两种山羊豆属植物花粉形态与花粉母细胞减数分裂行为观察

采用扫描电镜与1%醋酸洋红压片的方法,观察山羊豆属两种植物的花粉形态及花粉母细胞减数分裂行为,从而为山羊豆属的遗传学及分类学提供依据。两种山羊豆属植物立体形状为近球形或扁球形,均具三孔沟,穴—网状纹饰,极面观为近三角形,赤道面观为圆形。东方山羊豆的大小为:赤道轴×极轴=14.3（13.5～15.0）×12.6（12.1～13.7）μm,山羊豆的大小为：赤道轴×极轴=13.7（12.3～14.8）×12.4（10.6～13.0）μm.花粉母细胞减数分裂为同时型胞质分裂,终变期染色体构型为8Ⅱ,没有出现落后染色体及微核现象。小孢子四分体为四面体型。减数分裂过程有一定规律,与花蕾大小,颜色有密切关系。减数分裂进程存在一定差异,同一个花蕾中的不同花药花粉母细胞分裂速度基本相同,所处的时期及观察的分裂相一致,在同一个花药中的不同花粉母细胞中,可见各种不同时期的细胞。     

Cytological and morphology characteristics of natural microsporogenesis within Camellia oleifera

Camellia oleifera is believed to exhibit a complex intraspecific polyploidy phenomenon. Abnormal microsporogenesis can promote the formation of unreduced gametes in plants and lead to sexual polyploidy, so it is hypothesized that improper meiosis probably results in the formation of natural polyploidy in Camellia oleifera. In this study, based on the cytological observation of meiosis in pollen mother cells (PMCs), we found natural 2n pollen for the first time in Camellia oleifera, which may lead to the formation of natural polyploids by sexual polyploidization. Additionally, abnormal cytological behaviour during meiosis, including univalent chromosomes, extraequatorial chromosomes, early segregation, laggard chromosomes, chromosome stickiness, asynchronous meiosis and deviant cytokinesis (monad, dyads, triads), was observed, which could be the cause of 2n pollen formation. Moreover, we confirmed a relationship among the length–width ratio of flower buds, stylet length and microsporogenesis. This result suggested that we can immediately determine the microsporogenesis stages by phenotypic characteristics, which may be applicable to breeding advanced germplasm in Camellia oleifera.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-01002-5.     

Microsporogenesis and development of the male gametophyte in Allium senescens L. (Liliaceae) in China

Microsporogenesis and development of the male gametophyte of Allium senescens L. were studied by use of paraffin sections. The anther was found to be four-sporangia. Its wall includes four layers: epidermis, endothecium, middle layer, and secretory tapetum. Cytokinesis during meiosis of microspore mother cells (MMCs) is successive. Most mature pollen grains are two-celled, a few are three-celled. In addition, anomalies were observed during meiosis of MMC and in uninucleate microspores, including laggard chromosomes, lagging chromosomes, anaphase bridges, and micronucleus. These anomalies are possible reasons for abortive pollen.     

Impaired male meiosis, morphology and distribution pattern of different cytotypes of Bupleurum lanceolatum Wall. (Apiaceae) from the Western Himalayas

Detailed male meiosis, critical morphological observations and distribution pattern of diploid as well as tetraploid cytotypes of the Western Himalayan species, Bupleurum lanceolatum have been evaluated at present. A diploid (n = 8) cytotype is reported from Kashmir, whereas, both diploid (n = 8) and tetraploid (n = 16) cytotypes are available from two districts Kangra and Sirmaur of Himachal Pradesh. Out of these, the tetraploid cytotype makes new addition for the species on a worldwide basis. As per behavior, a tetraploid cytotype is characterized by abnormal meiosis leading to high pollen sterility and size variation of the pollen grains. Morphologically, tetraploids are noted to be luxuriant in comparison to the diploids.     

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.     

Microsporogenesis and pollen development in Leymus chinensis with emphasis on dynamic changes in callose deposition

Leymus chinensis is an economically and ecologically important grass that exhibits low seed production. To better understand the causes of its low sexual reproductivity, the microsporogenesis and pollen development of this species were investigated, with emphasis on dynamic changes in callose deposition. A variety of histochemical stains were employed, including Heidenhain's hematoxylin, decolorized aniline blue, DAPI, and acetocarmine, along with a temporary mount method. Microsporogenesis and pollen development generally took place from June 12 to 26. The meiosis of microspore mother cells (MMCs) was of the successive type and the tetrad was isobilateral in shape. Mature pollen grains comprised two sperms and a vegetative nucleus. Callose initially appeared in the center of the anther locule at the premeiotic phase, and then gradually and unevenly deposited around the MMC before the commencement of meiosis. At the onset of meiosis, the accumulation of callose enclosing the MMC peaked, accompanied by the disappearance of callose in the center of the locule. At the dyad and tetrad stages, the dyads and tetrads were surrounded by callose wall and the microspores in the tetrads were isolated by a crossed cell plate composed of callose. Microspores just released from tetrads were still enclosed in callose wall, and then callose gradually disappeared in the pollen wall. Ultimately, callose almost completely disappeared from the walls of mature pollen grains. In the large numbers of sections observed, most of the cases of meiosis of the MMCs, pollen development, and callose dynamics were normal, with only a few abnormities observed. The results suggest that microsporogenesis, male gametogenesis, and callose dynamics during these processes are generally normal in this species, and that the callose wall plays an important role in the production of functional pollen grains. The small numbers of abnormities of these processes that occurred likely do not adversely affect the production of viable pollen grains. Therefore, microsporogenesis and pollen development may not be factors in the low seed production of L. chinensis.     

A CENH3 mutation promotes meiotic exit and restores fertility in SMG7-deficient Arabidopsis

Meiosis in angiosperm plants is followed by mitotic divisions to form multicellular haploid gametophytes. Termination of meiosis and transition to gametophytic development is, in Arabidopsis, governed by a dedicated mechanism that involves SMG7 and TDM1 proteins. Mutants carrying the smg7-6 allele are semi-fertile due to reduced pollen production. We found that instead of forming tetrads, smg7-6 pollen mother cells undergo multiple rounds of chromosome condensation and spindle assembly at the end of meiosis, resembling aberrant attempts to undergo additional meiotic divisions. A suppressor screen uncovered a mutation in centromeric histone H3 (CENH3) that increased fertility and promoted meiotic exit in smg7-6 plants. The mutation led to inefficient splicing of the CENH3 mRNA and a substantial decrease of CENH3, resulting in smaller centromeres. The reduced level of CENH3 delayed formation of the mitotic spindle but did not have an apparent effect on plant growth and development. We suggest that impaired spindle re-assembly at the end of meiosis limits aberrant divisions in smg7-6 plants and promotes formation of tetrads and viable pollen. Furthermore, the mutant with reduced level of CENH3 was very inefficient haploid inducer indicating that differences in centromere size is not the key determinant of centromere-mediated genome elimination.     

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。     

Meiotic behavior and pollen morphology variation in Centaurium pulchellum (Gentianaceae)

Centaurium pulchellum is an annual herb native to Europe, but introduced in South America, where it is widely used in the preparation of digestive infusions and bitter drinks. In this species, a wide variation in the aperture pattern of pollen grains was reported and has been attributed to environmental factors or irregularities at meiosis. For this reason, cytological and palynological studies have been undertaken on two different populations. The pollen grain analysis showed that some types are more frequent within each population, but the most common forms were the typical 3-colporate and 4-colporate. The cytological analysis revealed that the analyzed populations of C. pulchellum have chromosome number 2n = 36. The presence of tetravalents at meiosis strongly suggests that these populations are autotetraploid based on x = 9. The meiotic behavior showed a significant percentage of irregularities in different phases: off-plate bivalents, precocious segregation, laggard chromosomes, bridges, and cytomixis. However these irregularities are not related to the variation in the aperture pattern of pollen grains. The heteromorphism in the pollen grains observed in C. pulchellum could be a normal condition to which the species is well adapted.     

Die ursachen der sterilität vonAllium sativum L.

A fully normal meiosis was found inAllium sativum L. by means of PMC analysis. Quadrivalents were formed only in several cultivars. The cultivation of inflorescence stem (after the exstirpation of bulblets) in a tetracycline solution of 900 i.u. induced a formation of fertile flowers. The first pollen mitosis was studied and the pollen parameters compared with those ofAllium cepa L. Evidence for the pollen sterility was given by crossings on sterile types. A microbiological factor was not isolated.     

Nucleic acids in relation to cell division in Lilium longiflorum

Methods are described for preparing cell suspensions of Lilium microsporocytes, microspores and pollen grains; for obtaining cell counts of these suspensions; and for their analysis for pentose nucleic acid (PNA) and desoxypentose nucleic acid (DNA).The results of these analyses have been calculated to nucleic acid content in μμg per microsporocyte, microspore or pollen grain, and the results related to logarithm of flower bud length, an index of the developmental status of the cells, and of their temporal relationship to meiosis, microspore mitosis and opening of the flower.DNA content per cell drops sharply at the end of meiosis, with the formation of four microspores from each microsporocyte. It then increases gradually during the microspore interphase between meiosis and the microspore mitosis. At microspore mitosis DNA content doubles rapidly. In the development of the resulting binucleate pollen grain, from microspore mitosis until the opening of the flower, there is a further gradual increase of DNA content. PNA content of these cells follows the same pattern up to microspore mitosis at a level about twice that of DNA, increases sharply at mitosis, and continues to increase rapidly at a rate nine times that for DNA in the maturing pollen grain.The absolute amounts of DNA and PNA are great. At the time of anthesis the two-celled pollen grain contains about 375 μμg of DNA and 1705 μμg of PNA.     

温度对斯托克通氏烟草雄配子体形成和发育的影响

为探究低温对斯托克通氏烟草(Nicotiana stocktonii)花粉母细胞(PMC)减数分裂及其雄配子体发育过程的影响,采用卡宝品红染色法,研究不同温度条件下该材料雄配子体形成和发育的过程。结果表明:种植于昼温(31±0.5)℃、夜温(11±0.5)℃人工气候箱中的Nicotiana stocktonii花粉母细胞减数分裂过程异常现象较少,出现微核的比率较低,用新鲜成熟的花粉做萌发实验花粉萌发率较高,为(71±3)%; 而种植于昼温(25±0.5)℃、夜温(3±0.5)℃条件下的Nicotiana stocktonii开花后花药大多干瘪,用新鲜成熟花粉做萌发实验花粉萌发率低,为(13.67±3)%,花粉母细胞减数分裂过程出现染色体桥、染色体不同步、染色体断片、落后染色体等现象,存在微核的细胞比率较高。因此,Nicotiana stocktonii花粉母细胞减数分裂与小孢子发育过程易受温度影响,从而影响花粉的可育性。     

Ultrastructural study of pollen and anther development in Luehea divaricata (Malvaceae,Grewioideae) and its systematic implications: Role of tapetal transfer cells,orbicules and male germ unit

Developmental processes of microsporogenesis and microgametogenesis in Luehea divaricata (Malvaceae, Grewioideae) were analyzed with transmission electron microscopy. The species studied has perfect flowers. The young anthers are bithecal and tetrasporangiate; microspore mother cells undergo simultaneous meiosis, forming tetrads with a tetrahedral arrangement. The development of the anther wall conforms to the basic type and the tapetum is secretory. The results highlight the presence of multinucleate tapetal cells, which acquire ultrastructural features characteristic of transfer cells at the young pollen grain stage, and which are associated with the presence of orbicules. Tapetal transfer cells have not been previously investigated in detail for other species of angiosperms; their function during pollen development is discussed. The present work is the first contribution to the knowledge of ultrastructural pollen development in the genus Luehea, as well as in the subfamily Grewioideae.     

Ultrastructure of microsporogenesis and microgametogenesis in Campsis radicans (L.) Seem. (Bignoniaceae)

In the present study, microsporogenesis, microgametogenesis and pollen wall ontogeny in Campsis radicans (L.) Seem. were studied from sporogenous cell stage to mature pollen using transmission electron microscopy. To observe the ultrastructural changes that occur in sporogenous cells, microspores and pollen through progressive developmental stages, anthers at different stages of development were fixed and embedded in Araldite. Microspore and pollen development in C. radicans follows the basic scheme in angiosperms. Microsporocytes secrete callose wall before meiotic division. Meiocytes undergo meiosis and simultaneous cytokinesis which result in the formation of tetrads mostly with a tetrahedral arrangement. After the development of free and vacuolated microspores, respectively, first mitotic division occurs and two-celled pollen grain is produced. Pollen grains are shed from the anther at two-celled stage. Pollen wall formation in C. radicans starts at tetrad stage by the formation of exine template called primexine. By the accumulation of electron dense material, produced by microspore, in the special places of the primexine, first of all protectum then columellae of exine elements are formed on the reticulate-patterned plasma membrane. After free microspore stage, exine development is completed by the addition of sporopollenin from tapetum. Formation of intine layer of pollen wall starts at the late vacuolated stage of pollen development and continue through the bicellular pollen stage.     

Impaired Male Meiosis due to Irregular Synapsis Coupled with Cytomixis in a New Diploid Cytotype of Dianthus angulatus (Caryophyllaceae) from Indian Cold Deserts

Dianthus angulatus (Caryophyllaceae) is cytologically examined here for the first time for the area of India. The diploid chromosome count of 2n?=?30, ascertained here, represents a new cytotype, supplementing the earlier report of a hexaploid cytotype with 2n?=?90 from outside of India. We report here the occurrence of two plants showing impaired meiosis due to irregular synapsis and cytomixis collected from Kinnaur district, Himachal Pradesh (India). The other plants of this species collected in Lahaul-Spiti region showed normal male meiosis (n?=?15) with a high (95%?100%) pollen fertility and normal seed set, and they reproduce sexually. Irregular synapsis in two plants from the Kinnaur region is characterized by the complete absence of chromosome pairing and the presence of 30 univalents at diakinesis and meta-anaphase. In addition, other meiotic irregularities were found, such as unoriented chromosomes, laggards, precocious movements of univalents at anaphase-I and micronuclei at telophase. Microsporogenesis was also abnormal, resulting in the formation of monads, dyads, triads, polyads, and tetrads with micronuclei. The occurrence of intra- and intermicrosporal chromatin material transfer during microsporogenesis was observed, which is a rather rarely observed phenomenon. The synaptic irregularities coupled with chromatin transfer in these plants seem to be responsible for the high pollen sterility (38%?42%) and heterogeneously sized pollen grains. In these plants no seeds were set, and plants reproduced vegetatively through root suckers.     

Diplospory and obligate apomixis in Miconia albicans (Miconieae, Melastomataceae) and an embryological comparison with its sexual congener M. chamissois

Apomixis, or asexual reproduction through seeds, has been reported for species of the tribe Miconieae, Melastomataceae, but details of the process have yet to be described. We analyzed and compared sporogenesis and gametogenesis in the apomictic Miconia albicans and the sexual M. chamissois. The results point to some differences between species, which were related to the apomictic process. In M. albicans microsporogenesis, problems during meiosis and degeneration of its products led to total pollen sterility, while M. chamissois presented normal bicellular pollen grains in the mature anther. The absence or abnormality of meiosis in M. albicans megasporogenesis led to the formation of an unreduced embryo sac and also to egg cell parthenogenesis, which gave rise to the apomictic embryo. Embryo and endosperm development were autonomous, resulting in seeds and fruits independent of pollination and fertilization. Thus, in this species, apomixis can be classified as diplosporic and obligate. In contrast, meiosis was as expected in the sexual M. chamissois, and led to the development of a reduced embryo sac. Despite the divergent pathways, many embryological characteristics were similar between the studied species and other Melastomataceae and they seem to be conservative character states for the family.     

Genes expressed in the male gametophyte of flowering plants and their isolation

Recombinant cDNA libraries to poly(A)RNA isolated from mature pollen of Zea mays and Tradescantia paludosa have been constructed. Northern blot analyses indicate that several of the clones are unique to pollen and are not expressed in vegetative tissues. The majority, however, are expressed both in pollen and vegetative tissues. Southern hybridizations show that the pollen specific sequences in corn are present in one or a very few copies in the genome. By using several of the clones as probes, it was found that there are at least two different groups of mRNAs with respect to their synthesis. The mRNAs of the first group represented by the pollen specific clones are synthesized after microspore mitosis and increase in concentration up to maturity. The second group, exemplified by actin mRNA, begins to accumulate soon after meiosis, reaches its maximum by late pollen interphase, and decreases thereafter. Although the actin mRNA and the pollen specific mRNAs studied show very different patterns of initiation of synthesis and accumulation during pollen development, the rates of decline of these mRNAs during the first 60 minutes of germination and pollen tube growth in Tradescantia are similar and reflect the previously observed declines in rates of protein synthesis during this period.     

Panicum shastense (gramineae), a sterile hybrid between P. Pacificum and P. Scribnerianum

Natural and synthetic hybrids are reported betweenPanicum, scribnerianum Nash and two other species,P. pacificum Hitchc. & Chase andP. occidentale Scribn. & Merr. The hybrids are morphologically intermediate between the parents, and those in whichP. pacificum is one parent strongly resemble the rareP. shastense Scribn. & Merr. All the parent plants studied cytologically had regular meiosis and a gametic number ofn = 9 chromosomes. The hybrids usually showed good pairing of chromosomes in meiosis, but irregularities such as univalents and lagging chromosomes were common. In the hybrids generally less than 7% of the pollen grains stained with cotton blue in lactophenol, and the plants set no seed, whereas the parent plants generally had over 90% pollen stainability and a high percentage of seed set. Pollen fromP. shastense, like that of the hybrids, was 3–4% stainable.Panicum shastense is believed to be based on a first generation sterile hybrid and the nameP. xshastense is proposed.     

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 %).