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.     

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.     

部分李属植物种间杂交不亲和性的研究

通过常规杂交及荧光显微镜观测等方法,对李、杏种间杂交不亲和性进行了研究.研究结果表明,铃铛花期授粉的坐果率显著高于初花期授粉的坐果率;同一杂交组合,正反交坐果率差异显著;母本对远缘杂交亲和性影响很大,父本对远缘杂交亲和性的影响较小;通过具备正常活力及萌芽率的花粉授粉后,李杏杂交存在一定程度的花粉和柱头不亲和及＂拒绝＂反应,以李为母本的杂交不亲和性部位在柱头表面和花柱上部,而以杏为母本的不亲和部位则在花柱中下部.研究结果可为阐释李、杏种间杂交不亲和机制提供理论依据.     

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.     

斑块开度对树木结实和交配格局的影响：以城市油松（Pinus tabulaeformis）种群为例

以城市种植油松斑块为例,探讨了斑块开度(作为林冠开度指标)对油松交配格局和结实的影响,发现斑块开度和油松自交率呈显著正相关,并与个体结实量具有一定的相关性.其原因可能是由于斑块开度增大导致自交种子败育率下降,或者是由于孤立木接收外源花粉相对较少.另外,本研究未发现斑块开度对花粉迁入有显著影响.     

Self-incompatibility triggers programmed cell death in Papaver pollen

Sexual reproduction in many angiosperm plants involves self-incompatibility (SI), which is one of the most important mechanisms to prevent inbreeding. SI is genetically controlled by the S-locus, and involves highly specific interactions during pollination between pollen and the pistil on which it lands. This results in the rejection of incompatible ('self') pollen, whereas compatible ('non-self') pollen is allowed to fertilize the plant. In Papaver rhoeas, S-proteins encoded by the stigma component of the S-locus interact with incompatible pollen, triggering a Ca2+-dependent signalling network, resulting in the inhibition of pollen-tube growth. Programmed cell death (PCD) is a mechanism used by many organisms to destroy unwanted cells in a precisely regulated manner. Here we show that PCD is triggered by SI in an S-specific manner in incompatible pollen. This provides a demonstration of a SI system using PCD, revealing a novel mechanism to prevent self-fertilization. Furthermore, our data reveal that the response is biphasic; rapid inhibition of pollen-tube growth is followed by PCD, which is involved in a later 'decision-making' phase, making inhibition irreversible.     

Identification and function analysis of spectrin-like protein in pollen tubes of lily （Lilium davidii Duch）

The elongation of pollen tube is an important process of sexual reproduction in higher plant. Cytoskeleton plays a major regulatory role in the elongation of pollen tubes. But whether membrane skeleton is involved in the pollen tube elongation is not clear. In this study, immunochemical detection of spectrin-like protein has been carried out in pollen tubes. By use of 2-dimensional electrophoresis(2DE) and western blotting, two spectrin-like proteins are found, one is 150 kD, and the other is 105 kD, with pl being 4.54 and 4.39, respectively. 150 kD spectrin-like protein is located in plasma membrane of pollen tube and 105 kD spectrin-like protein is located in cytoplasm, probably functioning as a subunit to form a dimmer (210 kD) in vivo. The elongation of pollen tubes is inhibited after spectrin antibody was injected into a growing pollen tube. These results suggest that spectrin-like proteins exist in pollen tube and play an important regulating role in the elongation process of pollen tubes from lily.     

不同授粉方式对祝光苹果受精过程的影响

祝光苹果自花授粉后2小时,花粉粒在柱头上萌发。授粉当日和次日,花粉管在花柱中生长。授粉后第三日,花粉管停止生长,最长花粉管平均长至花柱全长的26％。此时,花粉管先端明显膨大。而后,未见到有抵达花柱基部的花粉管,也未见到受精过程的发生。 异花授粉后4—6小时,大量花粉粒在柱头上萌发。授粉后1—2日,花粉管径直向花柱基部生长。授粉后50小时,花粉管穿过花柱基部,进入子室内。授粉后3—4日,观察到进入胚囊的花粉管将内含物释放到一个已退化的助细胞中。授粉后5—6日,观察到双受精过程。     

白头翁花部性状及花粉萌发特性研究

通过野外观察及室内试验,对白头翁花部性状及花粉萌发特征进行了观察研究.结果,白头翁花期在20 d左右,单花花期约10d;其花部性状存在着差异,开放时花冠的直径约3~5 cm;雌蕊和雄蕊数目在不同植株间不同花内都存在着差异,雄蕊数目为240~350枚,雌蕊数目为164~340枚;同一朵花内雄蕊长度变化较大,越接近基部越短小,雌蕊长度变化不大;大部分花其基部存在着花药发育不正常的雄蕊,内含花粉量少,介于正常花药和退化花药之间;每个花药中的花粉数目约(2145±515)个;体外萌发试验表明,该种花粉萌发率在开花后24 h达到最高值,开花后50 h(第3 d)花粉仍维持4.8%左右的萌发率,由此推测白头翁花粉具有远距离传播的潜能;白头翁苞片形态变异很大,共发现9种不同式样.     

北美鹅掌楸和中国鹅掌楸种间杂交胚胎学

１．中国鹅掌楸花粉在北美鹅掌楸柱头上萌发良好，速度快且整齐。２．花粉管生长速度最早在８０ｈ进入胚囊，大多数在１２０ｈ以后。个别花粉管在花柱中顶端膨大、破裂，或行至珠孔发生扭曲现象。３．花粉管通过柱头沟和花柱道时生长快，通过珠孔塞和珠孔以及珠柄时生长缓慢。４．控制授粉后５ｄ左右发生双受精，合子休眠约２０ｄ左右，３０ｄ左右是球形胚，９０ｄ子叶胚形成；１４０ｄ胚分化完成，种子成熟。５．连续三年的结籽率调     

麻疯树花部特征及其对传粉昆虫的影响

【目的】麻疯树为中国西南地区重要生物柴油树种,了解其花部特征对主要传粉昆虫的访花行为与访花频率可能产生的影响,有助于麻疯树的进一步繁殖成功。【方法】对麻疯树雌、雄花的形态特征进行测量,对花粉与柱头的微观特征进行扫描电镜观察,并对其花部构成、花冠尺寸及花部颜色对蝇类与蜜蜂类访花频率的影响进行观测。【结果】麻疯树雄花的花冠直径显著小于雌花,雄蕊的直径显著大于雌蕊,可能会影响传粉者对雌、雄花的偏好。雄、雌花的花蕊和花冠的相近,利于花粉的携带、转移与落置。麻疯树花粉粒表面具有瘤状突起,含水丰富,直径大; 雌花柱头有孔状凹陷,能分泌少量黏液,利于花粉在柱头上的粘附与萌发。去除花蕊与花瓣均会显著降低蝇类与蜜蜂对麻疯树花的访问频率,去除花萼对传粉者访问频率无影响。随着花冠尺寸变小,蝇类与蜜蜂对麻疯树花的访问频率均显著降低。花部颜色的改变显著降低蜜蜂类的访花频率,但对蝇类的访花频率无明显影响。【结论】麻疯树雌、雄花的花部形态特征有显著差别。蜜蜂类与蝇类传粉者对麻疯树的花部组成、花冠大小与花部颜色的视觉识别方式有显著不同。     

Distribution of a kinesinrelated protein on Golgi apparatus of tobacco pollen tubes

The conventional kinesin and kinesin-related proteins (KRPs) constitute a large family of microtubule-based molecular motors. Members of this family bind to micro-tubules in an ATP-dependent manner. They all have microtubule-activated ATPaes activities, and play essential roles in the transport of various vesicles and organelles in eukaryotic cells[1―3]. The first plant kinesin-like protein was detected in tobacco pollen tube with an antibody against animal kinesin[4]. Recently, a number …     

矮慈姑人工授粉后花粉管生长的荧光显微观察

用荧光显微术观察了矮慈姑人工授粉后雌蕊上花粉粒萌发及花粉管的生长过程与途径,结果表明：该种在授粉０．５ｈ后花粉粒全部萌发出花粉管,花粉管在授粉２ｈ后部分到达子房,４ｈ仙进入胚珠;花粉管在雌蕊之向心一侧的组织中行进,至子房基部后部分花部管转向胚珠,由珠孔进入珠心。     

From pollen actin to crop male sterility

Actin plays an important role in the life activity of animal and plant cells. Pollen cells have plenty of actin whose structure and characteristics are very similar to the animal actin. The nucleotide sequence and amino acid sequence of plant actin gene are very similar to those of the animal gene. The content of pollen actin from male sterile plants is much more lower than that from its maintainer plants. The expression of actin gene is organ-specific during the plant development. The expression quantity of actin gene in pollen is much more higher than those from root, stem and leaf. The expression plasmid of the anti-sense actin gene was constructed, transferred to the protoplasts of wheat and tomato to inhibit the expression of actin gene in pollen and thus the male sterile plants of wheat and tomato were obtained. The actin in pollens from the transgenic plants was reduced significantly, whereas the pistil was not affected. This study might pave a new way to breeding male sterile lines for the application of hybrid vigor of wheat and tomato.     

慈姑雌花在自然居群中的受粉过程--花粉粒落置、萌芽及花粉管生长的荧光显微观察

用荧光显微术观察了慈姑（Ｓａｇｉｔｔａｒｉａ ｔｒｉｆｏｌｉａ Ｌｉｎｎ）不同时期的雌花柱头上花粉粒的落置、萌发及花粉管的生长过程与途径。结果表明：该种雌花开花２ｈ 柱头受粉率为０,开花４ｈ 和８ｈ受粉率分别为５３８％ 和９８３％ ,柱头上花粉萌发率达１００％ ;花粉管沿雌蕊之向心一侧的组织中穿行,至子房基部后部分花粉管转向胚珠,由珠孔进入珠心。本研究发现该种花粉管常穿过子房基部至花托组织,并可进入其他雌蕊中,作者认为这种行为可能对保证雌花的结实率有一定意义     

番红花花器官形态结构的观察与自交试验

番红花 (CrocussativusL .)花器官形态结构正常 ,成熟同步 .自花授粉结果表明 :尽管授粉时期与子房见光生长对花粉管伸长长度有一定影响 ,但花粉管形状出现畸形 ,最终不能进入花柱和子房 .     

海甘蓝与芸苔属属间杂交的亲和性研究

对海甘蓝和芸苔属属间杂交的花粉粒萌发，花粉管生长以及珠孔受精情况进行了研究。结果表明，甘蓝型油采与海甘蓝杂交不亲和，表现在花粉管不能进入柱头或不能通过珠孔肥精，同时乳突细胞有强烈的胼胝质反应。白菜型油菜与海甘蓝杂交有少量花粉管萌发，花粉管顶端受阻于柱头乳突细胞中缠绕于乳突细胞表面，乳突细胞有大量的胼胝质产生。     

IAA对黄瓜花粉管伸长及花粉内源cAMP含量的影响

研究不同浓度的ＩＡＡ对黄瓜花粉管伸长及内源ｃＡＭＰ含量的影响。结果表明：５ｍｇ／ＬＩＡＡ明显促进花粉管伸长，而１００ｍｇ／ＬＩＡＡ则完全抑制花粉萌发。     

Gene flow from transgenic oilseed rape (Brassica napus L.) to cruciferous weeds under mentor pollen inducement

The alien gene flow between genetically modified glyphosate-resistant rapeseed variety Q3 (Brassica napus L.) and four cruciferous weeds was studied under mentor pollen inducement. The results showed that when Thlaspi arvense L., Capsella bursa-pastoris (L.) Medic, Cardamine hirsute L. and Rorippa palustris (L.) Besser were pollinated with mentor pollen, the mixed Q3 and the weed, pollen grains aggregated largely and germinated quickly, and the numbers of pollen tubes penetrating into the style and the ovary were greatly increased as compared with corresponding self-pollination groups. Twenty four to forty eight hours after pollination, several pollen tubes were observed to penetrate into the ovule via micropyle in each mentor combination. However, when the mentor progenies were analyzed by PCR, all of them showed negative for the Q3 herbicide-resistant gene. Collectively, these results indicated that crossing between T. arvense, C. bursa-pastoris, C. hirsuta, R. palustris (as female) and Q3 (as male) was highly incompatible and the herbicide-resistant gene could not flow from Q3 to these four weeds.     

Gene flow from transgenic oilseed rape (Brassica napus L.) to cruciferous weeds under mentor pollen inducement

The alien gene flow between genetically modified glyphosate-resistant rapeseed variety Q3 (Brassica napus L.) and four cruciferous weeds was studied under mentor pollen inducement. The results showed that when Thlaspi arvense L., Capsella bursa-pastoris (L.) Medic, Cardamine hirsute L. and Rorippa palustris (L.) Besser were pollinated with mentor pollen, the mixed Q3 and the weed, pollen grains aggregated largely and germinated quickly, and the numbers of pollen tubes penetrating into the style and the ovary were greatly increased as compared with corresponding self-pollination groups. Twenty four to forty eight hours after pollination, several pollen tubes were observed to penetrate into the ovule via micropyle in each mentor combination. However, when the mentor progenies were analyzed by PCR, all of them showed negative for the Q3 herbicide-resistant gene. Collectively, these results indicated that crossing between T. arvense, C. bursa-pastoris, C. hirsuta, R. palustris (as female) and Q3 (as male) was highly incompatible and the herbicide-resistant gene could not flow from Q3 to these four weeds.