Attractive and repulsive interactions between female and male gametophytes in Arabidopsis pollen tube guidance

Sexual reproduction in plants, unlike that of animals, requires the action of multicellular haploid gametophytes. The male gametophyte (pollen tube) is guided to a female gametophyte through diploid sporophytic cells in the pistil. While interactions between the pollen tube and diploid cells have been described, little is known about the intercellular recognition systems between the pollen tube and the female gametophyte. In particular, the mechanisms that enable only one pollen tube to interact with each female gametophyte, thereby preventing polysperm, are not understood. We isolated female gametophyte mutants named magatama (maa) from Arabidopsis thaliana by screening for siliques containing half the normal number of mature seeds. In maa1 and maa3 mutants, in which the development of the female gametophyte was delayed, pollen tube guidance was affected. Pollen tubes were directed to mutant female gametophytes, but they lost their way just before entering the micropyle and elongated in random directions. Moreover, the mutant female gametophytes attracted two pollen tubes at a high frequency. To explain the interaction between gametophytes, we propose a monogamy model in which a female gametophyte emits two attractants and prevents polyspermy. This prevention process by the female gametophyte could increase a plant's inclusive fitness by facilitating the fertilization of sibling female gametophytes. In addition, repulsion between pollen tubes might help prevent polyspermy. The reproductive isolations observed in interspecific crosses in Brassicaceae are also consistent with the monogamy model.     

中国科学家在植物受精过程中雌雄配子体信号识别机制研究中取得突破性进展

阐明植物雄配子体与雌配子体互作的分子机理一直是植物有性生殖研究的前沿和热点。但限于研究难度较大, 很多重要科学问题仍有待回答。关于花粉管如何感知雌配子体信号从而定向生长进入胚囊以投送精细胞就是悬疑多年的问题之一。最近, 中国科学家在解析雄配子体感知雌配子体引导信号的分子机制方面取得了突破性进展。     

Distinct expression of members of the LHT amino acid transporter family in flowers indicates specific roles in plant reproduction

Sexual plant reproduction necessitates proper development of pollen, pollen germination and tube growth through various tissues of the pistil, the female organ of the flower. Finally, sperm cells are released to fertilize the female gametophyte. These processes require high metabolic activities of all tissues involved and rely on the delivery of nitrogen assimilates for success. However, transporters mediating nitrogen fluxes are mostly unknown. The presented work provides an expression analysis of members of the LHT amino acid transporter family in relation to pollen development and pollen–pistil interaction. Expression of Arabidopsis LHTs was analyzed during flower development and the location of LHT function resolved by transporter-GFP and promoter-GUS studies. GFP-LHT localization in onion cells indicates that all LHTs analyzed are targeted to the plasma membrane. We further showed that LHTs are expressed in anthers and male gametophytes where they are proposed to function in transport of amino acids for pollen development and maturation. Expression in germinating pollen, pollen tubes and transmitting tissue of the pistil points to a role of LHTs in support of the fertilization process. Overall, our study suggests that LHT function in flowers is cell or tissue specific, developmentally regulated and highly coordinated between male and female tissue.     

花粉蛋白质组学研究进展

花粉是高度退化的生物体(雄配子体), 在植物有性生殖过程中具有重要作用。解析花粉发育、花粉-柱头识别、萌发和花粉管生长等细胞学过程的分子机制是当前研究的热点问题之一。近年来, 应用高通量的蛋白质组学技术平台, 对水稻、拟南芥和裸子植物花粉的蛋白质组学研究揭示了花粉中表达蛋白质的功能类群特征。花粉中参与细胞壁代谢、蛋白质代谢、细胞骨架动态和信号转导的蛋白质被高度代表, 并且近1/4蛋白质有多个同工型。本文综述了花粉蛋白质组学的研究进展。     

花粉蛋白质组学研究进展

花粉是高度退化的生物体（雄配子体）,在植物有性生殖过程中具有重要作用。解析花粉发育、花粉-柱头识别、萌发和花粉管生长等细胞学过程的分子机制是当前研究的热点问题之一。近年来,应用高通量的蛋白质组学技术平台,对水稻、拟南芥和裸子植物花粉的蛋白质组学研究揭示了花粉中表达蛋白质的功能类群特征。花粉中参与细胞壁代谢、蛋白质代谢、细胞骨架动态和信号转导的蛋白质被高度代表,并且近1／4蛋白质有多个同工型。本文综述了花粉蛋白质组学的研究进展。     

The Arabidopsis mutant feronia disrupts the female gametophytic control of pollen tube reception

Reproduction in angiosperms depends on communication processes of the male gametophyte (pollen) with the female floral organs (pistil, transmitting tissue) and the female gametophyte (embryo sac). Pollen-pistil interactions control pollen hydration, germination and growth through the stylar tissue. The female gametophyte is involved in guiding the growing pollen tube towards the micropyle and embryo sac. One of the two synergids flanking the egg cell starts to degenerate and becomes receptive for pollen tube entry. Pollen tube growth arrests and the tip of the pollen tube ruptures to release the sperm cells. Failures in the mutual interaction between the synergid and the pollen tube necessarily impair fertility. But the control of pollen tube reception is not understood. We isolated a semisterile, female gametophytic mutant from Arabidopsis thaliana, named feronia after the Etruscan goddess of fertility, which impairs this process. In the feronia mutant, embryo sac development and pollen tube guidance were unaffected in all ovules, although one half of the ovules bore mutant female gametophytes. However, when the pollen tube entered the receptive synergid of a feronia mutant female gametophyte, it continued to grow, failed to rupture and release the sperm cells, and invaded the embryo sac. Thus, the feronia mutation disrupts the interaction between the male and female gametophyte required to elicit these processes. Frequently, mutant embryo sacs received supernumerary pollen tubes. We analysed feronia with synergid-specific GUS marker lines, which demonstrated that the specification and differentiation of the synergids was normal. However, GUS expression in mutant gametophytes persisted after pollen tube entry, in contrast to wild-type embryo sacs where it rapidly decreased. Apparently, the failure in pollen tube reception results in the continued expression of synergid-specific genes, probably leading to an extended expression of a potential pollen tube attractant.     

被子植物有性生殖过程中的细胞程序死亡

细胞程序死亡是植物发育过程中的一种普遍现象。早就认识到高等植物生殖器官中一些细胞的死亡对植物有性生殖具有重要作用。这些细胞的死亡过程与动物组织中的细胞程序死亡基本相同。但植物体内诱导生殖细胞程序死亡的信号及其传导系统则显示出其特点 ,有些还表现出雌、雄性细胞的相互作用。探索植物生殖过程中的细胞程序死亡现象将有利于澄清植物生殖过程中的一些机理问题 ,使过去的细胞学研究结果深入到分子水平进行探讨     

Plotting a course: multiple signals guide pollen tubes to their targets

Pollen plays a critical role in the life cycle of all flowering plants, generating a polarized pollen tube that delivers sperm to the eggs in the interior of the flower. Pollen tubes perceive multiple extracellular signals during their extended growth through different floral environments; these environments discriminate among pollen grains, allowing only those that are appropriately recognized to invade. The phases of pollen tube growth include interactions that establish pollen polarity, entry of pollen tubes into female cell walls, and adhesion-based pollen tube motility through a carbohydrate-rich matrix. Recent studies have identified cells within the female germ unit as important sources of pollen guidance cues. Other signals undoubtedly exist, and their discovery will require genetic screens that target diploid tissues as well as haploid male and female cells.     

From pollination to fertilization in fruit trees

The phase that elapses from pollination to fertilization is re-examined giving special attention to pollen pistil interaction in compatible matings. Pollination induces an activation of the pistil. A number of changes take place in the different tissues of this organ that appear to support male gametophyte development and to assist fertilization. Thus pollination induces stigma secretion, the release of starch from the transmitting tissue and prolongs embryo sac viability. It appears that even those pollen grains that do not achieve fertilization have a synergistic role supporting others to do so.The pistil also has an effect on pollen tube growth. Pollen tube growth along the pistil is not continuous, accelerations and decelerations take place depending on the different tissues they traverse. The fact that pollen tube growth is heterotrophic, at the expenses of the pistil reserves, and that these reserves are not continuously produced confers the pistil with a role controlling pollen tube growth kinetics.     

Dynamics of pollen tube growth under different competition regimes

Pollen tube dynamics following different competition regimes were studied in sweet cherry (Prunus avium L.). In the process from pollination to fertilization, a constant reduction in the number of pollen tubes that travel along the style is observed. There could be two main causes of this reduction. One is a physical or physiological constraint consisting of the progressive decrease in the reserves and space available for pollen tube growth along the transmitting tissue of the style, and the other is genetic interaction both among the male gametophytes and between the male gametophytes and the female tissues of the flower. To evaluate the roles that these two forces play in reducing the number of pollen tubes that travel along the style, pistils were subjected to various pollen competition regimes by applying different mixtures of live and dead pollen onto the stigmata. The results obtained were similar when the experiment was repeated with different genotypes over 2 years, both in the laboratory and in the field. The role of stylar constriction is important, but it is not the only cause of pollen tube attrition because with low pollen loads fewer pollen tubes reach the different parts of the style than could fit therein. The fact that under different pollen competition regimes the number of pollen tubes is reduced by the same proportion in each stylar level indicates that genetic interactions play an important role in the control of pollen tube attrition.     

The Pollen Tube Pathway in Tasmannia insipida (Winteraceae): Homology of the Male Gametophyte Conduction Tissue in Angiosperms

Abstract: The pathway of the pollen tube in Tasmannia insipida (Winteraceae), an archaic angiosperm, is described. Visual observations and measurements of percentage sugar content from stigmatic exudate and the calyptra droplet were made. The droplet that forms on the calyptra of unopened flowers was artificially pollinated, but no pollen tubes appeared to grow through it to the gynoecium of female flowers. Calyptra droplets occur on both female and male (having reduced female structures) flowers of this dioecious plant. It is considered unlikely that a drop pollination mechanism, analogous to that found in some gymnosperms, is at work in T. insipida, and it is proposed that one possible droplet function is as a "reward" to potential pollinators in advance of flower opening. Pollen tubes were observed to grow almost exclusively along epidermal cells. Early thoughts on angiosperm transmission tissue are re-examined and built upon. It is proposed that the male gametophyte conduction tissue in angiosperms is homologous with epidermal tissue, and that pollen tube passage occurred originally solely along the surface of specialized epidermal cells, or possibly for short distances along unspecialized regions covered with exudate derived from these cells. Some earlier attempts to explain transmitting tissue homology and evolution are discussed.     

从柱头到胚囊——花粉管发育研究进展

花粉管的生长和发育是植物有性生殖过程中重要环节。本文首先介绍了花粉管的结构及其脉冲生长方式。然后介绍了花粉粒在柱头上萌发、花粉管在花柱道中生长及其进入胚囊的一些特点,同时还介绍了雌蕊在花粉管生长发育所起的重要作用。     

梅花个体内花柱长度的变异及其对繁殖成功的影响

植物个体内花型的变异影响繁殖成功, 雌性繁殖性状的变异可能影响雌性的繁殖成功, 也可能作为花粉的受体影响雄性的繁殖成功。然而, 植物个体内不同花柱长度的花产生的花粉是否影响植物的繁殖成功却少有研究。梅(Armeniaca mume)是原产我国的重要木本花卉和经济果树, 我们的野外观察发现在同一植株内, 不同花的花柱长度有变异, 存在长柱型、短柱型和雄花型(雌蕊败育)三种花型, 是比较雌性繁殖性状的变异对两性繁殖成功的影响的理想材料。本文主要测量了不同花型的花部特征, 统计花期, 并开展体外花粉萌发以及人工控制授粉实验。结果表明: 长柱型的花冠直径、雌蕊长、单花花粉数、花粉体积显著大于短柱型和雄花型。长柱型的单花期以及雌花期显著长于短柱型。长柱型、短柱型以及雄花型花粉在活体柱头上的萌发率没有显著性差异, 雄花型的花粉管长度显著高于长柱型和短柱型。长柱型为母本的花粉萌发率以及花粉管的长度要显著高于短柱型。长柱型、短柱型、雄花型花粉授粉与自然对照处理的坐果率没有显著性差异, 而长柱型为母本的坐果率要显著高于短柱型为母本的坐果率。这些结果表明野生梅花的长柱型为母本有利于花粉的萌发和花粉管的伸长, 有高的坐果率; 但其作为花粉供体的雄性功能与其他花型没有差异。     

Nuclear behavior, cell polarity, and cell specification in the female gametophyte

In flowering plants, the haploid gamete-forming generation comprises only a few cells and develops within the reproductive organs of the flower. The female gametophyte has become an attractive model system to study the genetic and molecular mechanisms involved in pattern formation and gamete specification. It originates from a single haploid spore through three free nuclear division cycles, giving rise to four different cell types. Research over recent years has allowed to catch a glimpse of the mechanisms that establish the distinct cell identities and suggests dynamic cell–cell communication to orchestrate not only development among the cells of the female gametophyte but also the interaction between male and female gametophytes. Additionally, cytological observations and mutant studies have highlighted the importance of nuclei migration- and positioning for patterning the female gametophyte. Here we review current knowledge on the mechanisms of cell specification in the female gametophyte, emphasizing the importance of positional cues for the establishment of distinct molecular profiles.     

Functions of neuromediators in plants

In plants, neurotransmitters play a major biological role in chemo-signaling, as regulating agents of growth and development, membrane permeability, etc. In the plant cells, there are also main elements of cholinergic and aminergic systems including the enzymes as well as functional analogues of cholino- and aminoreceptors. It is important that the systems should be taken into consideration in fertilizing and chemical interaction of the plant cells. Acetylcholine and cholinesterase were shown to be secreted in male sexual cells (the pollen). Depression of cholinesterase on the gene level correlates with male sterility. Preliminary treatment of a female gametophyte with antagonists of acetylcholine and histamine, prior to pollination, blocked the normal process of fertilizing. Cholinesterase was also found in secrets released from the pollen surface and pistil.     

Amborella trichopoda (Amborellaceae) and the evolutionary developmental origins of the angiosperm progamic phase

A remarkable number of the defining features of flowering plants are expressed during the life history stage between pollination and fertilization. Hand pollinations of Amborella trichopoda (Amborellaceae) in New Caledonia show that when the stigma is first receptive, the female gametophyte is near maturity. Pollen germinates within 2 h, and pollen tubes with callose walls and plugs grow entirely within secretions from stigma to stylar canal and ovarian cavity. Pollen tubes enter the micropyle within 14 h, and double fertilization occurs within 24 h. Hundreds of pollen tubes grow to the base of the stigma, but few enter the open stylar canal. New data from Amborella, combined with a review of fertilization biology of other early-divergent angiosperms, show that an evolutionary transition from slow reproduction to rapid reproduction occurred early in angiosperm history. I identify increased pollen tube growth rates within novel secretory carpel tissues as the primary mechanism for such a shift. The opportunity for prezygotic selection through interactions with the stigma is also an important innovation. Pollen tube wall construction and substantial modifications of the ovule and its associated structures greatly facilitated a new kind of reproductive biology.