高温对苹果花粉在花柱内萌发和生长的影响

以‘长富2号’红富士苹果为母本,‘红星’苹果为父本,研究35℃高温对苹果花粉在花柱内萌发和生长的影响。结果表明：温度影响花粉在柱头和花柱内的萌发及生长,与对照相比,高温促进花粉在柱头表面快速萌发,并加速花粉管在花柱内的伸长生长,但是在授粉72hN,高温处理下培养的花粉管形态出现花粉管变粗、弯曲并有瘤状小结的异化现象,同时,花柱发生褐变,并产生胼胝质,最终导致花粉管不能进入胚珠完成受精。     

‘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。     

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。     

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。     

青篱柴通过促进亲和花粉生长而提高传粉精确性

异型花柱植物的适应意义在于提高亲和花粉的传递, 从而促进异交, 但是有不少研究发现柱头上落置了大量不亲和花粉。目前, 很少有人研究柱头是否有利于亲和花粉的萌发和花粉管伸长, 以及通过去雄处理排除花内和植株内不亲和花粉干扰从而验证二型花柱植物是否可以促进异交。本研究以亚麻科青篱柴属(Tirpitzia)的二型花柱植物青篱柴(T. sinensis)为研究对象, 分居群调查了青篱柴长、短柱型的植株数量, 测量不同柱型的花粉与柱头大小以及观察其表面纹饰, 测定不同开花时间长、短柱型的柱头活性与花粉活力, 统计自然状态下柱头上落置的不同表型花粉比例, 进一步统计花内、株内去雄和自然对照下柱头上落置的不同表型花粉比例, 计算型间、型内和混合授粉花粉管长度和花粉萌发率, 人工授粉检测其是否型内异交不亲和与自交不亲和。结果表明, 自然居群中长、短柱型青篱柴植株数量没有显著性差异; 短柱型青篱柴花粉体积显著大于长柱型, 且二者花粉表面纹饰不同, 但不同柱型的柱头表面积与表面纹饰无差异; 青篱柴开花第一天的花粉活力显著高于开花第二天, 而柱头活性在开花前两天无显著性差异; 自然状态下柱头上落置的不亲和花粉比例显著高于亲和花粉; 在长柱型青篱柴中, 花内和株内去雄显著提高了柱头上亲和花粉的落置比例; 柱头上授型内花粉, 其花粉萌发率和花粉管的长度都显著低于授型间花粉; 授粉结果表明青篱柴为型内异交不亲和与自交不亲和。二型花柱植物通过促进亲和花粉的萌发和花粉管的伸长而有利于型间授粉, 从而显著提高其传粉精确性。     

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。     

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。     

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。     

细胞松弛素B和鬼笔环肽对梨花粉萌发和花粉管生长的影响

以砂梨(Pyrus pyrifoliaNakai)品种今村秋(Imamuraaki)和丰水(Hosui)为材料,分别用光学显微镜和荧光显微镜观察了离体和半活体条件下微丝骨架解聚剂细胞松弛素B(cytochalasin B,CB)和稳定剂鬼笔环肽(phalloidin)对梨花粉萌发和花粉管生长的影响.结果表明:(1)低浓度(10μg/mL)鬼笔环肽能促进花粉萌发和花粉管生长,但高浓度对花粉萌发和花粉管的生长具有抑制作用;CB抑制花粉萌发和花粉管生长,且抑制效应随其浓度的增加而增强.(2)鬼笔环肽处理柱头后进行自花授粉,可明显促进自花花粉萌发和花粉管的生长,而CB处理柱头后异花授粉则抑制异花花粉萌发及其花粉管生长.可见,微丝骨架参与了梨花粉萌发和花粉管生长过程,并参与了梨自交不亲和反应的调节.     

梨自交不亲和及其亲和突变品种花柱内S4（S4^SM）基因的表达与作用的比较

提取梨 (PyrusserotinaRehd .)自交不亲和品种“二十世纪”(基因型为S2 S4 )、自交亲和的突变品种“奥嗄二十世纪”(S2 SSM4 ,SM =Stylar_partmutant;花柱部分突变 )及其亲和后代花柱的可溶性蛋白。经等电聚焦电泳 (IEF_PAGE)分析表明 ,“奥嗄二十世纪”及其后代花柱仍存在SSM4 蛋白 ,但其含量逐代减少 ,同时发现“奥嗄二十世纪”的SSM4 基因仅在柱头表达 ,而“二十世纪”的S4 基因表达的部位除了柱头外 ,还包括花柱上部及花柱下部 ,且表达量呈现从柱头到花柱下部下降的趋势。S蛋白经等电聚焦电泳的凝胶板进行RNase活性染色处理 ,也得到相同的结果。从花柱 (包括柱头 )中纯化出的S蛋白经SDS_PAGE电泳后进行RNase活性染色的结果表明 ,S4 与SSM4 蛋白的分子量相近 (约 30kD) ,并且均具有RNase活性。进一步以酵母RNA为基质测定的比活性也基本相等 ,约为 2 75U·min-1·mg-1蛋白。在离体条件下 ,上述两种S蛋白 (S_RNase)也以相同的程度抑制S4 或SSM4 花粉发芽及花粉管伸长。研究证明 ,自交亲和突变品种“奥嗄二十世纪”的SSM4 基因也具有原始自交不亲和品种“二十世纪”S4 基因的功能。因此 ,其自交亲和的原因可归结为SSM4 基因的表达量较少及SSM4 基因仅在柱头中表达的缘故。     

梨花柱S-RNase对花粉管超微结构的影响

采用光学显微镜和透射电子显微镜研究了离体条件下不同品种梨花柱S—RNase对异花(亲和)及自花(不亲和)花粉萌发和花粉管生长及其超微结构的影响。结果表明,花柱S—RNase抑制不亲和花粉的萌发和花粉管的生长,对亲和花粉的萌发和花粉管的生长基本没有影响。花粉生长初期,亲和及不亲和花粉管超微结构相似;但培养24h以后,亲和花粉管中充满细胞质和细胞器,而不亲和花粉管中只有靠近花粉管前端有少量细胞质,细胞壁增厚,细胞壁与细胞质之间有一层胼胝质和电子透明区间隔。     

Post‐pollination process in a partially self‐compatible distylous plant,Primula sieboldii (Primulaceae)

Pollen germination and pollen‐tube growth under natural conditions were observed in a population of a distylous species, Primula sieboldii, in which partial self‐compatibility has been demonstrated in some long‐styled genets. We observed post‐pollination processes microscopically in styles collected after self‐morph and inter‐morph hand pollination (with standardized pollen load on the stigmas) in four genets each from the following three ‘genet types’: self‐incompatible long‐styled (SI), partially self‐compatible long‐styled (SC) and self‐incompatible short‐styled morph genets. Irrespective of the genet type, pollen germination began within 24 h after pollination and tubes of pollen reached to the style base with 48–96 h after inter‐morph pollination. Although pollen tubes germinated after self‐pollination in the SC genets, the number of germinated pollen tubes was significantly lower than in the case of inter‐morph pollination. Few pollen tubes germinated after self‐pollination of the SI or short‐styled genets. In SC genets, the rate of pollen‐tube growth did not differ between self‐morph and inter‐morph pollination (～1.9 mm/day). Therefore, differences in self‐compatibility between SC and SI genets in P. sieboldii are likely to be attributable to differential pollen germination rates rather than to differential pollen‐tube growth rates.     

Disentangling the components of pollen limitation in a widespread herb with gametophytic self-incompatibility

Premise

Seed production is frequently limited by the receipt of insufficient or low-quality pollen, collectively termed “pollen limitation” (PL). In taxa with gametophytic self-incompatibility (GSI), incompatible pollen can germinate on stigmas but pollen tubes are arrested in styles. This allows for estimates of pollen performance before, during, and after self-recognition, as well as insight into the factors underlying pollen quality limitation in GSI taxa.

Methods

We scored pollen performance following self and outcross pollinations in Argentina anserina to identify the location of self-recognition and establish the relationship between pollen tubes and seed production. We then estimated quantity and quality components of PL from >3300 field-collected styles. We combined our results with other studies to test the prediction that low pollen quality, but not quantity, drives higher PL in self-incompatible (SI) taxa than in self-compatible taxa (SC).

Results

Self and outcross pollen germinated readily on stigmas, but 96% of germinated self-pollen was arrested during early tube elongation. Reproduction in the field was more limited by pollen quality than by quantity, and pollen failure near the location of self-recognition was a stronger barrier to fertilization than pollen germination. Across 26 taxa, SI species experienced stronger pollen quality, but not quantity, limitation than SC species.

Conclusions

Evaluating pollen performance at multiple points within pistils can elucidate potential causes of pollen quality limitation. The receipt of incompatible pollen inhibits fertilization success more than insufficient pollen receipt or poor pollen germination in A. anserina. Likewise, pollen quality limitation drives high overall PL in other SI taxa.     

The effect of cherry leaf roll virus infection on the performance of birch pollen and studies on virus replication in germinating pollen

The rates of germ tube elongation in vitro of pollen from cherry leaf roll virus (CLRV)-infected birch (Betula pendula) did not differ significantly from those of pollen from virus-free trees. Differences in percentage germination of pollen collected from trees at different sites were significant but percentages of germination of pollen from virus-infected and virus-free trees did not differ greatly from one another. In vivo, pollen from infected trees germinated on healthy and CLRV-infected stigmas and callose plugs formed in both types of tissues. However, the extent of callose plug formation was greater in the pollen tubes of virus-free grains germinating in infected stigmas than in reciprocal crosses. CLRV coat antigen was detected by ELISA in stigmatic tissue, from healthy trees, on which virus-carrying pollen grains had germinated. Using fluorescein isothiocyanate conjugated to CLRV-specific γ-globulin, viral coat antigen was detected throughout germ tubes from virus-carrying but not virus-free pollen germinating in vitro. Protoplasts released following Driselase digestion of pollen germ tubes from virus-infected trees contained CLRV antigen detectable by ELISA. During germination of virus-infected pollen there was little synthesis of viral coat protein or nucleic acid but, following inoculation with purified virus particles, protoplasts made from healthy germinating pollen produced increasing amounts of CLRV-specific antigen implying that CLRV replicated in this system.     

Effects of prior self-pollination on outcrossed seed set in tristylous Pontederia sagittata (Pontederiaceae)

The potential inhibitory effects of incompatible pollen on outcrossed seed set were investigated in mass-flowering, self-incompatible, tristylous Pontederia sagittata. Prior application of self pollen, followed after 2, 4, or 6 h by compatible pollen, was conducted on five genotypes of each of the three style morphs under uniform glasshouse conditions. The greatest reductions in seed set occurred in pollinations of the long-styled (L) morph at the 6 h time interval. Smaller reductions were also found for this treatment in the mid-styled (M) morph. No significant reductions in seed set were observed in the short-styled (S) morph or in the other morphs at shorter time intervals. Observations of pollen germination and pollen tube growth indicated that the lack of inhibitory effects in the S morph may occur because relatively few pollen grains adhered to stigmas in selfpollinations. In the L and M morphs, early germination of self pollen may cause physical clogging of the stigma and style, resulting in a reduced number of compatible pollen tubes in styles. Observations of the structural integrity of styles indicated that prior germination of self pollen resulted in more rapid onset of pistil senescence, particularly in the L morph. These influences may contribute to the morph-specific differences in seed set observed following prior self-pollination of outcrossed flowers. The negative effects of incompatible pollen are likely to be most evident where ecological factors cause delays in the delivery of outcross pollen to stigmas.     

The genetics of self-incompatibility inOenothera rhombipetala

Pollen tubes were grown under controlled temperatures in stigmas and styles which had been cut from the flowers at the point of juncture of the style with the ovary. At constant temperatures the tubes compatible with the styles grew rapidly at uniform rates. They showed accelerated growth with increase in temperature within the range 10°–30°C. Growth of incompatible tubes was irregular and slow and was neither measurably accelerated nor depressed within this range of temperatures. Routine tests for compatibility were conducted at 27°C; at this temperature compatible tubes grew through the entire length of the styles (ca. 58 mm) in 9 hours. Much of the incompatible pollen failed to germinate; that which did, grew to a maximum length ofca. 2 mm inca. 4 hours. The incompatible tubes did not even enter the styles but remained confined to the stigmas. ThreeS alleles were involved and conformed in their behavior to the gametophytic system of incompatibility. Colchicine-induced tetraploid derivatives of these plants showed no increase in compatibility.     

沙田柚花粉低温贮藏的研究

研究了脱水时间 ,化冻方式和培养基对沙田柚 ( Citrus grandis var. shatinyu Hort. )花粉低温贮藏后萌发率的影响。结果表明 :贮藏 3个月 ,6个月期间 ,脱水 50 min的花粉萌发率最高 ;沙田柚花粉低温贮藏的最适含水量为 13.5%。沙田柚花粉适合快速化冻法。在 4种培养基中 ,萌发率无显著差异 ,但花粉管生长不一致     

砧木对梨叶芽休眠的影响及其与多胺代谢的关系

以分别嫁接在杜梨和豆梨上的砂梨品种‘丰水’为试材,研究了2008和2009年11～12月气温变化和不同砧木对‘丰水’梨叶芽休眠进程的影响,分析叶芽中游离态和束缚态内源多胺种类和含量的变化,结果表明：嫁接在豆梨上的‘丰水’叶芽自然休眠结束的时间要比嫁接在杜梨上的‘丰水’叶芽早10d左右,且游离态和束缚态腐胺（Put）、戊二胺（Cad）、己二胺（Hex）、亚精胺（Spd）和精胺（Spm）5种内源多胺含量开始升高的时间与供试材料叶芽自然休眠结束的时间一致,表明梨叶芽的休眠进程与砧木种类和多胺代谢有密切关系,尤其是与束缚态多胺含量变化的关系更为密切。