Cytomixis and associated meiotic abnormalities affecting pollen fertility in Clematis orientalis

Present cytological investigations from the cold desert regions of Lahaul-Spiti and Kinnaur (India) record the first ever tetraploid (2n=32) chromosome count and cytomixis in Clematis orientalis L. var. acutifolia Hook. f. et Thoms. The phenomenon of cytomixis (9.33–29.80 %) involving chromatin transfer among 2–3 proximate pollen mother cells (PMCs) during male meiosis occurs through narrow and broad cytoplasmic channels from early prophase to tetrad stage. However, frequency of its occurrence during the later meiotic stages is rather low. Chromatin transfer results into the formation of hypo-, hyperploid and enucleated PMCs. Various meiotic abnormalities associated with cytomixis such as chromatin stickiness, pycnotic chromatin, interbivalent connections, out of plate bivalents, late disjunction of bivalents, and laggards and bridges resulted into some pollen sterility (16.33–49.30 %) and heterogeneous pollen grains size.     

Persistent occurrence of meiotic abnormalities in a new hexaploid cytotype of <Emphasis Type="Italic">Thalictrum foetidum</Emphasis> from Indian cold deserts

Thalictrum foetidum L. (Ranunculaceae), a morphologically variable and widely distributed species of temperate and alpine Himalayas is worked out cytologically for the first time from India. Earlier studies from outside India were restricted to chromosome counts and karyotypic analysis. We studied the male meiosis, microsporogenesis and pollen viability in the wild accessions from the cold deserts of Lahaul-Spiti, Kinnaur and Pangi Valley of Himachal Pradesh. Present cytomorphological surveys in the species record the existence of two distinct morphotypes involving plant size; colour and size of leaf/leaflet; dentation of leaflet lobes; and degree of leaf pubescence. All the accessions in the two morphovariants share the same meiotic chromosome number (n = 21) and adds a new intraspecific hexaploid cytotype. The accessions show the phenomenon of cytomixis involving transfer of chromatin material among proximate pollen mother cells (PMCs) and associated meiotic abnormalities like, out of plate bivalents, interchromosomal connections, and laggards, bridges and micronuclei at anaphases/telophases. Microsporogenesis results into abnormal sporads (tetrads with micronuclei, dyads, triads and polyads). The products of such sporads resulted into some pollen sterility and pollen grains of heterogeneous sizes. The persistent occurrence of phenomenon of cytomixis and associated meiotic abnormalities and consequently pollen sterility and pollen grains of heterogeneous sizes in the hexaploid cytotype of T. foetidum seems to be under some genetic factors associated with the genome.     

Induction of cytomixis affects microsporogenesis in Sesamum indicum L. (Pedaliaceae)

Cytomixis was recorded during microsporogenesis in sesame (Sesamum indicum L.), a member of the family Pedaliaceae. The phenomenon of cytomixis was observed at various stages of meiosis in 0.5% sodium azide (SA) treated populations of Sesamum indicum L. Cytomixis was observed to occur through various methods, i.e. by forming cytoplasmic channels and direct fusion of pollen mother cells (PMCs), the former was more frequent than the latter. The migration of nuclear content involved all the chromatin/chromosomes or part of it from donor to recipient cell/cells. Some completely empty meiocytes were also observed. Stickiness, precocious movement, laggards, unorientation and micronuclei were observed in almost all the sets treated with various doses of SA. Increase in the doses of SA had a positive effect on the percentage of PMCs showing cytomixis and chromosomal abnormalities. The impact of cytomixis on meiotic behaviour, reduced pollen viability and heterogeneous sized pollen grains were observed.     

First detection of cytomixis and its consequences in <Emphasis Type="Italic">Thalictrum cultratum</Emphasis> Wall. (Ranunculaceae)

In current paper, we studied the meiotic chromosome number and details of cytomixis recorded for the first time in Thalictrum cultratum Wall. from the high altitude areas of northwest Himalaya (India). Cytomixis, the phenomenon of inter PMC (pollen mother cell) migration of chromatin material has been reported in many angiosperms and other groups of plants, but there is no published report in Thalictrum cultratum. All the presently studied accessions of the species existed at diploid level (x = 7). The present chromosome count of n = 14 has been reported for the first time from the study area. The cytological stability of any plant is an important consideration in view of its existence in nature. Meiotic disturbances can impose threat to the reproductive potential of plant. Current study reveals the occurrence of inter PMC migration of chromatin material. In present investigation, the phenomenon of cytomixis can be observed between 2 to 9 PMCs. Chromatin migration occurs through narrow and broad cytoplasmic channels or occasionally fused together during metaphase-I and anaphase-I. Cytomixis resulted in the formation of hypo- (n = 2, 2 + 1 small fragment, n = 3 + 3 small fragments, 4, 5 + extra mass of chromatin, 13) and hyperploid (n = 7 + 2 small fragments, 8 + 2 small fragments, 14, 14 + 2 small fragments, 15) PMCs, variable sized pollen grains and pollen sterility (20%). The current study reveals that cytomixis is responsible for formation of PMCs with variable chromosomes (hypoand hyperploid) and pollen grains of heterogeneous sizes and pollen sterility in T. cultratum. This is the first report of cytomixis in the species.     

Cytomixis impairs meiosis and influences reproductive success inChlorophytum comosum (Thunb) Jacq. — An additional strategy and possible implications

Spontaneous intercellular chromatin migration/cytomixis was observed to occur in the pollen mother cells (PMCs) of theChlorophytum comosum for the first time. The migration through cytomictic channels was more pronounced in meiosis-I and very rare in meiosis-II. The process was associated with erratic meiosis, which was characterized by defects in chromosome organization and segregation. Cytomixis was more intense in the month of April than in July and consequently the frequency of meiotic irregularities was much more pronounced during the month of April. As a consequence of abnormal meiosis, fertility was drastically reduced resulting in meager seed efficiency of 17% only. Recombination system also does not guarantee the release of sufficient variability. We view the phenomenon of cytomixis as genetically controlled mechanism involving meiotic genes and operating through signal transduction pathway triggered by the environmental stimuli. The evolutionary significance and tenable hypothesis in the backdrop of existing literature is also proposed.     

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.     

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 %).     

Cytomixis in pollen mother cells of Medicago sativa L

Cytomixis (i.e., chromatin migration between meiocytes) has been detected in many plant species, but not in Medicago sativa spp. In the present study we report the identification of a few cytomictic alfalfa plants. Those plants, the "mother plants," were selfed and crossed with a normal control plant. Microsporogenesis analysis was performed on the mother plants, on the S(1) and F(1) plants, and on controls. The S(1) and F(1) plants, like the mother plants, were found to be cytomictic. Single or multiple chromatin bridges between two or more meiocytes were observed almost exclusively in prophase I. Some completely empty meiocytes were also observed. In addition to cytomixis, other meiotic abnormalities were found. Control plants showed an almost regular meiosis. The highest values of cytomixis were observed in the mother plants, and the lowest in their F(1) progenies. Variability of cytomixis in the F(1) plants is probably due to a heterozygotic condition of the parents for this trait. No significant correlation was found between cytomixis and pollen viability, even if the cytomictic plants showed low values of pollen viability.     

The Relation between the Patterns of Cytomixis and Variation of Chromosome Numbers in Pollen Mother Cells of Jimsonweed ( Datura stramonium L.)

The phenomenon of cytomixis is a general occurenee in the pollen mother cells (PMCs) of certain angiosperms. Recently, we found this phenomenon also occurred in jimsonweed. According to our observations and a test of chi-square, it has been shown that the discrepancies between the patterns of intercellular chromatin migration and the variation of chromosome numbers in the PMCs of jimsonweed, as well as rye, are not significant. It means that the patterns of intercellulr chromatin migration and the variation chromosome numbers are related to each other.     

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.     

An ultrastructural study of cytomixis in tobacco pollen mother cells

Intercellular chromatin migration (cytomixis) in the pollen mother cells of two tobacco (Nicotiana tabacum L.) lines was analyzed by electron microscopy during the first meiotic prophase. The maximal manifestation of cytomixis was observed in the pachytene. As a rule, several cells connected with one another by cytomictic channels wherein the nuclei migrated were observable at this stage. In the majority of cases, nuclei passed from cell to cell concurrently through several closely located cytomictic channels. Chromatin migrated between cells within the nuclear envelope, and its disintegration was unobservable. The nucleus, after passing through cytomictic channels into another cell, can be divided into individual micronuclei or, in the case of a direct contact with another nucleus, can form a nuclear bridge. It has been demonstrated that the chromatin structure after intracellular migration visually matches the chromatin structure before it passed through the cytomictic channel. No signs of pyknosis were observable in the chromatin of the micronuclei formed after cytomixis, and the synaptonemal complex was distinctly seen. The dynamics of changes in the nucleoli during cytomixis was for the first time monitored on an ultrastructural level. Possible mechanisms determining cytomixis are discussed and the significance of this process in plant development is considered.     

Methomyl,imbraclaobrid and clethodim induced cytomixis and syncytes behaviors in PMCs of Pisum sativum L: Causes and outcomes

Cytomixis is a common phenomenon observed in meiotic cells such as anther which is influenced by various factors. Use of pesticides is a common practice in agriculture. However, it is not known whether pesticides can induce cytomixis in plant cells and induce genetic variation. To understand this, the present study was planned to assess the cytomixis and syncytes behaviors in PMCs of Pisum sativum L. Seeds of P. sativum (Family: Fabaceae) were treated with different concentrations of commonly used pesticides methomyl (ME), imbraclaobrid (IM) and clethodim (CL). Seeds were treated with various concentrations (0.1, 0.2, 0.3, 0.4 and 0.5% of ME, IM and CL prepared in water) for 1 and 3 h. Effect of pesticides on pollen fertility, frequency of cytomixis, and kind of cytomixis cells was assessed. In the cytomixis cells, the cytomictic channel (CC) and direct fusion (DF), and various stages of meiosis (PI, MI, AI and TI) with cytomixis cells were observed. In addition, frequency of syncytes cell and their various stages of meiosis I (PI, MI, AI and TI) in pollen mother cells (PMCs) was assessed. During the microsporogenesis in P. sativum, the occurrence of cytomixis and syncytes at various stages of meiosis I were seen. The formation of cytoplasmic channels and direct fusing of pollen mother cells (PMCs) were both seen to cause cytomixis, with the former being more common than the latter. The percentage of PMCs with cytomixis and syncytes cells increased with increase in the concentration of pesticides. The result of the present investigation indicates that commonly used pesticides ME, IM, and CL have a significant effect on pollen fertility, frequency of cytomixis, and kind of cytomixis cells, the cytomictic channel (CC) and direct fusion (DF), in addition, frequency of syncytes cell and their various stages of meiosis I (PI, MI, AI and TI) in pollen mother cells (PMCs) on P. sativum.     

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.     

Cytomixis and its possible evolutionary role in a Kuwaiti population of Diplotaxis harra (Brassicaceae)

During a cytotaxonomical study of a Kuwaiti diploid (2 n  = 26) population of Diplotaxis harra (Forssk.) Boiss (Brassicaceae), cytomixis and aneuploidy were found in 1.5% and 7.8% of the pollen mother cells (PMCs), respectively. Cytomictic cells revealed wide variability as far as the number of involved PMCs and their stage of division were concerned. The cytoplasmic channels, which were of different sizes, contained migrating chromosomes in a few cases. In some PMCs, the location of some of the chromosomes was clearly indicative of an imminent migration from a donor to its attached recipient cell. At metaphase I and anaphase I, PMCs showed variable degree of chromosomal transfer. In addition to a genetic control of the phenomenon, stress factors such as high temperature or drought in certain periods of the growing season may have contributed to the incidence of cytomixis in the material. A possible relationship between cytomixis and detected aneuploid PMCs, as well as a reconsideration of an evolutionary role of cytomixis is discussed.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 143 , 169–175.     

Male meiosis and behaviour of sex chromosomes in different populations of Rumex acetosa L. from the Western Himalayas, India

Detailed meiotic analysis in 28 North West Himalayan populations of dioecious plant Rumex acetosa L. was carried out. The species is generally discussed as an important plant having sex chromosomes. Male meiosis in all the studied populations clearly showed the formation of six bivalents and one trivalent during diakinesis and metaphase-I. The sex chromosomes in male plants exhibit a chain of trivalent (Y₁–X–Y₂). In addition, among the presently investigated populations ring-shaped trivalents were also observed for the first time in the species. Varied frequency of abnormal segregation of sex trivalent was also observed leading to XY:Y segregation instead of normal X:Y₁Y₂ segregation. A majority of the populations exhibit normal meiosis. Plants of six populations show meiotic abnormalities like cytomixis, laggards, bridges, chromatin stickiness, etc., leading to reduced pollen fertility. Translocation between an autosome and sex chromosomes was also observed in some of the populations. 0–1B chromosomes were noticed in one population. This is the first ever meiotic analysis of the species from India.     

Cytomixis in Lolium perenne

Cytomixis is a spontaneous process occurring through the formation of cytoplasmic bridges between adjacent pollen mother cells. This phenomenon was observed in the pollen mother cells of 3 genotypes of Lolium perenne which had been subjected to directional selection for productivity of green material. — The process has led to the formation of up to 34.8% of PMCs with chromosome numbers deviating from the normal diploid number 2n=14. The abnormal PMCs contained chromosome numbers ranging from 2–56 as observed at first metaphase. — Abnormal PMCs were also observed with approximately the same frequency in Meiosis II. This phenomenon is under genetic control. The evolutionary significance of cytomixis and the cytological consequences are discussed.     

Cytogeography and phenomenon of cytomixis in Silene vulgaris from cold regions of Northwest Himalayas (India)

Silene vulgaris is an important ethnobotanical species, but so far no thorough meiotic analysis has been performed, despite its wide occurrence in temperate and alpine Himalayas, this prompted us to undertake present study. We investigated original meiotic chromosome number and effect of cytomixis on meiotic course, microsporogenesis and pollen grain size and fertility. The paper also summarizes the previously published chromosome counts in the species and cytogeography of 2x and 4x cytotypes in the Indian Himalayas and elsewhere. Meiocytes preparations were made through standard squashed technique in 1 % acetocarmine. Two ploidy levels have been determined during the cytomorphological surveys, diploid (2n = 2x = 24) and tetraploid (2n = 4x = 48). Most of the presently scored populations were found to be diploid (2n = 2x = 24). Morphologically 2x and 4x individuals were indistinguishable from each other. Meiosis was perfectly normal in 4x cytotype. However, some of the 2x plants showed the phenomenon of cytomixis and associated meiotic abnormalities. Consequent to cytomixis, hypo-, hyperploid and enucleated pollen mother cells (PMCs) were resulted. Such PMCs combined with other meiotic abnormalities yielded sterile and variable sized pollen grains. On the basis of overall information, it is clear that 2x cytotype is the most widely distributed cytotype in the Indian Himalayas as well as elsewhere. On the other hand, the 4x cytotype seems to be less frequently present in the Indian Himalayas and outside of India.     

The Relation between Cytomixis and Variation of Chromosome Numbers in Pollen Mother ceils of Rye (Secale cereale L.)

Cytomixis is a spontaneous and normal process occurring through the formation of plasma channels between neighbouring pollen mother cells (PMCs). This phenomenon was observed in the PMCs of 2 variaties of Secale cereale which had been collected from Gausu and Shanxi Provinces. The process had led to the formation of up to 6.37% of PMCs with chromosome numbers deviating from the normal haploid number n=7. The abnormal PMCs contained chromosome numbers ranging from 5–9 in metaphase Ⅰ. Abnormal PMCs with decreased and increased chromosome numbers could be the result of cytomixis. The cytological consequences and the evolutionary significance of cytomixis are discussed.     

蚕豆花粉母细胞染色体畸变同染色质胞间转移的关系

采用压碎、石蜡切片及植物染色体分带技术观察蚕豆花粉母细胞减数分裂过程发现:1.蚕豆花粉母细胞减数分裂的凝线期也出现染色质胞间转移现象;2.在8.08%的花粉母细胞内,染色体偏离正常的数目(n=6);3.染色体结构也有显著改变,其中包括染色体断裂、桥和长度的增减;4.出现双核和无核的花粉母细胞。大量证据表明,这些畸变现象同凝线期的染色质胞间转移有关。     

烟草花粉母细胞细胞融合过程中细胞骨架的超微结构观察

应用普通电镜和DGD去包埋技术 ,研究了烟草花粉母细胞中的细胞融合现象及细胞融合过程中细胞骨架的变化。观察发现 ,处于凝线期的花粉母细胞 ,其内含物 ,包括细胞器和染色质 ,主要通过胞质通道向相邻细胞发生转移。DGD去包埋观察发现 ,花粉母细胞中核骨架与细胞质内及胞间连丝和胞质通道内胞质骨架连接成一个整体。在整个细胞融合过程中 ,均有核骨架纤维与染色质相连。本文讨论了细胞骨架在细胞融合过程中的作用