Cytological studies on meiosis and male gametophyte development in autotetraploid cucumber

With improved staining and chromosome preparation techniques, meiosis of pollen mother cells (PMCs) and male gametophyte development in autotetraploid cucumber (Cucumis sativus L.) was studied to understand the correlation between chromosomes behaviour and fertility. Various chromosome configurations, e.g. multivalent, quadrivalents, trivalents, bivalents and univalents were observed in most PMCs at metaphase I. Lagging chromosomes were frequently observed at anaphase in both meiotic divisions. In addition, chromosomes segregations were not synchronous and equal in some PMCs during anaphase II and telophase II. Dyads, triads, tetrads with micronuclei and polyads were observed at tetrad stage, and the frequencies of normal tetrad with four microcytes were only 55.4 %. The frequency of abnormal behaviour in each stage of meiosis was counted, and the average value was 37.2 %. The normal meiotic process could be accomplished to form the microspore tetrads via simultaneous cytokinesis. Most microspores could develop into fertile gametophytes with 2 cells and 3 germ pores through the following stages: single-nucleus early stage, single-nucleus late stage and 2-celled stage. The frequency of abnormalities was low during the process of male gametophyte development. The germination rate of pollen grains was 46.9 %. These results suggested that abnormal meiosis in PMCs was the reason for low pollen fertility in the autotetraploid cucumber.     

同一居群韭莲不同植株减数分裂行为差异的遗传分析

对韭莲(2n=48)小孢子母细胞减数分裂及小孢子发育进行研究。结果显示同一居群植株的减数分裂行为存在明显差异。多数韭莲植株小孢子母细胞减数分裂存在少量落后染色体、微核等现象,平均每株中具有异常分离行为的母细胞占14.02%,小孢子发育正常,但花粉无活力。并首次从减数分裂后期Ⅰ的特殊的细胞学形态证明韭莲是臂内倒位杂合体。而少数植株韭莲的小孢子母细胞减数分裂极其紊乱,后期Ⅰ出现多极分离、大量落后染色体,小孢子母细胞减数分裂总异常分离高达94.3%。四分孢子期多分孢子体高达73.4%。分析认为:前者减数分裂行为异常的原因主要由染色体结构变异所致,而后者的原因除染色体结构变异外,还可能与控制纺锤体形成的基因突变有关。     

鱼腥草花粉母细胞减数分裂特征及其育性研究

为深入了解鱼腥草花粉母细胞的减数分裂特征与花粉育性的关系,该研究采用卡宝品红染色法对2个鱼腥草居群花粉母细胞的减数分裂过程进行观察,并采用氯化三苯基四氮唑(TTC)染色法、I2-KI染色法、B-K培养基培养法及荧光显微镜观察法来检测鱼腥草花粉的活力及萌发率。结果发现:(1)鱼腥草减数分裂的进程与花序大小、花药颜色、花药长度均有密切的关系。(2)2个居群的鱼腥草中花粉母细胞减数分裂过程正常占88.2%,有11.8%的花粉母细胞减数分裂异常。(3)减数分裂异常表现在减数分裂过程中出现微核、落后染色体、染色体桥、不均等分离、多分体等现象,并发现在二分体阶段及单核花粉发育过程中存在细胞融合。(4)2个居群的鱼腥草花粉活力均不超过1.5%,花粉几乎不萌发。研究认为,鱼腥草花粉育性低的主要原因是单核花粉的发育过程异常,而非鱼腥草花粉母细胞减数分裂异常所致。     

Karyology and evolution inSesamoides (Resedaceae)

The meiotic behaviour abnormalities, fertility and size of pollen of 6 taxa ofSesamoides have been analysed. Besides diploids (2x), polyploids (4x, 6x, 8x) have been found. The chromosome base number is x = 10, but an origin from x = 5 is suggested.     

Abnormal male meiosis explains pollen sterility in the polyploid medicinal plant Pinellia ternata (Araceae)

Pinellia ternata is an important traditional Chinese medicinal plant. Its different populations in China have various ploidy levels, based on x = 13, as well as extensive aneuploid series. The microsporogenesis process was observed in specimens from three populations from three regions of Hubei Province; they were characterized by normal and abnormal meiotic divisions in pollen mother cells (PMCs) at all stages simultaneously. Meiotic abnormalities including univalents/multivalents, chromosomal laggards/bridges and micronuclei appeared in about 50% of the PMCs, together with abnormal cytokinesis. Chromatin/chromosome transfer between meiocytes occurred only during the first division, at low frequency; this might contribute to these meiotic abnormalities. Although the remaining 50% of the PMCs presented normal cytological behavior, pollen fertility was only about 2%. These results provide cytological explanations for its low seed-set and the general use of asexual reproduction through tubers and bulbils; it also explains the wide variations in chromosome number.     

Chromosome numbers and meiotic analysis in the pre-breeding of <Emphasis Type="BoldItalic">Brachiaria decumbens</Emphasis> (Poaceae)

A total of 44 accessions of Brachiaria decumbens were analysed for chromosome count and meiotic behaviour in order to identify potential progenitors for crosses. Among them, 15 accessions presented 2n = 18; 27 accessions, 2n = 36; and 2 accessions, 2n = 45 chromosomes. Among the diploid accessions, the rate of meiotic abnormalities was low, ranging from 0.82% to 7.93%. In the 27 tetraploid accessions, the rate of meiotic abnormalities ranged from 18.41% to 65.83%. The most common meiotic abnormalities were related to irregular chromosome segregation, but chromosome stickiness and abnormal cytokinesis were observed in low frequency. All abnormalities can compromise pollen viability by generating unbalanced gametes. Based on the chromosome number and meiotic stability, the present study indicates the apomictic tetraploid accessions that can act as male genitor to produce interspecific hybrids with B. ruziziensis or intraspecific hybrids with recently artificially tetraploidized accessions.     

Allelic interaction of F1 pollen sterility loci and abnormal chromosome behaviour caused pollen sterility in intersubspecific autotetraploid rice hybrids

The intersubspecific hybrids of autotetraploid rice has many features that increase rice yield, but lower seed set is a major hindrance in its utilization. Pollen sterility is one of the most important factors which cause intersubspecific hybrid sterility. The hybrids with greater variation in seed set were used to study how the F(1) pollen sterile loci (S-a, S-b, and S-c) interact with each other and how abnormal chromosome behaviour and allelic interaction of F(1) sterility loci affect pollen fertility and seed set of intersubspecific autotetraploid rice hybrids. The results showed that interaction between pollen sterility loci have significant effects on the pollen fertility of autotetraploid hybrids, and pollen fertility further decreased with an increase in the allelic interaction of F(1) pollen sterility loci. Abnormal ultra-structure and microtubule distribution patterns during pollen mother cell (PMC) meiosis were found in the hybrids with low pollen fertility in interphase and leptotene, suggesting that the effect-time of pollen sterility loci interaction was very early. There were highly significant differences in the number of quadrivalents and bivalents, and in chromosome configuration among all the hybrids, and quadrivalents decreased with an increase in the seed set of autotetraploid hybrids. Many different kinds of chromosomal abnormalities, such as chromosome straggling, chromosome lagging, asynchrony of chromosome disjunction, and tri-fission were found during the various developmental stages of PMC meiosis. All these abnormalities were significantly higher in sterile hybrids than in fertile hybrids, suggesting that pollen sterility gene interactions tend to increase the chromosomal abnormalities which cause the partial abortion of male gametes and leads to the decline in the seed set of the autotetraploid rice hybrids.     

Trisomics of ryegrass and their transmission

Summary Primary trisomics of perennial ryegrass, Lolium perenne L., were studied for meiotic behaviour, fertility, morphology and trisome transmission. Trisomics differed from each other in mean meiotic association, pollen fertility, seed set and morphology. The combined cytomorphological data suggested that the investigated trisomic plants included trisomes 2 to 7. No pollen transmission of trisomes was detected. Female transmission of trisomes ranged from 12% for tri 3 to 37% for tri 4 with a mean of 24% for the six trisomes. Trisome transmission was not related to either chromosome size or trivalent/univalent frequency, although the larger trisomes formed trivalents more frequently than the smaller trisomes.Part of M.Sc.Agr. Thesis, submitted by senior author to the National University of Ireland, Dublin     

Cytomorphological investigations in Oxyria digyna Hill. from the Kashmir Himalaya,India

In the present paper, detailed cytomorphological investigations in Oxyria digyna Hill. from Kashmir Himalaya—India have been reported for the first time. All the 14 investigated populations of O. digyna are diploid based on x = 7. Out of these in two populations 0–2B chromosomes have been recorded for the first time while 6 populations differed significantly in their meiotic characteristics. Meiotic abnormalities during male meiosis observed include inter PMC chromatin transfer (cytomixis). Non-synchronous disjunction of some bivalents, laggards and bridges at anaphases and telophases. Consequent to these meiotic anomalies, microsporogenesis in meiocytes is abnormal resulting in to dyads, triads and polyads with or without micronuclei. The overall effect is seen in reduced pollen fertility. Unreduced pollen grains were observed in some populations, which differed significantly in their size compared to the normal (reduced) pollen grains. It is observed that a smaller frequency of pollen grains differed morphologically in Aharbal and Yosmarg populations. The remaining eight populations showed regular meiotic course, normal microsporogenesis and high percentage of pollen fertility (95.09–99.09 %).     

Segregation for fertility and meiotic stability in novel <Emphasis Type="Italic">Brassica</Emphasis> allohexaploids

Key message

Allohexaploid Brassica populations reveal ongoing segregation for fertility, while genotype influences fertility and meiotic stability.

Abstract

Creation of a new Brassica allohexaploid species is of interest for the development of a crop type with increased heterosis and adaptability. At present, no naturally occurring, meiotically stable Brassica allohexaploid exists, with little data available on chromosome behaviour and meiotic control in allohexaploid germplasm. In this study, 100 plants from the cross B. carinata?×?B. rapa (A2 allohexaploid population) and 69 plants from the cross (B. napus?×?B. carinata)?×?B. juncea (H2 allohexaploid population) were assessed for fertility and meiotic behaviour. Estimated pollen viability, self-pollinated seed set, number of seeds on the main shoot, number of pods on the main shoot, seeds per ten pods and plant height were measured for both the A2 and H2 populations and for a set of reference control cultivars. The H2 population had high segregation for pollen viability and meiotic stability, while the A2 population was characterised by low pollen fertility and a high level of chromosome loss. Both populations were taller, but had lower average fertility trait values than the control cultivar samples. The study also characterises fertility and meiotic chromosome behaviour in genotypes and progeny sets in heterozygous allotetraploid Brassica derived lines, and indicates that genotypes of the parents and H1 hybrids are affecting chromosome pairing and fertility phenotypes in the H2 population. The identification and characterisation of factors influencing stability in novel allohexaploid Brassica populations will assist in the development of this as a new crop species for food and agricultural benefit.

     

不同倍性水稻亚种间杂种小孢子发生的细胞学观察

利用塑料半薄切片对水稻同源四倍体亚种间杂种F1及其对应的二倍体杂种F1小孢子母细胞减数分裂过程的细胞学变化进行观察研究.结果表明,同源四倍体水稻亚种间杂种造孢细胞期和小孢子母细胞期已经表现出较高频率的异常;减数分裂过程中,小孢子母细胞出现异常更加复杂,主要包括小孢子母细胞液泡化和退化两大类,这些异常是导致杂种花粉败育和花粉低育性的重要原因之一;此外,绒毡层异常也是导致杂种的花粉育性降低的因素.二倍体杂种小孢子母细胞在减数分裂过程的异常类型与水稻同源四倍体亚种间杂种基本相似,其绒毡层异常频率较低,对其花粉低育性影响不大.     

Reproductive biology of henequen (Agave fourcroydes) and its wild ancestor Agave Angustifolia (Agavaceae). i. Gametophyte development

The pathways of micro- and megagametophyte development in Agave fourcroydes (henequén) and A. angustifolia were studied. We used histology and light microscopy to observe anther ontogeny and ovary differentiation in relation to flower bud size. Both species have the same sexual reproductive strategies and gametophyte development that may be divided into three phases: (1) premeiotic, which includes the establishment of the megaspore mother cell and the pollen mother cell; (2) meiotic, the formation of mature microspores and functional megaspores; (3) postmeiotic, which encompasses the development of mature pollen grains and the formation of the embryo sac. A successive type microsporogenesis was found in both species with formation of T-shaped tetrads and binuclear pollen grains. In vitro germination tests revealed very low pollen fertility. The female gametophyte is formed from two micropylar megaspore cells after the first meiotic division (bisporic type). Male and female gametogenesis occur asynchronously with microsporogenesis finishing before macrosporogenesis. The results so far show that the formation of male and female gametophytes in henequén is affected at different stages and that these alterations might be responsible for the low fertility shown by this species.     

Impact of cytomixis on meiosis, pollen viability and pollen size in wild populations of Himalayan poppy (Meconopsis aculeata Royle)

We report the occurrence of cytomixis in wild populations of Himalayan poppy (Meconopsis aculeata Royle), which is considered to be an important and threatened medicinal plant growing in the high hills of the Himalayas. The impact of cytomixis on meiotic behaviour, reduced pollen viability and heterogeneous-sized pollen grains was also studied. Cytological studies in the seven wild populations from the high hills of Himachal Pradesh revealed that all the Himalayan populations exist uniformly at the tetraploid level (2n=56) on x=14. The phenomenon of chromatin transfer among the proximate pollen mother cells (PMCs) in six populations caused various meiotic abnormalities. Chromatin transfer also resulted in the formation of coenocytes, aneuploid, polyploid and anucleated PMCs. Among individuals that showed chromatin transfer, chromosome stickiness and interbivalent connections were frequently observed in some PMCs. The phenomenon of cytomixis in the species seems to be directly under genetic control; it affects the meiotic course considerably and results in reduced pollen viability.     

Fertility and meiotic chromosome behaviour in autotetraploid pearl millet

Summary Tetraploidy was induced in outbred pearl millet and selection for high and low seed set was started in the C₁ generation. Segregation in the C₃ generation was observed for fertility and also for meiotic features: per cent seed set in selfed earhead, chiasma frequency, chromosome association and chromosome distribution in pollen mother cells were all affected. However, variation in seed set was observed even between samples not differing in meiotic features. It is apparent that factors regulating seed set in autotetraploid pearl millet were genic as well as chromosomal.A high frequency of univalents and trivalents was the main cause of sterility; quadrivalent misdisjunction was not a significant factor. As univalency decreased with increased chiasma formation, the gain was in the form of quadrivalents. However, individuals not differing in chiasma frequency did differ in chromosome association frequencies, indicating that the dependence of chromosome pairing behaviour on chiasmata was subject to genotypic influence.     

Cytomixis and meiotic abnormalities during microsporogenesis are responsible for male sterility and chromosome variations in Houttuynia cordata

Houttuynia cordata (Saururaceae) is a leaf vegetable and a medicinal herb througout much of Asia. Cytomixis and meiotic abnormalities during microsporogenesis were found in two populations of H. cordata with different ploidy levels (2n = 38, 96). Cytomixis occurred in pollen mother cells during meiosis at high frequencies and with variable degrees of chromatin/chromosome transfer. Meiotic abnormalities, such as chromosome laggards, asymmetric segregation and polyads, also prevailed in pollen mother cells at metaphase of the first division and later stages. They were caused by cytomixis and resulted in very low pollen viability and male sterility. Pollen mother cells from the population with 2n = 38 showed only simultaneous cytokinesis, but most pollen mother cells from the population with 2n = 96 showed successive cytokinesis; a minority underwent simultaneous cytokinesis. Cytomixis and irregular meiotic divisions appear to be the origin of the intraspecific polyploidy in this species, which has large variations in chromosome numbers.     

Apomixis versus sexuality in blackberries (Rubus subgen.Rubus,Rosaceae)

InRubus L. a connection seems to exist between the degree of meiotic disturbances on the one hand, and the production of unreduced embryo sacs, pollen fertility and relative seed set on the other hand. Severe meiotic disturbances commonly encountered in apomictic taxa decrease pollen fertility and thereby seed set since pollen is necessary for endosperm development. By contrast interspecific hybrids between apomictic taxa appear to be sexual and exhibit high pollen fertilities, probably due to an improved meiosis. Thus, apomixis leads to a decreased fertility inRubus, not the opposite, as often discussed.     

Estimation of efficiency of seed irradiation by thermal neutrons for inducing chromosomal aberration in M2 of cotton Gossipium hirsutum L

Cytogenetic analysis of M2 plants after irradiation of cotton by thermal neutrons was performed in 56 families. In 40 plants of 27 M2 families, different abnormalities of chromosome pairing were found. These abnormalities were caused by primary monosomy, chromosomal interchange, and desynapsis. The presence of chromosome aberrations in some cases decreased meiotic index and pollen fertility. Comparison of the results of cytogenetics analysis, performed in M1 and M2 after irradiation, showed a nearly two-fold decrease in the number of plants with chromosomal aberrations in M2, as well as narrowing of the spectrum of these aberrations. The latter result is explained by the fact that some mutations are impossible to detect in subsequent generations because of complete or partial sterility of aberrant M1 plants. It was established that the most efficient radiation doses for inducing chromosomal aberrations in the present study were 15 and 25 Gy, since they affected survival and fertility of altered plant to a lesser extent.     

Development and cytology of pollen in Dahlstedtia Malme (Leguminosae: Papilionoideae)

Microsporogenesis, chromosome number, meiotic behaviour and meiotic index were investigated in Dahlstedtia pinnata and D . pentaphylla , two legume species occurring largely in Brazil, in order to ascertain whether the pollen could limit fertilization events. Archesporial cells originate primary sporogenous and anther wall precursor cells, the tapetum is uniseriate, uninucleate and glandular. Tetrads are tetrahedric or decussate, and cytokinesis is of the simultaneous type. Mature pollen grains are tricolpate and bicellular. No abnormalities in microsporogenesis were found. In both species the chromosome number is n = 11, a number not reported previously. The base number for Dahlstedtia is also 11, because cytological observations include both species of Dahlstedtia . D. pentaphylla has a higher meiotic index and lower individual variation values, and it is considered meiotically stable. Its pollen grains do not limit fertilization. D. pinnata has a lower meiotic index, and the pollen is one of the factors which limit fertilization. Furthermore, D. pinnata has numerous adventitious shoots, which suggest that vegetative propagation is important in its reproductive process. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 138 , 461–471.     

Cytotaxonomical studies in Tribulus terrestris and T. alatus (Zygophyllaceae)

The chromosome numbers and meiotic behaviour of twenty morphologically variable populations of 7. terrestris and four of T. alatus were studied and compared. All populations of /. terrestris were hexaploid (2n = 36). They fall in one of the three morphological–eytological groups: I) robust with normal meiosis and pollen grains, II) less–robust with abnormal meiosis and high frequency of smaller pollen grains, and III) mixture of type II and dwarf plants. Seed set was high in all but the dwarfs. It appears that group I reproduces sexually while the other two groups have a certain degree of apomixis, and that the dwarf plants probably resulted from the fusion of cytologically unbalanced gametes. Tribulus alatus populations were tetraploid (2n = 24). Bud material adequate for meiotic studies were unavailable, but the wide range in size of pollen grains indirectly signifies meiotic abnormalities. So far, only the tetraploid race has been examined, while other chromosome races may exist.     

Meiotic behavior and pollen fertility in triploid and tetraploid natural populations of Campuloclinium macrocephalum (Eupatorieae,Asteraceae)

Campuloclinium macrocephalum DC. is a perennial herb widely distributed in the New World and introduced in South Africa, where it is commonly called “pompom weed”. This species is considered one of the most important weeds of Brazil and one of the problematic invasive plants of South Africa. The meiotic system can be studied to assess the ability of a weed to spread, but only few studies on C. macrocephalum have been realized. In this study, we examined the meiotic behavior and pollen fertility of 14 natural populations of C. macrocephalum from Argentina and Uruguay. Meiotic analysis revealed 2 triploid (2n = 3x = 30), 11 tetraploid (2n = 4x = 40) and 1 mixed population (2n = 2x = 20, 2n = 4x = 40). Both, triploid and tetraploid specimens showed a widely variable meiotic behavior with irregular chromosome pairing showing univalents, bivalents, trivalents (in triploids) and tetravalents (in tetraploids) at diacinesis of first meiotic division. Different abnormalities were observed, such as: laggard chromosomes, chromatin bridges, and out of plate chromosomes at metaphase I. During meiosis I (prophase), some cells showed the phenomenon of cytomixis or chromatin transfer between pollen mother cells. The meiotic indexes suggest that only four populations were normally fertile (over 90 % of fertile pollen), indicating meiotically stable plants. The remaining populations share variable pollen fertility, with triploids ranging from 46.64 to 54.83 % and tetraploids varying from 3.54 to 45.30 %. We suggest that polyploidy seems to be recurrent in C. macrocephalum, promoting partial sterility of pollen grains, generating large numbers of individuals by apomixis promoting invasion of crop fields. This study presents the meiotic behavior of this weed, these could be useful for future studies of biological control in areas with no natural enemies.