Induced polyploidization in Brassica campestris L. (Brassicaceae)
Present experimental design has been made up to obtain crop with higher ploidy level via synthetic polyploidization. Since ploidy manipulation is generally associated with the obtainment of some increased enviable traits of the crop and also provides them greater adaptability to unfavorable or harsh...
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| Cite this: | Induced polyploidization in Brassica campestris L. (Brassicaceae) / G. Kumar, K. Dwivedi // Цитология и генетика. — 2014. — Т. 48, № 2. — С. 43-51. — Бібліогр.: 43 назв. — англ. |
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nasplib_isofts_kiev_ua-123456789-1266252025-02-23T17:13:30Z Induced polyploidization in Brassica campestris L. (Brassicaceae) Индуцированная полиплоидизация у Brassica campestris (Brassicaceae) Kumar, G. Dwivedi, K. Оригинальные работы Present experimental design has been made up to obtain crop with higher ploidy level via synthetic polyploidization. Since ploidy manipulation is generally associated with the obtainment of some increased enviable traits of the crop and also provides them greater adaptability to unfavorable or harsh circumstances as compared to its diploids counterparts. Thus, herein present research autotetraploids of Brassica campestris L. have been lucratively achieved by the application of colchicine. Two methods of treatment were utilized i.e. seed treatment and seedling treatment. No polyploidy could be obtained through seed treatment while seedling treatment responded well towards polyploidy. However, the status of autotetraploidy has been confirmed by cytomorphological investigations of treated plants as against its diploids counterparts. For the purpose, morphological parameters such as increased stomata size, pollen diameter, flower size, reproductive organs whereas reduction in plant height, leaf length, leaf breadth, stomata frequency, number of flowers/inflorescence etc. were appraised. Further, cytological observations were made that had clearly revealed the doubling of genome in the autotetraploids as compared to diploids. Meanwhile, pollen fertility and size of pollen grains were evaluated as well. Разработана экспериментальная схема получения сельскохозяйственных культур с более высокой плодиностью путем искусственной полиплоидизации. Изменение плоидности обычно связано с приобретением некоторых важных признаков у таких культур, а также обеспечивает их более высокую приспособляемость к неблагоприятным или суровым условиям внешней среды по сравнению с диплоидными аналогами. В настоящем исследовании представлено успешное получение автотетраплоидов Brassica campestris L. с помощью колхицина. Для этого применяли два метода обработки – семян и проростков. Никакой полиплоидии не обнаружено при обработке семян, тогда как после обработки проростков полиплоиды были получены. С помощью цитоморфологических исследований подтверждено наличие автотетраплоидности у обработанных растений по сравнению с их диплоидными аналогами. Для этого оценивали такие морфологические параметры, как увеличенные размер устьиц, диаметр пыльцы, размер цветка, репродуктивные органы, а также уменьшенные высота растений, длина листа, ширина листа, частота устьиц, число цветков и соцветий. Более того, в результате цитологических исследований четко установлено удвоение генома у автотетраплоидов по сравнению с диплоидами. Наряду с этим в работе оценивали фертильность пыльцы и величину пыльцевых зерен. We are grateful to N.B.P.G.R, New Delhi for providing inbred seeds of Brassica campestris L. accession number – IC363713. One of the authors (K. Dwivedi) is thankful to DST, New Delhi for providing financial assistance as Inspire Fellowship. 2014 Article Induced polyploidization in Brassica campestris L. (Brassicaceae) / G. Kumar, K. Dwivedi // Цитология и генетика. — 2014. — Т. 48, № 2. — С. 43-51. — Бібліогр.: 43 назв. — англ. 0564-3783 DOI: 10.3103/S0095452714020066 https://nasplib.isofts.kiev.ua/handle/123456789/126625 en Цитология и генетика application/pdf Інститут клітинної біології та генетичної інженерії НАН України |
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Оригинальные работы Оригинальные работы |
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Оригинальные работы Оригинальные работы Kumar, G. Dwivedi, K. Induced polyploidization in Brassica campestris L. (Brassicaceae) Цитология и генетика |
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Present experimental design has been made up to obtain crop with higher ploidy level via synthetic polyploidization. Since ploidy manipulation is generally associated with the obtainment of some increased enviable traits of the crop and also provides them greater adaptability to unfavorable or harsh circumstances as compared to its diploids counterparts. Thus, herein present research autotetraploids of Brassica campestris L. have been lucratively achieved by the application of colchicine. Two methods of treatment were utilized i.e. seed treatment and seedling treatment. No polyploidy could be obtained through seed treatment while seedling treatment responded well towards polyploidy. However, the status of autotetraploidy has been confirmed by cytomorphological investigations of treated plants as against its diploids counterparts. For the purpose, morphological parameters such as increased stomata size, pollen diameter, flower size, reproductive organs whereas reduction in plant height, leaf length, leaf breadth, stomata frequency, number of flowers/inflorescence etc. were appraised. Further, cytological observations were made that had clearly revealed the doubling of genome in the autotetraploids as compared to diploids. Meanwhile, pollen fertility and size of pollen grains were evaluated as well. |
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Article |
| author |
Kumar, G. Dwivedi, K. |
| author_facet |
Kumar, G. Dwivedi, K. |
| author_sort |
Kumar, G. |
| title |
Induced polyploidization in Brassica campestris L. (Brassicaceae) |
| title_short |
Induced polyploidization in Brassica campestris L. (Brassicaceae) |
| title_full |
Induced polyploidization in Brassica campestris L. (Brassicaceae) |
| title_fullStr |
Induced polyploidization in Brassica campestris L. (Brassicaceae) |
| title_full_unstemmed |
Induced polyploidization in Brassica campestris L. (Brassicaceae) |
| title_sort |
induced polyploidization in brassica campestris l. (brassicaceae) |
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Інститут клітинної біології та генетичної інженерії НАН України |
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2014 |
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Оригинальные работы |
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https://nasplib.isofts.kiev.ua/handle/123456789/126625 |
| citation_txt |
Induced polyploidization in Brassica campestris L. (Brassicaceae) / G. Kumar, K. Dwivedi // Цитология и генетика. — 2014. — Т. 48, № 2. — С. 43-51. — Бібліогр.: 43 назв. — англ. |
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Цитология и генетика |
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2025-11-24T02:28:41Z |
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1849637021143793664 |
| fulltext |
43ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 2
Present experimental design has been made up to obtain
crop with higher ploidy level via synthetic polyploidization.
Since ploidy manipulation is generally associated with the
obtainment of some increased enviable traits of the crop
and also provides them greater adaptability to unfavorable
or harsh circumstances as compared to its diploids
counterparts. Thus, herein present research autotetraploids
of Brassica campestris L. have been lucratively achieved
by the application of colchicine. Two methods of treatment
were utilized i.e. seed treatment and seedling treatment.
No polyploidy could be obtained through seed treatment
while seedling treatment responded well towards polyploidy.
However, the status of autotetraploidy has been confirmed
by cytomorphological investigations of treated plants as
against its diploids counterparts. For the purpose, morpho-
logical parameters such as increased stomata size, pollen
diameter, flower size, reproductive organs whereas re-
duction in plant height, leaf length, leaf breadth, stomata
frequency, number of flowers/inflorescence etc. were appraised.
Further, cytological observations were made that had clearly
revealed the doubling of genome in the autotetraploids as
compared to diploids. Meanwhile, pollen fertility and size of
pollen grains were evaluated as well.
Keywords: autotetraploid, Brassica campestris L., colchi-
cine, diploid.
Introduction. Polyploidy, i.e., the possession of
more than three complete sets of chromosomes has
been a pivotal factor in plant evolution [1]. Hence,
from ancient to contemporary polyploidization is
considered as an ongoing and recurrent process that
has also been acknowledged as an invaluable force
for characterization of evolutionary trends in plants.
Moreover the genome doubling (polyploidy) has
been regarded as a pervasive force in plant evolution
[2]. It might be responsible for adaptation and
variation among similar or different kind of plant
species. Recent estimates suggest that 70 % of all
angiosperms have experienced one or more episodes
of polyploidization [3] and most flowering plant
species have evolved through one or more round
of polyploidy either by doubling of chromosome
number with in a species (autopolyploidy) or by
combining chromosome sets from distinct but re-
lated species (allopolyploidy). Topical studies con-
vincingly demonstrate that polyploidy entails far
more than mere merger of two genomes who pas-
sively acquiescence to their sudden collaboration
but instead involve a whole spectrum of molecular
and physiological adjustments [2].
Moreover, induced polyploidy offers some bene-
fits for horticulture, pharmaceutical and agricultu-
ral improvement of plants [4]. For instance, higher
ploidy level was associated with increased flower
size in rhododendron and intensified flower colors
in carnation and cyclamen [5, 6] and this manipula-
tion in ploidy is also significant for genetic im-
provement of many plants [7] often generates
variants that may possess useful characteristics and
by doubling the gene products it also provides a
wider germplasm base for breeding studies. Many
researchers have induced polyploidy in several
plant species through the antimitotic chemical,
colchicine (C22H25NO6) [8–10] which is obtained
from the seeds and bulbs of the plant Colchicum
autumnale L. More importantly without damaging
any other life processes of the chromosomes,
colchicine effects a temporary inactivation of the
spindle mechanism, both the exterior part working
from the centrosomes and the interior centromeric
part [11]. Among various antimicrotubular drugs col-
chicine has proven to be the most useful to double
the chromosome numbers of numerous crop spe-
cies, including decorative flowers, medical and agri-
cultural plants [12].
Before going ahead we wish to mention that,
rapeseed is a group of oilseed crops which assumes
the significance by occupying third rank among the
oilseed crops after soybean and oil palm in pro-
duction of vegetable oils in the international oilseed
market [13]. It is among the oldest cultivated plant
in human civilization. The oilseed crop belongs to
the family Brassicaceae, cultivated throughout the
world with several species and types as oilseed crops,
vegetables and forage crops [14]. They are also used
in the production of condiments and recognized as
G. KUMAR, K. DWIVEDI
Plant Genetics Laboratory, Department of Botany, University of Allahabad, India
E-mail: kshama.dwivedi@gmail.com
INDUCED POLYPLOIDIZATION
IN BRASSICA CAMPESTRIS L. (BRASSICACEAE)
© G. KUMAR, K. DWIVEDI, 2014
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 244
G. Kumar, K. Dwivedi
a valuable source of dietary fiber and moreover,
Brassica vegetables contain little fat, and are sources
of vitamins, minerals, and fiber [14]. However,
its cake obtained after crushing of seeds is highly
nutritious for cattle due to containing some amount
of protein hence extensively used as cattle feed.
Thus, in view of considerable economic im-
portance of Brassica campestris L., the present
work has been aimed to obtain polyploid mutant
lines by the implementation of colchicine in Bras-
sica campestris L. which were verified on basis of
significant differences in various phenotypic traits
of autotetraploid over its diploid counterparts. The-
reby, resulting genome doubling or ploidy mani-
pulation could be correlated with the enhancement
of various economically important traits.
Materials and methods. Inbred seeds of culti-
var Brassica campestris L. (Accession number-
IC363713) were obtained from National Bureau of
Plant Genetic Resources (NBPGR), New Delhi.
Colchicine used in present work was manufac-
tured by Loba Chemie Pvt. Ltd. Mumbai (India).
Two methods were utilized for polyploidization
1-seed treatment and 2-seedling treatment. No
polyploidy could be obtained via seed treatment
however seedlings treatment responded well and
showed positive response towards polyploidy. To
assess polyploidy, healthy, dry and homogeneous
seeds of Brassica campestris were washed and pre-
soaked in distilled water for 2 hours, excess water
was removed by drying seeds on blotting paper,
sown in pots in triplicates at each dose. Concur-
rently, control was maintained. After 7 days’ of ger-
mination when seedlings reached 2–3 cm in length,
they were treated with different concentrations of
aqueous solution of colchicine (C22H25NO6) viz.,
0.3, 0.4, 0.5, 0.6 %. For this sterilized cotton balls
were dipped in 0.3 % concentration of colchicine
then placed on the growth apex of young seed-
lings for 12, 24 and 36 consecutive hours along with
the recovery of 12 hours and the similar methods
were repeated for each dose. The pots were covered
with perforated thin plastic sheets as a measure to
check evaporation. Cotton balls were removed and
apical tips were washed with tap water for proper
growth and recovery.
Deformed preliminary leaves and delayed growth
rate have been noticed just after colchinization.
Furthermore, length and breadth of stomata has
been documented and their frequency of occurrence
were measured since stomata based analyses has
been proved very significant to confirm induction of
polyploidy before the cytological observations. For
the purpose, young leaves were peeled off from its
dorsal surface, stained with safranin for few minu-
tes and washed thoroughly and mounted in glyce-
Table. 1. Colchicine treatment in seedlings of Brassica campestris L.
* Confirmed by meiotic observation.
Concentration, %
Duration
(hours)
Number of
seedlings
treated
Plants
survived
Expected
polyploids
Reverted
plants
Confirmed auto-
tetraploids *
No. %
Overnight treatment 6.00 pm – 6.00 am
0.3
0.3
0.3
0.4
0.4
0.4
0.5
0.5
0.5
0.6
0.6
0.6
12
24
36
12
24
36
12
24
36
12
24
36
30
30
30
30
30
30
30
30
30
30
30
30
29
25
29
27
24
27
27
21
21
27
21
27
9
15
27
12
9
15
12
21
18
18
24
24
9
15
25
12
9
15
12
21
17
15
24
23
–
–
2
–
–
–
–
–
1
3
–
1
–
–
6.66
–
–
–
–
–
3.33
10
–
3.33
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 2 45
Induced polyploidization in Brassica campestris L. (Brassicaceae)
rin gel, observed under optical microscope. Length
and breadth of stomata was measured using ocular
micrometer in different views and their mean values
were calculated. Morphological characters such as
plant height, leaf length and breadth etc. have been
documented. Large sized young flower buds and
remarkable increase in flower size with less number
of flowers per inflorescence were also marked.
For meiotic preparation, small sized floral buds
were fixed in glacial acetic acid : ethyl alcohol
(1 : 3, v/v) and then transferred to 70 % alcohol
and refrigerated at 4 ºC until use. After 24 hours
slides were prepared by using anther squash techni-
que with 2 % standard aceto-carmine stain with
traces of ferric chloride [15] and observed under
microscope (Olympus CH20i). Pollen viability was
also recorded on the basis of pollen stainability test
i.e. stained and filled pollen grains were considered
as fertile however empty, small and shriveled were
assumed as sterile. Photomicrographs were made by
using Pinnacle PCTV captured device and statistical
analyses have been done by using statistica-8 software
of stat-soft to measure mean values of variables.
Results. Genomic constitution of diploid plant
was absolutely normal (2n = 20) while, one fold
increment in genomic constitution of Brassica cam-
pestris L. was encountered in autotetraploid plants
(4n = 40). Out of 360 seedlings treated, only 23.32 %
Fig.1. a – deformed growth of seedling due to colchicine
treatment; b – normal seedling with newly formed ve-
getative (true) leaves; c – an autotetraploid plant; d – a
diploid plant; e – flowers of diploid (2n), mixoploid and
autotetraploid (4n); f – a mixoploid plant; g – a flower
having five petals; h – reproductive organs of autotetraplo-
id (4n) and diploid (2n) where 4n displays bigger repro-
ductive organs
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 246
G. Kumar, K. Dwivedi
of seedlings were confirmed as autotetraploids on
morphological as well as cytological basis however,
remaining percentage of treated seedlings either di-
ed or reverted (Table 1).
Morphological observations. Herein present case,
preliminary manifestations of colchinization were
delayed growth rate which was later found to be
accelerated though unable to exceed that of diploid
while diploid showed their regular way of growth
pattern with normal plant height (Table 2). Later,
treated juvenile seedlings showed elongated petioles
of cotyledonary leaves and, emergence of highly de-
formed true leaves showing swelling, curling and
rolling of lamina (Fig. 1, a) which were highly
thicker, dark green, leathery and quite smaller as
against diploids. However, some other phenotypic
characters such as flower (cruciform) size (Fig. 1, e),
stomata size (Fig. 1, k), pollen grains diameter
(Fig. 1, i) and reproductive organs (Fig. 1, h) have
been increased vigorously in autotetraploid plants.
Further 50 % flowering were registered that showed
delay of 11 days in autotetraploids as compared to
diploids. Autotetraploids also exhibited less number
of flowers per inflorescence; i.e., 14 ± 0.86, while
it was 40 ± 1.34 in diploids, with marked large
sized flowers in autotetraploid plants (Fig. 1, e).
While more interestingly, few flowers having more
than four petals (penta-petalous) with larger sized
reproductive organ has been documented in present
case (Fig. 1, g, h). Presence of five petals in Brassica
is in fact a rare phenomenon observed in present
case and never noticed earlier. Since Brassica crops
strictly bear four petals which are reminiscent of a
cross means «flowers with four petals in cross sec-
tion» known as crucifers. Some other morphological
features observed were plant height, leaf length, leaf
breadth and number of inflorescences/plant which
registered decline in its mean value over diploid.
Stomatal observation. Length of stomata in auto-
tetrapoloids was 52.48 ± 0.02 m, although it was
27.97 ± 0.21 m in diploids.
Similar difference in breadth has been observed
in stomata (Fig. 1, j, k). In spite of bigger sized
stomata, significant difference in the frequency of
stomata (number of stomata/mm2) of diploid and
autotetarploid has been administered (Table 2).
Palynological observation. Pollen diameter in
autotetraploid and diploid plants was 67.95 ± 0.02
and 26.05 ± 0.28 m, respectively. That clearly
reveals the difference between pollen sizes in treated
as well as non-treated plants. Thus, the majority of
pollen grains in autotetraploids were bigger com-
pared to diploids. A significant reduction in the
pollen fertility of autotetraploids has been recorded
(Table 2).
Thus, the above morphological observations
clearly verify that the induced autotetraploid plants
were robust, more compact bearing increased ve-
getative and reproductive organs, in short was su-
perior over its diploid counterparts.
Cytological observation. The somatic complement
of Brassica campestris L. is 2n = 20 while doub-
ling of chromosomes was found in induced auto-
tetraploid. Fig. 2, a–c clearly display the diaki-
Table 2. Comparative analyses of various morphological and physiological characters
of diploids and induced autotetraploids (mean ± SE)
Characteristics Diploid (2n = 20) Autotetraploid (4n = 40)
Plant height, m
No. of flowers/inflorescence
Pollen diameter, m
Stomata length, m
Stomata breadth, m
Frequency of stomata/mm2
50 % flowering
Maturity
Pollen fertility, %
Leaf length, cm
Leaf breadth, cm
Number of inflorescence/plant
1.46 ± 0.04
40.00 ± 1.34
26.05 ± 0.28
27.97 ± 0.21
15.08 ± 0.24
21.20 ± 0.50
80.0 ± 0.18
142.00 ± 0.45
98.66 ± 0.26
24.50 ± 0.16
8.44 ± 0.71
11.40 ± 0.86
0.96 ± 0.03*
14.00 ± 0.86
67.95 ± 0.02
52.48 ± 0.02
25.89 ± 0.33
6.80 ± 0.50
91.00 ± 0.47
135.00 ± 0.32
63.33 ± 3.33
16.91 ± 1.32
6.34 ± 0.14
9.21 ± 0.33
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 2 47
Induced polyploidization in Brassica campestris L. (Brassicaceae)
Fig. 2. a – Normal PMC showing diakinesis (n = 10); b – Normal PMC showing metaphase-I (2n = 20); c – Normal
PMC showing anaphase-I (10 : 10); d – PMC showing autotetraploid diakinesis; e – PMC showing autotetraploid
metaphase-I (4n = 40); f – PMC showing autotetraploid anaphase-I; g – PMC showing autotetraploid metaphase-II;
h – Typical tricolpate diploid pollen grains of diploid; i – Bigger sized fertile tetracolpate and pentacolpate pollen
grains of autotetraploid; j – Stomata of diploids; k – Stomata of colchicine induced autotetraploids (bigger sized
stomata with decreased frequency of stomata/mm2 over diploid). Scale bar: a–i – 4.2 m; j–k – 10 m
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 248
G. Kumar, K. Dwivedi
nesis, metaphase-I and anaphase-I, respectively in
diploids. Meiosis in PMCs of diploids was regu-
lar however, in treated plants, autotetraploid status
could be confirmed by chromosomal doubling i.e.,
4n = 40 that showed the increment in the number
of bivalents in PMCs of polyploids despite univa-
lents and other association in some of the PMCs
yet the frequency of their occurrence was almost
negligible. Fig. 2, d–g show diakinesis, metapha-
se-I, anaphase-I and metaphase-II, respectively in
autotetraploid. Thus, on the basis of above ob-
servations, percentage of induced autotetraploid
plants has been mentioned in Table 1. Notwithstan-
ding polyploids, low frequency of mixoploids was
also discernible in some case (Fig. 1, f).
Discussion. Polyploid plants may often possess
superior agronomic traits over their diploid coun-
terparts. For example, ployploids may have larger
leaves and flowers, thicker stems and roots, dar-
ker green leaves, an increased width-to-length ra-
tio of the leaves, increased cell size, leading to lar-
ger reproductive and vegetative organs a more com-
pact growth habit and a higher tolerance to en-
vironmental stress [16–18]. On the basis of above
mentioned facts, herein present study the utility of
various morphological, physiological and cytologi-
cal analyses in distinguishing autotetraploids from
diploids has been tested.
Just after colchinization, number of plants was
suspected for being polyploids though only few were
confirmed as autotetraploid after cytological study
and rest has been reverted and started to behaving
as controls. The retarded growth rate might be due
to the reduced rate of cell division that resulted from
the physiological disturbance caused by colchici-
ne [19]. Joshi et al. [20] recorded that polyploid
plants were slower in growth initially, but later
showed gigantism. They are generally larger than
their diploid counterparts and frequently develop
into thicker tissues, resulting in large sized plant
organs [21]. Present investigation fully agreed with
these findings i.e. thicker leaves, stem, increased
reproductive organs etc. however some traits were
found to be negatively correlated such as plant
height, leaf length, leaf breadth etc. Though among
them, decreased plant height of the autotetraplo-
id has been found to be enviable since it prevent
plant from being lodged. Lodging is a serious prob-
lem for oilseed crop and it reduces the yield capa-
city of crop. However increase in the number and
size of the chloroplast with quantitative increase in
the DNA content could be the considerable cause
of the dark green thicker leaves of autotetraplo-
ids [22].
The relationship between flower sizes with num-
ber of flowers per inflorescence was found to be
reciprocal. Differences in length of cruciform pe-
tals in polyploids and diploids were also promi-
nent i.e. flowers in autotetraploids were noticeably
larger over diploid which could be due to «gigas
effect» which was an increase in polyploid cell size
that often resulted in increased organ size through-
out the plant [23]. Jana et al. [24] found that the
autotetraploid individuals had larger flowers than
their diploid counterparts more often than expec-
ted by chance and also documented that autopoly-
ploid species and genera should have larger inflo-
rescences than diploid species and genera. Bigger
flowers in polyploids were also subsequently con-
firmed by various researchers [25–27]. The oc-
currence of five petals in some of the flowers of
autotetraploids is ambiguous however it might be
possible that colchicine induced disturbed male
meiosis may lead to some disturbance in particu-
lar gene(s) that governs the typical «cross-bearing»
four-petalous condition of the present crop thereby
resulting more than 4-petals.
In plants stomatal analysis and pollen diameter
provide an efficient mean for characterization of
polyploidy as these characters generally increased
in polyploids [28–31]. Cohen and Yao [32] pro-
duced tetraploid Zantedecsbia via in vitro colchi-
cine treatment and primarily verify the status of
tetraploid via stomatal analysis and found that
stomatal analyses could be the most efficient crite-
ria for ploidy confirmation. Similarly increase in
the size of pollen diameter has been documented
in present case which eventually concurred with
the observation of the Jaskani et al. [33] who ap-
plied colchicine to watermelon and found the
autotetraploid plants bearing bigger flowers with
increased number of pollen colpi over diploids.
Despite larger size of pollens, declining frequen-
cy of pollen viability has been discernible that in
turn reduced the seed yield. Reduced pollen fertil-
ity and increased cell and organ size (gigas-effects)
were considered as universal effects of artificial
polyploidization [34–36].
We delve further in to the cytological observa-
tion that indeed, became difficult because of very
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 2 49
Induced polyploidization in Brassica campestris L. (Brassicaceae)
small and poorly differentiated chromosomes of
Brassica and found that their identification through
the ordinary cytogenetical techniques was extremely
difficult [37]. Hence, it had been a difficult task to
explore the exact number of bivalents especially in
case of diploids. Yet, one fold increment of biva-
lent count was observed in case of autotetraploids
compared to diploids due to colchicine treatment.
The high frequency of bivalents in the autotetra-
ploids might be due to small size of chromosomes
or due to a gene favouring bivalents formation [38].
Treatments with 0.3 % for 24 hours and 0.6 %
for 12 hours also possessed some mixo-ploids am-
ong the population of diploids and auto-tetraplo-
ids which could be justified by the fact that the
antimitotic agent cannot reach all meristematic
cells at the time of treatment [39]. Occurrence of
mixoploids is a major problem in the recovery of
autotetraploids [40].
Meiotic behavior of autotetraploids was almost
normal except few instances of meiotic aberrations
viz. univalents, laggards and spindle malfunction-
ing that might have compromised with the pollen
fertility and resulted in the reduced fertility of crops.
According to Biswas [41] presence of univalents
in polyploids during reduction division indicated
failure of chiasma formation or due to loss of chi-
asmata with the progress of meiosis.
Lastly, remove hyphen interests in polyploids
have been renewed in the past decade following the
discovery of multiple origins and pattern of polyp-
loid formation [42].
Conclusion. The consequences of polyploidy ha-
ve been of long-standing interests in genetics,
evolution, and systemic [43]. Thus, from the abo-
ve observations, we simply wish to infer that the
application of colchicine to seedling is an effici-
ent method for polyploidization especially in Brassica
campestris L., Hence, it might be worthy to re-
commend that colchicine could be used to raise
possible fertile autopolyploid mutant lines. Howe-
ver, these mutant lines also provide broader germ-
plasm base for polyploid breeding and hence,
could be utilized by breeders for production of
economic important autotetraploid crops exhibi-
ting superiority in quantitative and qualitative traits
of crop compared to diploid parent.
We are grateful to N.B.P.G.R, New Delhi for pro-
viding inbred seeds of Brassica campestris L. accession
number – IC363713. One of the authors (K. Dwivedi)
is thankful to DST, New Delhi for providing financial
assistance as Inspire Fellowship.
G. Kumar, K. Dwivedi
ÈÍÄÓÖÈÐÎÂÀÍÍÀß ÏÎËÈÏËÎÈÄÈÇÀÖÈß
Ó BRASSICA CAMPESTRIS (BRASSICACEAE)
Ðàç ðàáîòàíà ýêñïåðèìåíòàëüíàÿ ñõåìà ïîëó÷åíèÿ
ñåëüñêîõîçÿéñòâåííûõ êóëüòóð ñ áîëåå âûñîêîé ïëîèä-
íîñòüþ ïóòåì èñêóññòâåííîé ïîëèïëîèäèçàöèè. Èç-
ìåíåíèå ïëîèäíîñòè îáû÷íî ñâÿçàíî ñ ïðèîáðåòåíè-
åì íåêîòîðûõ âàæíûõ ïðèçíàêîâ ó òàêèõ êóëüòóð,
à òàêæå îáåñïå÷èâàåò èõ áîëåå âûñîêóþ ïðèñïîñîá-
ëÿåìîñòü ê íåáëàãîïðèÿòíûì èëè ñóðîâûì óñëîâè-
ÿì âíåøíåé ñðåäû ïî ñðàâíåíèþ ñ äèïëîèäíûìè
àíàëîãàìè. Â íàñòîÿùåì èññëåäîâàíèè ïðåäñòàâëåíî
óñïåøíîå ïîëó÷åíèå àâòîòåòðàïëîèäîâ Brassica cam-
pestris L. ñ ïîìîùüþ êîëõèöèíà. Äëÿ ýòîãî ïðèìå-
íÿëè äâà ìåòîäà îáðàáîòêè – ñåìÿí è ïðîðîñòêîâ.
Íèêàêîé ïîëèïëîèäèè íå îáíàðóæåíî ïðè îáðàáîò-
êå ñåìÿí, òîãäà êàê ïîñëå îáðàáîòêè ïðîðîñòêîâ
ïîëèïëîèäû áûëè ïîëó÷åíû. Ñ ïîìîùüþ öèòîìîð-
ôîëîãè÷åñêèõ èññëåäîâàíèé ïîäòâåðæäåíî íàëè-
÷èå àâòîòåòðàïëîèäíîñòè ó îáðàáîòàííûõ ðàñòåíèé
ïî ñðàâíåíèþ ñ èõ äèïëîèäíûìè àíàëîãàìè. Äëÿ
ýòîãî îöåíèâàëè òàêèå ìîðôîëîãè÷åñêèå ïàðàìåò-
ðû, êàê óâåëè÷åííûå ðàçìåð óñòüèö, äèàìåòð ïûëü-
öû, ðàçìåð öâåòêà, ðåïðîäóêòèâíûå îðãàíû, à òàêæå
óìåíüøåííûå âûñîòà ðàñòåíèé, äëèíà ëèñòà, øèðèíà
ëèñòà, ÷àñòîòà óñòüèö, ÷èñëî öâåòêîâ è ñîöâåòèé.
Áîëåå òîãî, â ðåçóëüòàòå öèòîëîãè÷åñêèõ èññëåäî-
âàíèé ÷åòêî óñòàíîâëåíî óäâîåíèå ãåíîìà ó àâòî-
òåòðàïëîèäîâ ïî ñðàâíåíèþ ñ äèïëîèäàìè. Íàðÿäó
ñ ýòèì â ðàáîòå îöåíèâàëè ôåðòèëüíîñòü ïûëüöû è
âåëè÷èíó ïûëüöåâûõ çåðåí.
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Received 01.06.12
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/NLD (Gebruik deze instellingen om Adobe PDF-documenten te maken die zijn geoptimaliseerd voor prepress-afdrukken van hoge kwaliteit. De gemaakte PDF-documenten kunnen worden geopend met Acrobat en Adobe Reader 5.0 en hoger.)
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/UKR <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>
/ENU (Use these settings to create Adobe PDF documents best suited for high-quality prepress printing. Created PDF documents can be opened with Acrobat and Adobe Reader 5.0 and later.)
>>
/Namespace [
(Adobe)
(Common)
(1.0)
]
/OtherNamespaces [
<<
/AsReaderSpreads false
/CropImagesToFrames true
/ErrorControl /WarnAndContinue
/FlattenerIgnoreSpreadOverrides false
/IncludeGuidesGrids false
/IncludeNonPrinting false
/IncludeSlug false
/Namespace [
(Adobe)
(InDesign)
(4.0)
]
/OmitPlacedBitmaps false
/OmitPlacedEPS false
/OmitPlacedPDF false
/SimulateOverprint /Legacy
>>
<<
/AddBleedMarks false
/AddColorBars false
/AddCropMarks false
/AddPageInfo false
/AddRegMarks false
/ConvertColors /ConvertToCMYK
/DestinationProfileName ()
/DestinationProfileSelector /DocumentCMYK
/Downsample16BitImages true
/FlattenerPreset <<
/PresetSelector /MediumResolution
>>
/FormElements false
/GenerateStructure false
/IncludeBookmarks false
/IncludeHyperlinks false
/IncludeInteractive false
/IncludeLayers false
/IncludeProfiles false
/MultimediaHandling /UseObjectSettings
/Namespace [
(Adobe)
(CreativeSuite)
(2.0)
]
/PDFXOutputIntentProfileSelector /DocumentCMYK
/PreserveEditing true
/UntaggedCMYKHandling /LeaveUntagged
/UntaggedRGBHandling /UseDocumentProfile
/UseDocumentBleed false
>>
]
>> setdistillerparams
<<
/HWResolution [2400 2400]
/PageSize [612.000 792.000]
>> setpagedevice
|