Phylogenetic comparative analysis of microsporogenesis in angiosperms with a focus on monocots

This paper presents the first broad overview of three main features of microsporogenesis (male meiosis) in angiosperms: cytokinesis (cell division), intersporal wall formation, and tetrad form. A phylogenetic comparative approach was used to test for correlated evolution among these characters and to make hypotheses about evolutionary trends in microsporogenesis. The link between features of microsporogenesis and pollen aperture type was examined. We show that the pathway associated with successive cytokinesis (cytoplasm is partitioned after each meiotic division) is restricted to wall formation mediated by centrifugally developing cell plates, and tetragonal (or decussate, T-shaped, linear) tetrads. Conversely, much more flexibility is observed when cytokinesis is simultaneous (two meiotic divisions completed before cytoplasmic partitioning). We suggest that the ancestral type of microsporogenesis for angiosperms, and perhaps for all seed plants, associated simultaneous cytokinesis with centripetal wall formation, resulting in a large diversity in tetrad forms, ranging from regular tetrahedral to tetragonal tetrads, including rhomboidal tetrads. From this ancestral pathway, switches toward successive cytokinesis occurred among basal angiosperms and monocots, generally associated with a switch toward centrifugal intersporal wall formation, whereas eudicots evolved toward an almost exclusive production of regular tetrahedral tetrads. No straightforward link is found between the type of microsporogenesis and pollen aperture type.     

华北落叶松小孢子母细胞(PMC)减数分裂和花粉的发育

采用染色体压片法系统地观察了华北落叶松Larix principis-rupprechtii小孢子母细胞(PMC)减数分裂的细胞学特征和花粉发育过程。结果表明:华北落叶松PMC减数分裂始于当年秋季,以弥散双线期的特殊状态休眠越冬,次年春天又重新恢复其后续的减数分裂过程。主要特点是在同一花药中,减数分裂不同步,且各阶段分裂速度较快,3天内全部停留在四分体阶段;二价体构型主要为两臂共发生过两次以上交换的环形二价体,少数细胞中偶见一臂发生交换的棒状二价体;其平均构型为10.62II 1.38II;中期II核分裂相既有平行式样,又有互为垂直式样,形成左右对称型和四面体型四分体,兼具单、双子叶植物四分体主要类型特点。四分体持续一周后,细胞壁开始溶解,4个小孢子分离并游离在药室中,成为独立的单核花粉。其后细胞核开始有丝分裂,最后形成由2个原叶细胞、1个管细胞、1个柄细胞和1个体细胞共5个细胞组成的成熟花粉。此外,还观察到成熟花粉当中有13.6%是由4个细胞组成的。     

Frequent Changes in the Number of Reiterated Ribosomal RNA Genes Throughout the Life Cycle of the Basidiomycete Coprinus Cinereus

We have examined the stability of the tandemly repeated genes that encode the ribosomal RNA in Coprinus cinereus. These genes are contained within two linked HindIII fragments in a 3.0-Mb chromosome. We monitored the size of these fragments in both mitotic and meiotic segregants using the contour-clamped homogeneous electric field (CHEF) method. No length changes were observed in the smaller HindIII fragment (100 kb; 10 repeats) among the DNAs prepared from 46 asexual spore derivatives (oidia) or 128 meiotic segregants (basidiospores from 32 tetrads). However, the larger HindIII fragment (1100 kb; 120 repeats) did exhibit variability. Substantial changes, involving up to 40% of the larger HindIII fragment were recorded in 7 of 46 oidial isolates (including 4 of 22 transformed derivatives). To learn if the changes were confined to the vegetative portion of the life cycle, we examined transmission of HindIII variants through three crosses. In the first two crosses (16 tetrads total), no changes were observed in the large HindIII fragment. However, in the third cross (16 tetrads), each tetrad showed at least one alteration. In half of the tetrads from the third cross, the altered patterns segregated 2:2, suggesting that the changes occurred after mating but prior to premeiotic DNA replication. We conclude that breakage and rejoining reactions within the rDNA are frequent and are not confined to any particular stage of the life cycle. It also appears that certain repeats are sheltered from these events. Finally, marked differences in rDNA stability were observed in the crosses analyzed.     

Pteridophytic features in some Lower Carboniferous seed megaspores

Some Lower Carboniferous seed megaspores have triradiate sutures–unequivocal evidence of their being arranged in a tetrahedral configuration. This type of tetrad arrangement is unknown in the ovules of modern gymnospermS. Some of the fossil tetrads consist of one large functional megaspore and three smaller abortive spores attached at the apex of the large one. All four spores have an exinous covering.
Spores arranged in tetrahedral tetrads, and hence with triradiate sutures, are characteristic of many modern and fossil free-sporing pteridophyteS. In possessing this feature the fossil seed megaspores are more similar to the spores of these pteridophytes than to the megaspores of modern gymnosperms.     

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.     

脉孢霉两对基因顺序四分子分析

真菌和单细胞藻类四分子分析能够利用单次减数分裂的4个产物进行独特的遗传分析, 是帮助人们直观地理解遗传机制的重要手段, 已被用于高等生物遗传作图。文章运用孟德尔遗传规律、遗传重组机理与遗传作图原理, 分析了两对基因杂交的子囊、子囊孢子类型间关系; 推导出了两对基因顺序四分子分析的完整步骤。     

Intranuclear membranes and the formation of the first meiotic spindle in Xenos peckii (Acroschismus wheeleri) oocytes

The ultrastructure of spindle formation during the first meiotic division in oocytes of the Strepsipteran insect Xenos peckii Kirby (Acroschismus wheeleri Pierce) was examined in serial thick (0.25- micron) and thin sections. During late prophase the nuclear envelope became extremely convoluted and fenestrated. At this time vesicular and tubular membrane elements permeated the nucleoplasm and formed a thin fusiform sheath, 5-7 micron in length, around each of the randomly oriented and condensing tetrads. These membrane elements appeared to arise from the nuclear envelope and/or in association with annulate lamellae in the nuclear region. All of the individual tetrads and their associated fusiform sheaths became aligned within the nucleus subsequent to the breakdown of the nuclear envelope. Microtubules (MTs) were found associated with membranes of the meiotic apparatus only after the nuclear envelope had broken down. Kinetochores, with associated MTs, were first recognizable as electron-opaque patches on the chromosomes at this time. The fully formed metaphase arrested Xenos oocyte meiotic apparatus contained an abundance of membranes and had diffuse poles that lacked distinct polar MT organizing centers. From these observations we conclude that the apparent individual chromosomal spindles--seen in the light microscope to form around each Xenos tetrad during "intranuclear prometaphase" (Hughes-Schrader, S., 1924, J. Morphol. 39:157-197)--actually form during late prophase, lack MTs, and are therefore not complete miniature bipolar spindles, as had been commonly assumed. Thus, the unique mode of spindle formation in Xenos oocytes cannot be used to support the hypothesis that chromosomes (kinetochores) induce the polymerization of their associated MTs. Our observation that MTs appeared in association with and parallel to tubular membrane components of the Xenos meiotic apparatus after these membranes became oriented with respect to the tetrads, is consistent with the notion that membranes associated with the spindle determine the orientation of spindle MTs and also play a part in regulating their formation.     

Ordered Linear Tetrads Are Produced by the Sporulation of Newly Formed Zygotes of Saccharomyces cerevisiae

Diploid Saccharomyces cerevisiae strains normally sporulate to produce tetrahedral unordered asci containing four spores (tetrads). We report that when newly formed zygotes are subjected to the same sporulation conditions, they form predominantly linear ordered tetrads. We show that the two spores from each end of such a linear tetrad invariably contain nonsister centromeres. Spore viability, recombination and independence of centromere segregation appear unaffected.     

Mortality of pollen grains may result from errors of meiosis: study of pollen tetrads in Typha latifolia L

In the cattail Typha latifolia the four haploid products of meiosis remain attached and form the flat tetrad of pollen grains. Gametophytic lethals arisen de novo in diploid cells of sporophyte must manifest themselves as pollen tetrads with two dead grains. This could allow to estimate the rate of recessive lethals arresting pollen grain development. We studied pollen samples collected from 44 sprouts in two populations in the vicinity of Novosibirsk. The anomalous tetrads T1, T2, T3, and T4 carrying one, two, three, and four dead grains, respectively, were detected in each sampled individual. The mean frequency of all anomalous tetrads in the two populations was 3.4% and 8.7%. The frequencies of tetrad classes varied widely among the individuals with correlation coefficient up to 0.94, but their ratios remained nearly constant. The majority of anomalous tetrads were presented by T1 and T2 classes (their sum comprising 72.7 and 74.0% in two populations), T1 being a little more abundant. The observed pattern of frequencies of tetrads with dead grains can be explained by errors of male meiosis such as chromosome non-disjunction in both meiotic divisions. The tetrads with two dead pollen grains may result mostly from non-disjunction in anaphase I, and those with one pollen grain from non-disjunction in anaphase II, thus making tetrad analysis ineffective for estimating the rate of gametophytic lethals.     

Sporogenesis and Gametogenesis of Endangered Adenophora lobophylla Hong (Campanulaceae)

Reported in this paper are the microsporogenesis, megasporogenesis and the development of male and female gametophytes of the endangered species Adenophora lobophylla. The anther is four-sporangiate with its wall composed of four layers: epidermis, fibrous endothelium, middle layer and glandular tapetum of binucleate cells. The cytokinesis of microspore mother cell in meiosis is simultaneous, and tetrads are tetrahedral. Pollen grains are 2-celled at the dispersal stage. Ovule is anatropous, unitegmic and tenuinucellate with a hypodermal archesporial cell developing directly as the megaspore mother cell which undergoes meiotic division and then forms a linear tetrad. The embryo sac is of Polygonum type. The polar nuclei fuse before fertilization. An endothelium differentiates when the uninucleate embryo sac forms. Comparative studies were made with the closely related and widely distributed species A. potaninii. No differences between them were found. No obstacle to sexual reproduction in this endangered species was observed. As a conclusion, no endangering factors were found to influence the sporogenesis and gametogenesis of A. lobophylla. According to our observision and the references on embryological studies of the Campanulaceae(s. l. ), there is no evident differentiation in sporogenesis and gametogenesis in this family.     

濒危植物裂叶沙参的大小孢子发生及雌雄配子体发育

本文以近缘广布种泡沙参为对照,对濒危物种裂叶沙参进行了大小孢子发生和雌雄配子体发育的研究。裂叶沙参的药壁发育为双子叶型,绒毡层为腺质型,细胞含两核。小孢子母细胞在减数分裂过程中胞质分裂为同时型。小孢子四分体为四面体型,成熟花粉粒为2-细胞型。胚珠倒生,单珠被,薄珠心,大孢子四分体为线形排列,胚囊发育为蓼型。成熟胚囊中两极核在受精之前融合为一个大的次生核。当胚囊发育至单核胚囊时,珠被的最内层细胞发育为珠被绒毡层。濒危植物裂叶沙参在大、小孢子发生及雌、雄配子体的发育过程中,未见有败育及其它异常现象;与对照种泡沙参相比,也未见有差异,这说明裂叶沙参的致濒原因不在于有性生殖过程。     

Commitment of Mitotic Cells to Meiosis during the G2 Phase of Premeiosis

In the developing anther, archesporial cells that proliferateby mitotic division are converted into meiotic cells duringthe premeiotic interphase. Experiments with explanted microsporocytesof Lilium and Trillium were made to obtain evidence for theconversion of mitotic to meiotic cells during the premeioticperiod. Explanted premeiotic cells were cultured through thedivision cycle at relatively high division frequencies and showeda variety of division types with respect to chromosomal events.The type of division depended on the premeiotic stage at whichthe cells were explanted. Cells in the G₁, S and early G² phasesunderwent mitotic division and formed a diad or binucleate monad.Cells explanted at the late G₂ phase were cultured throughoutthe normal meiotic cycle, which resulted in typical tetrad configuration. In microsporocytes explanted during the main part of the G₂interval, centromere behavior was meiotic, but chromosome pairingand chiasma formation were disturbed. Thus, she G₂ intervalwas shown to be critical for the commitment of mitotic cellsto meiotic division. Detailed analysis showed that the intracellularchanges that commit the cells to meiosis begin shortly aftercompletion of premeiotic DNA synthesis and that these changesare progressive and cumulative. (Received February 2, 1982; Accepted May 24, 1982)     

Meiotic interallelic conversion at the human minisatellite MS32 in yeast triggers recombination in several chromatids

Tandem repetitive DNA sequences such as minisatellites include the most polymorphic loci yet identified in the human genome. The high mutation rates at many of these loci are driven by incompletely understood recombination-based mechanisms that operate in the germline. To analyse aspects of minisatellite mutation processes and general eukaryotic recombination in meiosis that cannot be studied in humans or other mammals, including crosstalk and interplay between all four chromatids, we have previously constructed a eukaryotic model system, enabling the analysis of all four products of meiosis. In this system we have integrated alleles of the human minisatellite MS32, flanked by synthetic markers, in the vicinity of a meiotic recombination hot spot in chromosome III of Saccharomyces cerevisiae. In the present study, tetrad analysis showed that gene conversion is the predominant and possibly the universal pathway leading to interallelic transfer of repeats, with or without exchange of flanking regions. The data also suggest a hyper-recombinogenic state, triggered by interallelic mutation processes which generate a cascade of mutant alleles in the same meiosis. A number of tetrads contained identical mutant alleles of meiotic origin. Several tetrads could not be explained by the current models for minisatellite mutation. Accordingly, we here present a modified model based on the successive repair of multiple double-strand breaks.     

濒危植物瑶山苣苔的大小孢子发生和雌雄配子体发育研究

利用石蜡切片技术对瑶山苣苔大小孢子发生、雌雄配子体发育及胚胎发育进行了细胞学观察,结果表明:瑶山苣苔胚珠倒生,单珠被,薄珠心,具珠被绒毡层。大孢子母细胞减数分裂形成的四分体呈直线排列,合点端的大孢子发育为功能大孢子,其余3个大孢子退化,胚囊为单孢子发生的蓼型胚囊发育方式。花药为四囊形,花药壁由外到内依次为表皮、药室内壁、中层和腺质绒毡层,小孢子形成时胞质分裂为修饰性同时型,小孢子四分体排列方式为四面体形,成熟花粉为2核细胞。胚乳发育为细胞型,在胚的发育过程中被吸收耗尽。瑶山苣苔大小孢子发生和雌雄配子体发育基本正常,不是导致其濒危的原因。但瑶山苣苔果实成熟时,胚仅发育至球形胚时期,需要经过一定时间休眠才能完成形态后熟,表明胚未发育完全可能是该物种天然更新困难的原因之一。     

Classic Weinstein: tetrad analysis, genetic variation and achiasmate segregation in Drosophila and humans.

A maximum-likelihood method for the estimation of tetrad frequencies from single-spore data is presented. The multilocus exchange with interference and viability (MEIV) model incorporates a clearly defined model of exchange, interference, and viability whose parameters define a multinomial distribution for single-spore data. Maximum-likelihood analysis of the MEIV model (MEIVLA) allows point estimation of tetrad frequencies and determination of confidence intervals. We employ MEIVLA to determine tetrad frequencies among 15 X chromosomes sampled at random from Drosophila melanogaster natural populations in Africa and North America. Significant variation in the frequency of nonexchange, or E(0) tetrads, is observed within both natural populations. Because most nondisjunction arises from E(0) tetrads, this observation is quite unexpected given both the prevalence and the deleterious consequences of nondisjunction in D. melanogaster. Use of MEIVLA is also demonstrated by reanalyzing a recently published human chromosome 21 dataset. Analysis of simulated datasets demonstrates that MEIVLA is superior to previous methods of tetrad frequency estimation and is particularly well suited to analyze samples where the E(0) tetrad frequency is low and sample sizes are small, conditions likely to be met in most samples from human populations. We discuss the implications of our analysis for determining whether an achiasmate system exists in humans to ensure the proper segregation of E(0) tetrads.     

Pollen of Ceratostema (Ericaceae, Vaccinieae): tetrads without septa

The pollen morphology of two species of the Neotropical genus Ceratostema (Ericaceae) was examined by light, scanning and transmission electron microscopy. The Ceratostema species examined have 3-colporate pollen grains united in permanent tetrahedral tetrads that show a common condition encountered in the Ericaceae. But the septal exine was absent between two neighboring grains in each pollen tetrad of Ceratostema. The pollen tetrads without septa are the first report for the Ericaceae as well as other angiosperm families.     

银杏大孢子形成的形态学研究

有关银杏（CinkgobitobaL．）生殖的研究,早在19世纪后期,Strasbuopr首先作了描述「'］,此后,在这一方面陆续有不少报道。到了本世纪50年代,对于银杏的整个生殖过程已有大致轮廓卜一'］,但是对于生殖各阶段的详细变化,无论在胚胎学或细胞学方面,都有待进一步的观?..     

ULTRASTRUCTURE OF SPOROGENESIS IN THE MOSS,AMBLYSTEGIUM RIPARIUM I. MEIOSIS AND CYTOKINESIS

Emphasis is placed on three aspects of meiosis in the moss Amblystegium riparium (Hedw.) BSG: 1***) nature of the sporogenous layer; 2) prophasic microtubules and polarity; and 3) cleavage pattern. Spore tetrads develop while still encased by archesporial cell walls. The cellular nature of the sporogenous layer differs from the more usual occurrence of free sporocytes released into a common spore sac. Two important events mark the establishment of sporocyte polarity during meiotic prophase: 1) migration of the four plastids to the distal tetrad poles (telophase II poles); and 2) ingrowth of the sporocyte wall in eventual cleavage planes between the tetrad poles. An extensive, plastid-based microtubule system is associated with organelle migration during the establishment of sporocyte polarity in meiotic prophase. Disruption of the nuclear envelope in prometaphase I occurs at sites opposite the four plastids where microtubules extend from plastid envelope to nuclear envelope. Formation of a cell plate following the first meiotic division results in a dyad, whereas in many mosses meiosis is completed in the undivided sporocyte and is followed by simultaneous cleavage into a spore tetrad. Spore cleavage is accomplished by vesicular coalescence resulting in septa that coincide with the prophasic wall ingrowths.