Self-incompatibility in Papaver targets soluble inorganic pyrophosphatases in pollen

In higher plants, sexual reproduction involves interactions between pollen and pistil. A key mechanism to prevent inbreeding is self-incompatibility through rejection of incompatible ('self') pollen. In Papaver rhoeas, S proteins encoded by the stigma interact with incompatible pollen, triggering a Ca2+-dependent signalling network resulting in pollen tube inhibition and programmed cell death. The cytosolic phosphoprotein p26.1, which has been identified in incompatible pollen, shows rapid, self-incompatibility-induced Ca2+-dependent hyperphosphorylation in vivo. Here we show that p26.1 comprises two proteins, Pr-p26.1a and Pr-p26.1b, which are soluble inorganic pyrophosphatases (sPPases). These proteins have classic Mg2+-dependent sPPase activity, which is inhibited by Ca2+, and unexpectedly can be phosphorylated in vitro. We show that phosphorylation inhibits sPPase activity, establishing a previously unknown mechanism for regulating eukaryotic sPPases. Reduced sPPase activity is predicted to result in the inhibition of many biosynthetic pathways, suggesting that there may be additional mechanisms of self-incompatibility-mediated pollen tube inhibition. We provide evidence that sPPases are required for growth and that self-incompatibility results in an increase in inorganic pyrophosphate, implying a functional role for Pr-p26.1.     

S-RNase uptake by compatible pollen tubes in gametophytic self-incompatibility

Many flowering plants avoid inbreeding through a genetic mechanism termed self-incompatibility. An extremely polymorphic S-locus controls the gametophytic self-incompatibility system that causes pollen rejection (that is, active arrest of pollen tube growth inside the style) when an S-allele carried by haploid pollen matches one of the S-alleles present in the diploid style. The only known product of the S-locus is an S-RNase expressed in the mature style. The pollen component to this cell-cell recognition system is unknown and current models propose that it either acts as a gatekeeper allowing only its cognate S-RNase to enter the pollen tube, or as an inhibitor of non-cognate S-RNases. In the latter case, all S-RNases are presumed to enter pollen tubes; thus, the two models make diametrically opposed predictions concerning the entry of S-RNases into compatible pollen. Here we use immunocytochemical labelling of pollen tubes growing in styles to show accumulation of an S-RNase in the cytoplasm of all pollen-tube haplotypes, thus providing experimental support for the inhibitor model.     

The S-locus receptor kinase is inhibited by thioredoxins and activated by pollen coat proteins

The self-incompatibility response in Brassica allows recognition and rejection of self-pollen by the stigmatic papillae. The transmembrane S-locus receptor kinase (SRK), a member of the receptor-like kinase superfamily in plants, mediates recognition of self-pollen on the female side, whereas the S-locus cysteine-rich protein (SCR) is the male component of the self-incompatibility response. SCR is presumably located in the pollen coat, and is thought to be the SRK ligand. Although many receptor-like kinases have been isolated in plants, the mechanisms of signal transduction mediated by these molecules remain largely unknown. Here we show that SRK is phosphorylated in vivo within one hour of self-pollination. We also show that, in vitro, autophosphorylation of SRK is prevented by the stigma thioredoxin THL1 in the absence of a ligand. This inhibition is released in a haplotype-specific manner by the addition of pollen coat proteins. Our data indicate that SRK is inhibited by thioredoxins and activated by pollen coat proteins.     

Identification of the pollen determinant of S-RNase-mediated self-incompatibility

Many flowering plants have adopted self-incompatibility mechanisms to prevent inbreeding and promote out-crosses. In the Solanaceae, Rosaceae and Scrophulariaceae, two separate genes at the highly polymorphic S-locus control self-incompatibility interactions: the S-RNase gene encodes the pistil determinant and the previously unidentified S-gene encodes the pollen determinant. S-RNases interact with pollen S-allele products to inhibit the growth of self-pollen tubes in the style. Pollen-expressed F-box genes showing allelic sequence polymorphism have recently been identified near to the S-RNase gene in members of the Rosaceae and Scrophulariaceae; but until now have not been directly shown to encode the pollen determinant. Here we report the identification and characterization of PiSLF, an S-locus F-box gene of Petunia inflata (Solanaceae). We show that transformation of S1S1, S1S2 and S2S3 plants with the S2-allele of PiSLF causes breakdown of their pollen function in self-incompatibility. This breakdown of pollen function is consistent with 'competitive interaction', in which pollen carrying two different pollen S-alleles fails to function in self-incompatibility. We conclude that PiSLF encodes the pollen self-incompatibility determinant.     

Direct ligand-receptor complex interaction controls Brassica self-incompatibility

Many higher plants have evolved self-incompatibility mechanisms to prevent self-fertilization. In Brassica self-incompatibility, recognition between pollen and the stigma is controlled by the S locus, which contains three highly polymorphic genes: S-receptor kinase (SRK), S-locus protein 11 (SP11) (also called S-locus cysteine-rich protein; SCR) and S-locus glycoprotein (SLG). SRK encodes a membrane-spanning serine/threonine kinase that determines the S-haplotype specificity of the stigma, and SP11 encodes a small cysteine-rich protein that determines the S-haplotype specificity of pollen. SP11 is localized in the pollen coat. It is thought that, during self-pollination, SP11 is secreted from the pollen coat and interacts with its cognate SRK in the papilla cell of the stigma to elicit the self-incompatibility response. SLG is a secreted stigma protein that is highly homologous to the SRK extracellular domain. Although it is not required for S-haplotype specificity of the stigma, SLG enhances the self-incompatibility response; however, how this is accomplished remains controversial. Here we show that a single form of SP11 of the S8 haplotype (S8-SP11) stabilized with four intramolecular disulphide bonds specifically binds the stigma membrane of the S8 haplotype to induce autophosphorylation of SRK8, and that SRK8 and SLG8 together form a high-affinity receptor complex for S8-SP11 on the stigma membrane.     

植物生殖与胚胎发育过程中的细胞程序性死亡

生殖与胚胎发育过程中植物细胞程序性死亡（ＰＣＤ）的典型特点在于它的细胞类型特异性。即在特定时期，只有特定的细胞才会发生ＰＣＤ。介绍了植物生殖和胚胎发育过程中各种类型的细胞在特定时期所发生的ＰＣＤ的形态和生化特征。这些ＰＣＤ类型包括大小孢子的退化、单性花的形成、性别决定、心皮通化、花粉发育及绒毡层的解体、胚胎发育过程中胚柄及糊粉层的消失以及体细胞胚胎发生等。同时对植物生长和发育过程中ＰＣＤ的作用及其     

紫薇受粉习性及花粉管生长的研究

用脱色苯胺蓝对紫薇花粉和花粉管进行染色,在荧光显微镜下观察其自然条件下的受粉习性及花粉管生长行为,以确定花粉管通道的形成时期．结果表明,盛花期的紫薇在开花后1.5h内完成受粉,花粉落量多,受粉率为100％;开花后24h左右进行受精;单果种子数为36～50粒,认为可以用花粉管通道法转化紫薇,导入外源DNA的最佳时机为开花后24h左右.     

植物有性生殖过程中的细胞程序性死亡

细胞程序性死亡(pmgrammed cell death,PCD)已成为当前生物学的研究热点之一,PCD是植物发育过程中的一种普遍现象,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,有性生殖过程中的植物细胞程序性死亡具有细胞类型特异性,即在特定时期,只有特定的细胞才会发生程序性死亡,本文介绍了植物有性生殖过程中各种类型的细胞在特定时期所发生的程序性死亡的形态和生化特征,这些PCD类型包括单性花的形成、性别决定、大小孢子的退化、花粉发育及绒毡层的解体、雌雄配子体的发育、花粉管的生长、胚胎发育过程中胚柄、胚乳以及糊粉层的消失等等。     

Programmed cell death during terminal bud senescence in a sympodial branching tree, Eucommia ulmoides

Eucommia ulmoides Oliv. is a typical sympodial branching tree. The apical bud of the branch ages and dies every year, replaced by the nearby axillary bud in the second year. Structural assays and a series of biochemical analyses were performed to analyze the senescence mechanism in the apical bud. It was revealed that most cells of the apical bud underwent the programmed cell death (PCD) during the senescence: the chromosomes were congregated and the nuclear contents were condensed, as shown by 4′,6-diamidino-2-phenylindole (DAPI) fluorescence. DNA fragmentation was detected during senescence using the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end in situ labeling (TUNEL) method, coincident with the appearance of a DNA ladder. Moreover, a 20 kD DNase related to fragmentation was found. PCD was initiated first in the young leaves, leaf primordia and peripheral zone cells, then in the central mother cells and initial layer cells in the apical meristem. The terminal buds remain in vegetative growth during senescence, in contrast to buds of many annual plants.     

细胞编程性死亡及其生物学意义

细胞编程性死亡(英文编写为PCD)是多细胞生物的自然属性,它按照一定的基因编码在时空上顺次表达使死亡细胞呈现一定的表观现象及生理生化特征。PCD受来自机体内其它细胞信号等内源性因素和药物、物理等外源性因素的调节。PCD是一种主动死亡方式且表现为单细胞性,与病理因素作用下的坏死不同,但又有一定联系。PCD在动、植物体的生长发育和成体新陈代谢中有重要的调节作用,PCD异常是某些疾病发病的重要原因     

聚晶金刚石复合片非脆性去除磨削机理研究

聚晶金刚石(polycrystalline diamond,PCD)的磨削是PCD刀具制造中的关键环节,磨削参数的变化影响PCD的去除方式,进而决定刀具的切削性能.通过扫描电镜观察分析腐蚀后的PCD磨削表面微观形貌,研究了金刚石砂轮磨削PCD中的刻划作用、滑擦作用和热化学反应等非脆性去除机理,及其与磨削参数的关系.结果表明:刻划作用仅在砂轮磨粒较锋利时产生作用,多存在于湿磨初期;砂轮磨钝后,滑擦作用和热化学反应是PCD的主要去除方式,且滑擦作用与热化学反应同时发生.通过设计一组特殊磨削试验,间接证明了磨削后的PCD表面存在由热化学反应产生的软化层.     

维管植物导管元素分化与细胞程序性死亡

通过遗传控制的细胞死亡是植物正常生长和发育的重要过程,是近年来国内外的研究热点之一。维管植物的导管和筛管由死亡的管导分子组成,本文着重讨论了维管植物的导分子形成过程中的细胞程序性死亡（ｐｒｏｇｒａｍｍｅｄ ｃｅｌｌｄｅａｔｈＰＣＤ）的形态和化特性以及参与找在因,并介绍了植物生长发育过程中ＰＣＤ存在的时空广泛性。虽然植物发育生殖过程中的ＰＣＤ与动物细胞通过遗控制的细胞死亡是植物正常生长和发育的重要过程,是近年来国内外的研究热点之一,维管植物的导管和筛管由天文馆的导管分子组成。本文着重讨论了维管植物的导管分子形成过程中的细程序性死亡（programmed cell death PCD)的形态和生化特征以及参与的基因,并介绍了植物生长发育过程中ＰＣＤ存在的时空广泛性。虽然植物发育和生殖过程中的ＰＣＤ与动物细胞凋亡存在许多相似的形太必生化特征,但细胞通过自溶或自自我吞噬来死亡,而并不形成动物细胞凋亡中典型的凋亡小体。说明这些ＰＣＤ类型本质上凋亡型ＰＣＤ。导管分子的死亡是一种典型的植物ＰＣＤ。近年来体外系统研究表明,导管元素分析中的细胞死亡程序与动物细胞凋亡不同。     

玉米中的细胞程序性死亡及其遗传机制

玉米是主要农作物之一 ,也是一种重要的模式研究植物。玉米生长发育过程中自始至终伴随着细胞程序性死亡 (programmedcelldeathPCD)。这些PCD大致分为三大类 ,一类是正常生长发育过程中的PCD ;一类是抗病过程中的PCD或突变模拟病斑 ;另一类是外界逆境因素诱导的PCD。在玉米中已发现许多细胞死亡突变体 ,导致这些突变的一般是一个已知的可转座元素 ,这样便可通过转座子标签法来分离这些突变体的相应基因。对这些突变体的分析将阐明玉米PCD的分子机制及遗传调控 ,从而为人工控制玉米PCD提供理论基础 ,并将为其它植物的PCD研究提供一种模型。     

Programmed cell death, mitochondria and the plant hypersensitive response.

The plant response to attempted infection by microbial pathogens is often accompanied by rapid cell death in and around the initial infection site, a reaction known as the hypersensitive response. This response is associated with restricted pathogen growth and represents a form of programmed cell death (PCD). Recent pharmacological and molecular studies have provided functional evidence for the conservation of some of the basic regulatory mechanisms underlying the response to pathogens and the activation of PCD in animal and plant systems. In animals, the mitochondrion integrates diverse cellular stress signals and initiates the death execution pathway, and studies indicate a similar involvement for mitochondria in regulating PCD in plants. But many of the cell-death regulators that have been characterized in humans, worms and flies are absent from the Arabidopsis genome, indicating that plants probably use other regulators to control this process.     

荔枝(Litchi chinensis Sonn.) 花芽分化过程的细胞超微结构观察

应用透射电镜对荔枝茎类生长锥由营养生长转变为生殖生长过程的细胞超微结构进行观察。结果表明，荔枝圆锥花序发育期细胞排列紧密,细胞质浓，液泡小，细胞核大并占据着细胞大部分位置；细胞质内有丰富的线粒体和膜状系统，整个膜状系统内连细胞核，细胞内囊泡大量形成、融合与迁移。这些均显示出细胞内结构物质在此期经历着活跃的周转和合成代谢，细胞内部在基因 的控制下有序地表达，为发育过程质的转变积累足够的信息和物质。生长锥由尖长变为宽平的花序原基时，其组织细胞结构也发生了相应的变化，最显著的特征是在此过程中，有一些细胞发生了程序性死亡。正在凋亡的细胞核染色质固缩、趋边、核膜破裂或皱缩、细胞质囊泡化并将降解的物质运转到周围细胞，从而保证了其他细胞正常发育以及生长的细胞对物质和能量的需求。     

植物细胞程序性死亡研究进展

植物细胞程序性死亡普遍存在于植物生长发育及环境相互作用过程中,是由基因调控的、主动的细胞死亡过程。植物PCD已成为当前生物学研究的热点之一。本文概述了植物细胞程序性死亡的形态学、分子生物学特征以及植物PCD在植物发育、环境胁迫、超敏反应中的存在,并就植物PCD的细胞形态学、分子生物学等检测方法以及植物PCD的基因、酶和信号分子的调控进行了综述。     

梯度升温降温条件下球形棕囊藻生长和生理生化响应

球形棕囊藻(Phaeocystis globosa)藻华频繁暴发,不仅会危害海洋生态系统和渔业生产安全,还会堵塞核电站冷源取水系统产生严重安全隐患。球形棕囊藻藻华的暴发有明显的季节性特征,本研究采用梯度升温降温实验,研究不同温度(高温、低温)条件下球形棕囊藻的生理生化响应及温度胁迫诱导的细胞程序性死亡(Programmed Cell Death,PCD)过程。结果显示：低温胁迫抑制球形棕囊藻细胞的正常生长繁殖,但不会激活其进入PCD响应;高温胁迫使球形棕囊藻出现氧化应激,从而导致其发生PCD响应,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性显著提高,半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)被激活后球形棕囊藻细胞出现Annexin V-FITC阳性反应。环境胁迫使球形棕囊藻细胞产生氧化应激,抗氧化酶活性快速升高,并激活Caspase-3使藻细胞发生PCD响应,表现出细胞磷脂酰丝氨酸(PS)外翻、细胞收缩和质膜从细胞壁分离等特征。本研究可为深入探讨球形棕囊藻藻华的生消机制与防治赤潮灾害提供一定的理论基础。     

Maize transformation via pollen tube pathway and the inheritance of transgene in progeny

The herbicide-resistant gene als was introduced into maize inbred line Qi319 through pollen-tube pathway. The inheritance of transgene was studied up to 33 generation. Transformation was performed by applying plasmid DNA carrying solution on the severed styles after self-pollination, or just prior to selfpoll/nation. After herbicide screening, PCR and Southern blot analysis, seventeen plants were confirmed positive in TO generation, and sixteen of them were self-bred to construct family to 33 progeny. Through PCR analysis and herbicide screening in offspring, four lines were confirmed following a 3 : 1 Mendelian segregation ratio. In other lines the numbers of positive plants were lower than expected. Therefore, it is necessary to select large population in progeny to produce stable inherited lines when transgene was delivered by pollen tube method.     

A P-type ATPase required for rice blast disease and induction of host resistance

To cause diseases in plants, pathogenic microorganisms have evolved mechanisms to deliver proteins directly into plant cells, where they suppress plant defences and facilitate tissue invasion. How plant pathogenic fungi, which cause many of the world's most serious plant diseases, deliver proteins during plant infection is currently unknown. Here we report the characterization of a P-type ATPase-encoding gene, MgAPT2, in the economically important rice blast pathogen Magnaporthe grisea, which is required for exocytosis during plant infection. Targeted gene replacement showed that MgAPT2 is required for both foliar and root infection by the fungus, and for the rapid induction of host defence responses in an incompatible reaction. DeltaMgapt2 mutants are impaired in the secretion of a range of extracellular enzymes and accumulate abnormal Golgi-like cisternae. However, the loss of MgAPT2 does not significantly affect hyphal growth or sporulation, indicating that the establishment of rice blast disease involves the use of MgApt2-dependent exocytotic processes that operate during plant infection.