UNIVERSITI PUTRA MALAYSIA
IN VITRO CULTURE AND SYMBIOTIC RELATIONSHIPS
OF PAPHIOPEDILUM NIVEUM, TAENIOPHYLLUM OBTUSUM AND
VANDA HOOKERIANA WITH THEIR ASSOCIATED FUNGI
IRAWATI
FP 1987 8
It is hereby cert i f ied that we have read th is thesis
ent itled
"In V itro Culture and Symb iotic Relat ionsh ips o f
Paphiopedilum niveum, Taeniophyllum obtusum and Vanda hookeriana
w ith the ir Assoc iated Fung i" by Irawati, and in our op in ion it is
satis factory in terms o f scope, qual ity, and presentation as
partial ful f ilment o f the requ irements for the degree o f Doctor
o f Ph ilosophv
. .-:-�.':'�:�: .
SULAlMAN
M.
YASSIN,
'":") �
Ph.D.
Professor
Dean of Graduate Studies
Un ivers it i Pertanian Malaysia
(Cha irman Board of Exam iners)
�
r\
I) 1 �
.. ..... ......... �jt/L rLtll-
\\
JOSE H AR ITTI, Ph.D.
Pro fessor
School o f B iolog ical Sc iences
Univers ity o f Cali forn ia, U.S.A
(External Exam iner)
Jg.<� i:t,.
�
GioF�Y . HABL�Y : . Ph:
(
C-'l
D. ---
Department o f Plant Sc ience
University o f Aberdeen
St. Machar Dr ive, Aberdeen
England
(External Examiner)
MOHD. ZAIN HJ. ABD. KARIM, Ph.D.
Pro fessor/Dean
Faculty o f Agr iculture
Un ivers iti Pertanian Malaysia
(Internal Examiner)
MOHAMMAD MD. AL I, Ph.D.
Assoc iate Pro fessor
Faculty o f Agriculture
Univers iti Pertan ian Malays ia
( Internal Examiner/Superv isor)
This thesis was submitted to the Senate o f Universiti
Pertanian Malaysia and was accepted as partial fulfilment of
the requirements for the Degree o f Doctor o f Philosophy.
Date: ..
,l
i J U L t:)��
SULAIMAN M. YASSIN, Ph. D.
Pro fessor
Dean of Graduate Studies
IN VITRO C UL TURE AND SY MBIOTIC RELATIONSHIPS
OF PAPHIOPEDILUM NIVEUM, TAENIOPHYLLUM OBTUSUM AND
VANDA HOO KERIANA WITH THEIR ASSOCIATED FUNGI
by
Irawati
A thesis submitted in partial fulfilment o f the
requirements for the degree o f Doctor o f Philosophy
in the Faculty o f Agriculture,
Universiti Pertanian Malaysia
September 1987
AC KNOWLEDGEMENT
To
my
individuals and institutions I would l ike to
express
appreciation and sincere gratitute for their contributions
in completion of this study .
*
My
supervi sors ,
Pro fessor )
and
Dr .
Mohammad
Dr . Hasan Mad
Md .
Ali
( Associate
for their guidance and
advi ce throughout the course of
this study and in the
preparation of this manuscript .
* Mr .
Saleh
Kadzimin ,
for
all
his
help
since
the
beginning of this study .
*
Mr . Mark A . Clements , for
the
valuable
discussions ,
his helpful comments and suggestions and also for
his
help in fungal determination .
*
Dr . Zaliha Christine Alang , for reading thi s thesis .
*
The
of
Faculty
the
of Agriculture , and the
Universiti
facilities
in
Pertanian
conducting
this
Computer Centre
Malaysia ,
research
for
and
the
in
preparing this manuscript .
*
Co mmonwealth
�ycological
identification .
ii
Institute
for
fungal
*
Mr
Ho O i Kuan,
Mrs. Am inah,
Mr
Mr
Yusof Suki,
How Yee Peng,
Prof. Yap Thoo Chai
for
Ms. Fadzlon Mohd. Juso f,
Mr
Mimbar
Haryadi,
Dr. La i Chao May
and
their assistance and also to
Mrs. Chairun Nisa Haris
and
Mrs. Kamariah Hamid for the ir kind help.
*
Public
of
a
Services
fellowship
Department o f Malaysia for the award
under
Assistance Programme.
iii
the
Malays ian
Technical
TABLE OF CONTENTS
Page
ACKNO WLEDGEMENT
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TABLE OF CONfEN'I'S
LIS T OF TABLES
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LIS T OF PLATES
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LIS T OF ABBREVIA TIONS
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CHAPTER I I .
xii
xvi
xvi i
1
INTRODUCTION
CHAPTER I .
ix
xv
LIS T OF APPENDICES
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LIS T OF FIGURES
ABS TRACT
ii
6
LI TERATURE REVIEW
8
A.
Plant Materials
B.
Tissue Culture of Orchids
13
C.
Symbiotic Germination of Orchid Seeds
17
D.
Fungal Myc orrhiza Associated with Orchids
23
E.
Symbiotic Development of Germinated Seeds
29
F.
Asymbiotic Germination of Orchid Seeds
34
G.
Establishment of Seedlings
37
CHAPTER III.
A.
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MATERIALS AND METHODS
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Plant Materials
1.
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Paphiopedilum niveum Rchb . f .
40
a.
In vitro culture
40
b.
Seed
41
c.
Protocorm-l ike b ody
42
iv
Seedling
e.
Fungus
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42
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42
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a.
In vitro culture
b.
Seed
c.
Protocorm-like body
d.
Seedling
e.
Fungus
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a.
In vitro culture
47
b.
Seed
47
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Protocorm-like body
47
d.
Seedling
48
e.
Fungus
48
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49
Media
2.
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Yanda hookeriana Rchb . f .
3.
1.
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Taeniophyllum obtusum Bl .
2.
B.
d.
Paphiopedilum n iveum Rchb . f .
50
a.
In vitro culture
50
b.
Seedling
51
Taeniophyllum obtusum Bl .
52
a.
In v itro culture
52
b.
Protocorm-like body
54
c.
Seedling
55
Yanda hookeriana Rchb . f .
56
a.
In vitro culture
56
b.
Protocorm-like body
56
c.
Seedling
57
v
c.
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57
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58
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59
4.
Fungus
5.
Symbiotic studies
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a.
Seed stage
b.
Protocor m-l ike b ody stage
59
c.
Seedling stage
60
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Techniq ue/Working Methods
1.
2.
60
In vi tro culture
60
a.
Paphiopedilu m niveu m Rchb . f .
b.
Taeniophyllum obtusu m Bl .
63
c.
Yanda hookeriana Rchb . f .
65
Preparation of seedling
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a.
Paphiopedilu m niveu m Rchb . f
b.
Taeniophyllu m obtusu m Bl .
66
c.
Yanda hookeriana Rchb . f .
67
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Isolation of fungi
68
4.
Methods of inoculation
70
a.
Seed s tage
70
b.
Protocor m-l ike bod y stage
72
c.
Seedling stage
73
5.
Environmental conditions
74
6.
Description of treatments
7.
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a.
Seed stage
b.
Protocor m-like bod y stage
76
c.
Seedling stage
78
Observation
a.
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74
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In vitro culture
vi
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79
b.
Symbiotic germination
c.
Symbio tic
development of protocorm-
like b o dy
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80
d.
Symbiotic development of see dling . .
80
e.
Observation under light microscope .
80
f.
Observation
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electron
un der
83
microscope
RESULTS
85
In Vitro Culture
86
C HAPTER IV.
A.
79
1.
Paphiope dilum niveum Rchb . f .
2.
Taeniophyllum obtusum Bl .
3.
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a.
Root culture
92
b.
Infloresc ence culture
95
c.
Shoot tip culture
96
Van da hookeriana Rchb . f .
99
B.
Fungal Culture
101
C.
See dl ing
121
D.
1.
Paphiope dilum niveum Rchb . f .
1 21
2.
Taeniophyllum obtusum Bl .
1 28
3.
Yan da hookeriana Rchb . f.
1 35
Symbiotic Germination
1 36
1.
Paphiope dilum niveum Rchb . f .
2.
Taeniophyllum obtusum Bl .
3.
Yan da hookeriana Rchb . f .
vii
1 37
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E.
F.
Symbiotic Development of Protocorm-like body .
161
1.
Taeniophyllum obtusum Bl .
162
2.
Vanda hookeriana Rchb . f .
1 66
Symbiotic Development o f Seedling
1 69
1.
Taeniophyllum
2.
Vanda hookeriana Rchb . f .
1 69
obtusum Bl .
175
DISCUS SION
1 80
A.
I n vitro culture
180
B.
Fungal culture
c.
Seedling
185
D.
Symbiotic germination
191
E.
Symbiotic development of protocorm-like body .
197
F.
Symbiotic development o f seedling
1 99
CHAPTER V .
CHAPTER VI .
CHAPTER VII .
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203
SU MMARY AND CONCLUSIONS
BIBLIOGRAPHY
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209
222
APPENDICES
viii
LIST OF TABLES
page
Table
1.
Envi ronmental conditions in various trials
75
2.
Treat ments o f the seed
77
3.
Treatments o f the protocor m-like body
78
4.
Dec ontamination
niveu m explants
procedures for
Paphiopedilum
88
5.
Deconta mination procedures for
obtusu m explants
Taeniophyllum
6.
Taen iophyllu m obtusu m root tip cultures on
modified Knudson's C mediu m at different pH
after two weeks . . . . . . . . . . . . . . . . . . . . . . . . . . . .
93
The
effect of sterile coconut water
or
hor mones added to Taeniophyllu m obtusu m root
tip cultures after two months
94
Dark treat ment on Taeniophyllum obtusu m root
tip cuI tures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
95
Ef fect of BA or NAA on inflorescence cultures
of Taeniophyllum obtusu m
96
Fungi isolated from Paphiopedilum niveu m,
Taenioph yllu m obtusu m and Vanda hookeriana at
dif fer ent ti mes . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
117
11.
Duration ( days) for the fungal hyphae to reach
one c enti meter in Fungi isolating mediu m
118
12.
The growth of P I , T l and VI fungi on different
media after six days in culture
119
Develop ment of fungi fr om three different
orch ids
on Oat mediu m and fungus P I on
mixtures of Oat mediu m
1 20
Ger mination of Paphiopedilum niveu m seeds at
different maturity
124
Germination of Taeniophyllum obtusu m seeds at
different medi a . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1 29
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ix
91
page
Tab le
16.
Ger mination of Taenioph yllum obtusu m seeds at
different maturity . . . . . . . . . . . . . . . . . . . . . . . . .
.
17.
Paphiopedilum niveu m
different treat ments
18.
Ti me of development of
part s during the
germination of Taeniophyllu m obtusu m seeds
19 .
Taeniophyl lu m obtusu m
different treatments
20 .
Ti me of develop ment of
parts during
germination of Vanda hookeriana seeds
seed
germination
seed
at
germination
•
seed
at
the
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germination
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130
1 39
1 46
1 47
152
21 .
Mean Vanda hookeriana
different treatments
22 .
Average
number of Vanda hookeriana leaves
and roots per seedling . . . . . . . . . . . . . . . . . . . . .
158
23 .
Develop ment of Taeniophyllum obtusu m protocor m
-like bodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
163
24 .
Develop ment
l ike bodies
167
25 .
Averag e
number of leaves
and roots
protocor m-like body of Vanda hookeriana
26.
Protocor m bud ding
seedlings
27.
Survival
seedlings
at
.
28 .
of
Vanda hookeriana
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Additional
weight
obtusu m seedling
.
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Taeniophylum obtusu m
of
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gra m Taeniophyllu m
per
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per
Taenioph yllu m obtusu m
in
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protocor m-
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1 53
168
1 70
172
1 74
29 .
Nitrog en , phosphorus , potassiu m, iron and zinc
content of Taenioph vllum obtusum seedlings
176
30 .
Survival ability of Vanda hookeriana seedlings
177
31 .
Nitrogen , phosphorus , potassiu m, iron and zinc
c ontent of Vanda hookeriana seedlings
1 79
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LIST OF FIGURES
Figure
1.
2.
page
Correlation between rate of germination and
perc entage of germination of Paphiopedilum
niveum seeds
125
Correlation between rate of germination and
age of Paphiopedilum niveum seeds
126
Correlation between percentage o f germination
and age of Paphiopedilum niveum seeds
1 27
Correlat ion between rat e of germination and
perc entage of germination of Taeniophyllum
obtusum seeds
132
Correlation bet\veen rate of germination
age of Taeniophyllum obtusum seeds
133
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and
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6.
Correlation between perc entage of germination
and age of Taeniophyllum obtusum seeds
1 34
7.
l eaf number per seedling
Mean
of Yanda
hookeriana at dif ferent treatments ( Tl , VI,
V4 , KCbc )
1 57
root number per seedling
Mean
of Yanda
hookeriana at different treatments ( Tl , VI,
V4 , KCbc )
1 59
8.
xi
LIST OF PLATES
PLATE
I.
II.
III .
IV .
page
9
Paphiopedilum niveum flower
Taeniophyllum obtusum
11
Yanda hookeriana
12
Young flower b ud
of Paphiopedilum
after seven months in culture
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VI .
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niveum
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Shoot tip cul ture of Paphiopedillum niveum
Section of Taeniophyllum obtusum inflorescence
culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
97
.
VII .
VII I .
obtusum
Multiple branches of Taeniophyllum
infl orescence culture ( NAA treatment )
Roots developing from Taeniophyllum obtusum
inflorescence culture ( BA treatment )
100
Yanda hookeriana protocorm-like bodies
100
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IX.
X.
Fungal
root
di stribution
XI .
Fungal
root
distribution in
XII .
XIII .
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Scanning electron microgram of digested fungal
clumps in Paphiopedilum niveum roo t
Digested fungal clump
root
Living
root
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fungal
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in Paphiopedi lum niveum
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102
Taeniophyllum obtusum
Fungal distribut ion in Vanda hookeriana root
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XV .
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Paphiopedilum niveum
in
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XIV .
97
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clumps in Paphiopedilum niveum
102
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103
104
104
105
XVI .
Thick fungus cell wall inside the host cell . .
1 07
XVII .
Thin fungus cel l wall ins ide the host cell . . .
107
Thick cel l wall of digested hyphae
108
XVI II .
xii
PLATE
XIX .
XX .
page
Cellulose layers at both s ides of host cell
wall . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
109
Transmission
clump
1 10
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electron
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microgram
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of a fungal
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XXla .
Fungus PI from Paphiopedilum n iveum
1 14
XXlb .
Fungus P2 from Paphiopedilum niveum
114
XXlc .
Fungus P3 from Paphiopedilum niveum
114
XXld .
Fungus P4 from Paphiopedilum niveum
1 15
XXle .
Fung us T l from Taeniophyl1um obt usum
1 15
XXlf .
Fung us VI from Yanda hookeriana
1 15
XXlg .
Fungus V2 from Yanda hookeriana
1 16
XXlh .
Fungus V3 from Yanda hookeriana
1 16
XXIi .
Fungus V4 from Yanda hookeriana
1 16
XXlla .
Paphiopedilum n iveum
seed
after
Swollen
six weeks in culture . . . . . . . . . . . . . . . . . . . . . . . .
141
XXllb .
contamination
Bacterial
Paphiopedilum niveum seed
14 1
XXIII .
Paphiopedilum n iveum protocorm grown from seed
inoculated with PI (Tulasnella sp . ) fungus
after three months in culture
142
obtusum protocorm grown from
Taeniophyllum
seed inoculated with Tl (Ceratobasidium sp . )
fungus after 20 weeks in culture
145
obtusum
seed
Culture
of
Taeniophyllum
inoculated
with
Tl
(Ceratobasidium sp . )
fungus after one week in culture
149
Bacterial
contaminations on
Taeniophyllum
seeds
inoculated
with
Tl
obtu sum
( Ceratobasidium sp . ) fungus after one week in
culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1 49
germinating
on
•
XXIV .
•
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XXV .
•
XXVI .
xiii
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PLATE
XXVII .
XXVIII .
XXI X .
xxx.
page
Multiple branche s of Taeniophyllum obtusum
protocorms in
complete
medium
(modified
Knudson ' s C medium + bean sprout extract )
1 50
Fungus
VI (Ceratobasidium s p . )
germinating Vanda hookeriana seed
156
Protocorm-like
bodies
of
obtusum in simple (Oat ) medium
covered the
Taeniophyllum
Protocorm-like
bodies
of
Taeniophyllum
obtusum in modified Knudson ' s C medium
16 5
Protocorm
seedling
1 71
.
XXXI .
165
budding
of
x iv
.
•
•
.
•
Taeniophyllum obtusum
LIST OF APPENDICES
APPENDIX
page
A
Modified Vacin & Went medium
B
Modified Knudson ' s C medium (Arditti , 1974)
C
Oat medium (Clements , 198 1 )
222
•
•
•
•
•
•
•
•
•
•
Fungi isolating medium (Clements , 1981 )
xv
•
•
•
•
•
•
•
•
•
223
224
224
LIST OF ABBREVIATIONS
BA
6-benzyladenine
2,4-D
2,4-dichlorophenoxyacetic acid
EDTA
ethylenediamine tetraacetic acid
FAA
formol acetic alcohol
IAA
indole-3-acetic acid
NAA
naphthaleneacetic acid
ppm
parts per million
TBA
tertiary butyl alcohol
xvi
An abstract of the thesis presented to the Senate o f
Universiti Pertanian Halaysia i n partial ful filment o f the
requirements for the Degree o f Doctor o f Philosophy.
IN VITRO CULTURE fu�D SYMBIOTIC RELA TIONSHIPS
OF PAPHIOPEDILUI'1 NIVEUM, TAENIOPHYLLUM OBTUSUM AND
VANDA HOOKERIANA WITH THEIR ASSOCIA TED FUNGI
by
Irawati
September, 1 987
Supervisor
Dr. Mohammad Hd. Ali
Co-supervisor
Dr. Hasan Mad
Faculty
Agriculture
In
vitro
Paphiopedilum
cultures
of
three
orchid
niveum,
Taeniophyllum
species,
obtusum
and
namely,
Yanda
hookeriana, were initiated from dif ferent parts o f the plants.
Proliferous
tips
protocorm-like bodies were obtained
and axillary
in florescence o f
P.
niveum
V. hookeriana
T. obtusum.
grew
In florescence
branches
buds o f
into
culture o f
shoot
and shoot tips and
Shoot tips and axillary buds o f
single
T.
o f in florescences,
from
or
mUltiple
plantlets.
obtusum developed into multiple
calli or produced
roots.
Young
lea f tip or leaf base o f P. niveum and root o f T. obtusum were
unable
to dif ferentiate into
young flower
bud o f
P. niveum
callus.
produced
months.
xvii
However,
leaf
culture
of
a fter seven
High
pH of t he medium was suitable for
Low
concentration
of modified
preferable to the standard
T . obtusum
Knudson's
concentration for
C
cultures .
medium
was
the culture
of
P. niveum .
Efforts to raise seedlings of the three di fferent orchids
140
to 160 -day
showed
that seeds
n iveum
and 60 to lIS-day old fruits of
better
than dry
from
seeds ,
o ld fruits
of
P.
T . obtusum germinated
while for V.
hookeriana dry
seeds
were able to germinate freely .
Myc orrhizal fungi isolated f rom P . niveum , T . obtusum and
V. hookeriana
their own
roots were
able to induce the
Fungal mycorrhiza from
seeds .
germination
T . obtusum showed a
greater abi lity to initiate germination and promote growth
V. hookeriana
Other
fungi
deleterious
Most
not
seeds
fungi
influenc e
and
species
were
seeds .
associated
the
of
own fungal mycorrhizas .
associated with the three orchids
to the
V. hookeriana
compared to i t s
of
with
these
three
development of protocorm-like
were not compatib le to
orchids
did
bodies
T. obtusum .
of
Their
influence in the establishment of V. hookeriana and T . obtusum
seedlings in the
nursery
was also not s ignificantly shown in
this study .
xviii
Abstrak tesis yang dikemukakan kepada Senat Universiti
daripada
Pertanian
Malaysia sebagai memenuhi sebahagian
keperluan untuk ijazah Doktor Falsafah.
KULTUR I� VITRO Dfu� P ERHUBUNG AN SIMBIOSIS
ANTARA PAPHIOPEDILUM NIV EUM, T AENIOPHY LLUM OBTUSUM DAN
VANDA HOOKERIANA D ENGAN KUL AT-KUL AT SEKUTUANNY A
oleh
Irawati
September, 1987
Penyelia
Dr. Mohammad Md. Ali
Penyelia bersama
Dr. Hasan Mad
Fakulti
Pertanian
Kultur
Paphiopedilum
hookeriana,
in
vitro
niveum,
telah
Struktur-struktur
cepat
bagi
spesies
Taeniophyllum
dibuat
dari
diperolehi dari pucuk
samping dari P. niveum
Kultur jambak
dan
bahagian
Pucuk dan tunas
T. obtusum.
tumbuh menjadi anak benih tunggal atau
bunga
dari T. obtusum
tumbuh
ganda, kalus atau menghasilkan akar.
pangkal daun
yang
muda dari
muda
P. niveum dan akar
kalus.
P. niveum
tanaman.
V. hookeriana
jambak bunga
dapat menghasilkan
Vanda
yang mampu �embiak dengan
dan tunas samping
bunga
iaitu,
orkid,
obtusum
beberapa
seperti protokom
dan dari pucuk dan jambak
ganda.
tiga
dapat
bulan.
xix
daun
Hujung dan
T. obtusum tidak
Akan tetapi kultur
menumbuhkan
menjadi
kuncup
bunga
setelah
tujuh
Medium
yang
T. obtusum.
mempunyai pH
Kepekatan
tinggi adalah
yang
rendah
sesuai untuk kultur
dari medium
Knudson's C
yang diubahsuaikan adalah lebih baik bagi membiakkan P. niveum
daripada yang berkepekatan biasa.
Usaha untuk menumbuhkan anak benih dari ketiga-tiga jenis
orkid yang berbeza didapati biji benih
140 hingga 160 hari dan biji benih
T. obtusum yang berumur 60
daripada biji keringnya,
hingga 115 hari bercambah lebih baik
manakala
biji
kering
P. niveum yang berumur
V. hookeriana pula
bercambah
dengan
bebas.
Kulat-kulat
mikoriza
P. niveum, T. obtusum dan
percambahan
T. obtusum
biji
baik
orkidnya.
menunjukkan keupayaan
dibandingkan
Kulat-kulat
V. hookeriana
benih
menggalakkan pertumbuhan
lain
dari
mampu
Kulat
memulakan
biji-biji
dengan
yang
diasingkan
yang
merangsangkan
mikoriza
dari
percambahan
dan
V. hookeriana
kulat-kulat
akar-akar
yang lebih
mikorizanya
bersekutu dengan ketiga
sendiri.
jenis
orkid
merosakkan biji-biji benihnya.
Kebanyrucan
jenis
orkid
ini
kulat-kulat yang bersekutu dengan ketiga-tiga
tidak
mempengaruhi
pertumbuhan
struktur­
struktur seperti protokom dari V. hookeriana dan tidak
bagi T. obtusum.
anak
benih
sesuai
Pengaruhnya terhadap penyesuaian tumbuh dari
V. hookeriana dan
T. obtusum pada
semaian
benih juga tidak tampak berkesan dalam penyelidikan ini.
xx
anak
CHAPTER I
I N T
Among
ornamental
U C T ION
R OD
plants, orchids are one
economically important in this region.
made
to
support the
propagation
practices
seedlings,
are
marketing
plants
over-collection
valuable
techniques
of
to ful fil
through
cultivation
the
demand
However, recently
(Hagsater
world
have
germplasms from
are
e f forts are
methods,
,dId orchids or land
measures
Conservation
studies
most
of
people
possible loss o f many orchid species due to
the
in
countries
breeding
or cut flowers.
aware o f the
There fore
the
development o f orchid industry
techniques,
and
of
to be
clearing
& Steward,
taken to
extinction.
in
1986 ) .
prevent
Suitable
needed to multiply them.
many
This
these
propagation
e f fort
also encourage further breeding and utilization o f the
would
orchid
species and also has a signi ficant role on their conservation.
There
them, three
they
are
are
species
but is
had
Malaysian
been
orchids, and among
chosen for this
Paphiopedilum niveum,
Yanda hookeriana.
in nature.
many important
Taeniophyl1um
research and
obtusum
Paphiopedilum niveum is restrictedly
and
found
This species is not only an attractive wild orchid
also a potential species for breeding.
1
Its commercial