梨远缘花粉原位萌发及生长特性

应用荧光标记方法对梨远缘花粉在‘丰水’和‘噢嗄二十世纪’柱头上萌发及花粉管生长特性进行观察,结果表明:(1)梨远缘花粉均能在柱头上萌发,但其萌发率不同,授粉后24 h,在‘丰水’柱头上‘红叶桃’花粉萌发率最高,达62.8%,而‘盖县大李’花粉萌发率仅为12.0%,各种远缘花粉在‘丰水’柱头萌发率均高于‘噢嗄二十世纪’柱头。(2)各种远缘花粉管在梨柱头或花柱内生长情况也有差异,‘红叶桃’等核果类花粉管在梨柱头上均表现为扭曲、盘绕等现象,不能穿过柱头;‘红星’和‘红富士’花粉管虽然有少量穿过柱头,但不能进一步在花柱内生长,表现为扭曲变形、先端膨大等不亲和性现象。因此,梨与远缘果树杂交不亲和在柱头上就已发生,这与梨自交不亲和反应发生在花柱内的现象不同。     

柚雌蕊不同部位抽提液对自交不亲和阻抑效果的研究

以‘土柚’（Citrus grandis ‘Tuyou’）作对照,采用荧光显微技术对‘琯溪蜜柚’（Citrus grandis‘Guanxi-miyou’）、‘度尾蜜柚’（Citrus grandis ‘Duweimiyou’）两个品种花粉在本品种和异品种雌蕊不同部位抽提液中花粉萌发和花粉管伸长状态进行了观察。结果表明：‘琯溪蜜柚’和‘度尾蜜柚’子房抽提液对自身花粉管伸长阻抑最明显,并产生严重的弯曲;柱头抽提液对自身花粉管伸长稍有影响,出现中度弯曲;花柱抽提液中自身花粉管伸长正常,仅有轻微弯曲。然而,对于异品种的花粉以及自交亲和的‘土柚’,其子房、花柱和柱头抽提液对花粉管伸长没有这种阻抑效应。     

‘晚大新高’梨授粉及受精过程的显微动态研究

应用荧光显微法和石蜡切片解剖法对‘晚大新高’梨授粉受精过程进行了系统观察研究。结果表明：‘晚大新高’梨自花授粉不结实;异花最佳授粉品种为‘黄花’,其次为‘翠冠’和‘丰水’。与选用‘黄花’为异花授粉品种相比,自花和异花的授粉受精过程存在明显差异,自花花粉在授粉后2h开始萌发,8h花粉管生长至离柱头约1／3处停止生长,顶端膨大呈球形,表现出自交不亲和性;异花花粉在授粉后1h开始部分萌发,8h花粉管生长至花柱中部,24h到达花柱基部并进入子房,48h进入胚囊,72h完成双受精过程。     

枇杷少核株系花粉管生长差异研究

以‘大五星’枇杷退化种子少核株系‘川农1号’(C1)为试材,以‘大五星’枇杷自花授粉为对照,采用荧光显微技术,对枇杷少核株系自花、异花授粉后花粉管生长情况进行观察。结果表明:(1)C1株系异花授粉后花粉能在柱头上萌发,并伸入中央花柱道,在授粉后48h左右到达花柱基部。(2)C1株系自花授粉后,花粉萌发与花粉管的伸长速度相对于异花授粉滞后,自花授粉后36h大多数花粉管到达花柱的中上部并停止生长,且伴随产生花粉管顶端形态异常、荧光异常明亮等现象,最终只有极少数花粉管能到达花柱基部。研究表明,C1株系为配子体自交不亲和,C1株系的自交不亲和性是引起其少核的重要原因之一。     

中国樱桃与甜樱桃花粉原位萌发及花粉管生长的差异

以‘垂丝’、‘东塘’(中国樱桃)和‘莫利’、‘拉宾斯’(甜樱桃)为试材,分别于自花、异花授粉后不同时间切取花柱,用FAA固定,荧光染色后压片观察。结果显示,中国樱桃和甜樱桃的自花、异花花粉均能在柱头萌发,且其花粉管在花柱中表现为“极快—慢—快—稳定”的动态变化过程。但甜樱桃花粉萌发率高于中国樱桃,异花高于自花。中国樱桃自花、异花都有花粉管到达花柱基部;甜樱桃‘莫利’ב拉宾斯’和‘拉宾斯’自交有花粉管到达花柱基部,而‘莫利’自交的花粉管在花柱中上部已停止生长。结果表明,中国樱桃表现自交亲和性,而甜樱桃除人工诱变导致自交亲和的‘拉宾斯’等品种外,表现典型的植物配子体自交不亲和性。     

不同倍性香石竹杂交花粉管生长荧光显微观察及结实研究

以香石竹四倍体材料‘紫蝴蝶’(2n=4x=60)为母本,二倍体材料‘珍珠粉’和‘NH6’(2n=2x=30)为父本,利用荧光显微镜观察其授粉后花粉管生长情况,统计其座果率、亲和指数及种子萌发率,并对杂交后代进行倍性鉴定。结果表明,在‘紫蝴蝶’柱头上,‘珍珠粉’和‘NH6’的花粉2h开始萌发,花粉管多处出现胼胝质塞,且花柱组织出现胼胝质反应,4h花粉管到达柱头中部并出现胼胝质塞,6h花粉管到达柱头基部,17h柱头基部的花粉管增多,花粉管进入子房组织且子房组织出现胼胝质反应,17～24h花粉管能与胚珠结合,但结合率低;‘紫蝴蝶’ב珍珠粉’杂交未获得植株,‘紫蝴蝶’בNH6’杂交获得3株植株,染色体倍性鉴定表明3株植株均为四倍体,这可能是‘NH6’产生2n配子的缘故。     

百合远缘杂交花粉萌发及花粉管生长过程观察

利用荧光显微镜对百合远缘杂交组合Bernini×Pollyanna花粉萌发及花粉管生长过程进行观察研究,结果显示,授粉后3~30 h内花粉萌发形成花粉管,且花粉管生长速度由快到慢,48~51 h内花粉管停止生长,花粉管最后深入到花柱的1/3处,并观察到一些异常的花粉管形态;花粉管生长过程中还伴随着一系列的胼胝质反应,出现的部位依次是乳突细胞、花粉管、花柱通道细胞、胚珠中的胚囊.结果表明该杂交组合不亲和.研究认为Berni-ni柱头乳突细胞是阻碍Pollyanna花粉管生长的第一道屏障,花柱通道细胞是抑制花粉管在花柱内生长的第二道屏障.     

高山杜鹃与大喇叭杜鹃种间杂交过程的观察研究

以高山杜鹃‘Nova Zembla’为母本,大喇叭杜鹃为父本进行人工杂交授粉,利用荧光显微镜对杂交组合花粉萌发及花粉管生长过程进行观察,并统计其杂交的田间坐果率。结果显示:(1)授粉后1d花粉开始萌发形成花粉管,其萌发率在授粉后1～5d内显著增长,其后增长缓慢,到第12天萌发率最高达37.36%。(2)花粉萌发后花粉管生长速度由慢变快,授粉后2d花粉管进入花柱,7d进入子房,10d进入胚珠;实验中杂交花粉管与胚珠虽有结合,但与胚珠结合率低,授粉后12d仅7.42条花粉管进入胚珠。(3)在花粉管生长过程中,伴有大量异常现象,表现为授粉后柱头细胞、花粉管、花柱引导组织、子房组织、胚珠中的胚囊等部位依次出现胼胝质沉积反应,花粉管生长中出现的膨大、先端沉积胼胝塞而中途停止生长、螺旋扭曲、粗细不均、杂乱生长或螺旋膨大且逆向生长等异常。(4)该实验杂交的田间坐果率为零。研究表明,高山杜鹃‘Nova Zembla’与大喇叭杜鹃杂交不亲和,杂交后花粉管生长的异常行为可能是种间杂交不亲和的主要原因,且受精前障碍与受精后障碍可能同时存在。     

类整合素在烟草花粉萌发和花粉管生长中的作用

本文研究了动物整合素VnR抗血清及动物整合素专一性抑制剂含RGD的多肽对体外及半体内培养条件下烟草花粉萌发及花粉管生长的影响。结果表明在体外培养条件下,VnR抗血清及GRGDSP肽对花粉的萌发及花粉管的生长没有明显的抑制作用,但可抑制钙调素促进的花粉萌发和花粉管的生长;两者对柱头上进行的花粉萌发及在花柱里进行的花粉管生长也有一定程度的抑制。对类整合素在花粉萌发及花粉管生长中的作用进行了讨论。     

小报春不同授粉组合亲和性研究

利用小报春‘红星’、‘紫霞’、‘罗兰香’3个品种分别设置8种授粉组合,统计了不同授粉组合的结实情况和单果种子数量,并对‘红星’品种部分授粉组合的花粉萌发与花粉管伸长过程进行荧光显微观察。结果显示:(1)3个品种的结实率和单果种子数量均表现为异型植株授粉>长花柱同型异株授粉、长花柱自交>短花柱同型异株授粉、短花柱自交,不同授粉组合间单果种子数量差异极显著,异型植株授粉类型结实率均达到100%,单果种子数量为44～181粒,显著高于自交组合和同型异株杂交组合;以异型致死花粉作为蒙导对短花柱同型异株授粉组合的结实有一定促进作用,但对长花柱同型异株授粉组合的结实没有一致的促进作用;3个品种中‘红星’的结实率和单果种子数量最高。(2)荧光显微观察表明,异型授粉组合花粉在柱头上大量萌发并在花柱中伸长,授粉144h后花粉管开始进入子房;同型授粉组合授粉12h后花粉开始萌发,但直到授粉后144h花粉管还没有进入花柱;自交授粉组合授粉后144h仍无花粉萌发。实验结果说明,小报春存在明显的自交不亲和性,且短花柱类型的自交不亲和性比长花柱类型强。     

S-RNases from self-incompatible and -compatible apple cultivars: purification,cloning, enzymic properties,and pollen tube growth inhibitory activity

Four S-RNases (RNase associated with self-incompatibility) were purified from the styles of two apple cultivars (Malus domestica), a self-incompatible cv., Starking Delicious (SD), and a self-compatible cv., Megumi (MG). Each cultivar produced two S-RNases and their enzymatic properties such as specific activity, pH optimum, thermal stability, and molecular mass, were characterized. The four S-RNases inhibited the tube growth of apple pollen in an in vitro bioassay at 25 microg/ml (1.0 microM), but did not distinguish self from non-self pollen. The cDNAs of four S-RNases were cloned, and the nucleotide and deduced amino acid sequences were analyzed. The nucleotide sequence of SD-Se RNase was a new one and the other was identical to that of Sc-RNase of cv. Fuji. In MG one was identical to the sequence of SD-Sc RNase and the other to that of Sa-RNase of cv. Golden Delicious except for one base. From results of the isolation amounts and the Western blot analysis for stylar crude extracts the amount of S-RNases in MG was apparently less than that in SD.     

花柱和花粉胞外钙调素对花粉萌发和花粉管伸长的影响

以烟草为材料,通过半体内实验,就花柱和花粉胞外钙调素对花粉萌发和花粉管伸长的影响进行了观察。发现用EGTA及钙调素抗血清处理柱头或花粉均可抑制花粉在柱头上的萌发;向花柱引导组织中显微注射纯化钙调素可促进花粉管束伸长,而注射钙调素抗血清可抑制花粉管束伸长;同时证实玉米花柱和花粉细胞壁中均存在钙调素及钙调素结合蛋白,而且花粉和花柱细胞壁中钙调素结合蛋白的种类有差异。结果表明存在于花粉和花柱细胞外的钙调素对花粉萌发和花粉管伸长均有促进作用。     

景宁木兰(Magnolia sinostellata)远缘杂交授粉过程障碍研究

以景宁木兰（Magnolia sinostellata）为亲本,分别与望春玉兰（M.biondii）、紫玉兰（M.liliiflora）、红运二乔（M.soulangeana ‘Red lucky’）、丹馨玉兰（M.soulangeana ‘Danxin’）进行种间远缘杂交,并以景宁木兰自交为对照,研究了景宁木兰自交和杂交亲和性。结果表明：（1）景宁木兰具有较高的花粉活力与柱头可授性,花粉管能正常进入杂交母本花柱,自交结实率为2%;（2）以景宁木兰为父本,紫玉兰柱头对其花粉具有较好的识别性,花粉萌发率和花粉管生长速度均高于以红运二乔和丹馨玉兰做母本的组合,花粉管生长较为正常,结实率为25%,而与红运玉兰杂交的结实率为4%,与丹馨玉兰杂交的结实率为0%;（3）以景宁木兰为母本,望春玉兰花粉在其柱头上的萌发比率高于其他4个组合,但花粉管生长过程中出现扭曲,景宁木兰花柱内出现胼胝质沉积,且最终并未获得种子。花粉管发育镜检结果表明：受精前障碍即花粉管进入花柱基部前生长异常是引起景宁木兰与丹馨玉兰、望春玉兰杂交不结实的主要原因;并进一步推测花粉管进入子房后的异常状态是导致景宁木兰自交,与红运二乔杂交结实率低的主要原因。     

Heterostyly inPrimula. 2. Sites of pollen inhibition,and effects of pistil constituents on compatible and incompatible pollen-tube growth

Summary In incompatible (intramorph) pollinations of the heterostylousPrimula vulgaris, pollen germination or tube growth may be partially inhibited in several sites associated with the stigma or style. Blockage may occur, a) on the stigma surface through the failure of germination or of pollen tube penetration after germination, b) in the stigma head during the passage of the tube through the specialized transmitting tissue of the head, or c) in the transmitting tract of the style. None of the barriers is complete, and the prohibition of selfing or intramorph crossing depends upon the cumulative screening effect of one following upon the other. In both morphs, the germination of incompatible pollen on the stigma is enhanced in high ambient relative humidity, but many tubes still fail to penetrate the stigma. Those that do are retarded or blocked in their growth in the transmitting tissues of the stigma head and style. Crude extracts from the tissues of the stigma head and style show some differential effect on the growth of pollen tubesin vitro, and dialysates of extracts containing high molecular weight fractions show a consistent differential effect, those from thrum tissues retarding thrum tubes while having a lesser effect on pin tubes, and those from pin tissues retarding pin tubes while having lesser effect on thrum. It is suggested that the factors influencing tube growth are present in the intercellular secretions of the transmitting tract.     

POLLEN TUBE DISTRIBUTIONS IN NICOTIANA GLAUCA: EVIDENCE FOR DENSITY DEPENDENT GROWTH

I hand sectioned styles of Nicotiana glauca at intervals along their length and counted the number of pollen tubes in each section using fluorescence microscopy. Evidence of density dependent growth was found for three stages of pollen growth. Pollen germination on the stigma increased with increasing pollen population size. Pollen tube penetration in the stigma was unaffected by increasing density from low to moderate levels but was reduced at high densities. Pollen tube penetration in the style was enhanced by increasing density. This enhanced growth in the style was apparently confounded by interference among pollen tubes growing at high densities. In particular, the area of tissue able to support pollen tube growth decreases from the stigma into the lower style, which could cause overcrowding of pollen tubes growing at high densities. Enhanced pollen tube penetration with increasing density combined with interference among pollen tubes growing at high densities resulted in greater mean pollen tube lengths for populations with moderate densities. The shift from density independent growth in the stigma to positively density dependent growth in the style may represent a shift from autotrophic to heterotrophic growth stages of pollen.     

Gametophyte–Sporophyte Interactions in the Pollen–Pistil System: 4. The Hormonal Status and the Mechanism of Self-Incompatibility

Following 4 and 8 h after self-incompatible pollination of Petunia hybrida plants, ethylene evolution and the contents of IAA, ABA, and cytokinins were measured in pistils and their parts (stigma, style, and ovary). The germination and initial growth of pollen tubes within the initial 4 h of the experiment were accompanied with an almost tenfold increase of the rate of ethylene production by the stigma and a twofold increase of the ABA content in the stigma and style. The inhibition of pollen tube growth in the style tissues during next 4 h coincided with a fivefold increase in the cytokinin content in the style, while high ABA content was maintained in the stigma and style. The authors conclude that phytohormones participate in the mechanism of gametophyte self-incompatibility.     

Comparative analysis of superfamilies of NBS-encoding disease resistance gene analogs in cultivated and wild apple species

Eleven distinct families of resistance gene analogs (RGAs) with the characteristic nucleotide-binding sequence (NBS) were identified in two wild apple species, Malus prunifolia and M. baccata, and two cultivated apple cultivars, M. domestica cv. Fuji and M. domestica cv. Hong-ok, using PCR approaches with degenerate primers based on two conserved motifs of known NBS-LRR resistance genes. These RGA families were found to be represented in all the apple species tested, including wild and cultivated species. However, their sequences are very divergent from each other. Furthermore, the low level of recombination detected within their RGA families supports the idea that the evolution of NBS-encoding sequences in apple species involves the gradual accumulation of mutations. Despite the high diversity of the RGA families found in all apple species, the apparent lack of differentiation between wild and cultivated forms suggests that other factors, such as the capacity to tolerate pathogens, might play an important role in the survival of wild-type species.     

AtROP1 negatively regulates potato resistance to Phytophthora infestans via NADPH oxidase-mediated accumulation of H2O2

Background

The characteristics of pollen tube growth are not constant, but display distinct patterns of growth within the different tissues of the pistil. In the stigma, the growth rate is slow and autotrophic, whereas in the style, it is rapid and heterotrophic. Very little is known about the interactions between these distinct maternal tissues and the traversing pollen tube and the role of this interaction on the observed metabolism. In this work we characterise pollen tube growth in the apple flower and look for differences in glycoprotein epitope localization between two different maternal tissues, the stigma and the style.

Results

While immunocytochemically-detected arabinogalactan proteins were present at high levels in the stigma, they were not detected in the transmitting tissue of the style, where extensins were abundant. Whereas extensins remained at high levels in unpollinated pistils, they were no longer present in the style following pollen tube passage. Similarily, while abundant in unpollinated styles, insoluble polysaccharides such as β-glucans, were depleted in pollinated pistils.

Conclusions

The switch from autotropic to heterotrophic pollen tube growth correlates spatially with a change of glycoprotein epitopes between the stigma and the style. The depletion of extensins and polysaccharides following pollen tube passage in the style suggest a possible contribution to the acceleration of heterotrophic pollen tube growth, which would imply an active contribution of female tissues on prezygotic male–female crosstalk.     

POLLEN LONGEVITY AND STIGMA PRE-EMPTION IN CLARKIA

The ability of pollen to effect seed set decreases with age when stored at room temperature or on the plant, but not when stored at 5 C. Seed set declines rapidly with pollen stored 10 days or more at room temperature. Seed set declines rapidly with pollen stored 4 days or more on the plant. The degree of pollen germination on the stigma and the number of pollen tubes in the style correlate positively with seed set. Pollen germination on the stigma and pollen tube growth in the style are strongly correlated with seed set for pollen stored at room temperature, but not, or only weakly, correlated for pollen stored at 5 C. In the latter case, seed sets are lower than expected, indicating a possible viability reduction associated with a post-pollination function. Inviable pollen grains temporarily interfere with normal pollen germination on the stigma. Essentially no seeds are produced when in viable pollen, immediately followed by viable pollen, is placed on a fresh stigma. However 27 to 65 seeds are produced when in viable pollen, followed in 1 to 6 days by viable pollen, is placed on fresh stigmas. New stigma surface is exposed at the center as expanding stigma surfaces carry the interfering inviable pollen toward the edge of the stigma.