Caryologia

International Journal of Cytology, Cytosystematics and Cytogenetics

ISSN: 0008-7114 (Print) 2165-5391 (Online) Journal homepage: http://www.tandfonline.com/loi/tcar20

Meiosis in Monoploid Asiatic Cotton (Gossypium

Arboreum L.)

S. S. Mehetre & M. V. Thombre

To cite this article: S. S. Mehetre & M. V. Thombre (1980) Meiosis in Monoploid Asiatic Cotton
(Gossypium�Arboreum L.), Caryologia, 33:3, 393-400, DOI: 10.1080/00087114.1980.10796852

To link to this article: https://doi.org/10.1080/00087114.1980.10796852

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 MEIOSIS IN MONOPLOID ASIATIC COTTON
(GOSSYPIUM ARBOREUM L.)

S. S. MEHETRE and M. V. THOMBRE

All India Co-ordinated Cotton Improvement Project,
Mahatma Phule Agricultural University, Rahuri, Dist. Ahrnednagar (M.S.), India

Received: lJih December 1979

INTRODUCTION

Haploids with gametic chromosome complements offer excellent op-

portunities for investigation of many fundamental problems concerned with
genetics and cytology. Excellent reviews covering the different aspects of
haploidy in flowering plants are available (KIMBER and RILEY 1963;
MAGOON and KHANNA 1963; KIRILLOVA 1966; CHASE 1969).
The genus Gossypium includes a diversity of species, some of which
have a high economic value belonging to both diploid (2n = 2x = 26) and
tetraploid (2n = 4x = 52) groups and designated to A to E genomes
(BEASLEY 1942; PHILLIPS and STRICKLAND 1966).
KIMBER and RILEY ( 1963) reviewed haploidy in Angiosperms including
cultivated G. hirsutum and G. barbadense cottons and wild diploids
(2n = 26) like American G. davidsonii Kell. (DtDt) and Australian G.
sturtii Muell. (CC) respectively. Monoploid in cultivated Asiatic cotton
G. arboreum (AtAt) (2n = 26) has been reported recently by THOMBRE
and MEHETRE (1977). SKOVSTED (1935, 1937) studied chromosome mor-
phology in G. arboreum, G. herbaceum, G. anomalum, G. sturtianum and
G. barbadense.
}ACOB (1941 and 1942-43) concluded seven as a basic number of
diploid Gossypium species and considered these as secondary allotetraploids
with 4b-2 constitution. He further inferred that these species might have
evolved through hybridization between species with n =
7 chromosomes.
SIKKA, KHAN and AFZAL (1944) also indicated an allotetraploid nature of
G. arboreum, G. herbaceum and G. hirsutum. According to IYENGAR (1947)

Caryologia, 33: 393-400, 1980

394 MEHTRE and THOMBRE

a set of thirteen chromosomes in polyploid cottons might have been derived

through lower basic number of six or seven. These conclusions are supported
by KRASICHKOV and SHEVITCHENKO (1972 ).
The study of chromosome pairing in monoploids is a significant
measure of the level of chromosome duplication within species (SADPSIVAIAH
197 4) and evidence for determination of basic number of the genus
(THOMPSON 1956). The details of chromosome behaviour during meiosis
in G. arboreum L. Monoploid (2n = 13) have been studied. The probable
origin of present diploid G. arboreum cotton has been discussed, in the
light of these results.

MATERIALS AND METHODS

A single conspicuous dwarf plant having smaller leaves and flowers with
fewer and sterile anthers were noticed in the field population of G. arboreum var.
LD 132, at this Cotton Research Station during October 1976. Its young flower
buds were fixed in Carnoy's fluid (6: 3: 1) and squashed in propionocarmine. The
PMCs were analysed from temporary preparations. Pollen fertility was tested with
1 per cent IKI solution and differential stain (ALEXANDER 1969).

RESULTS

The pamng between non-homologous chromosomes which normally

remains obscure in its corresponding diploid can be evidenced in different
stages of meiosis in monoploid plants.
a) Premetaphase and metaphase pairing.
Mean number of chromosome associations observed from pachytene to
metaphase I are presented in Table 1.

TABLE 1
Average chromosome a.rsociations observed in mezoszs
of G. arboreum var. LD 132 monoploid (2n = x = 13).

Chromosome associations Total PMCs

Meiotic stages X'ta/PMC examined
I II

Pachytene 12.60 0.2 0.002 10

Diakinesis 11.20 0.9 0.002 30
Metaphase· I 10.28 1.36 0.030 70

On an average 0.2 and 0.9 paired bivalents were observed at pachytene

and diakinesis respectively (Figs. 4 and 5).
An interesting configuration of 611 + II (Fig. 7) was observed in four
 MEIOSIS IN MONOPLOID ASIATIC COTTON 395

Figs. 1-12. - Meiosis and pollen development in diploid (2n = 2x = 26) and monoploid
(2n = x = 13) G. arboreum L. cotton. All chromosomes magnified at 1350x and teatrads and
pollens at 45x.
Figs. 1-3. - Meiosis in diploid cotton. 1) Diakinesis. 2) Metaphase-! with un. 3)
Fertile pollen grains.
Figs. 4-12. - Meiosis in monoploid cotton. 4) Pachytene with paired chromosomes. 5) Late
diakinesis with 5II + 3I. 6) and 7) Metaphase-! with 6II + II. 8) Metaphase-! with !II+ 121_
9) Metaphase-! with 13I. 10) Anaphase-! showing tripolar distribution. 11) Abnormal tetrads.
12) Sterile pollen grains with high size variation.
396 MEHTRE and THOMBRE

PMCs at metaphase I. Out of seventy PMCs showing metaphase-!, 23

showed paired bivalents. In these twenty-three PMCs, typical chromosome
associations of 611 + 11 (Fig. 7 ), 5 11 + 31 (Fig. 8 ), 111 + 12\ 3n + 71,
211 + 9\ 111 + 11 1 (Fig. 8) were observed in 4, 4, 10, 2 and 3 PMC
respectively. The paired chromosomes were true bivalents as these formed
at least one chiasma per bivalent. The average number of chiasma per
bivalent at metaphase-! being 0.99. In the remaining fortyseven PMCs
mostly thirteen univalents were observed. However, in rare cases presence
of pseudobivalents was noticed as side to side, end to end and side to
end associations observed giving an appearance of pseudobivalents. Thirteen
univalent chromosomes (Fig. 9) were also observed in remaining PMCs.
The chiasmata in the bivalents were observed from pachytene and
persisted upto metaphase-!. At pachytene and diakinesis, fold back type of
pairing of univalent chromosomes was also observed (Fig. 4 ).
b) Second meiotic division.
Second meiotic division was highly irregular, the frequency of number
of chromosomes distributed at each pole was 12-1, 11-2, 10-3, 9-4, 8-5, 7-6,
in 1, 4, 13, 8, 9 and 12 PMCs analysed.
The formation of bipolar or multipolar spindles (Fig. 10) was observed.
Chromosomes did not orient on equatorial plate and were irregularly
distributed. A few univalents instead of passing to either poles remained as
laggards giving rise to micronuclei. Contrary to observations of BROWN
( 1961) in other tetraploid Gossypium species in no cases division of uni-
valents was observed in this monoploid at this stage. Chromatid bridges
were also not observed.
Non-reduction with splitting of chromosomes at first division forming
dyads, containing normal set of 13 chromosomes was also observed, however,
frequency of such cells was lower than one per cent.
Clear second metaphases were not observed. The univalents distributed
at different poles during anaphase-! remained as such without orienting
to either poles. Thus presence of secondary associations could not be
detected.
Second anaphase formation was not observed. The distribution of
univalents and laggards gave rise to highly irregular tetrad formation at
the end of second anaphase. A number of dyads, triads, tetrads and even
higher polyads were observed (Fig. 11).
c) Pollen fertility.
Pollens formed were highly variable in size. The smallest pollen grains
were 25.10 !J. while largest being 117.30 p. in diameter. The average size
of pollen grain was 69.46 !J. as against 131.4 3 !J. in its diploid counterpart.
 MEIOSIS IN MONOPLOID ASIATIC COTTON 397

Pollen grains tested with one per cent IKI and differential stain (ALEXANDER
1969) indicated total sterility (Fig. 12).
d) Self and cross fertility.
The plant produced sufficient number of flowers, however, it was
completely male and female sterile and no boll setting was observed on this
plant when pollinated by pollens from its diploid counterpart and other
G. arboreum varieties.

DISCUSSION

Since the diploid organism has complete sets of its normal homologous
chromosomes, pairing affinities if any between nonhomologous chromosomes
remain obscure, as is normally evident from the study of meiosis in their
haploids or monoploids. On the contrary, a monoploids with only one set
of chromosomes serve as a important measure to study the degree of
chromosome duplication and also the basic chromosome number. The studies
in early stages of meiosis of monoploids have proved valuable guide lines
in the above respect particularly in the case of cereals (SADASIVAIAH 1974),
as chromosomes in these species are comparatively large and fairly deeply
stained at pachytene and onward stages.
Gossypium species however, are rather difficult for the study of the
early stages of meiosis as their chromosomes are comparatively short and
lightly stained. Efforts were however made to obtain pachytene and
succeeding stages for study of meiotic pairing.

Pachytene.
Paired structures representing non-homologous association in 36 out
of fifty pachytene cells were studied. In addition to the frequent occurrence
of single univalent, fold back type of pairing was observed. Similar pairing
has been reported in monoploids of maize (TING 1966; FoRD 1970; WEBER
and ALEXANDER 1972 ). The pachytene pairing might be due to the presence
of homologous regions in the non-homologous chromosomes. Thus segmental
duplication might be responsible for the non-homologous pairing. Preferential
pairing between identical segments of the non-homologous chromosomes
has been reported in monoploids of maize (TING 1966 ).

Chromosomes association at diakinesis and metaphase-!.

The occurrence of bivalent like structures at metaphase-! has been often
used as a criterion for predicting the basic chromosome number in species
with polyploid origin (KIMBER and RILEY 1963; MAGOON and KHANNA
1963; KIRILLOVA 1966). Based on these observations of 1-2 bivalent like
398 MEHTRE and THOMBRE

assoc1at1ons in monoploids of Zea mays (n = 10) and Sorghum vulgare

( n = 10) TING (1966) and REDDY {1968) respectively proposed an allopoly-
ploid origin for these species with their basic numbers as five. }AUHAR (1970)
and PRAKASH ( 197 3) also suggested an aneuploid origin and indicated basic
chromosome numbers of these species. An autopolyploid origin of Linum
africanum (n = 15) Ricinus communis (n = 10) was predicted from the
studies of meiosis in monoploids (YERMANOS and GILL 1967; NARAIN and
SING 1968 respectively).
The existence of autosyndesis has been pointed out in species hybrids
and polyploids of Gossypium (SKOVSTED 1935 and 1937). From the occasional
formation of trivalent or quadrivalents, ring bivalents and ring univalents
in a few cells at diakinesis of monoploid in Hordeum vulgare (n = 7)
KASHA and SADASIVAIAH (1971) indicated inter or intra chromosomes seg-
mental duplications. In the present studies chiasma formation was observed
beween non-homologous chromosomes at diakinesis which persisted upto
metaphase-I. These observations were similar to RIEGER ( 1957) who reported
chiasma in duplicated segments of monoploid Antirrhinum majus. The
formation of at least one chiasma in paired bivalent is certainly due to true
crossing over and unlikely to be due to chromosome stickiness. SADASIVAIAH
( 197 4) indicated five possibilities of bivalent formation between non-
homologous chromosomes in monoploid e.g., (a) breakage and reunion of
duplicated segments in the same order with respect to centromere (legitimate
recombination), (b) breakage and reunion of segments present in the reverse
side of centromere, (c) breakage and exchange between non-homologously
paired segments (illegitimate recombination), (d) breakage and reunion of
non-homologous segments in U type and (e) chromosome stickiness.
Of these only two possibilities (a) and (c) appear to be responsible
for formation of bivalents between non-homologous chromosomes in the
monoploid of G. arboreum as dicentric bridges with a centric fragments
at anaphase-! were absent. Thus bivalent formation in monoploids could be
explained on the basis of segmental duplications.
In PMCs where 611 + 11 were observed at diakinesis the univalent
showed, fold back type of pairing indicating the presence of duplications
in the same chromosome.
The bivalents observed in the monoploid plant under study indicated
the homologies which might be due to polyploid or an aneuploid origin
of contemporary chromosome number (JACOB 1941 and 1942-43). Thus
certain chromosomes of diploid parent of monoploid might have completely
or partially duplicated during the course of evolution. Alternatively such
pairing can be explained on the basis of duplicated segments which originally
arise through translocations being carried in diploid species.
 MEIOSIS IN MONOPLOID ASIATIC COTTON 399

Maximum number of bivalents observed in this species are indicative

that the basic chromosome number of this species might be seven and present
diploid G. arboreum (2n = 2x = 26) could be considered as an allotetraploid
in origin. JAcOB (1941 and 1942-43) concluded seven as a basic number
of diploid species, these species being secondary allotetraploids with 4b-2
constitution and evolved through hybridization between species with n = 7
chromosomes. SIKKA, KHAN and AFZAL (1944) and IYENGAR (1944) also
indicated allotetraploid nature of this species. KRASICHKOV and SHEVIT-
CHENKO (1972) concluded x = 6 as a basic number in the genus Gossypium.
The polyploid nature of G. arboreum indicated by IYENGAR (1947),
JAcOB ( 1941 and 1942-43) SIKKA, KHAN and AFZAL (1944) has been
confirmed from meiotic studies. From the maximum true bivalent formation
in this monoploid plant, the basic chromosome in the genus Gossypium
appears to be x = 7. Similar indications regarding basic chromosome number
(x = 5) were drawn from the bivalent formation in the monoploids of
Zea mays and Sorghum vulgare by TING ( 1966) and REDDY ( 1968) respectively.
Polyploid origin of existing species suggested by previous workers from
meiotic studies is also confirmed in the present studies. JACOB (1941)
suggested 4b-2 aneuploid origin of the diploid Gossypium species. The
present 2n = 26 chromosome number in G. arboreum, thus is likely due
to loss of a complete chromosome followed by duplication of the entire
aneuploid genome giving rise to secondary (allotetraploid) polyploid with
2n = 26 chromosomes (JACOB 1941 and 1942-43; IYENGAR 1947; SIKKA,
KHAN and AFZAL 1947).

REFERENCES
ALEXANDER D. E., 1964. - Spontaneous reciprocal translocations during megasporogenesis
in maize haploids. Nature, 201: 737-738.
ALEXANDER M. P ., 1969. - Differential staining of aborted and nonaborted pollen. Stain.
Tech., 44: 117-122.
BEASLEY J. 0., 1942. - Meiotic behaviours of species, species hybrids, haploids and induced
poly plaids of Gossypium. Genetics, 27: 25-54.
BROWN M. S., 1961. - Chromosome diffe;·entiation in Gossypium. Amer. J. Bot., 48: 532.
CHAsE S. S., 1969. - Mo11oploids and monoploid derivatives of maize (Zea mays L.).
Bot. Rev., 35: 117-167.
FoRD L., 1970. - Chromosome association in Zea mays monoploids. The Nucleus, 13: 99-105.
}AUHAR P. P., 1970. - Haploid meiosis and its bearing on the phylogeny of pearl millet,
Pennisetum t_vphoides Stafp. et Hubb. Genetica, 41: 532-540.
}ACOB K. T., 1941. - Preliminary observations on the chromosome morphology in Asiatic
cottons with special reference to their phylogeny and interrelationship. 2nd Con£. Cott.
Gr. Probl. India !CCC. pp. 42-45.
-, 1942-43. - Studies in cotton. IV. Morphology of somatic chromosomes in eight types
of Asiatic cotton. Trans. Bose Res. Inst., 15: 17-27.
IYENGAR N. K., 1944. - Cytological investigations on auto and allo-tetraploid Asiatic Cotton.
Indian J. Agric. Sci., 14: 30-40.
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- , 1947. - A review of chromosome con;ugation in allotetraploid cottons. 3rd Con£. Cott.

Gr. Probl. India ICCC. Bombay, pp. 58-69.
KASHA K. ]. and SADASIVAIAH R. S., 1971. - Genome relationship between Hordeum
vulgare L. and H. bulbosum L. Chromosoma, 35: 264-287.
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KIRILLOVA G. A., 1966. -The phenomenon on haploidy in angiosperms. Genetika, 2: 137-147.
KRASICHKOV V. P. and SHEVITCHENXO V. Yu., 1972. - On the original chromosome number
of the cotton plant. Cokl Akad. Fanhoi RSS TokiKISTON, 15: 57-59. Pl. Breed.
Ab. 45, No. 3726 {1975).
MAGOoN M. L. and KHANNA K. R., 1963. - Haploids. Caryologia, 16: 191-234.
NARAIN A. and SINGH D., 1968. - Haploid meiosis and its bearing to the constitution of
the castor oil plant. J. Hered., 59: 287-288.
PRAKASH S., 1973. - Haploidy in Brassica nigra Koch. Euphytica, 613-614.
PHILLIPS L. L. and STRICKLAND M. A., 1966. - The cytology of a hybrid between Gossypium
hirsutum and G. longicalyx. Canad. J. Genet. Cytol., 8: 91-95.
REDDY V. R., 1968. - Chromosome pairing in haploid sorghum. Cytologia, 33: 471-476.
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SUMMARY

The microsporogenesis was studied in monoploid plant {2n = x = 13) of Gossypium

arboreum cotton var. LD 132. The total pollen sterility observed in this plant is explained
on the basis of various meiotic irregularities observed during first and second meiotic division
caused due to loss of n = 13 chromosomes.
The pairing of non-homologous chromosomes is explained on the basis of segmental
duplications. Maximum of six bivalents and an univalent were observed which supported
seven {x = 7) as a basic number of the genus Gossypium. Secondary polyploid nature of
this species with the loss of one chromosome at diploid level 2n = 13 with subsequent
duplication of the entire aneuploid chromosome complement giving rise to 2n = 26 {4b-2)
chromosomes has been discussed.