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.     

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。     

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.     

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.     

The role of microtubular cytoskeleton and callose walls in the cytomixis process in tobacco (Nicotiana tabacum L.) pollen mother cells

We have studied the microtubule cytoskeleton structure and callose walls deposition in the course of meiosis at the cytomictic and normal tobacco (N. tabacum L.) PMCs. It was ascertained that microtubule cytoskeleton did not play an evident part in the process of cytomixis. Increased cytomixis frequency probably is determined by irregular callose walls deposition. The possible reasons of nuclear material passage between tobacco PMCs at the cellular level are discussed.     

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.     

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.     

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.     

小麦品种群体的细胞学研究

对９个小麦品种群体进行了细胞学观察。结果表明,４．５３％的细胞含有至少一对不配对的染色体,２．３１％的细胞里,染色体偏离正常的数目。显然,这种自然的不规则性可能导致非整倍体后代的产生。３．５７％的细胞发生了易位,群体的２．２２％为单体植株。     

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.     

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     

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.     

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.     

High efficiency production and genomic in situ hybridization analysis of Brassica aneuploids and homozygous plants

Inplantbreedingandgeneticresearch,karyotypicallystablecrosseswhichproducehybridplantshavebeenextensivelyusedtointroduceintocropsthetargettraitsandgenesfromrelatedwildorcultivatedspeciesortoconstructstocksforgeneticanalysis(alienchromosomeadditions,substitutionsandtranslocations)[1—3].Uniparentalgenomeeliminationinkaryotypicallyunstablehybridshasbeenutilizedforhaploidproduction[2,4].Becausetheartificiallysynthesizedallopoly-ploidscannotbeusedascropsformanyreasons,onepurposeofwidehybridizations…     

High efficiency production and genomic<Emphasis Type="Italic">in situ</Emphasis> hybridization analysis of<Emphasis Type="Italic">Brassica</Emphasis> aneuploids and homozygous plants

Interspecific and intergeneric hybridizations have been widely used in plant genetics and breeding to construct stocks for genetic analysis and to introduce into crops the desirable traits and genes from their relatives. The intergeneric crosses between Brassica juncea (L.) Czern. & Coss., B. carinata A. Braun and Orychophragmus violaceus (L.) O. E. Schulz were made and the plants produced were subjected to genomic in situ hybridization analysis. The mixoploids from the cross with B. juncea were divided into three groups. The partially fertile mixoploids in the first group (2n = 36-42) mainly contained the somatic cells and pollen mother cells (PMCs) with the 36 chromosomes of B. juncea and additional chromosomes of O. violaceus. The mixoploids (2n = 30-36) in the second and third groups were morphologically quite similar to the mother plants B. juncea and showed nearly normal fertility. The plants in the second group produced the majority of PMCs (2n = 36) with their chromosomes paired and segregated normally, but 1-4 pairs of the O. violaceus chromosomes were included in some PMCs. The plants in the third group produced only PMCs with the 36 B. juncea chromosomes, which were paired and segregated normally. The mixoploids (2n = 29-34) from the cross with B. carinata produced the majority of PMCs (2n = 34) with normal chromosome pairing and segregation, but some plants had some PMCs with 1-3 pairs of chromosomes from O. violaceus and other plants had only PMCs with the B. carinata chromosomes. The Brassica homozygous plants and aneuploids with complete or partial chromosome complements of Brassica parents and various numbers of O. violaceus chromosomes were derived from these progeny plants. The results in this study provided the molecular cytogenetic evidence for the separation of parental genomes which was previously proposed to occur in the hybridizations of these two genera.     

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.     

Efficiency of the induction of cytomixis in the microsporogenesis of dicotyledonous (<Emphasis Type="Italic">N. tabacum</Emphasis> L.) and monocotyledonous (<Emphasis Type="Italic">H. distichum</Emphasis> L.) plants by thermal stress

The efficiencies of the induction of cytomixis in microsporogenesis by thermal stress are compared in tobacco (N. tabacum L.) and barley (H. distichum L.) It has been shown that different thermal treatment schedules (budding tobacco plants at 50°C and air-dried barley grains at 48°C) produce similar results in the species: the frequency of cytomixis increases, and its maximum shifts to later stages of meiosis. However, the species show differences in response. The cytomixis frequency increase in tobacco is more pronounced, and its maximum shifts from the zygotene–pachytene stages of meiotic prophase I to prometaphase–metaphase I. Later in the meiosis, aberrations in chromosome structure and meiotic apparatus formation typical of cytomixis are noted, as well as cytomixis activation in tapetum cells. Thermal stress disturbs the integration of callose-bearing vesicles into the callose wall. Cold treatment at 7°C does not affect cytomixis frequency in tobacco microsporogenesis. Incubation of barley seeds at 48°C activates cytomixis in comparison to the control, shifts its maximum from the premeiotic interphase to zygotene, and changes the habit of cytomictic interactions from pairwise contacts to the formation of multicellular clusters. Thermal treatment induces cytomictic interactions within the tapetum and between microsporocytes and the tapetum. However, later meiotic phases show no adverse consequences of active cytomixis in barley. It is conjectured that heat stress affects callose metabolism and integration into the forming callose wall, thereby causing incomplete closure of cytomictic channels and favoring intercellular chromosome migration at advanced meiotic stages.     

Peculiarities of cytomixis in pollen mother cells of transgenic tobacco plants (Nicotiana tabacum L.) with mutant phenotype

The frequency characteristics and cytological picture of cytomixis in the course of male meiosis are described in transgenic tobacco plants (Nicotiana tabacum L.) with altered flower morphology and male sterility. Effects of cytomixis on qualitative composition of meiotic products are studied (formation of cytoplasts and polyads). Doubling of the chromosome number was established to increase frequency of cytomixis in the studied plants.     

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.     

萝卜与甘蓝属间杂种基因组原位杂交分析

用基因组原位杂交方法（Genomic in situ hybridization, 简称GISH）研究了萝卜( Raphanus sativus,2n=18,RR)和甘蓝（Brassica oleracea , 2n=18, CC）属间杂种F1减数分裂过程。结果表明杂种体细胞染色体组成为RC,2n=18,但花粉母细胞有三种不同类型：1. RC,2n=18, 终变期染色体平均配对构型为14.87Ⅰ+1.20Ⅱ+0.04Ⅲ+0.06Ⅳ, 染色体配对主要发生在萝卜和甘蓝染色体之间, 后期Ⅰ9条萝卜染色体主要以5/4和6/3的分离比移向两极, 所形成配子的染色体数目和组成均不平衡,配子败育; 2. RRCC,4n=36, 终变期染色体形成18个二价体,后期Ⅰ染色体均衡分离,形成RC不减数配子;3. RRCC缺体,4n=30－34, 少数萝卜染色体丢失,形成的配子具有全套的甘蓝染色体和部分萝卜染色体。