IJBRITISH-280
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Influence of chemical mutagens under variability
Of Lemon hybrid seedlings
Memarne G.1, Kashakashvili Ts.2, Khalvashi N.3, Gabaidze M.4
1
– PhD in Agriculture, Shota Rustaveli State University, Institute of Phytopathology and Biodiversity,
Kobuleti, Georgia, e-mail: [email protected]
2
– PhD in Agriculture, Institute of Tea, Subtropical Crops and Tea Industry, Ozurgeti, Georgia, e-mail:
[email protected]
3
– PhD in Agriculture, Shota Rustaveli State University, Institute of Phytopathology and Biodiversity,
Kobuleti, Georgia, e-mail: [email protected]
4
– PhD in Agrarian science, Shota Rustaveli State University, Institute of Phytopathology and Biodiversity,
Kobuleti, Georgia,
e-mail: [email protected]
Abstract
Pollens treated with chemical mutagen NEU and NMU in concentrations – 0.01%; 0.05%; 0.1% and
expositions – 1.5 h; 3 h; 6 h; then they were used in hybridization as a paternal component.
Specific reaction of useful knot, ability of germination has been determined. With increasing of
expositions (3-6 hrs) and concentrations (0.1%) of mutagen (NEU; NMU) the useful knot and seed
germination are decreased. This dose of exposition (6 h) and concentration (0.1%) of the both mutagen can
be considered to be critical, which causes lethality.
It should be underlined, that the appeared phenotypic variated hybrid seedlings of lemon differ
greatly from paternal forms and are received in a definite in case of NEU mutagen – 8.1%-65.5%; in case of
NMU -8.0%-32.1%.
Treatment of pollens with chemical mutagen (NEU and NMU) according to the power of influence
mutagen NEU appeared to be the most effective one; and its using in lemon crossing causes the increasing
of number hybrid seedlings variability (65.5%).
Key words: mutagenesis, lemon, chemical mutagen, induced mutations, pollen
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Chromosome alternation, also takes place and gives a good
INTRODUCTION
chance to realize wished locus combinations (11).
Mutation induction has a high potential for bringing about
genetic changes and improvements [1] and has contributed
Methodology
considerably to plant breeding. Mutations in nature are very
rare facts, and useful share for a person among them is not so
With the aim of increasing the lemon variability frequency,
much. Nowadays physical and especially chemical factors
the pollen of Mayer Lemon was treated with chemical
give a good opportunity to increase natural plant variability
mutagen, nitroethil-urea (NEU) and nitrozoethil-urea (NMU)
thousands times and obtain significant number of useful
in three concentration - 0.01%; 0.05%; 0.1% and expositions –
variations.
1.5 h; 3 h; 6 h and then they were used in hybridization as a
The number of mutant varieties officially released and
paternal component.
recorded in the FAO/IAEA Mutant Varieties Database
Lemon pollen seed (in gauzes sacs) were sunk into the
before the end of 2000 was 2252 among which many
solution of mutagen of different concentration. After washing
fruit trees [2]. Examples include mutants such as russet-
with water they were put on Petri dishes and placed into
free fruit in apple, seedless Citrus, disease resistance in
thermostat under 25°C for ripening.
Japanese pear and compactness in sweet cherry [3].
Control variant was treated with pure water. Treated in such a
The responsiveness of cells to physical and chemical
way the pollens were used in hybridization for obtaining inner
mutations is influenced, to a varying degree, by numerous
specific hybrids.
biological, environmental and chemical factors. These
The biometrical dates were treated using methods of dispersal
factors modify the effectiveness of mutagens in cells.
analyze of Dospekhov (12).
Moisture content was shown to be one of these factors. Low
moisture content seeds have been shown to improve the
tolerance of the seeds to radiation [4]. The effects of
gamma irradiation on Citrus seeds have been previously
studied, particularly in Shamouti orange (Citrus Sinensis)
seeds [5] and Satsuma (Citrus Unshiu) [6].
A radiation
dose of 0-120Gy was used on Shamouti, while 0-100 Gy was
used on Satsuma.
Recently great attention is paid spontaneous mutagens,
obtained artificially under influence of chemical factors on
vegetable and generative organs of plant. Plant pollen attracts
great attention as the latest stage of ontogenesis, where
heredity potential of the organisms is localized, and we
suggest that under chemical mutagen treatment the knot
appearing of seeds and seedlings will vary in high diapason
depending on chemical mutagen, its dose, exposition and
genotype. At the same time, according to literature data
(7,8,9,10), it is known that in treated pollen some changes take
place which, weaken heredity of symptoms of paternal plant,
but in some cases on the contrary – strengthen; and these
RESULTS AND DISCUSSIONS
Pollination results, treated by Mayer Lemon pollen with
Monackello Lemon in field conditions are given in Table 1.
As it is clearly seen from the Table, useful knotting varies
depending on concentration and exposition of mutagen in
comparison with the control. Under 1.5 h expositions in all
three concentrations, in case of NEU, the knotting varies
between 2.8% and 19.3%; but in case of NMU- 2.5%-15.2%
while in control it is – 3.7. Useful knot decreases with the
increasing exposition and concentration.
Seed germination studying showed (Table 1) that chemical
mutagen
stimulates
in
comparison
with
the
control.
Percentage of seed germination varies between 33.3%-83.3% in case of NEU and in case of NMU – 30.5%-69.6% on
concentration and exposition while in control it is – 52.3%.
Lower concentration (0.1%) of mutagen (NEU, NMU) was
decreasing seed germination power. This dose – concentration
0.1% and 6 h exposition of both mutagens can be considered
critical, that causes lethality.
changes will be realized as soon as zygote appears.
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0.01
0.05
0.1
3
3
3
5.2±0.3
3.8±1.5
2.5±1.8
69.6±3.7
50.0±4.4
30.5±4.1
NMU
NMU
NMU
0.01
0.05
0.1
6
6
6
4.4±0.8
3.3±1.0
-
32.3±3.3
31.1±3.3
-
NEU
NEU
NEU
0.01
0.05
0.1
1.5
1.5
1.5
19.3±2.1
18.3±2.1
6.4±1.3
80.0±3.3
61.5±4.3
66.6±4.2
NEU
NEU
NEU
0.01
0.05
0.1
3
3
3
6.3±2.1
4.1±2.3
2.8±1.9
83.3±2.2
61.5±3.3
33.3±3.5
NEU
NEU
NEU
0.01
0.05
0.1
6
6
6
5.4±0.8
4.3±1.0
-
66.6±4.0
60.0±4.0
-
Control
(water)
-
-
3.7±1.0
52.3±4.1
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
NMU
NMU
NMU
NMU
NMU
NMU
NMU
NMU
NMU
NEU
NEU
NEU
NEU
NEU
NEU
NEU
NEU
NEU
Control
(water)
0.01
0.05
0.1
0.01
0.05
0.1
0.01
0.05
0.1
0.01
0.05
0.1
0.01
0.05
0.1
0.01
0.05
0.1
-
1.5
1.5
1.5
3
3
3
6
6
6
1.5
1.5
1.5
3
3
3
6
6
6
-
18.1
22.5
22.7
16.9
22.4
21.5
15.8
13.6
19.6
17.5
22.6
21.2
16.4
26.5
23.2
15.3
17.2
64.4
54.0
45.2
74.6
63.5
67.6
75.7
73.4
35.9
20.3
11.9
37.0
42.1
23.6
63.5
76.6
79.6
Variability %
NMU
NMU
NMU
Seedlings deviated to
the maternal part
66.6±3.1
60.8±2.8
37.5±3.7
Seedlings deviated to
the parental
hybrid seedlings
germination %
15.2±2.0
13.3±1.4
6.2±1.2
Hybrid
Infructescense %
1.5
1.5
1.5
Monackello× Mayer
Exposition per
hour
0.01
0.05
0.1
N
Exposition per hour
Mutagen
concentration
NMU
NMU
NMU
Table 2. Variability and Domination of parental
features among hybrid seedlings of Lemon
Concentration
mutagen
Monackello× Mayer
Combination
Table 1. Influence of chemical mutagens on
infructescense and hybrid seedlings appearing
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Mutagen
IJBRITISH-280
17.5
23.6
32.1
8.5
14.1
10.9
9.2
8.0
44.5
62.2
65.5
41.3
41.5
49.9
12.3
8.1
3.2
The most attractive picture is obtained while combination
Monackello and Mayer. In case of pollen treatment with
mutagen (NEU and NMU) seedlings deviated to the paternal
part is being decreased up to 15.3% and 13.6%. The quantity
CONCLUSIONS
of seedlings is being increased, accordingly, deviated to the
As a result our experiments, we have the following
material part (Table 2).
1.
The table shows that seedlings variated phenotypically differ
effective mutagen is nitrozoethil-urea.
from the parent forms and are in some definite quantity; in
2.
case of mutagen NEU – 81%-65.5%, and in case NMU –
and 0.05% concentrations and 1.5-3 h exposition) the
8.0% - 32.1%.
percentage of knot quality and ability of germination increase.
Increasing of regeneration frequency with lemons while using
3.
the treated pollen in crossing, is probably connected with
lemons under using the treated pollen (NEU, NMU) in
direct mutagen influence on genetic pollen material, but
crossing is, probably, connected with direct mutagen influence
increasing of mutagen features of pollen plasma is not also an
on genetic pollen material, and it can explain the increasing of
exception.
hybrid seedlings variability frequency.
Among new formations the following ones can be pointed out;
4.
different forms of branching, shooting, power of growth, and
NMU) while using them in lemon crossing causes a
variation of a stem, leaf, and plants of dwarfish type appear.
significant increasing of mutations.
According to the power of influence, the most
In case of mutagen NEU and NMU under low (0.01%
Increasing of new formations frequency among
Treating of pollens with chemical mutagen (NEU and
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ACKNOWLEDGE
[7]
The authors would like to acknowledge scientists (Shota
on crossing effect. Chemical mutagenesis in
Goliadze, Lia Tikanadze and Aleksandra Vashalomidze) from
increasing of agricultural plant production, 1984.
Institute of Tea, Subtropical Crops and Tea Industry, Ozurgeti,
[8]
Georgia for their technical support during field work
mutagen on inseminating ability of citrus plants.
Diasamidze A.O. Influence of chemical
Subtropical crops. N. 3, 1970.
REFERENCES
[9]
[1]
Asadov Sh.I. Influence of cotton treatment
SIGURBJORNSSON,
B.,
Introduction:
Mutations in Plant Breeding Programmes, Manual
on Mutation Breeding, Joint FAO/IAEA Division
of Atomic Energy in Food and Agriculture 160,
1977.
Diasamidze A.O. Natural and Chemical
mutagenesis in citrus breeding. Batumi, 1994.
[ 10 ] Kerkadze
M.
et
al.,
Officially
released mutant varieties In: The FAO/IAEA
Database for Mutation Breeding 12, 2000.
Induced
mutation
of
subtropical crops. Subtropical crops.N2, 1985.
[ 11 ] Rapoport I.A. Perspectives of chemical
mutagenesis
[ 2 ] MALUSZYNSKI,
I.G.
using
in
breeding.
Chemical
mutagenesis and breeding, 1971.
[ 12 ] Dospechov B. Methods of field experience.
Kolos. Moskow, 1979.
[ 3 ] SANADA, T., AMANO, E. Induced mutation
in fruit trees In: Somaclonal Variation and
Author’s Profile and Image
Induced Mutations in Crop Improvement. JAIN,
S.M., et al., Eds., Kluwer Academic Publishers,
Dordrecht, 1998.
[4]
CONGER,
sensitivity and
B.V.,
al.,
Radiation
modifying factors
In: Plant
Breeding Programmes,
et
Manual
on Mutation
Breeding, Joint FAO/IAEA Division of Atomic
Energy in Food and Agriculture, 1977.
[5]
SPIELGEL–ROY,
Radiosensitivity
of
P.
shamouti
et
al.,
orange
(Citrus
Memarne Guram - Director of the Institute of
Phytopathology and Biodiversity, Kobuleti
sinensis) seeds and buds. Radiation Botany 13,
1973.
[ 6 ] SPIELGEL – ROY, P., PADOVA, P. Gamma
irradiation and pollen cultivar influence on
polyembryony of ‘Satsuma’ (Citrus unshiu Marc.).
Radiation Botany 12, 1972.
Kashakashvili Tsisana – main scientist of
Institute of Tea, Subtropical Crops and Tea
Industry, Ozurgeti
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Khalvashi Neli - main scientist of Department of
biodiversity monitoring and conservation of the
Institute of Phytopathology and Biodiversity,
Kobuleti
Gabaidze Mziuri - Senior Scientist of
Department of plant diseases monitoring,
diagnostics and molecular biology of the
Institute of Phytopathology and Biodiversity,
Kobuleti
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