Cytological Mechanism of Cytoplasmic Inheritance in Pinus tabulaeformis: II. Transmission of Male and Female Organelles During Fertilization and Proembryo Development

In an earlier report the ultrastructure and nucleoid organelles of male gamete in Pinus tabulaeformis Carr. have been described. Presently, the ultrastructure of the cytoplasm of the egg cell and pollen tube—immediately before fertilization and during cytoplasmic transmission of male gametophyte—has been described for the same species. The fate of parental plastids and mitochondria in the proembryo has also been followed. The mature egg cell contains a large amount of mitochondria, but seems to lack normal plastids. Most plastids have transformed into large inclusions. Apart from the large inclusions, there are abundant small inclusions and other organelles in the egg cell. During fertilization, pollen tube penetrates into the egg cell at the micropylar end and thereafter the contents are released. Plastid and mitochondrion of male origin are lacking near the fusing sperm-egg nuclei. The second sperm nucleus—not involved in karyogamy—remains at a site near the receptive vacuole. This nucleus is surrounded by large amount of male cytoplasm containing mixed organelles from the sperm cell, tube cell, and egg cell. At the free nuclear proembryo stage, organelles of male and female origin are visible in the perinucleus-cytoplasmic zone. Most of the mitochondria have the same morphological features as those in the egg cell. Some of the mitochondria appear to have originated from the sperm and tube cells. Plastids are most likely of male gametophyte origin because they have similar appearance as those of the sperm and tube cell. Large inclusions in the egg cell become vacuole-like. Paternal plastids have been incorporated into the neocytoplasm of the proembryo. In the cellular proembryo, maternal mitochondria are more abundant. Plastids resembling those of the sperm and tube cell are still present. These cytological results clearly show that in P. tabulaeformis , plastids are inherited paternally and mitochondria bipaternally. The cytological mechanism of plastid and mitochondrion inheritance in gymnosperm is discussed.     

The Cytological Mechanism of Biparental Cytoplasmic Inheritance in Pelargonium hortorum -Ultrastructural and DNA Fluorescence Studies of Male and Female

Electron microscopic investigation has demonstrated that plastids and mitochondria are conserved in the generative cell, sperm cells and egg cell of Pelargonium hortorum Bailey. The plastids in the generative cell which contain starch for a short period, gradually changed to proplastids during the maturation of generative cell. The plastids in the sperm cells are large and numerous the characteristics of dense matrix and an abundant endomembrane systems. These plastids always appear ringlike in cross section. In the generative cell and sperms, the spherical or rod-shaped mitochondria are smaller than the.plastids and remain unchanged during the development process from generative cell to sperm cells. DNA filaments are visualized in the transparent central zone of the mitochondria. In the egg cell, plastids are more abundant than mitochondria. The structures of the plastids and mitochondria are obviously different from those in the sperm cell. Most of the plastids are irregularly rod-shaped and contain starch, the mitochondria are about 3 times larger than those in the sperm cells. Most of them are cup-shaped as proved by successive sections. DNA epifluorescence study demonstrated that DNA nucleoids are present in both plastids and mitochondria of the egg, generative cell and sperm cells. In the sperm cells, there is no ringlike DNA nucleoid as is existed in the egg cell. This study has defined the characteristics of the plastids and mitochondria in both male and female gemates of P. hortorurn. The results are essential contributions for further investigation of the biparental organelle transmission in the zygote and proembryo.     

Cytological Studies of the Cytoplasmic Inheritance in Lilium regale and L. davidii

The distribution and characteristics of plastids and mitochondria in the generative and sperm cells of Lilium regale Wils. and L. davidii Duch. were described. In L. regale there were few plastids and abundant mitochondria in the newly formed generative cell. When the generative cell became free in the vegetative cytoplasm, the plastids degenerated completely within the generative cell. It was further proved by DAPI fluorescent technique that there was no organell DNA in the generative cell within the mature pollen grain or the pollen tube. However, distribution of the plastids was strictly polarizable during the division of the micmspore in L. davidii, resulting the lack of plastids in the newly formed generative cell. Data of RFLP analysis comparable between L. davidii, L. longifiorum and their interspecific hybrid have also proved the plastid inheritance in L. davidii to be of uniparental maternal transmission. Although the mitoehondria were observed both in the generative and sperm cells of L. regale and L. davidii but their DNA was decomposed in the male gametophyte stage. Therefore the mitochondda in the sperm cell could not be transmitted into the offspring. The results provided the detail, cytological evidence that organelles in the microgametophyte are incapable of genetic transmission in the two species of Lilium.     

Ultrastructural and Cytochemical Study on Mature Pollen of Pinus tabulaeformis

The pollen of Pinus tabulaeformis Cart. comprised two prothallial cells, a generative cell and a tube cell which degenerated at pollen maturation. The generative cell had its own cell wall, seperating from the intine of pollen, but with its side wall attached to the infine. Cytoplasmic channels were present on the side of the generative cell wall, which faced to the tube cell cytoplasm. The generative cell differed conspicuously from the tube cell. The main differences include: ( 1 ) The chromatin in the generative cell nucleus was condensed, but was dispersed and had numerous nueleare pores in the tube cell nucleus; (2)There was no microbody in the generative cell but many microbodies were present in the tube cell cytoplasm; (3)More inclusions were present in the tube cell than in the generative cell. Both the generative cell and the tube cells contained lipid bodies and amyloplasts in the cytoplasm, but there were more amyloplasts in the former. The tube cell also contained a few proteins which was absent in the generative cell. In addition, there were numerous mitochondria, polyribosomes, and a few endoplasmic reticulums and dictyosomes in the generative and tube cells. DAPI staining demonstrated numerous cytoplasmic DNA in both generative cell and tube cell. The mode of cytoplasmic inheritance, and the composition, structure and the nature of the pollen wall of P. tabulaefonnis are also discussed in this paper.     

Cytological Basis of Plastid Inheritance in Phaseolus vulgaris-Investigations of Plastids,Mitochondria and Their DNA Nucleoids During Pollen Development

Electron microscopic and DNA fluorescence microscopic observations of the plastids, mitochondria and their DNA in the developing pollen of Phaseolus vulgaris L. have demonstrated that the male plastids were excluded during microspore mitosis. The formed generative cell was free of plastids because of regional localization of plastids in early developing microspore and the extremely unequal distribution during division. The fluorescence observations of DNA showed that cytoplasmic (plastid and mitochondria) nucleoids degenerated and disappeared during the development of microspore/pollen, and were never presented in the generative cell at different development stages. These results provided precise cytological evidence of maternal plastid inheritance in Phaseolus vulgaris, which was not in accord with the biparental plastid inheritance identified from early genetic analysis. Based on authors' previous observations in a variety of common bean that the organelle DNA of male gamete was completely degenerated, the early genetic finding of the biparental plastid inheritance was unlikely to be effected by genotypic difference. Thus those biparental plastid inheritance might be caused by occational male plastid transmission, and plastid uniparental maternal inheritance was the species character of Phaseolus vulgaris.     

Microsporogenesis and the Mechanism of Cytoplasmic Male Sterility in Maize

Examination of the structural changes occurring during microsporogenesisin cms-T, -C and -S Zea mays L. with different nuclear backgroundsrevealed considerable variations in the stages at which abortionof the sporocytes occurred within each cytoplasmic genotype.This variation was not associated with the nuclear background.Significantly, we found that the mitochondria in cms-T plantsdid not always follow the pattern of degeneration describedpreviously. We conclude that these observations are not consistentwith a proposed mechanism of cytoplasmic male sterility involvingthe direct, antagonistic action of an anther produced substanceon an altered mitochondrion or plastid, that is functionallynormal in the rest of the plant. The value of microscopic studiesin detecting variation in apparently uniform material is emphasised,though it is recognized that other methods will be requiredto elucidate the mechanism of male sterility. male sterility, mitochondria, cytoplasmic inhertance, microsporogenesis, Zeamays L., maize, corn     

葫芦藓精子发生过程中胞间连丝与胞质桥的超微结构

从超微结构水平上对葫芦藓(Funaria hygrometrica Hedw.)精子发生过程中胞间连接系统的结构及其变化动态进行了研究.结果表明,同一区中的相邻生精细胞由大量胞质桥相连,而不同区的细胞之间则不存在胞质桥.胞间连丝存在于套细胞之间以及套细胞与生精细胞之间,但它在生精细胞间不存在.在精子器发生的后期,当精子细胞壁开始降解时,同一个精子器中所有的精子细胞似乎都由扩大的胞质桥相互连接.胞质桥一直保持到精子分化的后期,最终精子细胞同步分化成精子.胞间连丝与胞质桥具有不同的内部结、分布以及生物发生机制,这表明它们在精子器的发育过程中可能扮演着不同的角色.     

葫芦藓精子发生过程中胞间连丝与胞质桥的超微结构

从超微结构水平上对葫芦藓(Funaria hygrometrica Hedw.)精子发生过程中胞间连接系统的结构及其变化动态进行了研究。结果表明,同一区中的相邻生精细胞由大量胞质桥相连,而不同区的细胞之间则不存在胞质桥。胞间连丝存在于套细胞之间以及套细胞与生精细胞之间, 但它在生精细胞间不存在。在精子器发生的后期,当精子细胞壁开始降解时,同一个精子器中所有的精子细胞似乎都由扩大的胞质桥相互连接。胞质桥一直保持到精子分化的后期,最终精子细胞同步分化成精子。胞间连丝与胞质桥具有不同的内部结、分布以及生物发生机制,这表明它们在精子器的发育过程中可能扮演着不同的角色。     

不同坡向人工油松林生长状况与林下物种多样性分析

采用野外调查的方法,研究了绥德地区28年生不同坡向人工油松林林木个体生长、植物种类、相似系数和林下物种多样性特征,以明确坡向对人工油松林生长和群落结构的影响.结果显示,(1)阴坡油松的胸径、树高、冠高和冠幅均显著大于阳坡;(2)阴坡与阳坡林下分别有植物21和19种,共有种11种.群落相似系数为55%,说明坡向对人工油松林林下物种组成影响较大.(3)阴坡物种丰富度指数、均匀度指数和多样性指数均表现为草本层＞灌木层;而阳坡的Shannon-Wiener指数和Pielou指数(Jsw)则与阴坡相反,为灌木层＞草本层;群落总体多样性表现为丰富度指数和多样性指数均是阴坡稍大于阳坡,而均匀度指数表现为阳坡稍大于阴坡.研究表明,绥德地区人工油松林的生长状况阴坡显著优于阳坡,但与陕北南部地区人工油松林相比,该地区人工油松林林木生长各项指标还是较低,群落结构配置不合理.     

Genetics of Male Sterility in Gynodioecious Plantago Coronopus. I. Cytoplasmic Variation

Inheritance of male sterility was studied in the gynodioecious species Plantago coronopus using five plants and their descendants from an area of ~50 m(2) in each of six locations. The crosses were planned to test for cytoplasmic inheritance of male sterility. In four locations significant differences between reciprocal crosses were observed. The progenies of these reciprocal crosses were used in a crossing scheme designed to test whether these reciprocal differences were caused by different cytoplasmic types between the plants. In all four locations, the existence of at least two cytoplasmic types could be shown. Moreover, the results of the crosses between locations showed that the same two cytoplasmic types were present in all four locations. We therefore argue that there is only limited cytoplasmic variation in P. coronopus. In each cytoplasmic type a series of intermediate sex forms occurred. A marked difference in restoration level existed between the two cytoplasmic types. Plants with cytoplasmic type 2 hardly segregated male steriles, in contrast to plants with cytoplasmic type 1.     

普通白菜未减数雄配子形成的细胞学机制

以自然产生2n配子的二倍体普通白菜为材料,采用常规压片法研究2n雄配子发生的细胞学机制.结果表明,白菜未减数雄配子的形成途径有两条:(1)通过近平行型纺锤体形成二分体,从而产生2个未减数配子;(2)通过三极型纺锤体形成三分体,从而产生2个正常减数n配子和1个未减数2n配子,三极型纺锤体比平行型纺锤体能产生更高频率的未减数配子.2n花粉大小为n花粉的1.5倍,其频率不超过9%.     

Peculiarities of Inheritance of Pollen Fertility Restoration Trait in Sunflower with Cytoplasmic Male Sterility

Russian Journal of Genetics - The progenies from crosses of a sterile VIR116A line with PET1 CMS with pollen fertility restorer RIL130, VIR210, and VIR740 lines, differing in the type of cytoplasm...     

Cytological Characterization of Self Incompatibility in Gametes of the Ascidian, Ciona intestinalis

We have determined how many elements are involved in the regulation of self-fertilization in the solitary ascidian, Ciona intestinalis that is an incompletely self-sterile species. Animals collected in the field were repeatedly induced to spawn in order to examine their selfing ratios. About 20% of them were self-fertile, although the ratios fluctuated considerably among respective spawnings. Naturally or acid-induced self-fertile gametes required much longer time for selfing than that for crossing. Egg-suspending seawater (egg water) as such activated sperm motility, but it lowered conspicuously the self-fertilization ratio. Self-sterile spermatozoa could scarcely bind to the vitelline coat (VC) of glycerinated autologous eggs. or in case they bound well to it the sperm flagella ceased to beat within five min of the 'insemination'. The staining of sectioned gametes with DAPI, a fluorescent dye of DNA, showed that in selfing the spermatozoa could hardly penetrate the VC even though they bound well to it. The results of this study show that the block of self-fertilization can be classified into four elements from a phenomenal viewpoint, such as egg water, low affinity of sperm-VC binding, inactivation of bound sperm and difficulty in sperm penetration through the VC.     

黄龙山油松林和油松+辽东栎混交林物种组成及优势种群动态

根据黄龙山油松(Pinus tabulaeformis)林和油松+辽东栎(Quercus liaotungensis)混交林共6个群丛中27个典型样地调查资料,分析了松栎林群落物种组成、多样性指数和优势种群动态.结果表明:(1)松栎混交林中灌木层和草本层植物种数均大于油松林,且灌木和草本种类因群落生境不同而有差异.(2)物种多样性分析显示,油松林和松栎混交林中由于生境不同导致灌木层物种多样性差异最大.(3)优势种群分析表明,油松林中阴坡幼苗数量较多,种群最大的为Ⅳ和Ⅴ龄级个体;松栎混交林中油松和辽东栎幼苗充足,到Ⅱ、Ⅲ龄级二者均大量减少,此后油松个体增加而辽东栎减少,说明松栎林发育过程中各层次植物物种相互制约,优势种群之间也存在竞争,且受生境条件影响较大.(4)主成分分析表明,气温、土壤、光照和总郁闭度是主要影响因子.因此,在未来森林抚育中应适当减少优势种群密度,促进异龄化;采取有效措施合理搭配灌木和草本,强化对森林发育有利的因素,提高林分质量.     

Isolation of Viable Male and Female Gametes in Cunninghamia lanceolata

Viable male and female gametes were isolated from pollinated ovules of Cunninghamia lanceolata (Lamb.) Hook. Prior to their penetration into the female gametophyte, the pollen tubes were drawn out from the nucetlus. The isolated pollen tubes were branched and one of them became swollen. An enlarged spermatogenous cell and subsquently a pair of sperm cells were formed as the pollen tube reached the regions over and against the archegonia. The sperm cells were released from the pollen tubes manually with the use of a stereomicroscope. The positive FDA reaction gave evidence of the sperm cells viability, and the Fluorescent Brightener 28 positive demonstrated the presence of cell wall. The egg cells were enzymatically isolated from the female gametophytes. The isolated egg cells were spherical, contained 1 to 2 large and many small vacuoles. FDA test showed the egg cells were viable, and the viability sustained for 8 days in 2 to 4 ℃ without any protectants. Fusion between single pair of male and female gametic protoplasts was attempted with PEG method, but only adhesion of the two was obtained.     

Ultrastructural Aspects and Possible Origin of Cytoplasmic Channels Providing Intercellular Connection in Vegetative Tissues of Anthers1

Cytoplasmic channel represents a huge intercellular connection other than plasmodesma. They are proposed to play an essential role in controlling cell-to-cell trafficking of macromolecules. The present study ultrastructurally examined the occurrence, structure, and formation of this intercellular path in somatic tissues of wheat, tobacco, and onion anthers as well as their differences from those present in anther generative tissue. It was shown that cytoplasmic channels existed not only in the pollen mother cells, but also in both epidermis and vascular parenchyma of the anthers. In somatic tissues, they appeared as plasmallema-lined tubes 400–750 nm wide filled with nuclear or cytoplasmic material, the latter including cytoplasmic matrix, ribosomes, and filamentous structures. Their biogenesis seems to result from the thinning of the local portions of cell wall containing multiple plasmodesmata and the fusion of plasmodesmata in such regions induced by the wall-digesting enzymes released by nearby located vesicles. In contrast to the channels existing in the pollen mother cells of tobacco, the cytoplasmic channels in the epidermal or vascular parenchyma cells of wheat, onion, and tobacco anthers usually do not appear in groups, but are single. Perhaps this is the way for nuclear material to migrate from cell to cell via a single channel and then form a single chromatin spherical body in the adjoining cell. An individual cell could not accept the nuclear material from another cell while extruding its own to the third cell. Cell-to-cell migration of organelles through the cytoplasmic channels was not shown in the somatic tissues.     

Ultrastructural Localization of Calcium in the Acrosome and Jelly Coat of Starfish Gametes

The potassium pyroantimonate technique was employed to localize calcium ultrastructurally on both male and female starfish gamete regions that first interact at fertilization. In the spermatozoon of Marthasterias glacialis , antimonate precipitates in the peripheral dense component of the acrosomal vesicle, while in the oocyte it precipitates in the jelly coat and beneath the oolemma. Calcium was identified in the precipitates by testing the chelator-sensitivity and by X-ray microanalysis of the precipitates.     

油松雌球果的发生和发育研究

运用薄切片技术对油松（Ｐｉｎｕｓ ｌａｂｕｌａｅｆｏｒｍｉｓ Ｃａｒｒ．）雌球果的发生和发育进行了研究。结果表明：８月初,雌球果原基发生,共外部形态发生明显变化,但内部细胞组织学分区结构与营养茎端结构相似;１０月中旬,雌球果原基的鳞片叶腋处产生最裨的苞片原基,以后苞片原基由基部向顶端连续发生。此时球果原基的顶端结构姨生变化,顶端分生组织区、中央母细胞区和周围分生组织区衍化为套层,肋状分生组织衍化为