渐危植物鹅掌楸的授粉率及花粉管生长

对鹅掌楸Liriodendron chinense 3个自然居群连续3年直接统计落置柱头上的花粉量,其中 61.9％以上的柱头接受了花粉。每柱头上的平均花粉量为4.4～42.6粒,明显超过胚珠数2。授粉率是 结实率的6～8倍,鹅掌楸结实率低的主要原因可能不是由于花粉的限制。落置柱头的花粉几乎均可萌 发,少数花粉管穿过花柱道,经珠孔端进入胚囊,暗示花粉间存在着强烈的竞争。授粉率和柱头上的平 均花粉量与结实率呈正相关(R＝0.77,R=0.69)。在柱头上人工授同株和异株的花粉,花粉管生长速 率看不出明显差异,到达胚珠的时间在48～60 h之间。花开花药未裂时柱头上落置较高比例的花粉, 由于先期到达柱头的花粉最有可能受精,表明鹅掌楸有异交为主的繁育系统。人工处理包括套袋、套网、去除花被片,其结实率大大降低,而去雄后的结实率接近自然传粉结实率支持了这一观点。     

外来入侵植物胜红蓟的胚胎学观察及繁殖系统研究

以采自我国广东江门和广州两个种群的胜红蓟(Ageratum conyzoides L.)种子为材料,采用流式细胞种子筛选技术(FCSS)和人工控制授粉实验,对胜红蓟的繁殖系统进行研究,并结合整体透明技术和微分干涉差(DIC)显微镜观察法,对其胚珠发育过程和花药结构进行细胞胚胎学观察。种子筛选结果显示,胜红蓟的种子既可以通过有性生殖产生,又可以通过不需要假受精的无融合生殖产生,属于兼性无融合生殖类型。开放性授粉和套袋处理之间的结实率均较高,分别为88%±1.2%和86.2%±1.2%,两者之间无显著差异;而去雄处理的结实率和开放性授粉、套袋这两种处理之间差异显著。胚珠的细胞胚胎学观察结果发现,胜红蓟的有性生殖胚囊发育方式为蓼型,无融合生殖胚囊的发育方式为山柳菊型。胜红蓟的花粉粒在花药内就开始萌发出花粉管,具有闭花受精特性。研究结果表明闭花受精和兼性无融合生殖等繁殖特性保证了胜红蓟在各种生存环境下的结实量,提高其在新生境中归化和入侵的可能性。     

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。     

恶性入侵植物豚草的繁育系统特性

豚草(Ambrosia artemisiifolia L.)是原产于北美的一年生恶性入侵植物,目前已广泛分布在我国东北、华北、华中和华东等地,威胁农业、生态和民众健康。该种雌、雄头状花序同株,以种子繁殖,因而其有性繁殖特性对其扩散和爆发具有重要影响。运用套袋授粉和联苯胺-过氧化氢法等实验方法,对入侵江苏常熟的豚草种群的繁育系统特性进行了研究。结果表明,豚草花粉活力在开花后第4天开始出现,第8天到第10天花粉活力比较高;柱头可授性在开花第2天开始出现,第5天到第8天柱头可授性较高;同植株上同一时期开花的雌花的柱头可授性和雄花的花粉活力有5d左右的重叠期。不套袋处理(自然条件)和异株授粉处理下,豚草的结实率都比较高,分别达48.4%和44.4%,两者间无显著差异(P0.05);而同株授粉处理的结实率较低,仅3.4%,极显著低于不套袋处理和异株授粉的结实率(P0.01)但显著大于0(P0.05)。分析表明,豚草属于自交不亲和种,但又可部分自交亲和,不具有无融合生殖特性。因此,对在新分布区的零星豚草植株进行及时防除,将取得事半功倍的效果。     

野菊与菊花杂交中花粉活力和柱头可授性及胚胎发育研究

应用石蜡制片、活体压片、光学显微镜及扫描电子显微镜观察等方法,研究了四倍体河南云台山野菊(Dendranthema indicum)与栽培菊花'钟山金山'(D.grandiflorum 'Zhongshanjinshan')种间杂交中父本花粉活力、花粉在柱头萌发、花粉管生长及胚胎发育情况等.结果发现,父本云台山野菊的花粉活力在授粉时为12%左右.人工授粉后的不同时间,在柱头上都观察到正常萌发的花粉粒,且花粉管都能进入柱头,其中,在授粉后0.5 h时,平均每柱头有5.9粒花粉萌发;12 h时,为59.9粒;而24和48 h时,则分别降为47.1和35.7粒.此外,在授粉后8、10、12和15 d时,分别在49.1%、40.8%、39.7%和38.5%子房内观察到正常发育的胚胎,最终杂交结实率为44.8%,而母本自然开放结实率为52.3%.研究表明,授粉前其多数母本雌蕊发育良好、授粉后多数花粉能在柱头正常萌发和花粉管正常生长,在受精后大部分胚胎发育正常是野菊与栽培菊种间杂交较高结实率的重要保证,而授粉前父本较低的花粉活力对杂交结实率影响不大.     

濒危植物焕镛木的兼性无融合生殖

2001—2002年连续两年在广西环江木论和广西罗城大黄泥的2个焕镛木(Woonyoungia septentrionalis (Dandy) Law)自然种群中,对单性异株的濒危植物焕镛木进行繁育系统测定,对即将开花的雌花花蕾分别进行套袋、套网、人工授粉处理,并用自然授粉雌花作对照,其座果率和结实率统计结果表明：自然授粉、人工授粉、套袋和套网处理的花均能结实,但它们的座果率和结实率存在较大的差异。在两个种群中,人工授粉和自然授粉的总结实率(PERS)均比套袋和套网处理的高,其中人工授粉的最高,套网处理的最低。由此可见,焕镛木既能通过有性生殖方式结实,又能通过无融合生殖方式结实,而且这两种生殖方式获得的种子均能萌发成幼苗,由此断定,焕镛木的繁育系统为兼性无融合生殖。这是首次报道木兰科植物存在无融合生殖现象。     

丹东蒲公英专性无融合生殖特性

采用田间去雄套袋、花粉活性与花粉管萌发观测等交配系统实验,辅以染色体分析、胚囊发育观察以及流式细胞仪检测,对丹东蒲公英(Taraxacum antungense)生殖特性进行系统研究。结果表明,去雄套袋情况下,结籽率高达96%,由此证明交配系统具有极高的无融合发生率;自花授粉不能萌发,花粉活性仅为20.6%,异花授粉虽可萌发,但生长缓慢,花粉在到达子房前,花粉管已停止生长并消失;二分体时期,合子端大孢子发育为功能性大孢子,珠孔端大孢子退化消失,经蓼型发育,形成7细胞8核,卵细胞继续发育形成原胚,中央细胞形成胚乳,其它细胞退化消失,花未开放前,已形成球形胚。此外,丹东蒲公英为三倍体。流式细胞仪检测结果表明,丹东蒲公英为专性无融合生殖,胚乳自主发育。综合以上研究结果,说明丹东蒲公英具专性无融合生殖特性。     

草豆蔻传粉生物学的研究

比较研究了花柱卷曲性植物草豆蔻 Alpinia hainanensis两种表型(上举型和下垂型)的传粉生物学特性。结果表明:草豆蔻的花柱卷曲运动节律与其他已报道的山姜属Alpinia植物基本一致,而下垂型花的花柱卷曲运动明显滞后于上举型花约2 h。草豆蔻的花柱卷曲运动中存在一些不同步的现象,甚至在同一花序上的两朵花的花柱运动节律亦会表现出不一致的现象,但上举型花的花药开裂时间却严格同步,都发生在同类型个体的柱头全部位于花药上方之后进行。草豆蔻上举型花的花蜜分泌量、单花花粉量、花粉/胚珠比率(P/O)均明显比下垂型花多,而两种表型的胚珠数、花蜜糖含量、氨基酸含量无显著差异。在草豆蔻的单花期内不同时间段进行人工授粉,上举型花在其柱头位于花药下方时进行人工授粉,其结实率明显比柱头位于花药上方时人工授粉的处理高,下垂型花则没有显著差异。在自然居群中,草豆蔻的主要传粉者是无垫蜂Ameglla sp.和两种木蜂Xylocopa sp.,但存在传粉者不足而影响结实率的现象。完全套袋、去雄和去雌(去柱头)处理的均不结实,表明草豆蔻中不存在无融合生殖、主动自花授粉和滞后自交的生殖保障现象。而人工自交和异交均具有较高的结实率,表明草豆蔻为自交亲和植物。草豆蔻的繁育系统是具花柱卷曲性运动的异花授粉的交配系统。     

濒危植物焕镛木的兼性无融合生殖

2001～2002年连续两年在广西环江木论和广西罗城大黄泥的2个焕镛木(Woonyoungia septentrionalis(Dandy)Law)自然种群中,对单性异株的濒危植物焕镛木进行繁育系统测定,对即将开花的雌花花蕾分别进行套袋、套网、人工授粉处理,并用自然授粉雌花作对照,其座果率和结实率统计结果表明:自然授粉、人工授粉、套袋和套网处理的花均能结实,但它们的座果率和结实率存在较大的差异.在两个种群中,人工授粉和自然授粉的总结实率(PERS)均比套袋和套网处理的高,其中人工授粉的最高,套网处理的最低.由此可见,焕镛木既能通过有性生殖方式结实,又能通过无融合生殖方式结实,而且这两种生殖方式获得的种子均能萌发成幼苗,由此断定,焕镛木的繁育系统为兼性无融合生殖.这是首次报道木兰科植物存在无融合生殖现象.     

牛耳朵传粉生物学研究

苦苣苔科(Gesneriaceae)报春苣苔属(Primulina Hance)是一个近年来备受关注的类群,其纷繁复杂的物种多样性和属下种间的特有分布引起了分类学家和植物学研究者的极大兴趣。该属除了极少数的物种如牛耳朵[(Primulina eburnea(Hance)Y. Z. Wang)]以外,绝大部分的物种为狭域分布或地方特有种,其分布范围很窄。为了揭示牛耳朵的传粉生物学和繁育系统对其生殖过程和拓殖能力的影响机制,作者系统地研究了牛耳朵的开花物候、花粉与柱头活性、访花昆虫的种类和访花行为、花粉胚珠比、OCI指数和套袋实验结实率,探究其传粉等生殖过程对牛耳朵的广布是否有正面影响。结果表明:牛耳朵的自然花期是3—5月,全花期约45 d,其中盛花期约20 d,单花期6～8 d; 开花后1～2 d花粉活力最强,开花前柱头没有可授性; 花粉胚珠比为537; 杂交指数为5; 去雌套袋、去雄套袋均无法结实,说明本种不存在无融合生殖; 与自然授粉相比,自花授粉结实率略低,异花授粉结实率略高,说明自交亲和; 牛耳朵的主要传粉者是花条蜂(Anthophora florea)和熊蜂(Bombus sp.)。花蜜产量较高、花粉量较大、花粉活力较强等特点,有利于牛耳朵完成传粉和结实的整个繁殖过程。因此,这一结果显然有利于牛耳朵的拓殖进而广布在我国华南至西南地区的喀斯特地区。     

The cross-fertilization mechanism and reproductive interference of self-pollen of Medicago varia ‘Xinmu No. 1’

Some control tests and a technique employing the fluorescent properties of pollen and pollen tube stained with aniline blue dye were utilized to study the stigma receptivity, the optimum pollination period, the self-pollen clogging effect, pollen germination and the pollen tube growth state in alfalfa. The results can be summarized as follows: (1) The stigma receptivity and flower longevity barely overlap. On the first day after pollination, the pollination efficiency was the highest, with a pod-seting rate per raceme of 65.17%±2.01% (P<0.01). The period of the highest pollination efficiency was the same as the peak of stigma receptivity. When pollination was on the fourth day after flowering, the pod-setting percentage per raceme was still 34.25%±6.73%. (2) Self-pollination led to a lower pollen germination frequency and a lower pollen tube growth rate than cross-pollination. When artificial cross-pollination was applied after self-pollen grains on the stigma was re-moved, the pod-setting percentage per raceme was 72.31%±6.24%, and the number of seed per pod was 3.46±0.25, In contrast, these two values were 51.76%±5.37% (P<0.01) and 2.11%±0.18 respectively (P<0.01) when the self-pollen grains were not removbed in the artificial cross-pollination experiements. Hence, the clog-ging effect of self-pollen to stigma was significant. (3) In the breeding system of alfalfa, there are two mecha-nisms that help the plants to avoid self-fertilization: dichogamy and self-incompatibility. However, due to the structure of flowers, they were not effective enough to improve cross-fertilization rate, even if the stamen matured earlier than the stigma. The typical self-incompatibility system guaranteed the lower self-fertilization rate, but cannot avoid the reproductive interference of self-pollen.     

Calcium crystals in the anther of Petunia: the existence and biological significance in the pollination process

Using an X-ray microanalysis system fitted with variable-pressure scanning electron microscopy, we noted that many calcium crystals accumulated under the stomium in the anther of Petunia. When the anther was dehisced and pollen grains were released from the stomata, the calcium crystals adhered to pollen grains and moved to the stigma together with pollen grains. In contrast, an X-ray microanalysis of the stigma surface before pollination detected no calcium emission on the stigma surface. Furthermore, pollen germination and pollen tube growth in medium without Ca occurred as in complete medium. However, after the pollen grains had been washed with abundant germination medium without calcium, pollen germination in the medium without Ca was inhibited. These results show that the calcium crystals dissolved in the aqueous drop under the exudate on the stigma and supplied calcium ions for pollen germination. In addition, calcium crystals were produced not only in the anther of Petunia but also in Nicotiana, suggesting that calcium crystals supply pollen grains with the calcium ions required for pollen germination and serve to improve reproduction efficiency in Solanaceae.     

甜柿巨大花粉萌发特征及辐射敏感性研究

以‘禅寺丸’（Diospyros kakiL.f)为试材,对巨大花粉萌发率,花粉管伸长,亲和性及辐射敏感性进行了研究。结果表明：（1）巨大花粉在培养基和柱头上正常萌发,不存在萌发及亲和性障碍;（2）巨大花粉萌发迟缓及低萌发率造成其与普通花粉受精竞争中处于劣势;（3）巨大花粉和普通花粉对^60Coγ-射线辐射敏感性有差异。巨大花粉的敏感性低于普通花粉,1200Gy为刺激巨大花粉萌发的适合剂量,同时可抑制普通花粉萌发,从而可相对提高巨大花粉的受精竞争力;（4）辐射延迟效应造成巨大花粉的萌发率在一定期间内有下降趋势。但自身的修复机制可部分恢复其生活力。     

茜草科二型花柱植物滇丁香传粉精确性研究

异型花柱是一种受遗传因素控制的花型多态性现象,其适应意义在于提高不同花型间传粉精确性,从而促进异交。为检验二型花柱植物滇丁香（Luculia pinceana）是如何促进异交并确定有效传粉者的类型,实验统计自然种群中长、短柱型的植株数量,测量花部特征,统计自然种群植株柱头上的型间花粉落置的数量和比例,比较型内、型间花粉的萌发率以及花粉管长度,比较不同人工授粉处理结实情况,观察传粉昆虫类型并测量虫体特征。结果表明（1）滇丁香自然居群中长、短柱型植株的数量没有显著性差异。长柱型的筒深、筒直径、开口直径、雄蕊长和柱头长均显著小于短柱型,而长柱型的叶片长和宽、雌蕊长、花药体积、花粉粒体积、柱头厚均显著大于短柱型。（2）自然种群植株柱头上型间花粉所占比例显著低于型内花粉的比例。（3）型间授粉的花粉萌发率以及花粉管长度均大于型内授粉和混合授粉。（4）型间授粉的结实率显著高于型内授粉的结实率,型内授粉具有一定的结实。（5）夜行性长喙条背天蛾（Cechenena lineosa）是滇丁香的有效传粉者,其吻长和滇丁香的花筒深相适应。本研究表明二型花柱植物滇丁香主要是通过与其花部特征相匹配的长喙天蛾实现有效的传粉,通过促进型间花粉的萌发和花粉管的伸长而有利于型间授粉结实,提高传粉精确性。     

The effects of stigma age on receptivity in Silene alba (Caryophyllaceae)

Silene alba, a perennial, dioecious plant, produces flowers that open in the evening and can remain open and receptive to pollination for up to 5 d, though in hot and dry conditions the flowers will wilt during the day only to reopen night after night. In the field, it is visited by two different kinds of pollinators with differential success: moths visit the flowers at night, and their movements result in broad pollen dispersal and large seed production, whereas bees, wasps, and flies visit the flowers in the mornings and have decreased pollination effectiveness. However, this differential success may be due to a decrease in stigmatic receptivity soon after the flowers open. We performed controlled pollinations to determine the effect of stigma age on pollen germination and seed set. We pollinated flowers at 12-h intervals up to 120 h and divided these into two sets: from one set, we removed stigmas 24 h after pollination to examine percentage of pollen germination. The second set of flowers was allowed to produce fruits, and the seeds were counted and weighed. Pollen germination declined significantly with stigma age, but there was no significant effect of stigma age at pollination on the number or mass of resulting seeds. Thus, the decreased pollination success of bees is not due to a decrease in stigmatic receptivity but is most likely a result of pollinator inefficiency.     

Pollination Rates and Pollen Tube Growth in a Vulnerable Plant,Liriodendron chinense (Hemsl.) Sarg. (Magnoliaceae)

The limited seed production of insect-depended plant, Liriodendron chinense was once considered to be pollen-limited and insufficient cross pollination. In this study, we counted pollen grains deposited on stigmas in three populations in Guizhou, Hunan and Zhejiang provinces of China respectively. Over 61.9% stigmas were pollinated. From 1994 to 1996, the mean number of pollen grains on each stigma ranged from 4.4 to 42.6, much more than ovules(2) in each pistil. Based on observations of three years, both the pollination rate and pollen quantity on stigmas significantly affected seed set. When flowers opened without stamens dehiscencing at the early stage of anthesis, stigmas received considerable quantity of pollen grains. Pollen grains from different sources were able to germinate and pollen tube growth rates were not greatly variable. It is very likely that pollen grains arrived first would fertilize eggs. Since only several pollen tubes went through the stylar canal, the potential pollen competition may exist. In this case, there would be strong selection on floral syndrome which benefit early insect visits. Pollen grains from the early visits would have a greater chance to fertilize ovules than those from later visits, which implies that cross pollination is the predominant breeding system of this plant. The conclusion was also confirmed by following four artificial experiments. Three treatments, including flowers bagged, netted or with the perianth removed, all reduced seed set clearly, but flowers with the stamens removed (emasculation)had a higher seed production by open pollination. As the rates of deposited stigmas in three populations were 6～8 times more than full seed set, we considerthat lower seed production in this plant may not mainly be due to pollen limitation.     

Mating system,outcrossing distance effects and pollen availability in the wind-pollinated treeline species Polylepis australis BITT. (Rosaceae)

Fragmentation may negatively affect plant fitness through pollen limitation and increased levels of inbreeding. Effects of fragmentation may vary with regard to life form and breeding system, and few studies exist for wind-pollinated trees. We examined the effects of hand-selfing, varying outcrossing distances and pollen addition on seed mass and germination rate of Polylepis australis BITT. (Rosaceae), a wind-pollinated treeline species endemic to Argentina. We also investigated pollen germination on the stigma and pollen tube growth to determine compatibility resulting from selfing and outcrossing. Selfing reduced seed germination rates with significant differences between open pollination and outcrosses at 30 km. In addition, we found a tendency for pollen germination and pollen tube growth to decrease following selfing. Between-fragment crosses resulted in a trend of higher reproductive output than within-fragment crosses, whereas values were similar between open pollination and between-fragment crosses. Pollen addition did not increase reproductive success neither in small nor in larger fragments. Our results suggest that highly isolated P. australis forests have a potential for inbreeding depression through selfing and within-fragment crosses. However, the results also indicate that pollen flow between P. australis forest fragments is still effective at the current fragmentation level, counteracting negative effects on seed quality resulting from reproductive isolation.     

Relationship of stigma behaviors and breeding system in three Mazus (Phrymaceae) species with bilobed stigma

Sensitive stigma has been recognized to facilitate outcrossing. We hypothesized that species with different levels of sensitivity might have corresponding differences in components of their breeding system. In this study, three Mazus species with bilobed stigmas were used to test the hypothesis. We explored stigma behaviors of the species in reaction time, recovery time, permanent closing time, and the minimum pollen load causing permanent closure. We investigated floral traits, pollinator type and behavior, pollination intensity, and natural schedule of pollen deposition on stigma. Moreover, we evaluated the mating system of the species by checking seed set after controlled pollination treatments, namely, natural flowers with open pollination, enclosed flowers without pollination, and enclosed flowers with self and outcross hand pollination. Results indicated that stigma of M. pumilus (N. L. Burman) Steenis was not sensitive, whereas stigmas of M. miquelii Makino and M. stachydifolius (Turcz.) Maxim. closed and reopened quickly in response to pollination. Accordingly, hand pollination treatments revealed that seed set of self-spontaneous pollination in M. pumilus was similar to the other treatments. For M. miquelii, outcross pollen resulted in significantly higher seed set than self-pollen.Mazus stachydifolius was self-incompatible. Additionally, the corresponding characteristics in other components of the breeding system for each species were found. Our study indicated that the sensitivity of bilobed stigma might be linked with floral traits and the mating system in a given species. Sensitive stigma should be regarded as an evolutionary mechanism for enhancement of outcrossing.