࡚ࡡ▩ぜࢅ⩻࠻ࢂࡎࡾ࡛ࠉHap 々ྙమࡢࣀࢗࢪ࣭࢞ࣅࣤࢡ㐿ఎᏄࡡࡻ࠹࡝ᵋ
ᠺⓏ࡞Ⓠ⌟ࡊ࡙࠷ࡾ㐿ఎᏄ⩄࡚ࡢ࡝ࡂࠉరࡼ࠾ࡡㄇᑙࢅུࡄ࡙Ⓠ⌟ࡌࡾ㐿ఎᏄ⩄ࢅโᚒࡊ࡙࠷ࡾࡒࡴ࡚࠵ࡾ
࡛⩻࠻ࡼࡿࡒࠊୌ᪁ࠉ2 ಶ௧୕ᢒโࡈࡿࡾ㐿ఎᏄࢅぜ࡙ࡲࡾ࡛ࠉࣛ࣍ࢮ࣭࣏ࣜࢰࣤࣂࢠ㈻㐿ఎᏄ࡝࡜ࡡ⩳ゼ
࡞㛭౿ࡌࡾ㐿ఎᏄ⩄࠿ᢒโࡈࡿ࡙࠷ࡒࠊHap 々ྙమ࡛⩳ゼ࡛ࡡ㛭౿࡞ࡗ࠷࡙⩻ᐳࡌࡾࠊ
Comprehensive analysis of genes regulated by Aspergillus oryzae CCAAT-box binding factor.
Akemi Takahashi, Junya Sugiyama, Hideya Gouda, Motoaki Sano㸟, Tetuo Kobayashi, Masashi Kato
(Grad. Sch. of Bioagricultural Sci., Nagoya Univ.,
㸟
KIT)
68
P-67
㯜㯔Ⳟ⏍⫩Ⳟమහࣈࣞࢷ࢛࣭࣑ࡡよᯊ
♼ኯ㑳㸡➁හ㐠㞕 㸝᮶ா㎨ᕝኬ㝌㹺㎨㸞
A.oryzae RIB40 ࡡⳞమහࢰࣤࣂࢠ㈻࡞ࡗ࠷࡙ࠉ஦ḗඔ㞹ẴὃິࠉMALDI-TOF-MS ࢅ⏕࠷ࠉ◂✪ࢅ⾔ࡖ࡙ࡀ
ࡒࠊ๑ᅂࡡࢤࣤࣆ࢒ࣝࣤࢪ࡚ࡢࠉSP ᾦమᇰᆀ࡚ 1㹳3 ᪝㛣⏍⫩ࡈࡎࡒⳞమࡡ஦ḗඔ㞹Ẵὃິࣂࢰ࣭ࣤ࠾ࡼࠉ
⏍⫩᫤㛣࡞ᚉ࠷Ⓠ⌟㔖ࡡን໩ࡌࡾࢰࣤࣂࢠ㈻ࡡྜྷᏽࢅ⾔ࡖࡒࠊ௑ᅂࠉ㯔Ⳟ࠿ᾦమᇰ㣬࡛ᅖమᇰ㣬࡚ࡢࠉⓆ⌟
ࡌࡾࢰࣤࣂࢠ㈻ࡡ⛸㢦ࠉ㔖࠿␏࡝ࡾࡆ࡛࠾ࡼࠉᑚ㯇ࡨࡌࡱᇰᆀ࡚ᇰ㣬ࡊࡒⳞమ࡞ࡗ࠷࡙ࡡよᯊࢅ⾔ࡖࡒࠊⳞ
మࢅᅂ཭ࡌࡾ㝷ࠉᅖమᇰ㣬࡚ࡢⳞమ࡛ᇰᆀࡡฦ㞫࠿ᅏ㞬࡝ࡒࡴࠉᇰ㣬∸ࢅᇰᆀࡇ࡛ᾦమ✽⣪࡚◒◃ࡊࠉࢰࣤ
ࣂࢠ㈻ࢅ᢫ฝࡊࠉࢦࣤࣈ࡛ࣜࡊࡒࠊࡐࡡࢦࣤࣈࣜࢅ஦ḗඔ㞹Ẵὃິ࡚ᒈ㛜ࡊࡒ࡛ࡆࢀࠉࡨࡌࡱᇰᆀࡡࡲࢅὃ
ິࡊࡒ࡛ࡀ࡞⌟ࡿࡒࢪ࣎ࢴࢹ࠿࡝ࡂ࡝ࡖ࡙࠷ࡒࡒࡴࠉᇰᆀࡡࢰࣤࣂࢠ㈻ࡢฦよష⏕ࢅུࡄࡒࡵࡡ࡛⩻࠻ࡼࡿ
ࡒࠊSP ᇰᆀ࡚ 3 ᪝㛣ᇰ㣬ࡊࡒⳞమࡡὃິࣂࢰ࣭࡛ࣤࠉ3 ᪝㛣ࡨࡌࡱᇰ㣬ࡊࡒⳞమࡡࣂࢰ࣭ࣤࡢ᪺ࡼ࠾࡞␏࡝
ࡖ࡙࠷ࡒࠊࡨࡌࡱᇰ㣬ᚃ᢫ฝࡊࡒࢰࣤࣂࢠ㈻ࢅὃິࡊࡒࢣ࡚ࣜࠉ☔ヾ࡚ࡀࡒ 200 ᘽࡡࢪ࣎ࢴࢹ࡞ࡗ࠷࡙ྜྷᏽ
షᴏࢅ㐅ࡴࡒࠊࡐࡡ⤎ᯕࠉᶭ⬗ᮅ▩ࡡࢰࣤࣂࢠ㈻ࡡⓆ⌟࠿☔ヾࡈࡿࡾ࡝࡜ࠉᾦమᇰ㣬᫤ࡡࢰࣤࣂࢠ㈻ࡡⓆ⌟
࡛ࡢ␏࡝ࡖࡒലྡྷ࠿ぜࡼࡿࡒࠊᾦమᇰ㣬᫤࡞ࠉᇰ㣬᫤㛣࠿㛏ࡂ࡝ࡾࡆ࡛࡚௥ㅨ㛭㏻ࡡࢰࣤࣂࢠ㈻ࡡቌຊ࠿ぜ
ࡼࡿࡒ࠿ࠉᅖమᇰ㣬 3 ᪝┘ࡡሔྙࡵࡷࡢࡽ௥ㅨ㛭㏻ࡡࢰࣤࣂࢠ㈻࠿ኣࡂⓆ⌟ࡊ࡙࠷ࡒࠊ
Proteome analysis of intracellular proteins from A.oryzae RIB40.
Taro Ko, Michio Takeuchi
(Tokyo Univ. of Agriculture and Technology)
P-68
㯜㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ CPase 㐿ఎᏄࡡよᯊ
ಚ㔕㐡ᙢ㸡➁හ㐠㞕 㸝᮶ா㎨ᕝኬᏕ࣬㎨࣬ᚺ⏍⛁㸞
㯔Ⳟ㸝Aspergillus oryzae RIB40㸞ࡡࢣࢿ࣑よᯊ࠾ࡼὩᛮ୯ᚨࠉ࣓ࢲ࣭ࣆ࡝࡜࡞ࡻࡾ᳠⣬ࢅ⾔࡝ࡖࡒ࡛ࡆࢀࠉ㸦
㸧ࡡ࢜ࣜ࣍࢞ࢨ࣋ࣈࢲࢱ࣭ࢭ㸝CPase㸞ࡡ≁ᚡࢅᣚࡗ㐿ఎᏄ࠿ぜฝࡈࡿࡒࠊࢣࢿ࣑᝗ሒ࠿᪺ࡼ࠾࡞࡝ࡖࡒ⣊≟
Ⳟࡡ CPase 㐿ఎᏄ࡞ࡗ࠷࡙ࠉA. oryzae ࡞࠽ࡄࡾ CPase 㐿ఎᏄࡡሲᇱ㒼า࠾ࡼ᥆ᏽࡈࡿࡾ࢓࣐ࢿ㓗㒼า᝗ሒࢅ
ࡵ࡛࡞よᯊࢅ⾔࠷ CPase ࡡ⣌⤣ᶖࢅషᠺࡊࡒࠊࡐࡡ⤎ᯕࠉ⣊≟Ⳟࡡ CPase ࡢኬࡀࡂ㸩ࡗࡡࢡ࣭ࣜࣈ࡞ฦࡄࡼ
ࡿࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ㸩ࡗࡡࢡ࣭ࣜࣈࡢࠉCPase ࡡᑻᅹᛮࢅཬ᫆ࡊ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ㸩ࡗࡡࢡࣜ
࣭ࣈ࠾ࡼࡐࡿࡑࡿୌࡗࡡ CPase 㐿ఎᏄࢅཱིࡽ୕ࡅࠉࡆࡿࡼࡡ㐿ఎᏄ࠿ A. oryzae RIB40 ᰬ௧አࡡ A. oryzae ᰬ࡞
Ꮛᅹࡌࡾࡡ࠾ࢅㄢ࡬ࡾࡒࡴࠉA. oryzae RIB40 ࡡ྘㐿ఎᏄධ㛏ࢅࣈ࣭ࣞࣇ࡛ࡊ࡙ A. oryzae ᰬ㸩⛸㢦࡛ A. sojae
࡞ࡗ࠷࡙ࢦࢧࣤよᯊࢅ⾔࡝ࡖࡒࠊࡐࡡ⤎ᯕࠉ௙ࡡ A. oryzae ᰬࡷ A. sojae ࡞ࡵ A. oryzae RIB40 ࡡᣚࡗ CPase 㐿
ఎᏄࡢᏋᅹࡊ࡙࠷ࡒࠊA. oryzae ᰬࡡ CPase 㐿ఎᏄࡢ A. oryzae RIB40 ࡡ CPase 㐿ఎᏄ࡛ྜྷࡋన⨠࡞ࣁࣤࢺ࠿ฝ
⌟ࡊࡒࠊࡊ࠾ࡊࠉA. sojae ࡞Ꮛᅹࡌࡾ CPase 㐿ఎᏄࡡ୯࡞ࡢࠉA. oryzae RIB40 ࡡ CPase 㐿ఎᏄ࡛ࡢ␏࡝ࡾన⨠
࡞ࣁࣤࢺ࠿ฝ⌟ࡊࡒࡵࡡࡵᏋᅹࡊࡒࠊࡱࡒࠉ㒼า࠿ࡌ࡚࡞ሒ࿈ࡈࡿ࡙࠷ࡾ A. nidulans ࡞ࡗ࠷࡙ࡵྜྷᵕ࡞ࢦࢧ
ࣤよᯊࢅ⾔࡝ࡖࡒࠊ
Analysis of CPase genes from Filamentous fungi.
Tatsuhiko Matano, Michio Takeuchi
(Tokyo Univ. of Agriculture and Technology)
69
P-69
ࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ஦㔔◒ቪᰬࢅ⏕࠷ࡒࢬ࣭ࣜࣞࢪ⢶໩㯔ⳞࡡฦᏄ⫩⛸
୯஬ ႗ᙢ 1㸡᭯ᒱ Ꮥ 1㸡ᕖཾ๙ྒྷ 2㸡Ⲡ஬ᇱኰ 3㸡໪ᮇົࡥࡆ 1 㸝1 ᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸡2 㜨ᗋኬ㝌࣬
㎨⏍⛁㸡3 ୯㒂ኬ࣬ᚺ⏍㸞
࠘┘Ⓩ࠙㎾ᖳࠉ໩▴⇖ᩩ࡚࠵ࡾ▴ἔ࡞᭨ࢂࡾ㈠″࡛ࡊ᳔࡙∸ࣁ࢕࢛࣏ࢪࡡฺ⏕࠿Ἰ┘ࢅ㞗ࡴ࡙࠷ࡾࠊࡐࡡ୹
さᠺฦ࡚࠵ࡾ⤎ᬏᛮࢬ࣭ࣜࣞࢪࡢ㞬ฦよᛮࡡࡒࡴ࡞ฺ⏕࠿㐅ࢆ࡚࠷࡝࠷ࠊࡆࡡ⤎ᬏᛮࢬ࣭ࣜࣞࢪࢅฦよࡌࡾ
㓕⣪ࢅ⏍⏐ࡌࡾ᚜⏍∸࡛ࡊ࡙ Trichoderma reesei ࡷ Aspergillus aculeatus ࡝࡜࠿▩ࡼࡿ࡙࠷ࡾࠊᮇ◂✪ࡢ
A.aculeatus ࠿⏍⏐ࡌࡾ୹さ࡝ࢬ࣭ࣜࣚࢭ࡚࠵ࡾ FϨ-CMCaseࠉCellobiohydrolase ϨࠉBeta-glucosidase 1 ࢅࢤ࣭
ࢺࡌࡾ 3 㐿ఎᏄ(cmc1,cbhϨ,bgl1)ࢅ A.oryzae ࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ஦㔔◒ቪᰬ NS-tApE ᰬ(niaD,sC,ǼpepE,Ǽ
tppA)࡫ᑙථࡊࠉࢬ࣭ࣜࣞࢪ⢶໩㯔Ⳟࢅ⫩⛸ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙࠷ࡾࠊ
࠘᪁Ἢཀྵࡦ⤎ᯕ࠙A.aculeatus ⏜ᮮࡡࢬ࣭ࣜࣚࢭࢅࢤ࣭ࢺࡌࡾ 3 㐿ఎᏄ(cmc1,bgl1,cbhϨ)࡞ࡗ࠷࡙ niaD ࢅ㐽ᢝ
࣏࣭࣭࡛࢜ࡊ࡙⏕࠷ࠉtef1 ࣈ࣓࣭ࣞࢰ࣭ୖ࡚Ⓠ⌟ࡌࡾࣈࣚࢪ࣐ࢺࢅᵋ⠇ࡊࡒࠊࡆࡿࡼࢅ༟≺࡞ NS-tApE ᰬ࡞
ᑙථࡊࠉ྘ࠍࡡ㓕⣪Ὡᛮࢅㄢ࡬ࡒࠊcmc1 Ⓠ⌟ᰬ࡚ࡢᇰ㣬୕ΰ࡞࠽࠷࡙᭩ኬ࡚⣑ 120 unit/ml ࡡ endo-glucanase
Ὡᛮࢅ♟ࡊࠉbgl1 Ⓠ⌟ᰬ࡚ࡢᇰ㣬୕ΰཀྵࡦ⣵⬂ባ࡞ beta-blucosidase Ὡᛮ࠿ヾࡴࡼࡿࡒࠊࡱࡒࠉcbh ϨⓆ⌟ᰬࡢ
ཱིᚋ୯࡚࠵ࡾࠊࡈࡼ࡞ࠉࡆࡿࡼ 3 ࡗࡡࢬ࣭ࣜࣚࢭ㐿ఎᏄࢅྜྷ᫤࡞ NS-tApE ᰬ࡞ᑙථࡊࡒᰬࡡ⫩⛸ࢅゝ⏤ࡊ࡙
࠷ࡾࠊ
Molecular breeding of cellulose hydrolysing koji-mold using double protease gene disruptant.
Takahiko Nakai1, Manabu Arioka1, Takashi Kawaguchi2, Motoo Arai3, Katsuhiko Kitamoto1
(1Dept. of Biotechnol., Univ. of Tokyo, 2Dept.Appl.Life Sci., Osaka Pref.Univ., 3Dept.Env.Biol., Chubu Univ.)
P-70
ஜ⇩ᅰቫ⎌ሾ୯࡞࠽ࡄࡾ㐿ఎᏄ⤄ᥦ࠻㯔Ⳟ⬂Ꮔࡡ⏍Ꮛᛮࡡよᯊ
ᴃᮇ៿ୌ 1ࠉཿᕖ⫩௥ 1ࠉ໪ᮇ์⾔ 2 ࠉ㕝ᮄ⪵ 1ࠉ᯵ᮄ㇇ 1㸝1 ㎨◂ᶭᵋ࣬㣏⥪◂ࠉ2 យ▩⏐ᢇ◂࣬㣏ဗᕝᢇ㸞
࠘┘Ⓩ࠙㯔Ⳟࡡᐁ⏕Ⳟᰬ࡞ᑊࡌࡾ㐿ఎᏄ⤄ᥦ࠻ᢇ⾙࠿㐅ᒈࡌࡾ࡞ఔ࠷ࠉ⎌ሾ୯࡞࠽ࡄࡾ⤄ᥦ࠻Ⳟᰬࡡ⏍Ꮛᛮ
ࡡよ᪺࠿ịࡴࡼࡿ࡙࠷ࡾࠊⁿ⩽ࡼࡢࡌ࡚࡞ᐁ⏕㯔Ⳟ⏜ᮮᰬ࡛ࢬ࣭ࣜࣚࢭ㧏Ⓠ⌟ᛮ㐿ఎᏄ⤄ᥦ࠻ᰬࢅ⏕࠷࡙ࠉ
ᅧⰹ⏕ᅰቫ୯࡞࠽ࡄࡾ⏍ᏋᛮࢅẒ㍉᳠ゞࡊࡒ㸝ᖲᠺ 16 ᖳᗐᮇኬఌさ᪠㞗 p70㸞
ࠊࡐࡡ⤎ᯕࠉ⤄ᥦ࠻ᰬࡡᕖ▹
࡚ࡡ⏍Ꮛ⋙ࡢ෢Ꮢ࡞఩࠷ലྡྷ࡞࠵ࡖࡒࠊ⤄ᥦ࠻ᰬࡡ⏍Ꮛ⋙ࡡᏒ⟿ንິ࠿ᅰቫ୯ࡡỀฦྱ㔖࡞ᙫ㡢ࡈࡿࡾࡡ࡚
ࡢ࡝࠷࠾࡛⩻࠻ࠉᙁโஜ⇩ࡊࡒᅰቫ࡞࠽ࡄࡾ୦ᰬࡡ⏍Ꮛᛮ࡞ࡗ࠷࡙᳠ゞࢅ⾔ࡖࡒࠊ
࠘᪁Ἢཀྵࡦ⤎ᯕ࠙㓲ἔ⏕㯔Ⳟ⏜ᮮࡡ Aspergillus oryzae KBN616-39 ཀྵࡦࠉᮇᰬ࡞ࢬ࣭ࣜࣚࢭ㐿ఎᏄ celB ࢅᑙථ
ࡊࡒࢬ࣭ࣜࣚࢭ㧏⏍⏐ᰬ A. oryzae TB-1 ࢅ౐⏕ࡊࡒࠊᅧⰹ⏕ᅰቫ㸝ᕖ▹㸞90Υ࡚ୌᬄஜ⇩ࡈࡎࡒᚃࠉࡐࡡ 2g
ࢅࣆࢰ௛ࡀ࢙࣎ࣛࢲࣝࣤᐖჹ࡞ථࡿࠉKBN616-39 ࡱࡒࡢ TB-1 ࡡ⬂Ꮔ⣑ 106ࠉ0.01ml ࢅ᥃⛸ࡊࠉ25Υ࡚ᜇῺ㟴
⨠ᇰ㣬ࢅ⾔ࡖࡒࠊ⤊᫤Ⓩ࡞ᅰቫ᢫ฝᾦࢅㄢ⿿ࡊࠉᕵ㔐ᖲᯀἪ࡞ࡻࡽ⏍Ⳟᩐࢅ⟤ฝࡊࡒࠊࡐࡡ⤎ᯕࠉ᥃⛸ᚃ 7
᪝┘࡞࠽࠷࡙ࠉKBN616-39 ࡡ⏍Ꮛ⋙࠿ 12㸢24㸚㸝2 ཬᚗ㸞࡚࠵ࡖࡒࡡ࡞ᑊࡊ࡙ࠉTB-1 ࡚ࡢ 1.0㸢2.0㸚㸝2 ཬ
ᚗ㸞࡚࠵ࡖࡒࠊࡆࡡࡆ࡛࠾ࡼࠉTB-1 ࡡ⏍Ꮛ⋙ࡢ౩モᅰቫࡡᴗᗐ࡝఩Ềฦ᮪௲࡚ KBN616-39 ࡻࡽࡵ 10 ฦࡡ 1
௧ୖ࡞఩ୖࡌࡾࡆ࡛࠿ࢂ࠾ࡖࡒࠊ௑ᚃ TB-1 ᰬࡡ఩Ềฦୖ࡚ࡡ⏍Ꮛ⋙఩ୖさᅄ࠿よ᪺ࡈࡿࡾ࡛ࠉ㔕አ⎌ሾ୯࡚
Ṓ⁓㏷ᗐࡡ㏷࠷㐿ఎᏄ⤄ᥦ࠻㯔Ⳟࡡ⫩ᠺ࡞ࡗ࡝࠿ࡾ࡛⩻࠻ࡼࡿࡾࠊ
Analysis of the spore survivability of the recombinant koji mold in the dried soil environment
Ken-Ichi Kusumoto1, Ikuyo Furukawa1, Noriyuki Kitamoto2, Satoshi Suzuki1, Yutaka Kashiwagi1
(1Natl.Food.Res.Inst, 2Food Res. Center, Aichi Ind. Technol. Inst.)
70
P-71
㯔Ⳟ࡞࠽ࡄࡾ⠾౼ࢤࣞࢼ࣭PCR Ἢ࡛ࡐࡡᚺ⏕
㕝ᮄ ⪵㸡➁ㆺ༡Ꮔ, ᮄᮟኣỜ, ᮿୖ┷⏜⨶, ኣ⏛ຉ⏍, ᴃᮇ៿ୌ, ᯵ᮄ ㇇ 㸝㣏⥪◂㸞
㯔Ⳟࢣࢿ࣑᝗ሒ࠿ප㛜ࡈࡿࠉ᩺ぞ㐿ఎᏄᶭ⬗よᯊࡡࡒࡴࡡ㐿ఎᏄ◒ቪᐁ㥺࠿ᗀࡂ⾔ࢂࡿࡾࡻ࠹࡞࡝ࡖࡒࠊ
ࡐࡿ࡞ఔ࠷ࠉ◒ቪᰬࢪࢠ࣭ࣛࢼࣤࢡࡡࡒࡴࡡ⭶ኬ࡝ᩐࡡᙟ㈻㌹ᥦᰬࡡࢩ࢘ࢿࢰ࢕ࣅࣤࢡ࠿ᚪさ࡛࡝ࡾࠊࡆࡿ
ࡱ࡚ࠉPCR ࢷࣤࣈ࣭ࣝࢹ࡛࡝ࡾࢣࢿ࣑ DNA ࡡㄢ⿿ࡢࠉ⣊≟Ⳟࡡᙁᅖ࡝⣵⬂ባࡡࡒࡴ᫤㛣࡛ຘງ࠿ᚪさ࡚࠵
ࡖࡒࠊࡐࡆ࡚ࠉᠻࠍࡢ⠾౼࡝๑ฌ⌦࡚ࠉᐨኮࣈ࣭ࣝࢹ୕ࡡ㯔Ⳟࢤࣞࢼ࣭Ⳟమࢅ├᥃ PCR ࢷࣤࣈ࣭ࣝࢹ࡛ࡌࡾ
ࡆ࡛࠿ྊ⬗࡝ᡥἪࢅ㛜Ⓠࡊࡒࠊ
ᮇᡥἪࡢࠉFTA ࣭࢜ࢺ㸝Whatman㸞ࢅ⏕࠷࡙㯔Ⳟࢣࢿ࣑ࢅᅖᏽ໩ࠉᏭᏽ໩ࡈࡎࡾࡵࡡ࡚࠵ࡽࠉ᭯ᶭ⁈፳ࡷ
࢛࢜ࢹࣞࣅࢴࢠሲ࡝࡜ࡡ᭯ᐐ∸㈻ࠉ⣵⬂◒◃ᶭ➴ࡡ≁ื࡝ᶭ᲌ࢅᚪさ࡛ࡎࡍࠉᩐ mm3 ⛤ᗐࡡⳞమ࠾ࡼ PCR ࢷ
ࣤࣈ࣭ࣝࢹ࡞༎ฦ㔖ࡡ DNA ࢅ᢫ฝࡌࡾࡆ࡛࠿࡚ࡀࡾࠊࡱࡒࠉᮇᡥἪ࡚ࡢࠉ༖ᖳ㛣 4Υ࡞࡙ಕᏋࡈࡿࡒࢪࣚࣤ
ࢹᇰ㣬Ⳟᰬ࠾ࡼࡡ PCRࠉ࠵ࡾ࠷ࡢ࿝ძࠉ᪝ᮇ㒿࠾ࡼࡡ PCR ࡞ࡻࡾ⣊≟Ⳟࡡ᳠ฝ࡞ࡵᚺ⏕ࡌࡾࡆ࡛࠿࡚ࡀࡾࠊ
ᠻࠍࡢࠉᮇᡥἪࢅ⏕࠷࡙ࠉࢡࣜࢰ࣐ࢻ࣭ࢭ┞ྜྷ㐿ఎᏄࠉGPCR ┞ྜྷ㐿ఎᏄࡡ◒ቪᰬࢪࢠ࣭ࣛࢼࣤࢡࢅ⾔ࡖ
ࡒࠊࡱࡒࠉࣛ࣍ࢯ࣭࣑ DNA ࡡ ITS 㡷ᇡࢅቌᖕࡌࡾࣈࣚ࢕࣏࣭ࢬࢴࢹࢅ⏕࠷࡙ࠉ࿝ძ࠾ࡼࡡ⣊≟Ⳟ DNA ࡡ᳠
ฝࠉ㓕ẍ DNA ࡡ᳠ฝࠉཀྵࡦⳞᰬྜྷᏽࢅ⾔ࡖࡒࡡ࡚ሒ࿈ࡌࡾࠊ
Application of colony direct polymerase chain reaction of Aspergillus oryzae
Satoshi Suzuki, Hiroko Taketani, Tae Kimura, Mayumi Matsushita, Sawaki Tada, Ken-Ichi Kusumoto, Yutaka
Kashiwagi
(NFRI)
P-72
Fusarium oxysporum ⏜ᮮ⢶໩࢓࣐ࢿ㓗࢛࢞ࢨࢱ࣭ࢭࡡ㔘ᒌ࢕࢛ࣤ࡞ࡻࡾὩᛮ໩
⸠ཋ┷⣎ 1㸡⅛ㆺ㡨ୌ 1 㸡ཿ㈙᫬἖ 2㸡ⰶ㝘ୌ⏍ 2 㸡㧏ጏ༜ྒྷ 2㸡௑ᮟⱩ⾔ 2㸡ᕖཾ๙ྒྷ 1 㸡Ⲡ஬ᇱኰ 3 㸝1 ኬ㜨ᗋ
ኬ㝌࣬ᚺ⏍໩㸡2 ᪣໩ᠺࣆ࢒࣭࣏࣬チ᩷ⷾ◂㸡3 ୯㒂ኬ࣬⎌ሾ⏍∸㸞
࠘┘Ⓩ࠙⢶໩࢓࣐ࢿ㓗࢛࢞ࢨࢱ࣭ࢭ㸝FOD㸞ࡢ⢶ᑺ⑋࡞࠽ࡄࡾ⾉⢶ࢤࣤࢹ࣭࣏࣭࣭࡚ࣞࣜ࢜࠵ࡾ⢶໩ࢰࣤ
ࣂࢠࢅῼᏽࡌࡾチ࡛᩷ⷾࡊ࡙᭯⏕࡝㓕⣪࡚࠵ࡾࠊ⾉୯ࡡ୹࡝⢶໩ࢰࣤࣂࢠ࡞⢶໩࢓ࣜࣇ࣐࡛ࣤ⢶໩ࣉ࣓ࢡࣞ
ࣄࣤ࠿ᣪࡅࡼࡿࡾ࠿ࠉ๑⩽ࡢ⢶໩ࣛࢩࣤ㸝FK㸞ࠉᚃ⩽ࡢ⢶໩ࣁࣛࣤ㸝FV㸞ࢅྱࢆ࡚࠷ࡾࠊFusarium oxysporum
⏜ᮮ FOD ࡢࠉFKࠉFV ࡡ୦᪁࡞ష⏕ࡌࡾ࠿ࠉࣚࣤࢱ࣑ን␏ࢅᑙථࡌࡾࡆ࡛࡚ࠉFV ࡞࡮࡛ࢆ࡜ష⏕ࡊ࡝࠷
ን␏㓕⣪㸝K373 ⨠ᥦమ㸞ࢅཱིᚋࡊ࡙࠽ࡽࠉ௑ᅂࡢ WT ࡛ን␏㓕⣪࡞ᑊࡌࡾ㔘ᒌ࢕࢛ࣤࡡᙫ㡢࡞ࡗ࠷࡙ሒ࿈
ࡌࡾࠊ
࠘᪁Ἢཀྵࡦ⤎ᯕ࠙WT ࡛ 5 ⛸㢦ࡡ K373 ⨠ᥦమ࡞ࡗ࠷࡙ࠉᵕࠍ࡝㔘ᒌ࢕࢛ࣤࡡᏋᅹୖ㸝20 mM㸞࡚ࡐࡡ
Ὡᛮࢅῼᏽࡊࡒ࡛ࡆࢀࠉCa2+ࠉMn2+ࠉMg2+ ࡡᏋᅹୖ࡚ࠉ㓕⣪Ὡᛮࡡኬࡀ࡝୕᪴࠿ぜࡼࡿࡒࠊὩᛮࡡ୕᪴⋙ࡢ
㓕⣪ࡡ⛸㢦ࠉᇱ㈻࡞ࡻࡖ࡙ኬࡀࡂ␏࡝ࡖࡒࠊMg2+ ࡡሔྙࠉ㟸Ꮛᅹୖ࡛Ẓ㍉ࡊ࡙ WT ࡚ FK Ὡᛮ࠿ 113%ࠉ
FV Ὡᛮ࠿ 148% ࡚࠵ࡖࡒ࠿ࠉK373W ⨠ᥦమ࡚ࡢ FK Ὡᛮ࠿ 236%ࠉFV Ὡᛮࡢ 1410% ࡱ࡚୕᪴ࡊࡒࠊ
Mg2+ ⃨ᗐࡡን໩࡞ᑊࡌࡾ kinetic ࣂ࣒࣭ࣚࢰࡡን໩ࢅịࡴࡒ࡛ࡆࢀࠉWT ࡡ FK ࡞ᑊࡌࡾὩᛮࡢ Mg2+ ⃨ᗐ
ࡡᙫ㡢ࢅṜ࡜ུࡄ࡝࠾ࡖࡒ࠿ࠉࡐࡿ௧አࡡ㓕⣪㸢ᇱ㈻ࡡ⤄ࡲྙࢂࡎ࡚ࡢ Mg2+ ⃨ᗐ࠿㧏ࡂ࡝ࡾ࡮࡜ Km ࡢ఩
ୖࡊࠉkcat ࡢቌຊࡌࡾࡆ࡛࡚Ὡᛮ໩ࡈࡿࡾࡆ࡛࠿ࢂ࠾ࡖࡒࠊ
Activation of fructosyl amino acid oxidase from Fusarium oxysporum by metal ions.
Maki Fujiwara1, Jun-ichi Sumitani1, Shinji Koga2, Issei Yoshioka2, Takuji Kouzuma2, Shigeyuki Imamura2, Takashi
Kawaguchi1, Motoo Arai3
(1Grad. Sch. of Life & Env. Science, Osaka Pref. Univ. 2Diagnostics R&D Dept., Asahi Kasei Pharma Co. 3Dept. of
Env. Biol., Chubu Univ.)
71
P-73
Rhizopus microsporus ࡡ⣵⬂හභ⏍ࣁࢠࢷࣛ࢓ࡡᙟ㈻㌹ᥦ࡛␏⛸ࢰࣤࣂࢠ㈻Ⓠ⌟
ᮇᒜ㧏ᖶ 1㸡ⴘྙዄ⥬Ꮔ 1㸡᲻⏛┷⣟ 1,2㸡Ꮻఫ⨶ㄵ 2㸡ᕝ⸠ಆ❮ 1㸝1 ⌦◂࣬୯ኳ◂ࠉ2 ᮶Ὂኬ࣬ᕝ㸞
࠘┘Ⓩ࠙Rhizopus ࡢ᥃ྙⳞ㢦㸝ୖ➴Ⳟ㢦㸞࡞ᒌࡌࡾ⣊≟Ⳟ࡚࠵ࡽࠉⓆ㓕㣏ဗ⏍⏐➴࡞ฺ⏕ࡈࡿ࡙ࡀࡒࠊࡱࡒ
ୌ㒂ࡢ᳔∸ࡷິ∸ࡡ⑋ཋⳞ࡛ࡊ࡙ࡵ▩ࡼࡿ࡙࠷ࡾࠊ᭩㎾ࠉRhizopus microsporus ࡡୌ㒂࠿⣵⬂හභ⏍ࣁࢠࢷࣛ
࢓ࢅಕᣚࡌࡾࡆ࡛࠿ሒ࿈ࡈࡿࡒࠊᠻࠍࡢࠉR. microsporus ࡛⣵⬂හභ⏍ࣁࢠࢷࣛ࢓࡛ࡡභ⏍⣌࠿ࠉࢱ࢕࢛࢞ࢨ
ࣤ㢦࡝࡜ࡡࣁ࢕࢛࣒ࣝࢸ࢔࢙࣭ࢨࣘࣤ➴࡫ᚺ⏕࡚ࡀ࡝࠷࠾࡛⩻࠻ࠉභ⏍࣒࢜ࢼࢫ࣑ࡡよᯊࢅྱࡴࡒ◂✪ࢅ㐅
ࡴ࡙࠷ࡾࠊ
࠘᪁Ἢ࣬⤎ᯕ࠙⣵⬂හභ⏍ࣁࢠࢷࣛ࢓ࢅಕᣚࡌࡾ R. microsporus ᰬࡡⳞ⣊ࢅ◒◃ࡊࡒᚃࠉṹ㝭ᕵ㔐ࡊ࡙ࣈ࣭ࣝ
ࢷ࢔ࣤࢡࡌࡾࡆ࡛࡞ࡻࡽ⣵⬂හභ⏍ࣁࢠࢷࣛ࢓ࡡࢤࣞࢼ࣭ࢅᚋࡒࠊᙟ㈻㌹ᥦ⏕ࣈࣚࢪ࣐ࢺ࡛ࡊ࡙ࢡ࣑ࣚ㝔ᛮ
Ⳟࡡᗀᐙ୹ᇡ࣊ࢠࢰ࣭pMFY42 ࢅ⏕࠷࡙࢙ࣝࢠࢹ࣭ࣞ࣎ࣝࢨࣘࣤ࡞ࡻࡾᙟ㈻㌹ᥦ࡞ᠺຉࡊࡒࠊḗ࡞ࠉGFP ࡛
࣋ࣤࢰࢠࣞࣞࣆ࢘ࢿ࣭ࣜฦよ⣌㓕⣪ࢅྜྷ᫤࡞Ⓠ⌟ࡈࡎࡾࡒࡴࡡெᕝ࢛࣋ࣞࣤࢅ pMFY42 ࡞ࢠ࣭ࣞࢼࣤࢡࡊࠉ
ࣁࢠࢷࣛ࢓࡞ᑙථࡊࡒࠊᙟ㈻㌹ᥦమࡡ GFP ⺧කࢅ☔ヾࡊࡒᚃࠉᢘ⏍∸㈻࡞ࡻࡽࣁࢠࢷࣛ࢓ࢅ㝎Ⳟࡊࡒ R.
microsporus ࡞វ᯹ࡈࡎࡒ࡛ࡆࢀࠉⳞ㢦⣵⬂හ࡚ GFP ⺧කࢅⓆࡌࡾࣁࢠࢷࣛ࢓ࢅ☔ヾࡌࡾࡆ࡛࠿࡚ࡀࡒࠊ
Transformation of Rhizopus microsporus-endosymbiotic bacteria and expression of heterologous proteins in
transformants
Takayuki Motoyama1, Naoko Ochiai1, Masumi Morita1,2, Ron Usami2, Toshiaki Kudo1
(1Discovery Res. Institute, RIKEN, 2Fac. Technol., Univ. Toyo)
P-74
㯑Ⳟ(Aspergillus oryzae )ࡡᅖమᇰ㣬ิ᭿࡞Ⓠ⌟ࡌࡾ㐿ఎᏄ AOS13 ࡡよᯊ
ḯᓞ᜾ 1,2, ᒷୖ࿰⿩ 1,2, ᆊᮇ࿰ಆ 2, ㉝ᑹ೸ 2, ᒜ⏛ಞ 2, ⚽⏛ಞ 3, ୖ㣜ொ 1,2, ୔୕㔔᪺ 2㸝ᗀኬ࣬඙❻◂ 1, 㒿⥪
◂ 2, ᐁ㊰ዥᏄኬ 3 㸞
㯑Ⳟࡢฦ⏍Ꮔᙟᠺ࡝࡜ࡡฦ໩ࢅ⏍ࡋࡾࡆ࡛ࡷࠉᅖమᇰ㣬᮪௲ୖ࡚ኣ⛸ኣ㔖ࡡ㓕⣪ࢅ⏍⏐ࡌࡾࡆ࡛࠿▩ࡼࡿ
࡙࠷ࡾࠊᮇ◂✪࡚ࡢࠉ㯔Ⳟࡡᅖమᇰ㣬ิ᭿࡞Ⓠ⌟ࡌࡾ㐿ఎᏄ AOS13 ࡞ࡗ࠷࡙ࠉⓆ⌟โᚒᶭᵋࢅよ᪺ࡌࡾࡆ࡛
ࢅ┘Ⓩ࡛ࡊࡒࠊ
AOS13 ࡢᅖమᇰ㣬࡛ᾦమᇰ㣬ࡡࢦࣇࢹࣚࢠࢨࣘࣤἪ࡚ྜྷᏽࡈࡿࡒ㐿ఎᏄ࡚࠵ࡽࠉ5’-RACE, 3’-RACE ࡞ࡻࡽ
ORF ࢅỬᏽࡊࠉᵋ㏸よᯊࢅ⾔ࡖࡒ࡛ࡆࢀࠉN-acetyltransferase ࡡ motif ࠿ぜฝࡈࡿ࡙࠷ࡾࠊAOS13 ࡢࡨࡌࡱ࡚
ࡡᅖమᇰ㣬(20 ᫤㛣ࠉ27 ᫤㛣)࡚㧏Ⓠ⌟ࡌࡾࡆ࡛࠿ࢿ࣭ࢧࣤよᯊ࡞ࡻࡖ࡙᪺ࡼ࠾࡞ࡈࡿ࡙࠷ࡾࡆ࡛࠾ࡼࠉᅖమ
ᇰ㣬࡚ࡡ AOS13 ࡡⓆ⌟โᚒᶭᵋ࡞ࡗ࠷࡙よ᪺ࡌࡾࡒࡴ࡞ࣈ࣓࣭ࣞࢰ࣭㡷ᇡࡡ deletion よᯊࢅ⾔ࡖࡒ࡛ࡆࢀࠉ
⩳ゼ㛜ጙⅤ࠾ࡼ୕Ὦ-242bp~-484bp ࡱ࡚ࡡ㡷ᇡ࡞ᅖమᇰ㣬࡞࠽ࡄࡾㄇᑙࠉᾦమᇰ㣬࡞࠽ࡄࡾᢒโ࡞㛭୙ࡌࡾ
cis-element ࠿Ꮛᅹࡌࡾࡆ࡛࠿♟ြࡈࡿࡒࠊࡱࡒࠉAOS13 ࡢࣈ࣭ࣝࢹᇰ㣬࡚ࡵⓆ⌟ࡊ࡙࠽ࡽࠉ㯑Ⳟࡡฦ໩࡝࡜
࡞㛭㏻ࡌࡾ㐿ఎᏄ࡚࠵ࡾ࡛⩻࠻ࡼࡿࡾࡒࡴࠉAOS13 ࡡ GFP ⼝ྙᰬࢅషᠺࡊࠉࡐࡡᑻᅹᛮ➴ࡡよ᪺࡞ࡗ࠷࡙ࡵ
◂✪ࢅ㐅ࡴ࡙࠷ࡾࠊ
Analysis of AOS13 expressed in initial solid-culture in Aspergillus oryzae .
Y.Utashima1,2,K.Iwashita1,2,K.Sakamoto2,T.Akao2,O.Yamada2,O.Akita3,J.Shimoi1,2,S.Mikami2
(Hiroshima Univ. 1, NRIB2,Jissen Women’s Univ. 3)
72
P-75
㯜㯔Ⳟ Aspergillus oryzae ࡡ HSP30 㐿ఎᏄࣈ࣓࣭ࣞࢰ࣭ࢅ⏕࠷ࡒᾦమᇰ㣬࡞࠽ࡄࡾ 㓕
⣪㐿ఎᏄࡡⓆ⌟࡛ࢰࣤࣂࢠ㈻⏍⏐ࡡ᳠ゞ
ᮿୖ┷⏜⨶ࠉ㕝ᮄ⪵ࠉኣ⏛ຉ⏍ࠉ▴⏛༡ᶖ*ࠉᴃᮇ៿ୌࠉ᯵ᮄ㇇㸝㣏⥪◂ࠉ*᭮᰿෗⥪◂㸞
ࠔ ┘ Ⓩ ࠕ 㯔 Ⳟ Aspe rg illus oryzae ࡢ ࢂ ࠿ ᅗ ࡡ ఎ ⤣ Ⓠ 㓕 㣏 ဗ ࡚ ࠵ ࡾ ࡲ ࡐ ࠉ 㓲 ἔ ࠉ ΰ 㒿 ࡝ ࡜ ࡡ ⿿ ㏸
࡞୘ྊḖ࡚࠵ࡾ࡛࡛ࡵ࡞㓕⣪⏍⏐࡞ࡵ⏕࠷ࡼࡿࠉ⏐ᴏ୕㔔さ࡝⣊≟Ⳟ࡚࠵ࡾࠊᠻࠍࡢࠉ㯔Ⳟࡡ
hsp30 ࣈ ࣞ ࣓ ࣭ ࢰ ࣭ ࢅ ⏕ ࠷ ࡙ ࠉ ᅖ మ ᇰ 㣬 ≁ ␏ Ⓩ ࡞ Ⓠ ⌟ ࡌ ࡾ 㯔 Ⳟ ࢡ ࣜ ࢤ ࢓ ࣐ ࣚ ࣭ ࢭ 㐿 ఎ Ꮔ glaB 㐿
ఎ Ꮔ ࢅ ࣓ ࢸ ࣜ ࡛ ࡊ ࡙ ࠉ ᾦ మ ᇰ 㣬 ࡞ ࡙ ࠉ 㧏 Ὼ ㄇ ᑙ Ⓠ ⌟ ࡈ ࡎ ࡾ 㓕 ⣪ ⏍ ⏐ ࢅ ⾔ ࡖ ࡙ ࡀ ࡒ 㸝 1㸞ࠊ ᮇ ◂ ✪
࡚ ࡢ hsp30 ࣈ ࣞ ࣓ ࣭ ࢰ ࣭ ࢅ ⏕ ࠷ ࡒ 㓕 ⣪ ⏍ ⏐ ࡞ ࡗ ࠷ ࡙ ࡈ ࡼ ࡞ ࠉ レ ⣵ ࡝ よ ᯊ ࢅ モ ࡲ ࡒ ࠊ
ࠔ ᪁ Ἢ ࡛ ⤎ ᯕ ࠕhsp30 ࣈ ࣞ ࣓ ࣭ ࢰ ࣭ ࡡ ୖ Ὦ ࡞ glaB 㐿 ఎ Ꮔ ࡡ ORF ࢅ ㏻ ⤎ ࡊ ࡒ ⼝ ྙ 㐿 ఎ Ꮔ ࢅ ࠉn iaD
ࢅ ࣏ ࣭ ࢜ ࣭ ࡛ ࡊ ࡙ 1 ࢤ ࣅ ࣭ ᑙ ථ ࡊ ࡒ ᙟ ㈻ ㌹ ᥦ ᰬ ࢅ ᚋ ࡒ ࠊhsp30promoter ࡡ -388㹳 -1 㡷 ᇡ ࡡ ࡲ ⏕ ࠷
ࡾ ࡛ ࠉ 30Υ ࡞ ࠽ ࠷ ࡙ ㌹ ෕ ࡢ ᢒ โ ࡈ ࡿ ࠉ 㧏 Ὼ ㄇ ᑙ ࡞ ࡻ ࡽ ㌹ ෕ Ὡ ᛮ ࠿ ୕ ᪴ ࡊ ࡒ ࠊ ᚋ ࡼ ࡿ ࡒ ᙟ ㈻ ㌹ ᥦ
ᰬ ࢅ PYD ᾦ మ ᇰ ᆀ ࡚ 30Υ ୌ ᬄ ᇰ 㣬 ᚃ ࠉ40Υ 2 ᫤ 㛣 ࡡ 㧏 Ὼ ㄇ ᑙ ࢅ ⾔ ࡖ ࡙ ࠉᇰ 㣬 ୕ ΰ ࡞ ⏍ ⏐ ࡈ ࡿ ࡾ
ࢡ ࣜ ࢤ ࢓ ࣐ ࣚ ࣭ ࢭ Ὡ ᛮ ࢅ ῼ ᏽ ࡊ ࡒ ࠊࡐ ࡡ ⤎ ᯕ ࠉᅖ మ ᇰ 㣬 ≁ ␏ Ⓩ ࡞ ⏍ ⏐ ࡈ ࡿ ࡾ G laB ࢅ ࠉ㧏 Ὼ ㄇ ᑙ
ᚃ ࡡ ᇰ 㣬 ୕ ΰ ୯ ࡞ ⏍ ⏐ ࡈ ࡎ ࡾ ࡆ ࡛ ࠿ ࡚ ࡀ ࡒ ࠊ ࡊ ࠾ ࡊ ࠉ 㸼 㹊 㹈 㐿 ఎ Ꮔ ࢅ ⏕ ࠷ ࡒ D eletion よ ᯊ ࡡ ⤎
ᯕ㸝 㸧 㸞࠾ ࡼ ஢ ῼ ࡊ ࡙ ࠷ ࡒ ࡻ ࡽ ࡵ ࠉ㧏 Ὼ ㄇ ᑙ ࡞ ࡻ ࡾ 㓕 ⣪ Ὡ ᛮ ࡡ ୕ ᪴ ࠿ ぜ ࡼ ࡿ ࡝ ࠾ ࡖ ࡒ ࠊࡐ ࡆ ࡚ ࠉ
⌟ ᅹ ࠉⓆ ⌟ 㔖 ࡷ ᇰ 㣬 ᮪ ௲ ࡡ ᳠ ゞ ࡝ ࡜ よ ᯊ ࢅ ⾔ ࡖ ࡙ ࠷ ࡾ ࠊ (1) ᮿ ୖ ࡼ ࠉ᪝ ᮇ ㎨ ⰹ ໩ Ꮥ ఌ 2006 ᖳ
ᗐ ኬ ఌ ㅦ ⁿ さ ᪠ 㞗 ࠉ 㹥 116
(2) ᮿ ୖ ࡼ ࠉ ➠ ஫ ᅂ ⣊ ≟ Ⳟ ࢤ ࣤ ࣆ ࢒ ࣝ ࣤ ࢪ さ ᪠ 㞗 ࠉ 㹥 79
Analysis of a promoter region of HSP30 gene from Aspergillus oryzae and expression of GUS gene
Mayumi Matsushita, Satoshi Suzuki, Sawaki Tada, Hiroki Ishida*, Ken-Ichi Kusumoto, Yutaka Kashiwagi
㸝Natl. Food Res. Inst., * Research Institute, Gekkeikan Sake Co. Ltd)
P-76
A.oryzae ࡡ MAL ࢠࣚࢪࢰ࣭㐿ఎᏄࡢ࢓࣐࣭ࣚࢭ⣌㓕⣪㐿ఎᏄⓆ⌟࡞ࡵ㛭୙ࡌࡾ
㛏ㆺᕖ⚀Ꮔ㸡஫࿝ົஒ 㸝᮶໪ኬ㝌㎨࣬⏍∸⏐ᴏ๭ᠺ㸞
[┘Ⓩ]⏐ᴏⓏ࡞㔔さ࡝㯔Ⳟࡡ࢓࣐࣭ࣚࢭ⣌㓕⣪ࡡ⏍⏐ࡢࠉᇰᆀ୯ࡡࢸࣤࣈࣤࡷ࣏ࣜࢹ࣭ࢪ࡝࡜ࡡ࣏ࣜࢹ࢛ࣛࢥ
⢶ࡡᏋᅹ࡞ࡻࡽ㌹෕ᅄᏄ AmyR ࢅ௒ࡊ࡙ḿࡡโᚒࢅུࡄ࡙࠷ࡾࠊୌ᪁ࠉ㯔Ⳟ࡞࠽ࡄࡾ࣏ࣜࢹ࣭ࢪ㈠໩ࡢ MAL
ࢠࣚࢪࢰ࣭࠿୯ᚨⓏ࡝ᶭ⬗ࢅᯕࡒࡊ࡙࠽ࡽࠉࡆࡡ MAL ࢠࣚࢪࢰ࣭㐿ఎᏄࡡ࠹ࡔࠉ࣏ࣜࢹ࣭ࢪࣂ࣭࣐࢓࣭ࢭ㐿
ఎᏄ malPࠉⳞమහ࣏ࣜࢰ࣭ࢭ㐿ఎᏄ malT ࡢ࣏ࣜࢹ࣭ࢪ࡞ࡻࡽⓆ⌟࠿ㄇᑙࡈࡿࠉࡐࡡⓆ⌟ࡢ࢓࣐࣭ࣚࢭ⣌㓕
⣪㐿ఎᏄࡡ㌹෕ᅄᏄ AmyR ࡚ࡢ࡝ࡂࠉMAL ࢠࣚࢪࢰ࣭හ࡞ᏋᅹࡊᵋᠺⓏ࡞Ⓠ⌟ࡊ࡙࠷ࡾ malR ࡡㄇᑙୖ࡞࠵
ࡾࡆ࡛࠿♟ြࡈࡿ࡙࠷ࡾࠊᮇ◂✪࡚ࡢࠉࢸࣤࣈࣤࡷ࣏ࣜࢹ࣭ࢪ࡝࡜ࡡ࣏ࣜࢹ࢛ࣛࢥ⢶ࡡฦよ㈠໩࡞㛭ࢂࡾ㐿
ఎᏄࡡ⥪ྙⓏ࡝Ⓠ⌟โᚒᶭᵋࢅ᪺ࡼ࠾࡞ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙ࠉ㌹෕ᅄᏄ MalR ࢅࡢࡋࡴ࡛ࡌࡾ MAL ࢠࣚࢪ
ࢰ࣭㐿ఎᏄࡡᶭ⬗よᯊࢅ⾔ࡖࡒ࡛ࡆࢀࠉMAL ࢠࣚࢪࢰ࣭㐿ఎᏄ࠿࢓࣐࣭ࣚࢭ⣌㓕⣪㐿ఎᏄⓆ⌟࡞ᐞ୙ࡊ࡙࠷
ࡾࡆ࡛ࢅヾࡴࡒࡡ࡚ሒ࿈ࡌࡾࠊ
[⤎ᯕ]malP ◒ቪᰬࡷ malR ◒ቪᰬࢅష⿿ࡊࡒ⤎ᯕࠉȘѸ࢓࣐࣭ࣚࢭὩᛮ࠿΅ᑛࡊࡒࡆ࡛࠾ࡼࠉࡆࡿࡼࡡ㐿ఎᏄ
࠿࢓࣐࣭ࣚࢭ⣌㓕⣪⏍⏐࡞రࡼ࠾ࡡᶭ⬗ࢅᯕࡒࡊ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊࡱࡒࠉȘ-࢓࣐࣭ࣚࢭ㐿ఎᏄ amyB
Ⓠ⌟࡞ࡵᙫ㡢࠿ぜࡼࡿࡒࡆ࡛࠾ࡼࠉMalR ࡞ࡻࡽㄇᑙࡈࡿࡾ MalP ࡡ࣏ࣜࢹ࣭ࢪⳞమහཱིࡽ㎲ࡲᶭ⬗ࢅ㏳ࡊ࡙
AmyR ࠿Ὡᛮᆵ࡛࡝ࡽࠉὩᛮᆵ AmyR ࠿࢓࣐࣭ࣚࢭ⣌㓕⣪㐿ఎᏄⓆ⌟ࢅㄇᑙࡌࡾࡵࡡ࡛ᛦࢂࡿࡾࠊࡈࡼ࡞ࠉ
ᵋᠺⓏ࡞Ⓠ⌟ࡊ࡙㌹෕ᅄᏄ࡛ࡊ࡙ᶭ⬗ࡊ࡙࠷ࡾ MalR ࡷ AmyR ࡡὩᛮ໩ࢅᘤࡀ㉫ࡆࡌㄇᑙᅄᏄࢅ᪺ࡼ࠾࡞ࡌ
ࡾࡒࡴࠉȘ-ࢡࣜࢤࢨࢱ࣭ࢭὩᛮ㜴ᐐ๠ࢅ⏕࠷࡙࣏ࣜࢹ࣭ࢪ࡛࢕ࢮ࣏ࣜࢹ࣭ࢪᏋᅹୖ࡚ࡡȘ-࢓࣐࣭ࣚࢭὩᛮࠉ
amyBࠉglaA ࡡⓆ⌟ࢅㄢ࡬ࡒ࡛ࡆࢀࠉ࣓ࢸࣜ࢜ࣄ࡚࠵ࡾ A.nidulans ࡚ࡢ AmyR ࡢ࢕ࢮ࣏ࣜࢹ࣭ࢪ࡞ࡻࡽὩᛮ໩
ࡈࡿࡾࡡ࡞ᑊࡊࠉA.oryzae ࡚ࡢ࣏ࣜࢹ࣭ࢪ࡚ࡵὩᛮ໩ࡈࡿࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊࡈࡼ࡞ࠉA.nidulans ࡚ࡢ
A.oryzae ࡡ MAL ࢠࣚࢪࢰ࣭㐿ఎᏄࡡ࢛࣭ࢮࣞࢡࡢ AmyR ࡞ࡻࡽโᚒࡈࡿ࡙࠷ࡾ࡛ሒ࿈ࡈࡿ࡙࠷ࡾࡆ࡛࠾ࡼࠉ
௧୕ࡡ⤎ᯕࡢࢸࣤࣈࣤࡡฦよ㈠໩࡞ࡗ࠷࡙ࡢ A.oryzae ≺⮤ࡡᶭᵋ࠿Ꮛᅹࡌࡾࡆ࡛ࢅ♟ြࡌࡾࡵࡡ࡚࠵ࡾࠊ
The MAL cluster in Aspergillus oryzae is involved in production of amylolytic enzymes.
Sachiko Hasegawa, Katsuya Gomi (Div.Biotechnol.Future Bioind., Grad.Sch.Agric.Sci., Univ. of Tohoku)
73
P-77
Aspergillus nidulans ◢㓗㑇ඔ㓕⣪㐿ఎᏄࡡ᎒Ẵ᮪௲≁␏Ⓩ࡝㌹෕โᚒᶭᵋࡡよ ᪺
᱕ᑹಆ௒㸡⸠஬㐡ஒ, 㧏ㆺ├ᶖ 㸝➻ἴኬ㝌࣬⏍࿤⎌ሾ㸞
⣊≟Ⳟ Aspergillus nidulans ࡢࠉ࢙ࢰࢿ࣭ࣜࡡ㓗໩࡞ఔࡖ࡙◢㓗ࢅ࢓࣓ࣤࢼ࢓࡫࡛㑇ඔࡊࠉ᎒ẴⓏ࡞ ATP ࢅ
⏍ᠺࡌࡾ(࢓࣓ࣤࢼ࢓Ⓠ㓕)ࠊࡆࡡ㝷ࠉ◢㓗ࡢ◢㓗㑇ඔ㓕⣪(Nar)࠽ࡻࡦல◢㓗㑇ඔ㓕⣪(Nir)࡞ࡻࡽ࢓࣓ࣤࢼ࢓
࡫࡛㑇ඔࡈࡿࡾࠊይẴ᮪௲ୖ࡚ࡢࠉNar 㐿ఎᏄ(niaD)ࡡⓆ⌟ࡢࠉ◢㓗࡞ࡻࡖ࡙ㄇᑙࡈࡿࠉ࢓࣓ࣤࢼ࢓࡝࡜ࡡฺ
⏕ࡊࡷࡌ࠷✽⣪″࡞ࡻࡖ࡙ᢒโࡈࡿࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾ(࢓࣓ࣤࢼ࢓ᢒโ)ࠊୌ᪁ࠉᠻࠍࡢࠉ᎒ẴⓏ࡝࢓࣓ࣤ
ࢼ࢓Ⓠ㓕᮪௲ୖ࡚ࡢࡐࡡᢒโ࠿㉫ࡆࡼ࡝࠷ࡆ࡛ࢅ♟ࡊ࡙ࡀࡒࠊࡐࡆ࡚ࠉᮇ◂✪࡚ࡢࠉniaD ࡡࣈ࣓࣭ࣞࢰ࣭㡷
ᇡ࡛ኬ⭘Ⳟࡡș-࢝ࣚࢠࢹࢨࢱ࣭ࢭ㐿ఎᏄࡡ⼝ྙ㐿ఎᏄࢅష⿿ฺ࣬⏕ࡌࡾࡆ࡛࡞ࡻࡖ࡙ࠉniaD ࡡ✽⣪″࠽ࡻࡦ
㏳Ẵ᮪௲࡞ࡻࡾ㌹෕โᚒᶭᵋࡡよ᪺ࢅモࡲࡒࠊࡐࡡ⤎ᯕࠉ᎒Ẵ᮪௲ୖ࡚ࡢ niaD ࡡⓆ⌟ࡢࠉ✽⣪″࡞ࡻࡾḿ࣬
ㇿࡡโᚒࢅུࡄ࡝࠷ࡆ࡛ࠉ⬲ㄇᑙ࣬⬲ᢒโ᮪௲࡚ࡢይẴ᮪௲ୖࡻࡽࡵᙁ࠷ࡆ࡛࠿♟ࡈࡿࡒࠊࡈࡼ࡞ࠉࣈ࣓ࣞ
࣭ࢰ࣭㡷ᇡ࡞ 10 ⟘ᡜᏋᅹࡌࡾ㌹෕โᚒᅄᏄ AreAp ࡡヾㆉ㒼าࡡ࠹ࡔࠉይẴ᮪௲ୖ࡚ࡡ㌹෕࡞᭩ࡵ㔔さ࡛ࡈ
ࡿ࡙࠷ࡾ㒼า࠿ࠉ᎒Ẵ᮪௲ୖࡡ㌹෕㔖ࡡቌຊ࡞࡛ࡖ࡙ࡵ㔔さ࡚࠵ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊࡱࡒࠉAreAp 㐿
ఎᏄ◒ቪᰬ࡚ࡢࠉ᎒Ẵ᮪௲ୖ࡚ࡡ㌹෕㔖ࡡቌຊ࠿ぜࡼࡿ࡝ࡂ࡝ࡖࡒࠊ࢓࣓ࣤࢼ࢓ᢒโࡢ AreAp ࡡ഼ࡀ࠿㜴ᐐ
ࡈࡿࡾࡆ࡛࡞ࡻࡽ㉫ࡆࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾ࠿ࠉ᎒Ẵ᮪௲ୖ࡚ࡢ AreAp ࡡ഼ࡀ࠿㜴ᐐࡈࡿ࡝࠷ࡒࡴ࡞ࠉ࢓ࣤ
࣓ࢼ࢓ᢒโࢅ࠹ࡄ࡝࠷ྊ⬗ᛮ࠿⩻࠻ࡼࡿࡾࠊ
Gene expression of nitrate reductase under anaerobic conditions in Aspergillus nidulans
Shunsuke Masuo, Tatsuya Fjii, Naoki Takaya
(Graduate School of Life and Environmental Sciences, Univ. of Tsukuba)
P-78
Trichoderma reesei ࡞࠽ࡄࡾ࢞ࢨࣚࢻ࣭ࢭ III 㐿ఎᏄࡡㄇᑙⓆ⌟ᶭᵋࡡよᯊ
ཿᕖ 㝧⣎, ᑚ➗ཋ ῦ, ᒱ⏛ Ᏻᩝ, ᲻ᕖ ᗛ 㸝㛏ᒱᢇ⛁ኬ࣬⏍∸㸞
࠘┘Ⓩ࠙Trichoderma reesei PC-3-7 ࡞࠽࠷࡙࢞ࢨࣚࢻ࣭ࢭ III 㐿ఎᏄ(xyn3) ࡡⓆ⌟ࡢ࢞ࢨࣚࣤ⣌ࡡㄇᑙ∸㈻࡞
ࡻࡖ࡙ㄇᑙࡈࡿࡍࠉࢬ࣭ࣜࣞࢪ࡛ࡐࡡㄇᑙమ࠽ࡻࡦ༟⢶࡚࠵ࡾ L-sorbose ࡞ࡻࡖ࡙ࡡࡲㄇᑙࡈࡿࡾࠊࡆࡡࡒࡴ
xyn3 ࡡⓆ⌟ࡢࢬ࣭ࣜࣚࢭ㐿ఎᏄ⩄࡛ྜྷᵕࡡㄇᑙⓆ⌟ᶭᵋ࡞ࡻࡖ࡙โᚒࡈࡿ࡙࠷ࡾ࡛᥆ᏽࡈࡿࡾࠊᠻࠍࡢ T.
reesei ࡞࠽ࡄࡾࢬ࣭ࣜࣚࢭࡡㄇᑙ⏍⏐࡞㛭ࡌࡾ᩺ࡒ࡝▩ぜࢅᚋࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙ xyn3 ୕Ὦ㡷ᇡ(⣑ 1.1 kb)
ࡡよᯊࢅ㐅ࡴ࡙ࡀࡒࠊࡆࡿࡱ࡚ࡡ◂✪࡞ࡻࡽࠉxyn3 ࡡㄇᑙⓆ⌟࡞ࡢ᪜▩ࡡ㌹෕Ὡᛮ໩㒼าࢅྱࡱ࡝࠷-686㹳
-673 㡷ᇡ࠿᭩ࡵ㔔さ࡝ᙲ๪ࢅᯕࡒࡊ࡙࠷ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖ࡙࠷ࡾࠊᮇ◂✪࡚ࡢࡆࡡ㡷ᇡࢅレ⣵࡞よᯊࡊࠉ
㌹෕Ὡᛮ໩㒼าࡡྜྷᏽࢅモࡲࡒࠊ
࠘᪁Ἢ࡛⤎ᯕ࠙㛜ጙࢤࢺࣤࡡ୕Ὦ-686㹳-673 㡷ᇡහࢅ㏻⤾Ⓩ࡞ን␏ࡈࡎࡒን␏ᆵࡡ୕Ὦ㡷ᇡࢅష⿿ࡊࠉࣝ࣎
࣭ࢰ࣭よᯊ࡞ࡻࡽࡐࡿࡑࡿࡡㄇᑙᛮࢅフ౮ࡊࡒࠊࡐࡡ⤎ᯕࠉ-684ࠤ-673 㡷ᇡහࡡን␏࡞ఔ࠷ㄇᑙᛮࡡ఩ୖ࠿
ふᐳࡈࡿࡒࠊ≁࡞-681 AATTTTGCC-673 㡷ᇡࡡን␏࠿ⴥࡊ࠷఩ୖࢅᘤࡀ㉫ࡆࡊࡒࡆ࡛࠾ࡼࠉࡆࡡ㒼า࠿㔔さ
࡝㌹෕Ὡᛮ໩㒼า࡚࠵ࡾ࡛᥆ᐳࡈࡿࡒࠊT. reesei ࡡࢬ࣭ࣜࣚࢭ㐿ఎᏄ⩄ࡡ୕Ὦ㡷ᇡ࡞ࡢ 5’-GGCAAA-3’ 㒼า
࠿々ᩐᏋᅹࡊ࡙࠷ࡾࡆ࡛࠾ࡼࠉࡆࡡ㒼า࠿ xyn3 ௧አࡡ㐿ఎᏄ࡞࠽࠷࡙ࡵ㌹෕Ὡᛮ໩㒼า࡛ࡊ࡙ᶭ⬗ࡊ࡙࠷ࡾ
ྊ⬗ᛮ࠿♟ြࡈࡿࡒࠊୌ᪁ࠉ-684 AATTTTGCC -673 㒼าࡢࡐࡡ୕Ὦ࡞Ꮛᅹࡌࡾ-701 GGCTATATA -692 㒼า
࡛㏣ྡྷࡀཬᚗ㒼าᵕࡡᵋ㏸ࢅ࡛ࡖ࡙࠽ࡽࠉࡐࡡ㒼า࠽ࡻࡦన⨠㛭౿࠿ T. reesei ࡡ XlnR ࣓࣌ࣞࢡ࡚࠵ࡾ Xyr1
ࡡ⤎ྙ㒼า(5’-GGCTAAN10TTAGCC-3’)࡛ࡻࡂ㢦జࡊ࡙࠷ࡒࠊࡆࡡࡒࡴࠉ஦ࡗࡡ GGC-AT rich ࣓ࢲ࣭ࣆ࠿ୌࡗ
ࡡ㌹෕Ὡᛮ໩㒼า࡛ࡊ࡙ᶭ⬗ࡊ࡙࠷ࡾྊ⬗ᛮࡵ⩻࠻ࡼࡿࡒࠊ⌟ᅹࠉ-701㹳-692 㒼าࡡᶭ⬗࡞ࡗ࠷࡙よᯊࢅ㐅
ࡴ࡙࠷ࡾࠊ
Transcriptional regulation of xyn3 in Trichoderma reesei PC-3-7
Takanori Furukawa, Wataru ogasawara, Hirofumi Okada, Yasushi Morikawa
(Dept.of BioEng., Nagaoka Univ. of Tech.)
74
P-79
Trichoderma reesei ࡞࠽ࡄࡾࢬ࣭ࣜࣚࢭ㐿ఎᏄࡡㄇᑙᶭᵋࡡよᯊ
ᚷ⏛Ὂ௒㸡↻ㆺගᖶ㸡ᑚ➗ཋῦ㸡ᒱ⏛Ᏻᩝ㸡᲻ᕖᗛ㸝㛏ᒱᢇ⛁ኬ࣬⏍∸㸞
[┘Ⓩ] Trichoderma reesei ࡞࠽࠷࡙⛸ࠍࡡࢬ࣭ࣜࣚࢭ㐿ఎᏄࡢㄇᑙ᮪௲ୖ࡚ྜྷㄢࡊ࡙Ⓠ⌟ࡌࡾࠊࡐࡿࡑࡿࡡ
㌹෕㔖ࡢኬࡀࡂ␏࡝ࡾ࠿ࡐࡡ㔖Ẓࡢ⏕࠷ࡒㄇᑙ∸㈻࡞࠾࠾ࢂࡼࡍ࡮࡛ࢆ࡜ୌᏽ࡚࠵ࡾࠊࡐࡡࡒࡴභ㏳ࡡㄇᑙ
Ⓠ⌟ᶭᵋ࠿Ꮛᅹࡌࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊࢬࣞࣄ࢛ࣀ࢕ࢺ࣭ࣞࣚࢭ㐿ఎᏄ (cbh1,cbh2) ࡞㛭ࡊ࡙ᩐ⛸ࡡ㌹෕ㄢ
⟿ᅄᏄ࠿ሒ࿈ࡈࡿ࡙࠷ࡾ࠿ࠉࡌ࡬࡙ࡡࢬ࣭ࣜࣚࢭ㐿ఎᏄࢅྱࡳㄇᑙᶭᵋࡡධᐖよ᪺࡞ࡢ⮫ࡖ࡙࠷࡝࠷ࠊᮇ◂
✪ࡢࢬ࣭ࣜࣚࢭ㐿ఎᏄ⩄ࡡ୯࡚ࡵ㌹෕㔖ࡡᑛ࡝࠷࢙ࣤࢺࢡࣜ࢜ࢻ࣭ࢭ㐿ఎᏄ III(egl3)ࢅ◂✪ᑊ㇗࡛ࡊ࡙ࠉࢬࣜ
࣭ࣚࢭ㐿ఎᏄࡡㄇᑙⓆ⌟ᶭᵋ࡛㌹෕㔖࡞ᙫ㡢ࢅ୙࠻ࡾᅄᏄ࡞ࡗ࠷࡙᩺ࡒ࡝▩ぜࢅᚋࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙࠷ࡾࠊ
[᪁Ἢ࡛⤎ᯕ] cbh1,2, egl1ࠤ5 ࡡ୕Ὦ㡷ᇡࢅẒ㍉よᯊࡊࡒ࡛ࡆࢀࠉT. reesei ࡡࢬ࣭ࣜࣚࢭ㐿ఎᏄࡡ㌹෕Ὡᛮ
໩ᅄᏄ࡚࠵ࡾ ACEII ࡡ⤎ྙ㒼าࡢ㌹෕㔖ࡡኣ࠷ cbh1,2 ࡡ୕Ὦ㒼า࡞ኣࡂᏋᅹࡊ࡙࠷ࡒࠊࡊ࠾ࡊ egl ࡡ୯࡚
㌹෕㔖ࡡኣ࠷ egl1 ࡡ୕Ὦ㡷ᇡ࡞ࡢࡐࡡ┞㛭㛭౿࠿ぜࡼࡿࡍࠉACEII ࡓࡄ࡚ࡢ㌹෕㔖ࡡẒࢅㄕ᪺ࡌࡾࡆ࡛࠿
࡚ࡀ࡝࠷ࡒࡴ௙ࡡᅄᏄࡡᏋᅹ࠿⩻࠻ࡼࡿࡒࠊegl3 ࡡ୕Ὦ㡷ᇡࡡ๎㝎よᯊ࠽ࡻࡦን␏よᯊ࡚ࡢࠉACEII ⤎ྙ㒼
า (GGCTAA) ࡞ຊ࠻࡙㢦జࡊࡒ㒼า (GGCTAT) ࠿ egl3 ࡡㄇᑙⓆ⌟࡞㛭୙ࡊ࡙࠷ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖ
ࡒࠊࡆࡡ ACEII ᵕ㒼าࡢ྘ࢬ࣭ࣜࣚࢭ㐿ఎᏄࡡ୕Ὦ㡷ᇡ࡞ࡵᏋᅹࡊ࡙࠷ࡒࡆ࡛࠾ࡼࠉegl3 ௧አ࡚ࡵᶭ⬗ࡊ
࡙࠷ࡾྊ⬗ᛮ࠿⩻࠻ࡼࡿࡒࠊ࡜ࡡࡻ࠹࡝ᅄᏄ࠿ࡆࡡ ACEII ᵕ㒼า࡞⤎ྙࡌࡾࡡ࠾ࡢ⌟ᅹよᯊ୯࡚࠵ࡾࠊࡱࡒࠉ
egl3 ࡛௙ࡡ㐿ఎᏄࡡ࠷ࡂࡗ࠾ࡡ ACEII ⤎ྙ㒼า࡞ࡢࡐࡡ୕Ὦᩐሲᇱ࡞ࢂࡒࡖ࡙භ㏳ᛮ࠿ぜ࠷ࡓࡈࡿࡒࠊ௑ᚃ
ࡆࡡභ㏳㒼าࢅよᯊࡊ࡙࠷ࡂᚪさ࠿࠵ࡾࠊ
Analysis of induction mechanism of Trichoderma reesei cellulase genes
Yosuke Shida, Katsuyuki Kumagai, Wataru Ogasawara, Hirofumi Okada, Yasushi Morikawa
(Dept. of Bioeng, Nagaoka Univ. of Technol.)
P-80
㯔Ⳟ Aspergillus oryzae ࡞ࡻࡾ cellobiose phosphorylase ࡡ⏍⏐
ኣ⏛ ຉ⏍, 㕝ᮄ ⪵, ᴃᮇ ៿ୌ, ኮ㔕 Ⰳᙢ 1, ఫࠍᮄ ሐ, ᯵ᮄ ㇇ 㸝㣏⥪◂, 1 ಘᕗኬ㝌࣬ᕝ㸞
㸝┘Ⓩ㸞⣵ⳞCellvibrio gilvus⏜ᮮࡡcellobiose phosphorylase㸝CBPase㸞ࡢࢬࣞࣄ࢛࣭ࢪࢅ㐛ࣛࣤ㓗ฦよࡊࠉ
glucose-1-phosphate࡛glucoseࢅ⏍ᠺࡌࡾཬᚺࢅり፳ࡌࡾࠊྜྷ᫤࡞㏣ཬᚺࡵり፳ࡌࡾࡡ࡚ࠉࣉࢷ࢛ࣞࣛࢥ⢶ࢅ⏍
ᠺࡌࡾࡆ࡛࠿ྊ⬗࡚࠵ࡽࠉ᭯⏕࡝㓕⣪࡚࠵ࡾࠊࡊ࠾ࡊࠉC. gilvusᇰ㣬࡚ࡢ⏍⏐ᛮ࠿఩࠷ࡒࡴࠉ㯔ⳞAspergillus
oryzaeࢅ⏕࠷ࡒCBPase⏍⏐⣌ࢅᵋ⠇ࡊࠉ㓕⣪⏍⏐ᛮཀྵࡦࡐࡡᛮ㈻࡞ࡗ࠷࡙᳠ゞࡊࡒࠊ
㸝᪁Ἢ࡛⤎ᯕ㸞C. gilvusࡡCBPaseࡢࠉ⣑90kDaࡡฦᏄ㔖ࢅࡵࡗ㟸ᖏ࡞ኬࡀ࡝㓕⣪࡚࠵ࡽࠉࡱࡒ⣵⬂හ࡞࠽࠷࡙
஦㔖మ࠵ࡾ࠷ࡢᄿ㔖మ࡚Ꮛᅹࡌࡾࡆ࡛࠿ሒ࿈ࡈࡿ࡙࠷ࡾࠊࡐࡆ࡚ࠉamyBࣈ࣓࣭ࣞࢰ࣭ୖὮ࡞CBPase㐿ఎᏄࢅ
㏻⤎ࠉA. oryzae࡞ᑙථࡊࠉ⣵⬂හ⏍⏐ᆵࡡᙟ㈻㌹ᥦᰬࢅཱིᚋࡊࡒࠊᙟ㈻㌹ᥦᰬࡡ⣵⬂᢫ฝᾦ࡞ࡢC.gilvusࡡࡐ
ࡿ࡛Ẓ㍉ࡊ࡙༟న⺦Ⓣ㈻࠵ࡒࡽ⣑10ಶࡡCBPaseὩᛮ࠿Ꮛᅹࡊࠉ⣵⬂හ࡞࠽ࡄࡾCBPaseࡡ⏍⏐࠿☔ヾࡈࡿࡒࠊ
⮫㐲⏍⏐᮪௲࡞ࡗ࠷࡙ࡢࠉ⌟ᅹ᳠ゞ୯࡚࠵ࡾࠊྜྷ᫤࡞ࠉฦἢ⏍⏐ࡡྊ⬗ᛮ࡞ࡗ࠷࡙ࡵ᥀ࡖ࡙࠷ࡾࠊ
Production of cellobiose phosphorylase in Aspergillus oryzae
Sawaki Tada, Satoshi Suzuki, Ken-Ichi Kusumoto, Yoshihiko Amano1, Takashi Sasaki, Yutaka Kashiwagi (NFRI,
1
Grad.Sch.Engneering, Shinshu Univ.)
75
P-81
ᢰᏄⳞ Phanerochaete chrysosporium ࡡࢬࣜ ࣭ࣞࢪฦよ㓕⣪㐿ఎᏄⓆ⌟㔖࡞ࢡࣜࢤ࣭ ࢪ
⃨ᗐ࠽ࡻࡦᇰ㣬᫤㛣࠿୙࠻ࡾᙫ㡢
㕝ᮄୌྍ㸡஫༎ᔋᅽ᪝Ꮔ, 㩢ᓞḿᾀ 㸝᮶ኬ㝌࣬㎨⏍⛁㸞
ᢰᏄⳞ Phanerochaete chrysosporium ࡞ࡻࡾࢬ࣭ࣜࣞࢪฦよᶭᵋ࡞࠽࠷࡙ࡢࠉࢬࣞࣄ࢛ࣃࢺ࣭ࣞࣚࢭࠉࢬࣞ
ࣄ࢛࣭ࢪ⬲Ề⣪㓕⣪ࠉș-ࢡࣜࢤࢨࢱ࣭ࢭ࡛࠷ࡖࡒⳞమአ㓕⣪ࡡ㛭୙࠿♟ࡈࡿ࡙࠷ࡾࠊࡆࡿࡼࡡ㓕⣪ࡡ㐿ఎᏄ
Ⓠ⌟ࣂࢰ࣭ࣤࡢⳞࡡᇰ㣬࡞⏕࠷ࡾ⅛⣪″࡞ࡻࡖ࡙␏࡝ࡾࡆ࡛࠿▩ࡼࡿ࡙࠽ࡽࠉ⅛⣪″ࡡ⛸㢦࠿୙࠻ࡾᙫ㡢࡞
㛭ࡊ࡙ࡢᵕࠍ࡝◂✪࠿⾔ࢂࡿ࡙࠷ࡾࠊ≁࡞ࢡࣜࢤ࣭ࢪࡡᏋᅹ࡞ࡻࡖ࡙ࡆࡿࡼࡡ㐿ఎᏄⓆ⌟ࡡࣛࣈࣝࢴࢨࣘࣤ
࠿⏍ࡋࡾࡆ࡛࠿ࡻࡂ▩ࡼࡿ࡙࠷ࡾࠊࡊ࠾ࡊࠉࢡࣜࢤ࣭ࢪࡡ⃨ᗐ࠿ࣛࣈࣝࢴࢨࣘࣤ࡞୙࠻ࡾᙫ㡢ࡷࠉࢡࣜࢤ࣭
ࢪࢅ⅛⣪″࡛ࡊࡒሔྙࡡᇰ㣬᫤㛣࡞ࡻࡾⓆ⌟㔖ࡡᕣ࡞㛭ࡌࡾ᳠ゞࡢࡆࡿࡱ࡚༎ฦ࡞࡝ࡈࡿ࡙࠷࡝࠷ࠊࡐࡆ࡚
ᮇ◂✪࡚ࡢࠉP. chrysosporium ࡞␏࡝ࡾ⃨ᗐࡡࢡࣜࢤ࣭ࢪࢅ୙࠻ࡒሔྙࡡࠉࢬ࣭ࣜࣞࢪฦよ㓕⣪㐿ఎᏄࡡⓆ⌟
㔖ࡡ⤊᫤Ⓩን໩ࢅࣛ࢓ࣜࢰ࢕࣑ᏽ㔖 PCR ࡞ࡻࡖ࡙よᯊࡊࡒࠊࡱࡒ P. chrysosporium ࡡࢣࢿ࣑᝗ሒࢅฺ⏕ࡊࠉ
㐿ఎᏄⓆ⌟ㄢ⟿ᶭᵋ࡞ࡗ࠷࡙ࡡ▩ぜࢅᚋࡾࡒࡴ྘㐿ఎᏄࡡࣈ࣓࣭ࣞࢰ࣭୕࡞࠽ࡄࡾ㌹෕ㄢ⟿ᅄᏄ⤎ྙ㒂నࡡ
よᯊࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉ5000ȣM ࠵ࡾ࠷ࡢ 500ȣM ┞ᙔࡡࢡࣜࢤ࣭ࢪࢅ῟ຊࡊࡒሔྙ࡞ࡢࠉ῟ຊᚃ 6 ᫤㛣
ࡱ࡚࡞㐿ఎᏄⓆ⌟࠿ᢒโࡈࡿࡾ࠿ࠉ50ȣM ┞ᙔࡡࢡࣜࢤ࣭ࢪࢅ῟ຊࡊࡒሔྙ࡞ࡢࠉ῟ຊᚃ 6 ᫤㛣ࡱ࡚ࡢⓆ⌟
㔖࠿ᑊᩐⓏ࡞ቌຊࡌࡾࡆ࡛࠿♟ࡈࡿࡒࠊࡱࡒ྘㐿ఎᏄࡡࣈ࣓࣭ࣞࢰ࣭୕࡞ᏄᄙⳞࡡࢡࣜࢤ࣭ࢪࣛࣈࣝࢴࢦ࣭
⤎ྙ㒂న࡛┞ྜྷ࡝㡷ᇡࢅぜฝࡊࡒࠊ
Glucose concentration and cultivation time affected gene expression of cellulolytic enzymes in the basidiomycete
Phanerochaete chrysosporium
Hitoshi Suzuki, Kiyohiko Igarashi, and Masahiro Samejima
(Univ. of Tokyo)
P-82
Cluster analysis of microarray gene expression data during growth of Aspergillus oryzae in
cellulose substrate medium
Avid SHIAU, Praveen Rao JUVVADI, Jun-ichi MARUYAMA, Manabu ARIOKA, Katsuhiko KITAMOTO (Dept. of
Biotechnology, Univ. of Tokyo)
The filamentous fungus, Aspergillus oryzae, has been widely recognized as a safe organism and its importance for the
food industry, such as for the production of useful enzymes has been well established. The recent release of its complete
genome sequence has attracted fungal biologists and prompts investigations into its potential to be utilized for wider
biotechnological applications. We have designed custom array (Affymetrix Inc.) consisting of 12,050 genes from A.
oryzae. To understand the cellular machinery of biomass degradative processes/potential in A. oryzae, we studied the
genome wide expression data during its growth on cellulose as a substrate. A 24 h pre-grown culture of A. oryzae
RIB40 strain in Czapek’s Dox (CD) medium was shifted to CD medium supplemented with 2% Avicel and cultured for
a further period of 8 h.
The labeled cRNA prepared from the total RNA was hybridized onto the A. oryzae microarray.
The genome wide expression data obtained was processed and compared to similar data originating from RIB40 grown
on CD-Glucose medium. Of the 12,050 genes examined, a positive expression profile consisting of 3,454 upregulated
genes was produced. At the same time, data was retrieved using the comprehensive glucoside hydrolase database at
CAZY and 52 predicted cellulolytic genes (cellobiohydrolases, endoglucanases and beta-glucosidases) were identified
in A. oryzae. Fifteen of these hydrolases were found to be at least 2x upregulated in the positive expression profile.
Within this subset, the promoter of each gene will be analyzed for the cis-element binding sequences that are commonly
found in promoter regions of fungal hydrolytic enzymes.
76
P-83
Aspergillus oryzae ㌹෕ᅄᏄ AoXlnR ᨥ㒼ୖ࡞࠵ࡾኣ⢶ฦよ㓕⣪㐿ఎᏄ⩄ࡡྜྷᏽ
㔕ཾ♰ྒྷ㸡ఫ㔕ඔ᫓*㸡ຊ⸠㞖ኃ㸡ᑚᯐူኰ 㸝ྞኬ㝌⏍࿤㎨࣬⏍∸ᶭᵋ㸡*㔘Ἁᕝኬ㸞
࠘┘Ⓩ࠙ 㯔Ⳟ Aspergillus oryzae ࡢ᳔∸ᛮኣ⢶ฦよ⬗࡞ඁࡿ㸡ᙁງ࡝࢞ࢨࣚࢻ࣭ࢭ㸡ࢬ࣭ࣜࣚࢭ㐿ఎᏄ⩄ࢅ᭯
ࡊ࡙࠷ࡾ㸣ᙔ◂✪ᐄ࡚ࡢ㸡A. oryzae ࠿᭯ࡌࡾࡆࡿࡼ㐿ఎᏄ⩄ࡡᕝᴏⓏฺ⏕ࢅ┘Ⓩ࡛ࡊ࡙ AoXlnR ࢅ༟㞫ࡊࡒ㸣
ࡆࡿࡱ࡚࡞ AoXlnR ࡢ㸡࢞ࢨࣚࣤࡷ࢞ࢨ࣭ࣞࢪ࡞⏜ᮮࡌࡾࢨࢡࢻࣜ࡞ᚺ➽ࡊ࡙࢞ࢨࣚࢻ࣭ࢭ㸡ࢬ࣭ࣜࣚࢭ㐿
ఎᏄ⩄ࡡ㌹෕ࢅὩᛮ໩ࡌࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖ࡙࠷ࡾ㸣௑ᅂ㸡DNA ࣏࢕ࢠࣞ࢓ࣝ࢕よᯊ࡞ࡻࡖ࡙㸡AoXlnR
࠿㌹෕โᚒࡊ࡙࠷ࡾ㐿ఎᏄ⩄ࡡᏰධྜྷᏽࢅモࡲࡒ㸣
࠘᪁Ἢ࠙ DNA ࣏࢕ࢠࣞ࢓ࣝ࢕よᯊ࡞౩ࡌࡾⳞᰬ࡞ࡢ㸡ᵋᠺⓏ㧏⏍⏐ࣈ࣓࣭ࣞࢰ࣭࡚࠵ࡾ TEF1 ࣈ࣓࣭ࣞࢰ࣭
ࢅ⏕࠷ࡒ AoXlnR 㧏⏍⏐ᰬ࡛ AoXlnR ◒ቪᰬࢅ㐽ᢝࡊࡒ㸣྘Ⳟᰬࢅᇰ㣬ࡊ㸡Ⳟమࢅᅂ཭࣬ὑὯᚃ㸡0.1㸚⃨ᗐ
࢞ࢨ࣭ࣞࢪࢅྱࡳㄇᑙ᮪௲ୖࡡᾦమᇰᆀ࡚᭞࡞ 30 ฦ㛣ᇰ㣬ࡊࡒ㸣ᚋࡼࡿࡒⳞమ࠾ࡼ mRNA ࢅ᢫ฝࡊ㸡ㄇᑙ
᮪௲ୖ࡚ࡡ㐿ఎᏄⓆ⌟ࣈࣞࣆ࢒࢕ࣜࢅ DNA ࣏࢕ࢠࣞ࢓ࣝ࢕࡞ࡻࡽよᯊࡊࡒ㸣DNA ࣏࢕ࢠࣞ࢓ࣝ࢕よᯊࡡ⤎
ᯕ࠾ࡼ㸡AoXlnR 㧏⏍⏐ᰬ࡚Ⓠ⌟㔖࠿ኣ࠷㓕⣪㐿ఎᏄ⩄ࢅ᢫ฝࡊࡒ㸣ࡐࡡ⤎ᯕ㸡᪜࡞ AoXlnR ᨥ㒼ୖ࡚࠵ࡾࡆ
࡛࠿♟ࡈࡿ࡙࠷ࡾ 4 ಴ࡡ࢞ࢨࣚࢻ࣭ࢭ㸡4 ಴ࡡࢬ࣭ࣜࣚࢭ㐿ఎᏄ࡞ຊ࠻㸡᩺ࡒ࡞㸡6 ಴ࡡ࢞ࢨࣚࢻ࣭ࢭ㐿ఎᏄ㸡
9 ಴ࡡࢬ࣭ࣜࣚࢭ㐿ఎᏄ࠿ྜྷᏽࡈࡿࡒ㸣ࡈࡼ࡞㸡11 ಴ࡡ࢞ࢨࣚࣤഁ㙈ฦよ㓕⣪㐿ఎᏄ㸡3 ಴ࡡ࢞ࢨ࣭ࣞࢪ௥
ㅨ⣌㓕⣪㐿ఎᏄࡡⓆ⌟㔖ࡵቌኬࡊ࡙࠷ࡒ㸣௧୕࠾ࡼ㸡࢞ࢨࣚࣤ㹺ࢬ࣭ࣜࣞࢪฦよ࡞ᚪさ࡝ධ࡙ࡡ㓕⣪㐿ఎᏄ࠿
AoXlnR ࡡโᚒୖ࡞࠵ࡾࡆ࡛࠿☔ヾࡈࡿࡒ㸣
Identification of xylanolytic and cellulolytic enzyme genes under control of AoXlnR.
Yuji Noguchi, Motoaki Sano,* Masashi Kato, and Tetsuo Kobayashi
(Grad. Sch. of Bioagricultural Sci., Nagoya Unv., and *Kanazawa Inst. Tech.)
P-84
Trichoderma reesei ⏜ᮮࢬ࣭ࣜࣚࢭ ࡡᶭ⬗ᨭⰃ
୯⃕ ක㸡ᒱ⏛ ࿰ኃ, ⴘྙ ఘᗽ, ᑚ㔕ᑈ ᭰Ꮔ, ᑚ➗ཋ ῦ, ᒱ⏛ Ᏻᩝ, ᲻ᕖ ᗛ
㸝㛏ᒱᢇ⛁ኬ࣬⏍∸㸞
ᠻࠍࡢࠉࢬ࣭ࣜࣚࢭ㧏⏍⏐Ⳟ Trichoderma reesei ࡡࢬ࣭ࣜࣞࢪฦよ⬗ງࢅࡈࡼ࡞㧏ࡴࡾࡒࡴ࡞ࠉT.reesei ࡞
ࡻࡽ⏍⏐ࡈࡿࡾ಴ࠍࡡࢬ࣭ࣜࣚࢭ㸝Cellobiohydrolase㸝CBH㸞I,II,࠽ࡻࡦ Endoglucanase(EG) I,II,III㸞࡞ࡗ࠷࡙
り፳⬗ງ࠽ࡻࡦᏭᏽᛮࢅྡྷ୕ࡈࡎࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙㐅໩ฦᏄᕝᏕⓏ࡞ᨭንࡊ࡙࠷ࡾࠊ
ࡆࡡᨭንᡥἪ࡚ࡢ
ࣚࣤࢱ࣑ን␏㐿ఎᏄࣚ࢕ࣇ࣭࡚ࣚࣛᐙ୹ࢅᙟ㈻㌹ᥦࡊࠉᚋࡼࡿࡒኣᩐࡡን␏ᰬ࠾ࡼᨭⰃ㓕⣪Ⓠ⌟ᰬࢅ㐽ᢜࡌ
ࡾࡆ࡛࠿ᚪ㡪࡛࡝ࡾࠊࡐࡡࡒࡴ㧏࠷ᙟ㈻㌹ᥦຝ⋙ࢅ᭯ࡌࡾ Escherichia coli ࠽ࡻࡦ Saccharomyces cerevisiae ࢅ
ᐙ୹࡞⏕࠷࡙ࡱࡍࡢⓆ⌟ࡡྊྫྷࢅ☔ヾࡊࡒࠊ
ࡆࡡ࠹ࡔ E.coli ࡚ࡡⓆ⌟࡞ᠺຉࡊࡒ EG II ࠽ࡻࡦ EG III ࡞ࡗ࠷࡙
ᨭንࢅ⾔ࡖࡒ࡛ࡆࢀࠉEG III ࡞࠽࠷࡙ᇰ㣬ᾦᙔࡒࡽࡡὩᛮೋ࡚⣑ 2-5 ಶࡡྡྷ୕࠿ぜࡼࡿࡒࠊS.cerevisiae ࡚Ⓠ
⌟ࡌࡾ EG I ࡞ࡗ࠷࡙ࡢ⣑ 2 ಶࡡὩᛮྡྷ୕࠿ぜࡼࡿࡒࠊ⌟ᅹὩᛮ࠿୕᪴ࡊࡒᨭⰃ㓕⣪࡞ࡗ࠷࡙ẒὩᛮࠉᏭᏽᛮ
࠽ࡻࡦᙟ㈻㌹ᥦమࡡⓆ⌟㔖ࡡレ⣵࡝よᯊࢅ⾔ࡖ࡙࠷ࡾࠊ
Directed evolution of cellulases from Trichoderma reesei
Hikaru Nakazawa, Katsunori Okada, Nobuhiro Ochiai, Tomoko Onodera, Wataru Ogasawara, Hirofumi Okada,
Yasushi Morikawa. (Bioeng., Nagaoka Univ.Tech.)
77
P-85
Aspergillus aculeatus ⏜ᮮ aviIII 㐿ఎᏄࡡ A.aculeatus ࢅᐙ୹࡛ࡊࡒ㧏Ⓠ⌟
ᯀ᮶ኚᾇ㸡㔘ᨳ┷㸟ࠉㆺಞ἖ࠉ⅛ㆺ㡨ୌࠉⲠ஬ᇱኰ㸟㸟ࠉᕖཾ๙ྒྷ 㸝㜨ᗋኬ㝌࣬ᚺ⏍⛁ࠉ㸟㟴ᒱኬ㎨࣬ᚺ⏍⛁ࠉ
㸟㸟
୯㒂ኬ࣬ᚺ⏍㸞
㎾ᖳࠉ࢙ࢾ࣭ࣜ࢟༱ᶭ࡞ఔ࠷ࣁ࢕࢛࣏ࢪฺ⏕࠿Ἰ┘ࡈࡿ࡙࠷ࡾࠊᮇ◂✪ᐄ࡚༟㞫ࡈࡿࡒ A. aculeatus ࡢᙁ࠷
ࢬ࣭ࣜࣚࢭὩᛮࢅᣚࡔࠉࣁ࢕࢛࣏ࢪ᭯ຝฺ⏕࡞ኬࡀࡂ㈁⊡ࡌࡾࡵࡡ࡛ᛦࢂࡿࡾࠊࣁ࢕࢛࣏ࢪฺ⏕࡞ࡢ୹ᠺฦ
࡚࠵ࡾࢬ࣭ࣜࣞࢪࡡྱࡱࡿࡾࠉᙁᅖ࡞Ề⣪⤎ྙࡊࡒ⤎ᬏᛮࢬ࣭ࣜࣞࢪࡡฦよ࠿ၡ㢗࡛࡝ࡖ࡙࠷ࡾࠊࡐࡆ࡚௧
๑ࠉ⤎ᬏᛮࢬ࣭ࣜࣞࢪฦよ࡞᭯⏕࡝࢙࢞ࢮࢡࣜ࢜ࢻ࣭ࢭ㐿ఎᏄ㸝aviIII㸞ࢅྜྷᒌࡡ A. oryzae ࠉ㓕ẍ࡚㧏Ⓠ⌟
ࡌࡾ஥ࢅモࡲࡒ࠿᭯ណࡡ⤎ᯕࢅ♟ࡈ࡝࠾ࡖࡒࡒࡴࠉ௑ᅂࠉᙔ◂✪ᐄ࡚㛜ⓆࡊࡒඔⳞᰬ࡚࠵ࡾ A. aculeatus ࡡᐙ
୹࣊ࢠࢰ࣭⣌ࢅ⏕࠷࡙㧏Ⓠ⌟ࢅ⾔࠷ࠉᛮ㈻Ửᏽࢅ⾔ࡖࡒࠊ
⅛⣪″࡞ᙫ㡢ࡈࡿࡍᵋᠺⓏ࡝Ⓠ⌟ࢅྊ⬗࡞ࡌࡾࠉࣈ࣓࣭ࣞࢰ࣭P-No8142 ࡡୖὮ࡞ aviIII 㐿ఎᏄࢅ㏻⤎ࡊ A.
aculeatus ࡫ᑙථࡊࡒ⤎ᯕࠉaviIII 㧏Ⓠ⌟ᰬࢅཱིᚋࡌࡾࡆ࡛࡞ᠺຉࡊࡒࠊࡱࡒࠉᚋࡼࡿࡒᙟ㈻㌹ᥦమࡡᇰ㣬᮪௲
ࢅ᳠ゞࡊࠉOctyl-Sepharose ࢅ⏕࠷ࡒ␧Ềࢠ࣏ࣞࢹࢡࣚࣆ࢔࣭࡞౩ࡊࠉࡈࡼ࡞ࠉSP-TOYOPEARL 650M ࢅ⏕࠷
ࡒ㝟࢕࢛ࣤஹᥦࢠ࣏ࣞࢹࢡࣚࣆ࢔࣭࡞౩ࡌ஥࡚⢥⿿ࢅ⾔ࡖࡒࠊᚋࡼࡿࡒ⢥⿿ࢦࣤࣈࣜࢅ⏕࠷࡙ࠉ⮫㐲Ὼᗐࠉ
ῺᗐᏭᏽᛮࠉ⮫㐲 pH ࠉpH Ꮽᏽᛮࠉࡱࡒࠉᇱ㈻≁␏ᛮ࡞ࡗ࠷࡙ࡵㄢ࡬ࡒࠊ⌟ᅹࠉTLC ࡷ HPLC ࡞ࡻࡽཬᚺ
⏐∸ࢅよᯊࡊࠉࡆࡡ㓕⣪ࡡレࡊ࠷ᛮ㈻ࢅㄢ࡬࡙࠷ࡾࠊ
Overexpression of aviIII gene from Aspergillus aculeatus in A.aculeatus
Natsumi Bando, Shin Kanamasa*, Syuji Tani, Jun-ichi Sumitani, Motoo Arai**, Takashi Kawaguchi
(Graduate School of Life & Environmental Sociences, Osaka Prefecture Univ. , *Faculty of Agriculture, Univ. of
Shizuoka , and **Dept. of Biological Chemistry, Univ. of Chubu)
P-86
Aspergillus aculeatus ⏜ᮮș-࢞ࢨࣞࢨࢱ࣭ࢭ㐿ఎᏄࡡ A. oryzae ࡞࠽ࡄࡾⓆ⌟
ᑚこ 㐡ஒ㸡⏛ᑹ⸓Ꮔ㸡ㆺ ಞ἖㸡⅛ㆺ 㡨ୌ㸡Ⲡ஬ ᇱኰ 1㸡ᕖཾ ๙ྒྷ㸝㜨ᗋኬ㝌࣬ᚺ⏍⛁㸡1:୯㒂ኬ࣬ᚺ⏍㸞
㸱┘Ⓩ㸳ᙔ◂✪ᐄ࡚༟㞫ࡈࡿࡒ A. aculeatus ࡢࣉ࣐ࢬ࣭ࣜࣞࢪࢅຝ⋙ࡻࡂฦよࡌࡾࠊᮇⳞᇰ㣬୕ΰ୯࠾ࡼ࢞
ࢨࣚࣤฦよ㓕⣪࡛ࡊ࡙ 3 ⛸ࡡ࢞ࢨࣚࢻ࣭ࢭࠉཀྵࡦ 1 ⛸ࡡ࢞ࢨࣞࢨࢱ࣭ࢭ࠿⢥⿿ࡈࡿ࡙࠷ࡾࠊࡆࡡ࠹ࡔࠉ࢞ࢨ
ࣞࢨࢱ࣭ࢭ࡞ࡗ࠷࡙ࡢࠉಕᏋ㡷ᇡ࠾ࡼスゝࡊࡒࣈࣚ࢕࣏࣭ࢅ⏕࠷ࡒ PCR ࠾ࡼࠉ
2 ⛸ࡡ␏࡝ࡖࡒ㒼าࢅᣚࡗ DNA
᩷∞࠿ᚋࡼࡿࠉ2 ⛸ࡡ࢞ࢨࣞࢨࢱ࣭ࢭ㐿ఎᏄ࠿ࢠ࣭ࣞࢼࣤࢡࡈࡿ࡙࠷ࡾࠊᮇ◂✪࡚ࡢ 2 ⛸ࡡ㐿ఎᏄ࠿ᶭ⬗Ⓩ
࡝࢞ࢨࣞࢨࢱ࣭ࢭࢅࢤ࣭ࢺࡊ࡙࠷ࡾࡡ࠾࡜࠹࠾ࢅ᪺ࡼ࠾࡞ࡌࡾ࡛භ࡞ࠉ⢥⿿ῥࡲࡡ㓕⣪࠿࡜ࡔࡼࡡ㐿ఎᏄ࡞
ࢤ࣭ࢺࡈࡿ࡙࠷ࡾ࠾ࢅྜྷᏽࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊࡒࠊ㸱᪁Ἢ࣬⤎ᯕ㸳2 ⛸ࡡ࢞ࢨࣞࢨࢱ࣭ࢭ㐿ఎᏄ㸝xyl 1, xyl 2㸞
ࢅࡐࡿࡑࡿ⣊≟Ⳟ⏕㧏Ⓠ⌟࣊ࢠࢰ࣭ pNAN8142 ࡞᣼ථࡊ niaD ࢅ㐽ᢝ࣏࣭࣭࡛࢜ࡊ࡙ A. aculeatus ࡞ᑙථࡊࡒࠊ
ᚋࡼࡿࡒᙟ㈻㌹ᥦᰬ࡞ࡗ࠷࡙ࠉ4MU-xyl ࢅ⏕࠷ࡒࣈ࣭ࣝࢹ࢓ࢴࢬ࢕ࡷࠉᇰ㣬୕ΰ୯ࡡ࢞ࢨࣞࢨࢱ࣭ࢭὩᛮࢅ
pNP-xyl ࢅ⏕࠷࡙ῼᏽࡌࡾࡆ࡛࡞ࡻࡽ୦㐿ఎᏄ࡛ࡵ࡞ᶭ⬗Ⓩ࡝࢞ࢨࣞࢨࢱ࣭ࢭࢅࢤ࣭ࢺࡊ࡙࠷ࡾࡆ࡛ࢅ᪺ࡼ
࠾࡞ࡊࡒࠊࡱࡒࠉSDS-PAGE ࡞ࡻࡖ࡙Ⓠ⌟⏐∸ࡡ᳠ฝࢅ⾔ࡖࡒ࡛ࡆࢀࠉXYL 1 ࠽ࡻࡦ XYL 2 ୦⩽࡛ࡵ᪺☔
࡝Ⓠ⌟⏐∸ࡡࣁࣤࢺ࠿᳠ฝࡈࡿࡒ࠿ࠉ௛ຊࡌࡾ⢶㙈ࡡ㐢࠷࠾ࡼ࠾ࠉࡐࡡฦᏄ㔖ࡢඔⳞᰬ࠾ࡼ⢥⿿ࡈࡿ࡙࠷ࡾș-࢞ࢨࣞࢨࢱ࣭ࢭࡡฦᏄ㔖࡛ୌ⮬ࡊ࡝࠾ࡖࡒࠊࡐࡆ࡚ࠉ୦Ⓠ⌟⏐∸ࢅ⢥⿿ࡊࠉ಴ࠍࡡᛮ㈻ࢅỬࡴ࡙Ẓ㍉ࡌࡾ
ࡆ࡛࡞ࡊࡒࠊ⮫㐲Ὼᗐࠉ⮫㐲 pH ࠉ⇍ᏭᏽᛮࠉpH ᏭᏽᛮࠉẒὩᛮ࡞ࡗ࠷࡙ㄢ࡬ࡒ࡛ࡆࢀࠉXYL 2 ࡡ᪁࠿ඔⳞ
ᰬ࡚⢥⿿ࡈࡿ࡙࠷ࡾ㓕⣪࡛㎾࠷ೋࢅ♟ࡊࡒࠊࡻࡖ࡙ࠉ4 ࡗࡡᛮ㈻࡛ẒὩᛮ࠾ࡼ XYL 2 ࠿ඔⳞᰬ࡚⢥⿿ࡈࡿࡒ
ș-࢞ࢨࣞࢨࢱ࣭ࢭ࡚࠵ࡾྊ⬗ᛮ࠿ᙁࡂ♟ြࡈࡿࡒࠊ
Expression of the ș-Xylosidase Gene from Aspergillus aculeatus in A. oryzae
Tatsuya Konishi, Aiko Tao, Shuji Tani, Jun-ichi Sumitani, Motoo Arai, Takashi Kawaguchi
(Dept. of Applied Biological Chemistry, Univ. of Osaka Prefecture, and Dept. of Biological Chemistry, Univ. of chubu)
78
P-87
ࣃࢣ࢜ࣄ࢞ࢲࣤࢸ࢓ࢬࢲ࣭ࣚࢭ㐿ఎᏄࡡᕝᏕⓏฺ⏕
⡷ᮟ᫥Ꮔ㸡ᮟᒜ⫍Ꮔ 㸝㛭᮶Ꮥ㝌ኬ࣬ᕝ࣬∸㈻⏍࿤⛁Ꮥ㸞
ິ∸ᛮ㣏∸⦼⥌ࡡࡥ࡛ࡗ࡚࠵ࡾ࢞ࢲࣤࡢᆀ⌣᭩ᚃࡡࣁ࢕࢛࣏ࢪ࡛ゕࢂࡿࠉࣁ࢕࢛࣏࣭࡛࣎ࣛࡊ࡙ࡢࢬࣜࣞ
࣭ࢪ࡞ࡗ࠷࡚ኣ࠷ࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾ࠿ࠉ࢞ࢲࣤࢅ⬲࢓ࢬࢲࣜ໩ࡊ࡙ᚋࡼࡿࡾ࢞ࢹࢦࣤࡢࠉ࢞ࢲࣤࡻࡽࡵỏ
⏕ᛮ࠿㧏ࡂࠉ༈⒢ᮞᩩࡷᶭ⬗ᛮ㣏ဗࠉ໩⢕ဗࠉᅰቫᨭⰃ๠࡛ࡊ࡙⏕࠷ࡼࡿࠉᖳࠍࡐࡡ㞺さ࠿㧏ࡱࡖ࡙࠷ࡾࠊ
࢞ࢹࢦࣤࡢ⏍∸㈠″⏜ᮮࡡཋᩩࡻࡽ⏍⏐ࡈࡿࡾࡒࡴࠉ↋ᐐ࡚Ꮽධᛮ࠿㧏ࡂࠉ㈠″ᯜῤࡡྊ⬗ᛮ࠿఩ࡂࠉ⏍
∸ฦよᛮ࡚࠵ࡾ࡛࠷࠹≁ᚡ࠿࠵ࡾࠊ⌟ᅹ࢞ࢹࢦࣤࡡᕝᴏⓏ⏍⏐࡞ࡢࠉ࢞ࢲࣤࢅ㧏⃨ᗐࡡ࢓ࣜ࢜ࣛฌ⌦࡞ࡻࡖ
࡙⬲࢓ࢬࢲࣜࡌࡾ᪁Ἢ࠿⏕࠷ࡼࡿ࡙࠷ࡾ࠿ࠉࡐࡡ࡛ࡀ࡞⏍ࡋࡾ㧏⃨ᗐ࢓ࣜ࢜ࣛᗣᾦࡢ⎌ሾ◒ቪࡡཋᅄ࡛࡝ࡖ
࡙࠷ࡾࠊࡐࡡࡒࡴࠉ㧏⃨ᗐ࢓ࣜ࢜ࣛฌ⌦Ἢ࡞௥ࢂࡾ᪁Ἢ࠿⎌ሾࢅ㒼៎ࡌࡾ୕࡚㔔さ࡚࠵ࡾ࠿ࠉᙔ◂✪ᐄ࡚ࡢ
࢞ࢲࣤࢅ⬲࢓ࢬࢲࣜ໩ࡊ࡙࢞ࢹࢦࣤࢅ⏍ᠺࡌࡾ㓕⣪ࠉ࢞ࢲࣤࢸ࢓ࢬࢲ࣭ࣚࢭ࡞╌┘ࡊ࡙࠷ࡾࠊ
᥃ྙⳞ㢦ࡡୌ⛸࡚࠵ࡾࣃࢣ࢜ࣄࡢ⣵⬂ባࡡ୹さᠺฦ࡛ࡊ࡙࢞ࢲ࡛ࣤ࢞ࢹࢦࣤࢅྱࡲࠉ࢞ࢲࣤ࠾ࡼ⬲࢓ࢬࢲ
ࣜ໩ࢅ⾔࠹㓕⣪ࠉ࢞ࢲࣤࢸ࢓ࢬࢲ࣭ࣚࢭࢅᣚࡗⳞ㢦ࡡୌࡗ࡚࠵ࡾࠊࡊ࠾ࡊࣃࢣ࢜ࣄࢅኬ㔖࡞ᇰ㣬ࡊ࡙ࡵᚋࡼ
ࡿࡾ㓕⣪ࡢࢂࡍ࠾࡚࠵ࡾࡒࡴࠉࣃࢣ࢜ࣄࡡ࢞ࢲࣤࢸ࢓ࢬࢲ࣭ࣚࢭ㐿ఎᏄࢅࢠ࣭ࣞࢼࣤࢡࡊ࡙ࠉࡆࡡࢠ࣭ࣞࣤ
ࢅࢤࢗࢩ࢜ࣄ࡞ᑙථࡊࠉࢤࢗࢩ࢜ࣄ࡞࢞ࢲࣤࢸ࢓ࢬࢲ࣭ࣚࢭࢅኬ㔖࡞Ⓠ⌟ࡈࡎࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙◂✪ࡊ࡙
࠷ࡾࠊ
Industrial application of chitin deacetylase gene from Phycomyces blakesleeanus
Akiko Yonemura, Tadako Murayama
(Dept.Mater.Life Sci.,Kanto Gakuin Univ.)
P-88
⣊≟Ⳟ Aspergillus nidulans ࡡ࣐࢛ࢨࣤᵕࢺ࣒࢕ࣤࢅᣚࡗ࢞ࢲࣤྙᠺ㓕⣪
CsmAࠉCsmB ࡡࢺ࣒࢕ࣤ㛣ࡡᶭ⬗Ⓩᕣ␏ ࡡよᯊ
ᑘᓧ┷ᴃࠉ➁ୖ඼⏠ࠉᇷහ⿩஄ࠉኯ⏛᪺ᚠ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
࢞ࢲࣤࡢ⣊≟Ⳟ⣵⬂ባࡡ୹さᵋᠺᠺฦࡡୌࡗ࡚࠵ࡽࠉࡐࡡ⏍ྙᠺࡢᙟឺᙟᠺࠉฦ໩࡞㔔さ࡝ᙲ๪ࢅᣚࡗࡆ
࡛࠿♟ࡈࡿ࡙࠷ࡾࠊA. nidulans ࡞ࡢࠉN ᮆ❻ഁ࡞࣐࢛ࢨ࡛ࣤ┞ྜྷᛮࢅ♟ࡌࢺ࣒࢕ࣤ (MMD) ࠉC ᮆ❻ഁ࡞࢞
ࢲࣤྙᠺ㓕⣪ࢺ࣒࢕ࣤ (CSD) ࢅᣚࡗ࢞ࢲࣤྙᠺ㓕⣪ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ࠿ csmAࠉcsmB ࡡ஦⛸Ꮛᅹࡌࡾࠊࡆ
ࡿࡱ࡚࡞ࠉࡐࡿࡑࡿࡡ༟≺◒ቪᰬࡡ⾪⌟ᆵࡡ㢦జᛮࠉ஦㔔◒ቪࡡྙᠺ⮬Ṓᛮ࠿♟ࡈࡿ࡙࠷ࡾࠊୌ᪁ࠉcsmB ◒
ቪᰬ࡞࠽࠷࡙ csmA ࢅ㧏Ⓠ⌟ࡈࡎ࡙ࡵࠉcsmB ࡡ◒ቪ࡞ࡻࡾ␏ᖏ࠿ᅂᚗࡊ࡝࠷ࡆ࡛࠾ࡼࠉ୦⩽ࡡᶭ⬗Ⓩᕣ␏ࡵ
♟ြࡈࡿ࡙࠷ࡾࠊࡱࡒ CsmAࠉCsmB ࡢ࡛ࡵ࡞Ⳟ⣊඙❻ࡷ㝰ባᙟᠺ㒂న࡞ᑻᅹ໩ࡊ࡙࠽ࡽࠉࡐࡿࡑࡿࡡ MMD
࡛࢓ࢠࢲࣤ⣵⬂㦭᰹࡛ࡡ┞பష⏕࠿♟ြࡈࡿ࡙࠷ࡾ 1,2)ࠊ௑ᅂࠉ୦⩽ࡡᶭ⬗Ⓩᕣ␏࡞㛭ࢂࡾࢺ࣒࢕ࣤࢅ᳠ゞࡌ
ࡾࡒࡴࠉcsmB ◒ቪᰬ࡞࠽࠷࡙ࠉCsmA ࡡ MMD ࡛ CsmB ࡡ CSD ࡡ⼝ྙࢰࣤࣂࢠ㈻ MACBࠉ࠵ࡾ࠷ࡢ CsmB
ࡡ MMD ࡛ CsmA ࡡ CSD ࡡ⼝ྙࢰࣤࣂࢠ㈻ MBCA ࢅ㔕⏍ᆵ CsmB ࡡ௥ࢂࡽ࡞Ⓠ⌟ࡈࡎࡒ࡛ࡆࢀࠉ๑⩽࡚ࡢ
csmB ࡡ◒ቪ࡞ࡻࡾ⏍⫩ࡡ㐔ᘇࡷᙟឺ␏ᖏ࠿ᅂᚗࡊࠉᚃ⩽࡚ࡢᅂᚗࡊ࡝࠾ࡖࡒࠊࡆࡡࡆ࡛࠾ࡼࠉCsmA ࡡ MMD
ࡢ CsmB ࡡ MMD ࡡᶭ⬗ࢅ௥᭨࡚ࡀࡾ࠿ࠉCsmA ࡡ CSD ࡢ CsmB ࡡ CSD ࡡᶭ⬗ࢅ௥᭨࡚ࡀ࡝࠷ࡆ࡛࠿♟ြ
ࡈࡿࡒࠊ⌟ᅹࠉMACBࠉMBCA ࠿ csmA ࡡ◒ቪ࡞ࡻࡾ␏ᖏࢅ┞⿭ࡌࡾ࠾➴࡞ࡗ࠷࡙᳠ゞ୯࡚࠵ࡾࠊ
1) Takeshita, N. et al (2005) Mol. Biol. Cell 16:1961-1970
2) Takeshita, N. et al (2006) Mol. Microbiol. 59:1380-1394
Functional difference between the domains of two chitin synthases, CsmA and CsmB, in Aspergillus nidulans
Makusu Tsuizaki, Norio Takeshita, Hiroyuki Horiuchi, and Akinori Ohta
(Dept. Biotechnol., Univ. Tokyo)
79
P-89
⣊≟Ⳟ Aspergillus nidulans ࡡࢠࣚࢪ III ࢞ࢲࣤྙᠺ㓕⣪㸝ChsB㸞ࡡ⣵⬂හᑻᅹ㒂న ࡡ
よᯊ
⚗⏛ୌ἖ 1ࠉᒜୖಞୌ 2ࠉᇷහ⿩஄ 1 ࠉኯ⏛᪺ᚠ 1㸝᮶ኬ࣬㎨⏍⛁࣬1 ᚺ⏍ᕝ࣬2 ⏍⎌⏍㸞
࢞ࢲࣤࡢ⣊≟Ⳟࡡ⣵⬂ባࡡ୹さᵋᠺᠺฦࡡ㸦ࡗ࡚࠵ࡽࠉࡐࡡ⏍ྙᠺࡢᙟឺࡡᙟᠺࠉ⥌ᣚ࡞㛭ࢂࡾ࡛⩻࠻ࡼ
ࡿࡾࠊA. nidulans ࡡ࢞ࢲࣤྙᠺ㓕⣪ࡡࡥ࡛ࡗ࡚࠵ࡾ ChsB ࡢ 916 ࢓࣐ࢿ㓗࠾ࡼ࡝ࡽࠉࢠࣚࢪ III ࡞ᒌࡊ࡙࠷ࡾࠊ
ࢠࣚࢪ III ࡡ࢞ࢲࣤྙᠺ㓕⣪ࡢ㓕ẍ Saccharomyces cerevisiae ࡷ Shizosaccharomyces pombe ࡝࡜࡞ࡢᏋᅹࡎࡍࠉ
Ⳟ⣊≟ࡡᙟឺ࡚⏍⫩ྊ⬗࡝Ⳟ࠾ࡼࡊ࠾༟㞫ࡈࡿ࡙࠷࡝࠷ࠊchsB ࡡ㐿ఎᏄ◒ቪᰬ࡚ࡢⳞ⣊⏍㛏࠿ኬࡀࡂ㜴ᐐࡈ
ࡿࠉࡐࡡⳞ⣊ࡢฦᒪ࠿ኣࡂࠉ㟸ᖏ࡞ᑚࡈ࡝ࢤࣞࢼ࣭ࢅᙟᠺࡌࡾࠊᮇ◂✪࡚ࡢ ChsB ࡡ⣵⬂හ࡚ࡡᏋᅹ≟ឺࡷ
ᑻᅹ㒂నࢅよᯊࡌࡾࡒࡴࠉࢣࢿ࣑୕ࡡ chsB ࡡⓆ⌟࠿โᚒྊ⬗࡚ࠉࡈࡼ࡞ ChsB ࡡ N ᮆ❻࡞㸫xFLAG ࡱࡒࡢ
EGFP ࢅ㏻⤎ࡊࡒ FLAG-ChsB ࡱࡒࡢ EGFP-ChsB ࢅ argB 㒂న࡚Ⓠ⌟࡚ࡀࡾ FB-3 ᰬࠉEB-5 ᰬࢅࡐࡿࡑࡿష⿿
ࡊࡒࠊFLAG-ChsBࠉEGFP-ChsB ࠿ࡐࡿࡑࡿḿᖏ࡞ᶭ⬗ࡌࡾࡆ࡛ࢅ☔ヾࡊࡒࠊEB-5 ᰬ࡚ EGFP-ChsB ࡡⳞ⣊හ
࡚ࡡᑻᅹ㒂నࢅ᳠ゞࡊࡒ࡛ࡆࢀࠉᖏ࡞Ⳟ⣊ࡡ඙❻㒂ฦ࡞⺧ක࠿ぜࡼࡿࡒࠊࡱࡒࠉᙟᠺ᫤ࡡ㝰ባ࡚ࡵ⺧ක࠿ふ
ᐳࡈࡿࠉⳞ⣊ࡡአባ࠾ࡼ୯ᚨ㒂࡫཭⦨ࡌࡾᣪິࢅ♟ࡊࡒࠊFB-3 ᰬࢅ⏕࠷ࠉඞ␷㞹㢟࡞ࡻࡾ FLAG-ChsB ࡡ⣵
⬂හᑻᅹふᐳࢅ⾔ࡖࡒ࡛ࡆࢀࠉFLAG-ChsB ࠿㝰ባࡡ୯ᚨ㒂ࡷᑚ⬂࡞Ꮛᅹࡊ࡙࠷ࡾᵕᏄ࠿☔ヾࡈࡿࡒࠊࡆࡿࡼ
ࡡ⤎ᯕ࠾ࡼࠉChsB ࠿Ⳟ⣊ࡡ඙❻⏍㛏ࡷ㝰ባᙟᠺ࡞㔔さ࡝ᙲ๪ࢅᯕࡒࡌࡆ࡛࠿♟ြࡈࡿࡒࠊ
Localization of ClassIII Chitin Synthase (ChsB) in Aspergillus nidulans
Kazuharu Fukuda1, Shuichi Yamashita2, Hiroyuki Horiuchi1, and Akinori Ohta1 (1Dept. Biotechnol., Univ. of Tokyo,
1
Dept. Agric. and Envir. Biol., Univ. of Tokyo)
P-90
⣊≟Ⳟ Aspergillus nidulans ࡡ࢞ࢲࢻ࣭ࢭ㐿ఎᏄ chiHࠉchiR ࡡᶭ⬗よᯊ
ᒜᓧᬍ୓ࠉ⏛୯ᙤࠉᇷහ⿩஄ࠉኯ⏛᪺ᚠ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
⣊≟Ⳟ Aspergillus nidulans ࡡᙟឺᙟᠺ࡞ࡢࠉ⣵⬂ባࡡ୹さᵋᠺᠺฦࡡୌࡗ࡚࠵ࡾ࢞ࢲࣤࢅຊỀฦよࡌࡾ㓕⣪
࡚࠵ࡾ࢞ࢲࢻ࣭ࢭࡡᶭ⬗࠿୘ྊḖ࡚࠵ࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊA. nidulans ࡞Ꮛᅹࡌࡾ࡛᥆ᏽࡈࡿࡾ 18 ಴ࡡ࢞ࢲ
ࢻ࣭ࢭ㐿ఎᏄ୯ࠉchiH ࡡ㐿ఎᏄ⏐∸ࡢࠉRACE よᯊ࡞ࡻࡽࠉ558 ࢓࣐ࢿ㓗࠾ࡼ࡝ࡾ⺦Ⓣ㈻࡚࠵ࡽࠉN ᮆ❻࡞
ฦἢࢨࢡࢻࣜࠉὩᛮ୯ᚨࢅྱࡳ㡷ᇡࠉࡐࡿ࡞⤾ࡂ⣑ 180 ࢓࣐ࢿ㓗ࡡ Gln ࡞ᐣࡳ㡷ᇡ࠾ࡼ࡝ࡾ࡛᥆ᏽࡈࡿࡒࠊ
ࡱࡒྜྷᵕ࡞ chiR ࡡ㐿ఎᏄ⏐∸ࡢ 305 ࢓࣐ࢿ㓗࠾ࡼ࡝ࡾ⺦Ⓣ㈻࡚࠵ࡾ࡛᥆ᏽࡈࡿࠉὩᛮ୯ᚨࢅྱࡳ㡷ᇡ௧አ࡞
≁ᚡⓏ࡝㒼าࡢぜฝࡈࡿ࡝࠾ࡖࡒࠊchiH Ḗ኶ᰬࡢ㔕⏍ᆵᰬ࡛ྜྷᵕࡡ⏍⫩ࢅࡊࡒ࠿ࠉchiR Ḗ኶ᰬࡢฦ⏍Ꮔᙟᠺ
࡞␏ᖏ࠿ぜࡼࡿࡒࠊࡱࡒ chiH ࡢ㌹෕࡚ࣝ࣊ࣜฦ⏍Ꮔᙟᠺ᭿࡞Ⓠ⌟࠿ㄇᑙࡈࡿ࡙࠽ࡽࠉࡈࡼ࡞ ChiH ࡛ EGFP
࡛ࡡ⼝ྙ⺦Ⓣ㈻ࡢࣆ࢔࢓ࣚ࢕ࢺ࡛ࢤࢼࢸ࢔࢓ࡡ㛣࡞ࣛࣤࢡ≟࡞ᑻᅹࡌࡾࡆ࡛࠿ฦ࠾ࡖࡒࠊ௧୕ࡡࡆ࡛࠾ࡼ
chiHࠉchiR ࡢฦ⏍Ꮔᙟᠺ᭿࡞ᶭ⬗ࡊ࡙࠷ࡾྊ⬗ᛮ࠿⩻࠻ࡼࡿࡒࠊࡈࡼ࡞ࠉchiH ࡛ chiR ࡡ஦㔔Ḗ኶ᰬࡢࠉ≁ᏽ
ࡡ᮪௲ୖ࡚㔕⏍ᆵᰬࡷࡐࡿࡑࡿࡡ༟≺Ḗ኶ᰬ࡞Ẓ࡬ⴥࡊࡂ⏍⫩࠿㐔ࡿࠉ␏ᖏ࡝ᙟឺࢅ♟ࡊࡒࠊࡆࡡࡆ࡛࠾ࡼࠉ
chiHࠉchiR ࡢⳞ⣊⏍㛏᭿࡞࠽࠷࡙༝ㄢⓏ࡞ᶭ⬗ࡊ࡙࠷ࡾྊ⬗ᛮ࠿⩻࠻ࡼࡿࡒࠊ
Functional analysis of chitinase genes, chiH and chiR in Aspergillus nidulans
Harutake Yamazaki, Aya Tanaka, Hiroyuki Horiuchi and Akinori Ohta
(Dept. of Biotechnol., Univ. of Tokyo)
80
P-91
Rhizopus oryzae ࡡ࢞ࢲࣤ ྙᠺ㓕⣪㐿ఎᏄࡡⓆ⌟よᯊ
㜷㒂 Ṅ㸡ὰ㔕 ⾔ⶮ㸡᭧᰷ ㍜㞕 㸝໪ኬ㝌㎨࣬ᚺ⏕ⳞᏕ㸞
⣊≟Ⳟࡡ୯࡚ࡵ᥃ྙⳞ㢦ࡡ⣵⬂ባࡢ࢞ࢲࣤ࡞ᐣࢆ࡚࠽ࡽ㸡⣵⬂ࢅᙟషࡾ୕࡚࢞ࢲࣤྙᠺ㓕⣪ࡢ㔔さ
࡝ᙲ๪ࢅᯕࡒࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡾ࠿㸡Ꮔࡡ࠹Ⳟ㢦ࡷᢰᏄⳞ㢦࡞Ẓ࡬ࡾ࡛ࡐࡡ▩ぜࡢᑛ࡝࠷㸣ᠻࠍࡢ㸡
Rhizopus oryzae ࡞࠽ࡄࡾ࢞ࢲࣤྙᠺ㓕⣪ࡡᙲ๪ࢅ᪺ࡼ࠾࡞ࡌࡾࡒࡴ㸡R. oryzae CBS 112.07 ᰬ࠾ࡼ
degenerate primer ࢅ౐⏕ࡊ࡙㸡࢞ࢲࣤྙᠺ㓕⣪ࡡ┞ྜྷᛮ㒼าࡡቌᖕࢅ⾔࠷㸡࠷ࡂࡗ࠾ࡡ㒼าࡡ␏࡝ࡾ
࢞ࢲࣤྙᠺ㓕⣪(CHS)㐿ఎᏄ࠿࠵ࡾࡆ࡛ࢅ௧๑ሒ࿈ࡊࡒ㸣௑ᅂࡢ㸡ࡆࡿࡼࡡ㐿ఎᏄ࡞ࡗ࠷࡙㸡ප㛜ࡈ
ࡿ࡙࠷ࡾ R. oryzae ࡡࢣࢿ࣑㒼า࡛Ẓ㍉ࡊ࡙㸡㐿ఎᏄධ㛏ࢅཱིᚋࡊ㸡ࡐࡡⓆ⌟ࢅ RT-PCR ࢅ⏕࠷࡙☔
ヾࡌࡾ࡛࡛ࡵ࡞࢕ࣤࢹࣞࣤࡡỬᏽࢅ⾔ࡖࡒ㸣
degenerate PCR ࡛ࢣࢿ࣑᝗ሒ࡞ࡻࡖ࡙ class II CHS 㐿ఎᏄ࠿㸫⛸㸡class IV CHS 㐿ఎᏄ࠿㸧⛸㸡class
V CHS 㐿ఎᏄ࠿㸦⛸ᚋࡼࡿࡒ㸣ᾦమᇰ㣬࠽ࡻࡦᖲᯀᇰ㣬Ⳟమࡻࡽ RNA ࢅ᢫ฝࡊ㸡RT-PCR ࢅ⾔ࡖࡒ
࡛ࡆࢀ㸡class II ཀྵࡦ class V CHS 㐿ఎᏄࡢᾦమᇰ㣬ཀྵࡦᖲᯀᇰ㣬࡚Ⓠ⌟࠿☔ヾࡈࡿࡒ࠿㸡class IV CHS
ࡢ㸧⛸࡛ࡵⓆ⌟ࢅ☔ヾ࡚ࡀ࡝࠾ࡖࡒ㸣
Class I ཀྵࡦ III CHS 㐿ఎᏄ࠿ᚋࡼࡿ࡝࠾ࡖࡒࡆ࡛࠾ࡼ㸡࢞ࢲࣤྙᠺ㓕⣪ࡡಕᏋ㒼าࢅ⏕࠷࡙㸡R.
oryzae ࢣࢿ࣑㒼า࡚ Blast ᳠⣬ࢅ⾔ࡖࡒ⤎ᯕ㸡R. oryzae ࡞ࡢ class I ཀྵࡦ III CHS ࡢᏋᅹࡊ࡝࠷࡛⩻࠻
ࡼࡿࡒ㸣ࡱࡒ㸡class IV ཀྵࡦ V ࡞ࡗ࠷࡙ࡢ㸡௑ᅂཱིᚋࡊࡒ㐿ఎᏄࡡ௙࡞ࡵ┞ྜྷᛮࢅ♟ࡊࡒࡵࡡ࠿ᩐኣ
ࡂᏋᅹࡊࡒࡆ࡛࠾ࡼ㸡⌟ᅹࡆࡿࡼࡡ㐿ఎᏄࡡⓆ⌟よᯊࢅ⾔ࡖ࡙࠷ࡾ㸣
Expression analysis of chitin synthase genes in Rhizopus oryzae
Ayumi Abe, Kozo Asano, and Teruo Sone
(Lab. of Appl. Microbiol, Research Faculty of Agr. Hokkaido Univ.)
81
ெྞ⣬ᘤ㸝ゖḿ∟㸞
Truong Binh-Nguyen Suhn-Kee Chae Dong-Min Han Praveen Rao Juvvadi Hye-Ryoun Kim James R Kinghorn Sun-Hee Noh Berl R. Oakley Shinichi Oide Avid SHIAU Naimeh Taheri-Talesh B. Gillian Turgeon Shiela E Unkles Le Xuan-Tham ㉝ᑹ೸ ⣒᭰ᾀ ⚽⏛ಞ 㯖⏛᜜ᙢ ὰ㔕⾔ⶮ ῞㔕♰ᗀ ὰぜ⾔ᘧ Ⱙ⃕Ṃெ ᮶ಘᏳ ㊂❟ᾀ⨶ 㜷㒂Ṅ 㜷㒂ᩏᝃ ኮ㔕Ⰳᙢ Ⲡ஬ᇱኰ ᭯Ờງ ᭯ᒱᏕ ᭯㤷ᑋⱝ ᭯ᮇ⿩ 㣜⏛♰ୌ㑳 ஫༎ᔋᅽ᪝Ꮔ ờ⏛೸ୌ ௿⃕┷⃀ ▴ᕖ⤦⌦ ▴⏛༡ᶖ ☶㒂༐ᑖ ୌ▴᫓ᙢ ௿⸠㟱ኰ ௿⸠ᖶᠺ ⛼ᇄ㈶஦ ⛼ෞ⪵ ஬୕ᘧୌ ௑ᮟⱩ⾔ ᒷᓧ೸ኯ㑳 ᒷୖ࿰⿩ Ꮻఫ⨶ㄵ ḯᓞ᜾ හ⏛༡ᩅ හᐣ஁⨶Ꮔ ᱭ⃕ಆ᪺ ᾇ⩹ሪ㇇ ཀྵᕖⱝ⚽ ⩹⏛Ⱪ ኬ▴㯖⏜ ኬ໪⏜౅ ኯ⏛᪺ᚠ ኬ㔕⤚Ꮔ ኬ⟺ಘୌ ᒱᓧᏏ᫆ ᒱᓧක㞕 ᑚ➗ཋ༡ಘ ᑚ➗ཋῦ ᒱ⏛࿰ኃ ᒱ⏛Ᏻᩝ ᒱ㒂᪺Ꮔ ᒱᮇᬓඖ 㛏⏛⿩஄ ᑹ㛭೸஦ ⧂⏛೸ ᑹㆺᾀ ⴘྙዄ⥬Ꮔ ⴘྙఘᗽ ⴘྙ์ᖶ ᑚ㔕ᑈ᭰Ꮔ ᑚ⏷ᾀ ᑚཋᨼ᪺ ᰘⷭࢱࣚࣚࢴࢹ みㆺ├ᶖ ᤻ㆺ⚵᫓ ຊ἖ఫᖲ ᯵ᮄ▩Ꮔ ᯵ᮄ㇇ ຊ⸠├ᶖ ຊ⸠㞖ኃ 㔘ᨳ┷ 82
㔘୷ாᏄ 㙂ಲ㧏ᚷ ᕖཾ๙ྒྷ ᕖᓧ᮶ᙢ ᕖᮟ┷ᚷ ♼⏛㮒஁ ⳝờ㡝ᙢ ⳝ㛣㝧ᚷ ໪ᕖ἖ᜠ ໪ᔪຩ඼ ໪ᮇົࡥࡆ
໪ᮇ์⾔ 㨛㢄ⱝᶖ ᮄᮟኣỜ ᮄᮟಆ஄ ᮄᮟ┷ ᴃᮇ៿ୌ ᕝ⸠ಆ❮ ᕝ⸠Ὂᖲ ❉ᑈ㝧ᩝ ↻ㆺගᖶ ᱋㔕ူ▦ ྙ⏛⚵▦ ♼ኯ㑳 ἑ㔕ຩெ ཿ㈙᫬἖ ඡ⋚ᇱୌ᭹ ᑚこ㐡ஒ ᑚᯐல⣎Ꮔ ᑚᯐူኰ ᑚᯐ㟱඼ ஫࿝ົஒ ᩟⸠ᙤ⳱ ᆊ㔕┷ᖲ ᆊᮇ࿰ಆ 㜨ᮇ㱗ྒྷ ▹ᕖᨳⱝ ఫࠍᮄሐ ఫ⸠ฺḗ ఫ⸠ዄ὘Ꮔ ఫ⸠ḿொ ఫ㔕ඔ᫓ ఫ㔕⿩ᇱ 㩢ᓞḿᾀ ⏐ᮇᘧ஄ 83
ሲ⃕࠵ࡍࡈ ᚷ⏛Ὂ௒ ᓞ⏛ᖷኰ ୖ㣜ொ ୖᆊㄌ ḿ㊪ῗஒ ᩺ㆺᑠᘧ 㡪㈙ᬍ஁ ᮙ὾❟ᶖ ᮙᮇ├ᕤ ᮙᒜ⣟ஒ 㕝ᮄᙪ 㕝ᮄ㯖Ꮔ 㕝ᮄୌྍ 㕝ᮄ⪵ ⅛ㆺ㡨ୌ ເࠎᗛ௥ ᭧᰷㍜㞕 ᑘᓧ┷ᴃ ⏛ᑹ⸓Ꮔ 㧏ᮄᚰ 㧏ᮄᾇ 㧏ㆺ├ᶖ 㧏ጏ༜ྒྷ 㧏ᶣ᪺⌌ 㧏ᶣ▹⧂ ➁හ㐠㞕 Ṃᕖ἖ ➁ୖ඼⏠ ➁ㆺ༡Ꮔ ➁㔕ḿὊ ⏛ᓞㅅ ኣ⏛ຉ⏍ ㎦ᕦᬏ⣎ ⏛୯ᙤ ⏛୯⍖ᕤ ㆺཾ㞕သ ㆺಞ἖ ⠇㔕༜ኰ ᘿ᪺Ễ ᯼᳔ᑠᚷ ᑈᒱᚥ ᇸ๑├ ᇸᮇකᏄ ᫤ᮇᬊீ ᚠ஬⨶㔓 ᚠᒱ᪸ᩝ ᅰఫᖶ㞕 ෞ⏛ᡛ⏠ ཪ㔕⌦⏍ ୯஬႗ᙢ ୯⏿Ⰳ஦ ୯㤷࠷ࡘࡲ ୯⃕ක ୯ᓞᨼᙢ 㛏ᓞ⏜ኚ ୯㔕ḿ᪺ ୯ᒁᩔᆍ ༞㒂ኚᕵ こᬏᏄ こᮟ㢟 こᮟ㯖㔓Ờ ᰷ᮇᓤ 㔕ཾ♰ྒྷ 㔕ᓧຉୌ 㔕ᓧಘ⾔ ⴏཋኬ♰ ᮌᮠඒ 㛏ㆺᕖ⚀Ꮔ ⛑Ὂ஦ ᭱㒂Ṃᩝ ᧓ᮇ౅᪺ ᯀ᮶ኚᾇ ఔක༡ ᵵཾ⿩ḗ㑳 ஁⏛༡ඔ Ⓤ⏣‫‮‬᭹ ⋖ᗛὒ ᖲ஬ᾀᩝ ᖲᒱⰃ㍜ ᖲඳ㔔ୌ஄ ᗀᖏḿெ ⚗⏛ୌ἖ ⚗᲻ᩝᗛ ⸠஬໌ ⸠஬㐡ஒ ⸠ᒱᬓ์ ⸠ᮟ┷ ⸠ཋ┷⣎ ཿᕖ⫩௥ ཿᕖ೸ኯ㑳 ཿᕖ㈏Ꮔ ཿᕖ㝧⣎ ᆎᇄ㝧஄ ᮌ㱗ὒ ᫅⏜⣎Ꮔ ᇷහ⿩஄ ᇷහ᱀Ꮔ ᇷᑹူஒ ᮇᒜយ ᮇ⏛୙ୌ ๑ᕖ஦ኯ㑳 ๑⏛ඁ௒ 㤷᭨⏜⨶ ᱕ᑹಆ௒ ಚ㔕㐡ᙢ ⏣⏛㞖஄ ᮿ஬ᖶኯ㑳 ᮿ὾▩ᕤ ᮿᑹୌ㑳 ᮿ㮭᫓์ ᮿୖ┷⏜⨶ ᮿᯐⰃ༡ ┶ᒜ⁘ᚷ ୷஬ῗୌ㑳 ୷ᒜ₮ୌ ୔୕㔔᪺ ୔ᮄួ௒ Ềㆺ἖ Ề㔕⊓ Ề㔕ḿᾀ ‹୕㇇ ༞ᰜୌ ୔Ꮹ๙ྍ ᐋᓧಆ ᐋᮇᖶᜠ ୔ይኬ㍔ ᮟᒜ⫍Ꮔ Ⱪἠᖶኯ ᮇᒜ㧏ᖶ ᲻ᕖᗛ ᲻ཾᬓ⨶ ᲻⩟྿ ᲻⏛ᐰெ ᲻⏛┷⣟ Ꮼᒁᬓ༡ ▦ᓧୌྍ ᒜහ㝧ᐰ 84
ᒜᓃ㈶἖ ᒜᓃኬ㍔ ᒜᓧᬍ୓ ᒜୖ࿰Ᏻ ᒜୖಞୌ ᒜୖఘ㞕 ᒜ⠓㈏ྍ ᒜ⏛ಞ ᒜ⏛㞖ெ ᒜ୯ᓤ ᒜᮇᖷ༡ ᶋ஬ᑋ㑳 ྚ⏛㝧ᘇ ྚぜၤ ⰶ㝘ୌ⏍ ྚᮟ᪸ᚠ ౪⏛ᗛᏏ ⡷ᮟ᫥Ꮔ ῳ㑋ᙪ ῳ㑌⟿Ꮔ ῳ㎰㝧ྒྷ ῳ㑋▩ᶖ ῳ㎰஁ᩏ ῳ㑌ᓤெ ῳ㎰ὀ♰ 85
⣊≟ⳞฦᏄ⏍∸Ꮥ◂✪ఌ ఌ์
ᮇఌࢅ⣊≟ⳞฦᏄ⏍∸Ꮥ◂✪ఌ㸝+XQJFO2ROHHXOFU'LRORJ^8RHLHW^RI/FSFQ㸞࡛࿣ࡩࠊࡱࡒᮇఌ࠿
㛜ࡂ◂✪ఌࢅ⣊ ≟ⳞฦᏄ⏍ ∸Ꮥࢤࣤࣆ ࢒ࣝࣤࢪ㸝(RQIHUHQHH RQ +XQJFO ,HQHWLHV FQI 2ROHHXOFU
'LRORJ^㸞࡛࿣ࡩࠊ
ᮇఌࡢ⣊≟ⳞࡡฦᏄ⏍∸Ꮥࠉ⣵⬂⏍∸Ꮥࠉ⏍໩Ꮥࠉ⏍⌦Ꮥࠉ㐿ఎᏕ࡝࡜ࡡᬉཀྵⓆᒈࢅ┘Ⓩ࡛ࡌࡾࠊ
ᮇఌࡢࡐࡡ┘Ⓩࢅ㐡ᠺࡌࡾࡒࡴ࡞ḗࡡ஥ᴏࢅ⾔࠹ࠊ
◂✪ఌཀྵࡦ⥪ఌࡡ㛜തࠊ
ఌሒࡡⓆ⾔ࠊ
㛭㏻◂✪ᅆమ࡛ࡡ༝ງ஥ᴏࠊ
ࡐࡡ௙ࠉᚪさ࡛ᛦࢂࡿࡾ஥ᴏࠊ
ᮇఌࡢࡐࡡ┘Ⓩ࡞㈰ྜྷࡊ࡙ථఌࡊࡒ಴ெఌဤཀྵࡦ⥪ఌ࡞࠽࠷࡙ᢆヾࡈࡿࡒྞヽఌဤࢅᣚࡖ࡙ᵋᠺࡌࡾࠊ
ᮇఌථఌᕵ᭻⩽ࡢᡜᏽࡡථఌ⏞㎲᭡ࢅᥞฝࡊࠉื࡞ᏽࡴࡾථఌ㔘ࢅ⣙ථࡌࡾࡵࡡ࡛ࡌࡾࠊ
ᮇఌࡢࡐࡡ㐘ႜࡡࡒࡴࠉఌ㛏ࠉ㐘ႜጟဤⱕᖱྞ࠽ࡻࡦఌゝ┐ᰕ ࠤ ྞࢅ࠽ࡂࠊ௴᭿ࡢ ᖳ࡛ࡊࠉᨭ㐽ࡢ㐘
ႜጟဤࡡ᥆࡛ⷸ⥪ఌࡡᢆヾ࡞ࡻࡾࠊ
ఌ㛏ࡢᮇఌࢅ௥⾪ࡊࠉఌຸࢅ⤣ᣋࡌࡾࠊ
㐘ႜጟဤࡢ㐘ႜጟဤఌࢅᵋᠺࡊఌຸࢅᐼ㆗ࡌࡾࠊ㐘ႜጟဤ࡞ࡢᗚຸࠉఌゝࠉ⥽㞗ᢰᙔࠉᗀሒᢰᙔ
ࢅ࠽ࡂࠊ
ఌゝ┐ᰕࡢᮇఌࡡఌゝࢅ┐ᰕࡌࡾࠊ
ᮇఌࡢ஥ᴏ㐘ႜ࡞ᚪさ࡝ᐁ㈕ࢅᖳఌ㈕࡛ࡊ࡙಴ெఌဤ࠾ࡼᚡ཭ࡌࡾࠊ
ᮇఌࡡ஥ຸᖳᗐࡢ◂✪ఌࡡ㛜ത‵ങ㛜ጙ࠾ࡼࠔḗ᭿ࠕ◂✪ఌࡡ㛜ത‵ങ㛜ጙ├๑ࡱ࡚࡛ࡌࡾࠊ
๑஥ຸᖳᗐࡡᗚຸࠉఌゝ࡞ࡗ࠷࡙ࡢࠉࡆࡿࢅ⥪ఌ࡞࠽࠷࡙ሒ࿈ࡊࠉᢆヾࢅᚋࡾࡵࡡ࡛ࡌࡾࠊ
ᮇఌ์ࡡᨭᏽ࡞ࡢ⥪ఌฝᖆ⩽㐛༖ᩐࡡ㈰ᠺࢅᚪさ࡛ࡌࡾࠊ
௧୕
⿭์
ᮇఌ์ࡢ ᖳ ᭮ ᪝ࡻࡽⓆຝࡌࡾࠊ
ᮇఌථఌ㔘ࡢ ළ࡛ࡌࡾࠊ
ᖳఌ㈕ࡢୌ⯙ఌဤ ළࠉᏕ⏍ఌဤ ළ࡛ࡌࡾࠊ
◂✪ఌࡡ㏳▩ཀྵࡦఌሒࡢࠉᙔルᖳᗐࡱ࡚ࡡఌ㈕ࢅ⣙ථࡊࡒఌဤ࡞㏞௛ࡌࡾࠊ
ᖳᗐ࡞ࢂࡒࡖ࡙ఌ㈕⣙ථࡡ࡝࠷ఌဤࡢࠉࡐࡡ㈠᰹ࢅ኶࠹ࡵࡡ࡛ࡌࡾࠊ
◂✪ఌࡡⓆ⾪⩽ࡢࠉఌဤ࡞㝀ࡾࡵࡡ࡛ࡌࡾࠊ᩺ථఌဤࡡⁿ㢗⏞ࡊ㎲ࡲࡢఌ㈕⣙ථࡡ☔ヾࢅᣚࡖུ࡙⌦ࡌ
ࡾࠊ
86
⣊≟ⳞฦᏄ⏍∸Ꮥ◂✪ఌ㐘ႜጟဤఌྞ⡑
ఌ 㛏
໪ᮇ ົࡥࡆ
᮶ாኬᏕኬᏕ㝌㎨Ꮥ⏍࿤⛁Ꮥ◂✪⛁㸝ࠚ ᮶ா㒌ᩝா༇ᘲ⏍ 㸞
㐘ႜጟဤ
⚽⏛ ಞ
ᐁ㊰ዥᏄኬᏕ⏍Ὡ⛁Ꮥ㒂㸝ࠚ᮶ா㒌᪝㔕ᕰኬᆊ୕ 㸞
஫࿝ ົஒ
᮶໪ኬᏕኬᏕ㝌㎨Ꮥ◂✪⛁㸝ࠚ ௜ྋᕰ㟯ⴝ༇ላ㏳㞭ᐋ⏣ 㸞
㩢ᓞ ḿᾀ
᮶ாኬᏕኬᏕ㝌㎨Ꮥ⏍࿤⛁Ꮥ◂✪⛁㸝ࠚ ᮶ா㒌ᩝா༇ᘲ⏍ 㸞
⛑ Ὂ஦
᭮᰿෗ᰬᘟఌ♣⥪ྙ◂✪ᡜ㸝ࠚ ா㒌ᕰఄぜ༇ୖ㫵⩒ᑚᰏ⏣␊ᆀ㸞
ఌゝᢰᙔ
᭯Ờ ງ
᮶ா㎨ᕝኬᏕ㎨Ꮥ㒂㸝ࠚ ᮶ா㒌ᗋ୯ᕰᖶ⏣ 㸞
➁හ 㐠㞕
᮶ா㎨ᕝኬᏕ㎨Ꮥ㒂㸝ࠚ ᮶ா㒌ᗋ୯ᕰᖶ⏣ 㸞
⥽㞗ᢰᙔ
ᑚᯐ ူኰࠉຊ⸠ 㞖ኃ
ྞཿᒁኬᏕኬᏕ㝌⏍࿤㎨Ꮥ◂✪⛁㸝ࠚ ྞཿᒁᕰ༐⛸༇୘⩹⏣㸞
ᗀሒᢰᙔ
ᕖཾ ๙ྒྷ
ኬ㜨ᗋ❟ኬᏕኬᏕ㝌㎨Ꮥ⏍࿤⛁Ꮥ◂✪⛁㸝ࠚ ኬ㜨ᗋሗᕰᏕᅧ⏣ 㸞
ᗚຸᢰᙔ
ᇷහ ⿩஄
᮶ாኬᏕኬᏕ㝌㎨Ꮥ⏍࿤⛁Ꮥ◂✪⛁㸝ࠚ ᮶ா㒌ᩝா༇ᘲ⏍ 㸞
87

食糧と機能性食品生産のためのバイオテクノロジー

平成 18 年度報告書