钙在有花植物受精过程中的作用

钙作为第二信使在植物信号转导中的作用一直是植物生理学、细胞生物学和发育生物学研究的热点。近年已有不少综述和专著从不同角度对此作了详细评论［1～5］。虽然这些文章中只有部分内容涉及本文的主题———钙在植物受精中的作用,但是它们所论述的关于钙信使系统在植物...     

钙在被子植物受精过程中的作用

近年来,花粉管中的钙信号和生理功能的研究取得了明显的进展,同时在雌蕊系统中有关钙分布的研究也初步显示了其时、空特征与被子植物的受精作用密切相关。该文总结了花粉萌发和花粉管生长过程中外源钙和内源钙的作用机制,结合雌蕊组织中钙分布的特征,进一步探讨了钙在被子植物受精过程中的功能。     

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.     

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.     

Heat stress-induced limitations to reproductive success in Gossypium hirsutum

Using in vitro systems, numerous authors have cited the sensitivity of pollen tube growth to high temperature as a major cause of low yields for crops with valuable reproductive structures. We investigated the hypothesis that in vivo fertilization efficiency would be negatively affected by heat stress-induced changes in energy reserves and calcium-mediated oxidative status in the pistil. Gossypium hirsutum plants exposed to optimal (30/20°C) or high day temperature (38/20°C) conditions during flowering were analyzed for fertilization efficiency via UV microscopic observation of pollen tube-containing ovules and for soluble carbohydrates, adenosine triphosphate (ATP), calcium, antioxidant enzyme activity and NADPH oxidase (NOX; EC 1.6.3.1) activity in the pistil. Leaf measurements included gas exchange, chlorophyll content, quantum efficiency and ATP content of the subtending leaf on the day of anthesis. In the pistil fertilization efficiency, soluble carbohydrates, ATP content and NOX activity declined significantly, whereas water soluble calcium and glutathione reductase (EC 1.8.1.7) activity increased, and superoxide dismutase (EC 1.15.1.1) activity remained unchanged. In leaves, heat stress decreased photosynthesis, quantum efficiency and chlorophyll content, but increased stomatal conductance. We conclude that decreased source leaf activity either inhibits pollen development, tube growth through the style or guidance to the ovules as a result of an insufficient energy supply to the developing pistil. We further conclude that a calcium-augmented antioxidant response in heat-stressed pistils interferes with enzymatic superoxide production needed for normal pollen tube growth and fertilization of the ovule.     

The pollen receptor kinase LePRK2 mediates growth-promoting signals and positively regulates pollen germination and tube growth

In flowering plants, the process of pollen germination and tube growth is required for successful fertilization. A pollen receptor kinase from tomato (Solanum lycopersicum), LePRK2, has been implicated in signaling during pollen germination and tube growth as well as in mediating pollen (tube)-pistil communication. Here we show that reduced expression of LePRK2 affects four aspects of pollen germination and tube growth. First, the percentage of pollen that germinates is reduced, and the time window for competence to germinate is also shorter. Second, the pollen tube growth rate is reduced both in vitro and in the pistil. Third, tip-localized superoxide production by pollen tubes cannot be increased by exogenous calcium ions. Fourth, pollen tubes have defects in responses to style extract component (STIL), an extracellular growth-promoting signal from the pistil. Pollen tubes transiently overexpressing LePRK2-fluorescent protein fusions had slightly wider tips, whereas pollen tubes coexpressing LePRK2 and its cytoplasmic partner protein KPP (a Rop-GEF) had much wider tips. Together these results show that LePRK2 positively regulates pollen germination and tube growth and is involved in transducing responses to extracellular growth-promoting signals.     

INFLUENCE OF THE PISTIL ON POLLEN TUBE KINETICS IN PEACH (PRUNUS PERSICA)

Pollen tube growth has been studied in peach and has been related to changes in the pistil structures which the pollen tube has to traverse in its way from the stigma down to the ovule. Growth of the pollen tubes along the pistil is not continuous. While pollen tubes reach the base of the style 7 days after pollination, fertilization does not take place until 12 days later. Pollen tubes stop for 5 days at the top of the obturator and they further stop for 3 days before entering the ovule. The pollen tube growth is heterotrophic; starch, present all along the pistilar tract at anthesis, vanishes as the pollen tubes pass by. Discontinuous pollen tube growth appears to be controlled by the pistil. At anthesis the pistil is not fully matured. Maturation of the pistil implies a number of secretory processes that occur in a basipetal way starting from the stigma down to the style and ending in the ovule. Some of these secretions at the stigma and the style are triggered by pollination; others appear to be a maturative stage of the pistil and are produced in a discrete way. The fact that the pollen tube depends on these secretions together with the fact that these secretions are not continuously produced confer upon the pistil a role of controlling pollen tube kinetics and point out that, for a successful fertilization, male gametophyte development and pistil maturation need to by synchronized.     

High temperature limits in vivo pollen tube growth rates by altering diurnal carbohydrate balance in field-grown Gossypium hirsutum pistils

It has recently been reported that high temperature slows in vivo pollen tube growth rates in Gossypium hirsutum pistils under field conditions. Although numerous physical and biochemical pollen-pistil interactions are necessary for in vivo pollen tube growth to occur, studies investigating the influence of heat-induced changes in pistil biochemistry on in vivo pollen tube growth rates are lacking. We hypothesized that high temperature would alter diurnal pistil biochemistry and that pollen tube growth rates would be dependent upon the soluble carbohydrate content of the pistil during pollen tube growth. G. hirsutum seeds were sown on different dates to obtain flowers exposed to contrasting ambient temperatures but at the same developmental stage. Diurnal pistil measurements included carbohydrate balance, glutathione reductase (GR; EC 1.8.1.7), soluble protein, superoxide dismutase (SOD; EC 1.15.1.1), NADPH oxidase (NOX; EC 1.6.3.1), adenosine triphosphate (ATP), and water-soluble calcium. Soluble carbohydrate levels in cotton pistils were as much as 67.5% lower under high temperature conditions (34.6 °C maximum air temperature; August 4, 2009) than under cooler conditions (29.9 °C maximum air temperature; August 14, 2009). Regression analysis revealed that pollen tube growth rates were highly correlated with the soluble carbohydrate content of the pistil during pollen tube growth (r² = 0.932). Higher ambient temperature conditions on August 4 increased GR activity in the pistil only during periods not associated with in vivo pollen tube growth; pistil protein content declined earlier in the day under high temperatures; SOD and NOX were unaffected by either sample date or time of day; pistil ATP and water soluble calcium were unaffected by the warmer temperatures. We conclude that moderate heat stress significantly alters diurnal carbohydrate balance in the pistil and suggest that pollen tube growth rate through the style may be limited by soluble carbohydrate supply in the pistil.     

Characterization of the pollen growth transition in self-incompatible <Emphasis Type="Italic">Petunia inflata</Emphasis>

In Petunia inflata, as in other species that shed bicellular pollen, early pollen tube growth in the pistil is slow, then increases 2- to 5-fold depending on the genotype of the female parent. We refer to the time point at which pollen tubes enter the accelerated phase of growth as the pollen growth transition (PGT). Here, we present evidence that pre-PGT and post-PGT growth are quantitatively and qualitatively different, and that the PGT is triggered when pollen tubes reach the transition zone (TZ) below the stigma. The capacity of various pistil zones to precipitate the PGT was tested through 'stump' pollinations: varying lengths of the pistil apex were excised, the cut surface of the remaining pistil (the stump) coated with stigmatic exudates then dusted with compatible pollen. Pollen applied to TZ tissues entered the PGT earlier than pollen growing in intact control pistils; the PGT was delayed in stylar stumps, largely because of delayed germination and reduced pre-PGT growth. In immature pistils, the PGT was delayed by several hours relative to its onset in mature pistils. The PGT fails to occur in pollen cultured in vitro. Collectively, the data suggest that pollen tubes become competent to enter the PGT when they reach a critical size, but the physicochemical environment of the transmitting tissue is necessary for triggering the cellular changes that result in accelerated growth. An analysis of the distribution of pollen tube tips before and after the PGT suggests that pollen competition is most intense during the pre-PGT phase.     

Imaging elongating pollen tubes by green fluorescent protein

Pollen tube elongation is a dynamic process in which pollen tubes navigate and respond to female tissues to accomplish their mission of delivering the sperm cells for fertilization. The tube growth process itself is driven by regulated intracellular conditions that maintain the appropriate ionic environment, actin dynamics and a balance level of exocytosis and endocytosis to support growth at the tube apex. However, the interactive process within the pistil has not rendered itself accessible for direct observation. The contribution by individual cytosolic constituents of the pollen tube growth machinery remains to be determined. With the development of the green fluorescent protein reporter system, many of these questions can be addressed in live pollen tubes that elongate within the pistil and inchemically defined media. Analyses of the mechanisms that underlie pollen tube growth will be significantly facilitated. Received: 15 March 2001 / Accepted: 30 May 2001     

The Arabidopsis CrRLK1L protein kinases BUPS1 and BUPS2 are required for normal growth of pollen tubes in the pistil

In flowering plants, the interaction of pollen tubes with female tissues is important for the accomplishment of double fertilization. Little information is known about the mechanisms that underlie signalling between pollen tubes and female tissues. In this study, two Arabidopsis pollen tube‐expressed CrRLK1L protein kinases, Buddha's Paper Seal 1 (BUPS1) and BUPS2, were identified as being required for normal tip growth of pollen tubes in the pistil. They are expressed prolifically in pollen and pollen tubes and are localized on the plasma membrane of the pollen tube tip region. Mutations in BUPS1 drastically reduced seed set. Most of the bups1 mutant pollen tubes growing in the pistil exhibited a swollen pollen tube tip, leading to failure of fertilization. The bups2 pollen tubes had a slightly abnormal morphology but could still accomplish double fertilization. The bups1 bups2 double mutant exhibited a slightly enhanced phenotype compared to the single bups1 mutants. The BUPS1 proteins could form homomers and heteromers with BUPS2, whereas BUPS2 could only form heteromers with BUPS1. The BUPS proteins could interact with the Arabidopsis pollen‐expressed RopGEFs in the yeast two‐hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays. The results indicated that the BUPSs may mediate normal polar growth of pollen tubes in the pistil.     

Ethylene response to pollen tube growth in Nicotiana tabacum flowers

In flowers of Nicotiana tabacum L., pollination induces a transient increase in ethylene production by the pistil. The characteristic dynamics of the increase in ethylene correspond to the main steps of the pollen-tube journey into the pistil: penetration into the stigma, growth through the style, entry into the ovary and fertilization. Ethylene is synthesized de novo in the pistil, and its production is reduced in the dark. Ethylene production was monitored in tobacco flowers after pollination with incongruous pollen from three different Nicotiana species, N. rustica, N. repanda and N. trigonophylla, and with pollen from Petunia hybrida. Pollen from all of these different sources can germinate on the stigma surface but each pollen type shows a different behavior and efficiency in penetrating the pistil tissues. Thus, these different crosses provided a model with which to study the response of the pistil to pollination and fertilization. Ethylene evolution upon pollination in tobacco differed in each cross, suggesting that ethylene is correlated with the response to pollen tube growth in the tobacco flower.     

Mode of pollen tube growth in pistils of Ticodendron incognitum (Ticodendraceae,Fagales) and the evolution of chalazogamy

Ticodendron incognitum is the sole species of the Ticodendraceae, which was established as a new family in the Fagales less than 20 years ago. Considering the diverse modes of pollen tube growth observed in other Fagales, we investigated the growth of pollen tubes in the pistil of Ticodendron. At the time of pollination, T. incognitum had four immature ovules in a bilocular ovary, thus exhibiting delayed fertilization, as in other Fagales. During the period when fertilization was delayed, pollen tube growth in the pistil was intermittent, consisting of five steps associated with development of the ovules and embryo sacs. Four cessation sites occurred: in the style, in the tissue of the upper part of the ovary, inside and outside of the funicle and at the chalaza. A single pollen tube eventually reaches a mature embryo sac through the chalaza in one of the four ovules. While both delayed fertilization and intermittent pollen tube growth play a role in male and female gametophyte selection, as in other Fagales, the five‐step process of pollen tube growth through the chalaza (i.e. chalazogamy) is characteristic of lineages of the Casuarinaceae, Ticodendraceae and Betulaceae (the latter with the loss of one step). © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 157 , 621–631.     

Pistil strategies controlling pollen tube growth

The progamic phase appears especially well suited for pollen-pistil interaction. During this phase the pistil supports pollen germination and tube growth, and provides an adequate environment, nutrition and directional cues. However, this support does not occur indiscriminantly and some mechanisms operating in the pistil constrain pollen tube growth. An active, regulated constraint is the self-incompatibility reaction, but moderate restrictions of pollen tube growth also occur in compatible matings. These moderate restrictions involve reduced support by the pistil and they operate through two main strategies; one is by decreasing the amount of support and the other is by varying the time at which this support is provided. In this minireview, we examine the evidence that is accumulating for both support and constraint of pollen tube growth by the pistil and discuss the benefits of this dual system.     

Signaling in pollen-pistil interactions.

A complex set of cell interactions is required to achieve fertilization. The pollen grain must be recognized by the pistil, take up water, and grow a pollen tube directionally through the style in order to deliver the sperm to the ovule. In many families of flowering plants, self-fertilization can be prevented by recognition mechanisms that allow self-pollen rejection by the pistil. The self-incompatibility response is under the genetic control of a single multi-allelic locus, the (Self-incompatibility) locus. There are two major classes of self-incompatibility systems. Gametophytic self-incompatibility has been well characterized in the Solanaceae and in the Papaveraceae, while sporophytic self-incompatibility has been well characterized in the Brassicaceae. In this review article, we present recent advances in understanding the signals mediating pollen recognition and pollen tube growth, in both compatible and incompatible interactions.     

POLLEN TUBE COMPETITION AND SELECTION FOR METAL TOLERANCE IN SILENE DIOICA (CARYOPHYLLACEAE) AND MIMULUS GUTTATUS (SCROPHULARIACEAE)

An experiment was conducted to test whether metal tolerance expressed in pollen would provide a competitive advantage during pollen tube growth and fertilization. Copper or zinc was introduced into the pistil by growing metal tolerant plants of Silene dioica or Mimulus guttatus in a nutrient solution to which metals were added. Flowers from treated and control plants were pollinated with either metal tolerant or metal sensitive pollen. The rate of pollen tube growth, the number of seeds set, and the number of viable seeds produced were measured. In general, no effects were found on the rates of pollen tube growth. However, the number of fertilizations and viable seeds were affected. When toxic metals were present, the relative success of pollen from the nontolerant parent was reduced.     

Sulfinylated azadecalins act as functional mimics of a pollen germination stimulant in Arabidopsis pistils

Polarized cell elongation is triggered by small molecule cues during development of diverse organisms. During plant reproduction, pollen interactions with the stigma result in the polar outgrowth of a pollen tube, which delivers sperm cells to the female gametophyte to effect double fertilization. In many plants, pistils stimulate pollen germination. However, in Arabidopsis, the effect of pistils on pollen germination and the pistil factors that stimulate pollen germination remain poorly characterized. Here, we demonstrate that stigma, style, and ovules in Arabidopsis pistils stimulate pollen germination. We isolated an Arabidopsis pistil extract fraction that stimulates Arabidopsis pollen germination, and employed ultra‐high resolution electrospray ionization (ESI), Fourier‐transform ion cyclotron resonance (FT‐ICR) and MS/MS techniques to accurately determine the mass (202.126 Da) of a compound that is specifically present in this pistil extract fraction. Using the molecular formula (C₁₀H₁₉NOS) and tandem mass spectral fragmentation patterns of the m/z (mass to charge ratio) 202.126 ion, we postulated chemical structures, devised protocols, synthesized N‐methanesulfinyl 1‐ and 2‐azadecalins that are close structural mimics of the m/z 202.126 ion, and showed that they are sufficient to stimulate Arabidopsis pollen germination in vitro (30 μm stimulated approximately 50% germination) and elicit accession‐specific response. Although N‐methanesulfinyl 2‐azadecalin stimulated pollen germination in three species of Lineage I of Brassicaceae, it did not induce a germination response in Sisymbrium irio (Lineage II of Brassicaceae) and tobacco, indicating that activity of the compound is not random. Our results show that Arabidopsis pistils promote germination by producing azadecalin‐like molecules to ensure rapid fertilization by the appropriate pollen.     

异叶苦竹花粉管生长及双受精过程

以异叶苦竹为材料,采用扫描电镜、荧光显微镜技术及传统的石蜡制片技术,解剖观察其花粉管生长途径及双受精过程。结果表明:(1)授粉后,花粉在柱头上吸水膨胀,约30 min即可萌发。(2)授粉1～2 h后花粉管可达到花粉长度的5～10倍,花粉管在柱头分支中进一步伸长,并开始伸入花柱中生长。(3)授粉后5 h,大量花粉管沿引导组织进入花柱基部与子房顶部之间的子房壁,有少量花粉管在子房壁与外珠被之间的缝隙中生长。(4)授粉后8 h,少量花粉管到达珠孔端。(5)授粉后15～18 h,精核与极核融合,形成初生胚乳核;精、卵核融合,形成合子。(6)授粉后20～30 h,仍可在花柱中见到大量呈束状的花粉管。(7)授粉后48 h,子房内的大部分花粉管出现解体,大多数花粉死亡。研究认为,精细胞到达胚珠的时间为8 h。