被子植物受精作用的分子和细胞生物学机制

被子植物双受精包括精-卵、精子-中央细胞两个融合过程。由于双受精深藏于母体组织中进行, 长期以来一直是植物有性生殖研究中的难点。近年来, 随着各种植物配子体cDNA文库的构建, 各种离体研究系统的建立和突变体分析的兴起, 极大地推动了被子植物受精作用研究的快速发展, 增进了人们对被子植物受精过程的分子和细胞生物学机制的深入了解。本文着重讨论受精作用的若干重要发育事件, 包括受精前卵器细胞对花粉管向胚珠定向生长的近距离引导信号, 精子的靶向运动,精、卵细胞相互作用和配子融合后卵细胞的激活与中央细胞发育的启动等。     

Calcium function and distribution during fertilization in angiosperms

Calcium has an essential signaling, physiological, and regulatory role during sexual reproduction in flowering plants; elevation of calcium amounts is an accurate predictor of plant fertility. Calcium is present in three forms: (1) covalently bound calcium, (2) loosely bound calcium typically associated with fixed and mobile anions (ionic bonding); and (3) cytosolic free calcium-an important secondary messenger in cell signaling. Pollen often requires calcium for germination. Pollen tube elongation typically relies on external calcium stores in the pistil. Calcium establishes polarity of the pollen tube and forms a basis for pulsatory growth. Applying calcium on the tip may alter the axis; thus calcium may have a role in determining the directionality of tube elongation. In the ovary and ovule, an abundance of calcium signals receptivity, provides essential mineral nutrition, and guides the pollen tube in some plants. Calcium patterns in the embryo sac also correspond to synergid receptivity, reflecting programmed cell death in one synergid cell that triggers degeneration and prepares this cell to receive the pollen tube. Male gametes are released in the synergid, and fusion of the gametes requires calcium, according to in vitro fertilization studies. Fusion of plant gametes in vitro triggers calcium oscillations evident in both the zygote and primary endosperm during double fertilization that are similar to those in animals.     

In vitro fertilization as a tool for investigating sexual reproduction of angiosperms

In vitro fertilization (IVF) of isolated male and female gametes of flowering plants was first accomplished in the last decade. Successful isolation of male and female gametes, and culturing of in vitro zygotes to form new plants, is a prelude to the use of IVF for research into the cellular and molecular control of fertilization in higher plants and its application as a tool in biotechnology. Genes unique to male and female gametes and zygotes of higher plants, although currently incompletely characterized, are expected to permit direct molecular dissection of fertilization. By applying IVF and microculture to zygotes and endosperm obtained by both in vivo and in vitro methods, newly activated fusion products may be observed and manipulated in media where they are directly accessible to the techniques of molecular cell biology. IVF and zygote culture may also offer potential for creating new hybrid plants by fusing isolated gametes from different species to produce unique zygotes and ultimately plants that would be impossible to obtain using typical crossing techniques. Transformation and regeneration frequencies using IVF may also be high enough to avoid the necessity of adding controversial antibiotic and herbicide resistant genes to screen transformed products. This review describes advances using IVF in plant sexual reproduction and discusses its potential in the genetic improvement of flowering plants.     

Synergid degeneration in Nicotiana: a quantitative,fluorochromatic and chlorotetracycline study

Summary Synergid degeneration was examined in the isolated embryo sac and egg apparatus of Nicotiana tabacum using quantitative cytology, fluorochromatic reaction (FCR) and chlorotetracycline (CTC). Most synergid degeneration occurs after pollen tubes (PT) arrive in the ovarian chamber between 42 and 48 h after pollination; synergid degeneration was precluded when PT were prevented from entering the ovary by stylar excision indicating that the signal that triggers synergid degeneration travels only relatively short distances in this plant. There was no evidence for any preferentiality between right or left synergids with regard to cell size or degeneration pattern. FCR staining confirms that synergid degeneration involves the loss of membrane integrity and is a reliable indicator of the onset of degeneration. CTC labeling of the degenerated synergid reveals that a concentrated reserve of membrane-bound calcium is present in the receptive synergid, possibly aiding in the attraction, arrest and discharge of the PT, releasing the sperms into the receptive ES.     

Presence of a partially-coated reticulum in angiosperms

Summary The form and distribution of partially-coated membrane systems and coated vesicles in the leaf glands ofPhaseolus and the root cortical cells ofZea were investigated. Partially-coated membranes exist as a reticulum which is either sparsely branched or extensively anastomosed. Coated vesicles and coated membrane regions may occur at all points that are in the vicinity of such a reticulum. Golgi stack membranes, though associated with the partially-coated reticulum in many cases, show no consistent orientation to it. Occasionally, the reticulum and the associated coated vesicles are located away from any Golgi stack. We examined other plant tissues from such species asAllium cepa, Beta vulgaris andNicotiana tabacum and the partially-coated reticulum was observed in all material. We suggest that membrane flow may be occurring from the partially-coated reticulum to either the Golgi stack or to other intracellular compartments.     

Genome size and recombination in angiosperms: a second look

Despite dramatic differences in genome size--and thus space for recombination to occur--previous workers found no correlation between recombination rate and genome size in flowering plants. Here I re-investigate these claims using phylogenetic comparative methods to test a large data set of recombination data in angiosperms. I show that genome size is significantly correlated with recombination rate across a wide sampling of species and that change in genome size explains a meaningful proportion ( approximately 20%) of variation in recombination rate. I show that the strength of this correlation is comparable with that of several characters previously linked to evolutionary change in recombination rate, but argue that consideration of processes of genome size change likely make the observed correlation a conservative estimate. And finally, although I find that recombination rate increases less than proportionally to change in genome size, several mechanistic and theoretical arguments suggest that this result is not unexpected.     

Seed fertilization, development, and germination in Hydatellaceae (Nymphaeales): Implications for endosperm evolution in early angiosperms

New data on endosperm development in the early-divergent angiosperm Trithuria (Hydatellaceae) indicate that double fertilization results in formation of cellularized micropylar and unicellular chalazal domains with contrasting ontogenetic trajectories, as in waterlilies. The micropylar domain ultimately forms the cellular endosperm in the dispersed seed. The chalazal domain forms a single-celled haustorium with a large nucleus; this haustorium ultimately degenerates to form a space in the dispersed seed, similar to the chalazal endosperm haustorium of waterlilies. The endosperm condition in Trithuria and waterlilies resembles the helobial condition that characterizes some monocots, but contrasts with Amborella and Illicium, in which most of the mature endosperm is formed from the chalazal domain. The precise location of the primary endosperm nucleus governs the relative sizes of the chalazal and micropylar domains, but not their subsequent developmental trajectories. The unusual tissue layer surrounding the bilobed cotyledonary sheath in seedlings of some species of Trithuria is a belt of persistent endosperm, comparable with that of some other early-divergent angiosperms with a well-developed perisperm, such as Saururaceae and Piperaceae. The endosperm of Trithuria is limited in size and storage capacity but relatively persistent.     

Isolated generative cells in some angiosperms: a further study

Summary Generative cells were isolated from the pollen grains of three angiosperm species by a method similar to that previously reported for Haemanthus katherinae (Baker). Both the external appearance and the internal structure of the isolated generative cells were observed by light and scanning electron microscopy. The dynamic changes occurring in the cells after they had been liberated from the pollen grains were recorded by video-enhanced microscopy. The distribution of microtubules in the isolated cells was revealed by immunofluorescence.This work was done during the senior author's sabbatical leave at the Department of Biological Sciences, Dartmouth College, Hanover, N.H. (USA) and this paper is dedicated to Professor R.D. Allen, who did not live to see this work completed but who was directly involved in supporting this project.     

Incompatibility in angiosperms

 Since Darwinian times considerable knowledge has accumulated on the distribution, physiology and genetics of self-incompatibility (SI) in higher plants. In the second half of this century the first attempts were made to identify the biochemical bases of SI. These included thediscovery that cutinase enables pollen tube penetration at the surface* of the stigma in Cruciferae, sorting of segregation pollen S-phenotypes by serological techniques, a lock-and-key model of the SI reaction, the first detection and characterisation of SI proteins and the discovery of the role of the tapetum in the determination of pollen phenotypes in homomorphic sporophytic SI. This pioneering work was followed by a worldwide effort to identify and understand the cellular and molecular processes which lead to the recognition and rejection of SI pollen. The present review article summarizes briefly the current state of knowledge in areas essential for the understanding and exploitation of SI and outlines new information that has become available during recent years. Received: 14 March 1997 / Revision accepted: 10 June 1997     

Dichogamy in angiosperms

We obtained information on dichogamy and other aspects of the biology of over 4200 species of angiosperms from several hundred published and unpublished sources. We used this information to describe patterns of occurrence of dichogamy and to test specific hypotheses relating dichogamy to other characteristics of plants or their environments. Protandry was more common than protogyny at the intrafloral level, but the reverse was true at the interfloral level. Patterns of dichogamy varied significantly among major taxa, with protogyny more common among monocotyledons and primitive dicotyledons, and protandry expecially common in the Asteridae. Arctic species tended to be less dichogamous and more protogynous than temperate and tropical species. Aquatic and alpine species were especially protogynous. Patterns of dichogamy varied among sexual systems, with gynomonoecious and gynodioecious species especially protandrous, and monoecious species highly protogynous. Autogamous and self-compatible species were disproportionately protogynous. Flowers of intraflorally dichogamous species were slightly larger than those of adichogamous species, owing to the presence of many autogamous species in the latter group. Species with interfloral protogyny bore much smaller flowers than did species with interfloral protandry. Early-blooming species in north-temperate and polar regions were disproportionately protogynous. Sexual structures that abscised, shriveled or moved after completion of their function tended to be presented first, and those that facilitated the other sexual function were presented second. A negative association existed between type of intrafloral and interfloral dichogamy in diclinous species. Most animal-pollinated flowers were protandrous, except beetle-pollinated and refuge and trap blossoms. Wind pollination was markedly associated with protogyny. Vertical inflorescences visited by upwardly-moving vectors were protandrous.     

被子植物助细胞的发育和功能

助细胞是被子植物雌配子体的组成细胞之一,是大多数被子植物完成受精作用的关键环节之一。在受精过程中,助细胞吸引花粉管向雌配子体生长,并接受花粉管长入细胞程序死亡助细胞中。接下来的花粉管停止生长和花粉管顶端破裂释放出2个精细胞的过程可能也依赖于助细胞。另外,雄性生殖单位的解体、提供精细胞的运动轨道和启动精细胞合成DNA的生物学事件也可能与助细胞有关。助细胞的形态结构已研究得比较清楚,但有关助细胞的发育机理和它在受精过程中的作用则仍不明确。近年来对助细胞的研究又取得了一些新结果,尤其是一些分子生物学的研究结果为揭示助细胞的功能提供了重要线索。该文总结了这方面的研究成果,并对助细胞的生殖功能进行了分析。     

The age of the angiosperms: a molecular timescale without a clock

The age of the angiosperms has long been of interest to botanists and evolutionary biologists. Many early efforts to date the age of the angiosperms and evolutionary divergences within the angiosperm clade using a molecular clock have yielded age estimates that are grossly inconsistent with the fossil record. We investigated the age of angiosperms using Bayesian relaxed clock (BRC) and penalized likelihood (PL) approaches. Both of these methods allow the incorporation of multiple fossil constraints into the optimization procedure. The BRC method allows a range of values for among-lineage rate of substitution, from a nearly clocklike behavior to a condition in which each branch is allowed an optimal substitution rate, and also accounts for variation in molecular evolution across multiple genes. A topology derived from an analysis of genes from all three plant genomes for 71 taxa was used as a backbone. The effects on age estimates of different genes, single-gene versus concatenated datasets, and the inclusion and assumptions of fossils as age constraints were examined. In addition, the influence of prior distributions on estimates of divergence times was also explored. These results indicate that widely divergent age estimates can result from the different methods (198-139 million years ago), different sources of data (275-122 million years ago), and the inclusion of temporal constraints to topologies. Most dates, however, are between 180-140 million years ago, suggesting a Middle Jurassic-Early Cretaceous origin of flowering plants, predating the oldest unequivocal fossil angiosperms by about 45-5 million years. Nonetheless, these dates are consistent with other recent studies that have used methods that relax the assumption of a strict molecular clock and also agree with the hypothesis that the angiosperms may be somewhat older than the fossil record indicates.     

被子植物助细胞的发育和功能

助细胞是被子植物雌配子体的组成细胞之一, 是大多数被子植物完成受精作用的关键环节之一。在受精过程中, 助细胞吸引花粉管向雌配子体生长, 并接受花粉管长入细胞程序死亡助细胞中。接下来的花粉管停止生长和花粉管顶端破裂释放出2个精细胞的过程可能也依赖于助细胞。另外, 雄性生殖单位的解体、提供精细胞的运动轨道和启动精细胞合成DNA的生物学事件也可能与助细胞有关。助细胞的形态结构已研究得比较清楚, 但有关助细胞的发育机理和它在受精过程中的作用则仍不明确。近年来对助细胞的研究又取得了一些新结果, 尤其是一些分子生物学的研究结果为揭示助细胞的功能提供了重要线索。该文总结了这方面的研究成果, 并对助细胞的生殖功能进行了分析。