珍稀濒危植物陕西羽叶报春的开花及传粉生物学研究

陕西羽叶报春为中国珍稀濒危植物,研究其开花物候、传粉者及访花行为、花粉活力、柱头可授性、繁育系统,以明确陕西羽叶报春的传粉特性及制濒机制。以勉县陕西羽叶报春野生居群为研究对象,通过野外观察和人工授粉实验首次对陕西羽叶报春的传粉生物学进行研究。结果表明:(1)陕西羽叶报春种群花期为2-4月中旬,单花花期一般为14~17 d;单花开花进程中花部形态有6个明显发育变化:全部被萼片包裹、花苞伸出并为深粉色、花朵打开为淡粉色、花冠为白色、花朵枯萎、花冠脱落花萼宿存。(2)陕西羽叶报春为异花授粉植物,蜜蜂属为其主要传粉者。(3)柱头在花粉未开裂前就具有可授性,可授性逐渐增强;花药在花瓣打开后的第3天才开始开裂、散粉,在第4天活力最高,为98.18%,此后逐渐减弱。(4)繁育系统检测显示,长、短柱型花的花粉/胚珠比分别为902.26和831.48,杂交指数为4;套袋实验显示,陕西羽叶报春异型交配可育,同型交配不可育。由此表明,陕西羽叶报春的繁育系统为异交型,自交不亲和,需要传粉者。综上所述,陕西羽叶报春传粉者种类少、自交不亲和、胚珠败育现象可能是导致陕西羽叶报春濒危的重要原因。     

欧报春二型花柱的繁育特性研究

二型花柱植物通常具有自交不亲和性,在不同植物中所表现的可育性不同。欧报春(Primula vulgaris)是典型的二型花柱植物,为了探究欧报春的繁育特性,通过温室栽培,对欧报春的长花柱和短花柱的花部特征、花粉和花柱形态、花粉活力、柱头可授性、花粉/胚珠比、杂交指数、杂交亲和性、花粉管观察进行研究。结果表明,(1)长花柱和短花柱的花冠直径、花冠筒长度、柱头高度、花药高度和花筒中部直径均表现出两型性;(2)花粉和柱头观察发现花粉极轴长、花粉赤道轴长、花柱直径、柱头乳突细胞和花粉数量均有差异;(3)长花柱的花粉/胚珠比为384.20,短花柱为369.70,属于兼性异交类型;长花柱和短花柱的花粉活力和柱头可授性能在较长时间内维持较高活力;(4)长花柱杂交指数值为5,短花柱为4,表明繁育系统类型为异交,部分自交亲和,需要传粉者;6种授粉组合均能结实,异型花间授粉的结实数明显高于同型自花授粉和异株同型授粉;短花柱为母本的异型花间授粉组合亲和性优于其他组合;欧报春存在自交不亲和性,长花柱的自交不亲和性低于短花柱。     

濒危植物安徽羽叶报春两种花型的繁育特性及其适应进化

二型花柱的维持机制和自然选择压力多年来一直是生态学和进化学研究领域的热点之一。通过实验室栽培和野外观察统计相结合的方法,对安徽羽叶报春两种花型(长柱花和短柱花)的形态特征、花粉活力、柱头可授性、花粉/胚珠比、自然授粉及结籽能力、自交亲和性等繁育特性进行了比较研究。结果表明:长、短柱花的花冠直径和裂片宽无明显差异,而花冠筒、雌蕊和雄蕊高、花粉数目及大小、P/O比均有显著差异。在自然条件下,长柱花所接受的总花粉数要明显高于短柱花的总花粉数,但所接受的异型花花粉数和平均每果结籽数两者无显著差异。长柱花和短柱花的花粉和柱头活力相似,均能在较长时间内维持较高活力,仅在开花末期显著下降。两种花型的花在自花授粉、同型异花授粉、异型花授粉条件下均能结籽,但异型花授粉的结籽数均明显高于自花授粉和同型异花授粉结籽数。在长柱花各种授粉方式中,花粉萌发率无明显差异,但异型花花粉管的生长速度明显比同型异花花粉和自花花粉的快,而在短柱花柱头上表现为异型花授粉的萌发率最高,但只要萌发后在花柱中的生长速度无显明差异。此外,综合上述结果,对二种花型花部综合征的维持机制及自然选择压力进行了探讨。     

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。     

绣球茜的二型花柱及其传粉生物学初步研究

首次对绣球茜(Dunnia sinensis Tutch)的花药和柱头高度、花粉和柱头形态,以及人工辅助授粉的结实情况等进行了研究.结果表明:绣球茜在同一居群中同时具有长花柱(Pin 型)和短花柱(Thrum)两种花型,长花柱植株的柱头和雄蕊高分别为9.61±0.36 mm和6.79±0.38 mm,短花柱则为6.11±0.56 mm和8.96±0.59 mm,表现出互补式雌雄异位的花部特征.通过人工辅助授粉,两种花型植株的自交和同型植株间异交都不结实,仅在型间异交方能正常结实,说明绣球茜中存在极强的自交不亲和和型内不亲和现象.在花粉与柱头形态方面:长柱型花的单花花粉量(38555.6±6258.4)显著地小于短柱型花(52145.45±8924.5)(t=4.009,P＜0.01),长柱花和短柱型花的胚珠数分别为38.73±1.85和39.1±2.25(t=-4.13,P＞0.01);长柱型花花粉粒的大小(极径21.27±0.70 μm)显著地小于短柱花(21.92±0.79μm)类型(t=3.890,P＜0.01),而柱头乳突大小长柱类型要大于短柱类型,表现出明显的二型性.研究结果表明绣球茜是一种典型的二型花柱植物.此外,对其传粉昆虫及昆虫访花行为的观察表明,蝶类(青风蝶属Graphium)和蜂类(熊蜂Bombus sp.和木蜂Xylocopa magnifica Smith)为绣球茜的主要访花昆虫和传粉昆虫,它们能够在两种不同花型植株之间有效地传粉.     

异质生境对二型花柱植物雪地报春花性状分化的影响

该研究以生长在新疆北部的二型花柱植物雪地报春(Primula nivalis)为研究材料,对7个低海拔(1 657~2 013m)森林居群和7个高海拔(2 400~2 700m)草原居群的花型比例进行调查,对花部特征进行检测,并对长、短花柱植株在不同海拔居群的花性状分化进行了比较研究,以揭示该类群植物的演化与环境的关系。结果表明:(1)在高海拔草原居群中长花柱植株比例高于短花柱植株。(2)比较草原和森林生态系统中的雪地报春居群,长花柱花形态特征间存在显著差异,但短花柱花形态特征间的差异不显著。(3)高海拔草原居群中长花柱花的花冠筒长度、雄蕊长度、雌雄蕊距离、雌雄异位程度以及雌雄蕊空间位置是导致花性状分化的主要因素。(4)在草原居群中,长花柱花的雄蕊、雌雄异位程度及花冠口至雄蕊距离的变化减少了长短花柱花间的互补式对应性。研究认为,在异质生境中长花柱花性状的变化是导致该物种长短花柱花性状分化的主要因素。     

安徽羽叶报春核心和边缘居群的形态变异比较研究

为了解濒危二型花柱植物安徽羽叶报春(Primula merrilliana)居群的形态变异,对其4个核心居群和3个边缘居群的14个形态性状进行了比较研究。结果表明,核心居群形态性状的变异系数为0.09~0.50,边缘居群为0.05~0.56,变异幅度两者间差异不明显;核心居群有8个性状的均值显著高于边缘居群(P0.05),仅有3个性状(花冠直径、花瓣宽和叶宽)为边缘居群高于核心居群。核心居群两型花间的互惠指数较高也较稳定(0.83~0.90),花型间互惠程度较高,而边缘居群的互惠指数差异较大(0.60~0.93),不同居群的互惠程度变化较大。因此,两型花柱植物安徽羽叶报春的形态性状变异基本支持"中心丰富模型"假说,并对其保护策略进行了探讨。     

胭脂花野生居群花粉形态多样性研究

胭脂花是分布于中国北部的报春花科报春花属花柱二型植物,为了阐明胭脂花花粉形态的多样性,对胭脂花分布范围内的7个野生居群的花粉形态进行了测定。结果表明,胭脂花花粉属N3P4C5型,极面观多为三角形,具三孔沟,少数为四边形,具4孔沟;短花柱植株的花粉粒较大(16.53×17.58μm),花粉粒外壁粗糙,表面纹饰多,网眼大;长花柱植株的花粉粒较小(12.95×13.72μm),花粉粒外壁较光滑,表面纹饰少,网眼小。不同居群间花粉粒在大小、网眼直径、外壁纹饰等方面存在显著差异,花粉形态变异系数在4.88%~35.48%之间;短花柱类型各居群的平均变异系数为16.18%,居群间差异较大,其中庞泉沟居群变异系数最大(24.05%),海坨山居群最小(10.13%);长花柱类型各居群的平均变异系数为14.89%,庞泉沟居群的变异系数最大(19.75%),五台山与海坨山居群最小(12.50%),长花柱类型的花粉性状比短花柱植株更为稳定。UPGMA聚类分析发现,7个居群聚为两大类,除庞泉沟居群外,其他居群聚为一组,地理距离相近的居群一般聚在一起。研究结果为胭脂花种质资源遗传多样性及育种研究提供了依据。     

入侵植物酢浆草在中国的花型分布特点及自花授粉

通过野外调查和馆藏标本(IBSC)的查阅,对广泛分布于中国的入侵植物酢浆草Oxalis corniculata的花型、开花物候、访花昆虫和花粉胚珠比(P/O)进行了观测.通过人工辅助授粉实验检测了其繁育系统;使用扫描电子显微镜(SEM)观察了自花花粉在柱头上的萌发情况并使用荧光显微镜(FM)观察了花粉管在花柱中的生长情况.结果表明:酢浆草在中国有3种花型,半等长花柱型、准等长花柱型和长花柱型,大部分的居群只有半等长花柱型,分布于人为干扰较大的生境(如路边);云南安宁松树林下的自然居群和采自广东的两份标本为长花柱型花:少部分居群内兼有半等长花柱型和准等长花柱型植株.酢浆草花期从2月到11月.其中3_9月为开花高峰期:单花花期2天,花开当天傍晚花瓣会合拢,第二天早上花瓣再次打开,到花开第二天傍晚花瓣萎蔫.花蕾开放前花药已开裂,柱头有活性,大部分花粉散落到柱头上,且花粉已开始萌发;花粉胚珠比为83.3±33.08.有少量花蜜分泌.唯一的访花昆虫为一种小灰蝶Pseudozizeeria sp.,但并不是有效的传粉者.开花前套袋处理结实率(88.12±3.38)与自然结实率(88.03±2.48)基本没有差异:荧光显微镜观察证明自花花粉的花粉管可在花开第一天生长到达胚珠,说明酢浆草半等长花柱植株是自花授粉.     

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。     

Heterostylous morph differences in pollen transfer and deposition patterns in Primula sieboldii on a visitation by a queen bumblebee, measured with a semi-natural experimental system

Abstract Under a semi-natural setting the between-morph pollen exchange patterns were studied in distylous Primula sieboldii flowers by measuring pollen removal from the anthers on a single visit by a Bombus diversus tersatus queen, and stigmatic pollen deposition along the sequence of the visitation of the opposite-morph flowers by the bee. Despite the twofold larger number of pollen grains produced in a single flower of the long-styled morph compared to that of the short-styled morph, no significant difference in pollen removal from a flower was found between the morphs. The stigmas of the long-styled morph received significantly more opposite-morph pollen grains than those of the short-styled morph on a single visit by the bee. Sufficient legitimate pollen grains, surpassing the ovule number, were loaded on the stigmas of 27% and 17% of visited flowers of the long- and short-styled morphs, respectively. The short-styled morph could more efficiently donate pollen to the opposite morph stigmas than the long-styled morph.     

Pollen transfer patterns in a distylous plant with overlapping pollen-size distributions

In this study we quantify the relative rates and amounts of between-morph (legitimate) cross-pollination, within-morph (illegitimate) cross-pollination and self-pollination in a natural population of distylous Gaertnera vaginata (Rubiaceae) endemic to the oceanic island of La Réunion in the Indian Ocean. Since pollen size distributions of each morph overlap we develop a method, based on chi-squared comparisons of observed to expected pollen size distributions, to quantify pollen transfer proficiencies. This method, unlike previous methods, does not assume an equal chance of pollen transfer for pollen grains in the region of overlap. The probability that a single pollen grain from a short-styled plant undergoes legitimate pollen transfer to long-styled flowers (4.19×10⁻²) was more than twice that for illegitimate transfer to other short-styled plants (1.56×10⁻²). In contrast, the probability that a single pollen grain from a long-styled flower undergoes legitimate pollen transfer to short-styled flowers (1.65×10⁻²) was no greater than that of illegitimate pollen transfer to other long-styled plants (2.15×10⁻²). The transfer proficiencies of individual pollen grains are thus highly asymmetric. However, because long-styled flowers produce significantly more pollen, the ratio of legitimate to illegitimate pollen on the stigmas of short-styled flowers (1.85) markedly exceeded that for long-styled flowers (1.11). Short-styled flowers incurred significantly higher levels of self-pollination than long-styled plants, emphasizing the importance for self-pollination to be removed before comparing stigmatic pollen loads. The method we propose could be applied to other distylous species with pollen size overlap between morphs and thus allow for a more widespread study of the pollination ecology of heterostylous species.     

Elevation-related variation in the population characteristics of distylous Primula nivalis affects female fitness and inbreeding depression

The population characteristics of distylous species are highly sensitive to stochastic natural selection pressure.Therefore,populations growing under different environmental conditions may vary in floral morph ratios,potentially affecting female fitness and leading to inbreeding depression.However,the variation in offspring quality among populations as a result of inbreeding depression is poorly understood in distylous species.This study investigates variations in plant density,seed mass,seed viabilityfemale fitness,and post-dispersal inbreeding depression in both sexual morphs(long-styled and shortstyled plants)of the distylous Primula nivalis that were subjected to different pollination treatments along an elevational gradient from 1657 to 2704 m a.s.l.Population characteristics(morph plant density and ratio)and fruit set were significantly affected by sexual morph and elevation.Plant density and fruitset frequencies were lower for short-styled than for long-styled plants at 2704 m a.s.l.The seeds from the cross-pollinated flowers of both morphs were higher in quality than those of self-pollinated flowers.The female fitness of seeds from cross-pollinated flowers of both morphs was higher than that of seeds from open-pollinated and self-pollinated flowers.The female fitness of seeds from long-styled flowers was higher than that of seeds from short-styled flowers at all elevations.Inbreeding depression increased with elevation among plants with short-styled flowers but not among those with long-styled flowers.Variation in the elevation-dependent mating system might influence female fitness and affect inbreeding depression in both floral morphs.In conclusion,the low quality of seeds from short-styled flowers at high elevations might decrease short-styled flower frequency,affecting population characteristics.     

Reproductive ecology of distylous Palicourea Padifolia (Rubiaceae) in a tropical montane cloud forest. I. Hummingbirds' effectiveness as pollen vectors

The adaptiveness of distyly has been typically investigated in terms of its female function, specifically pollen receipt. However, pollen loads on stigmas can only provide moderate support for Darwin's hypothesis of the promotion of legitimate crosses. To determine the effectiveness of hummingbirds as pollen vectors between floral morphs and the consequences in terms of male (pollen transfer) and female function (pollen receipt) in Palicourea padifolia (Rubiaceae), floral visitors, their foraging modes, and temporal patterns of floral visitation were observed and documented. Differences in pollen and stigma morphology, pollen flow, rates of pollen deposition, and/or stigmatic pollen loads were then evaluated for their contribution toward differences in reproductive output between floral morphs. A pollination experiment with stuffed hummingbirds that varied in bill size was done to evaluate the contribution of bill variation toward differences between floral morphs in pollen receipt and pollen transfer and female reproductive output. Anthers of long-styled flowers contained significantly more and smaller pollen grains than those of short-styled flowers, independently of corolla and anther lengths. The shape and orientation of the stigma lobes differed between morphs and were significantly longer among short-styled flowers. Hummingbird visitation rates did not differ significantly between floral morphs, and foraging movements from focal plants towards neighboring plants were independent of floral morph. Stigmatic pollen loads under field conditions and those after controlled hummingbird visitation, along with rates of pollen accumulation through the day indicated that stigmas of short-styled flowers receive proportionately more legitimate (intermorph) pollen grains than did those of long-styled flowers. However, the species of hummingbird was marginally significant in explaining variation in pollen deposition on stigmas. Lastly, intermorph pollinations of P. padifolia resulted in significant differences in fruit production between floral morphs, independent of pollination treatment and pollinator species; short-styled flowers proportionately developed almost twice the number of fruits developed by long-styled flowers.     

Distyly and variation in heteromorphic incompatibility in Gaertnera vaginata (Rubiaceae) endemic to La Reunion Island

Documenting the floral biology of species throughout the Rubiaceae family is of particular interest since heterostyly and dioecy may have evolved more than once in this large family. Unfortunately many species in several tropical regions remain unstudied. The purpose of this paper is to describe the floral biology, the nature of self-incompatibility, morph ratios, and fecundity in natural populations of Gaertnera vaginata, a small tree endemic to the island of La Reunion in the Indian Ocean. Measurements of floral characters in populations across the entire distribution of this species showed that G. vaginata exhibits a reciprocal stigma height and anther height dimorphism characteristic of a distylous species. Pollen grain size and corolla tube length are consistently greater in short-styled plants and long-styled plants produce more pollen per flower. Controlled pollinations in a natural population showed that 25% of the short-styled plants gave at least one fruit on intramorph (illegitimate) pollination, whereas no long-styled plants set fruit on illegitimate pollination. In total, 19.4% of illegitimate pollinations produced fruit on short-styled plants. No self-pollination gave fruit on either morph and between-morph pollinations produced 92.2 and 92.8% for short and long-styled plants, respectively. Overall, short-styled plants were significantly more abundant than long-styled plants. Short-styled plants outnumbered long-styled plants in 16 of the 19 populations. In three of these populations the morph ratio was significantly different from 1:1. In two natural populations, fruit set was significantly higher on long-styled plants, although the number of seeds per fruit was not significantly different between the two morphs. The possible effect of variation in the strength of heteromorphic incompatibility on observed variation in morph abundance and the possible causes for the variation in fruit set are discussed.     

艳丽耳草的二型花柱及异型自交不亲和系统

异型花柱是一种受遗传因素控制的花型多态性现象,包括二型花柱和三型花柱两种类型.本文以茜草科艳丽耳草(Hedyotis pulcherrima)为实验材料,通过对其野外居群的花型、花部形态及花粉特征等观察,发现艳丽耳草野外居群同时存在长花柱型花和短花柱型花,长/短花柱型花的数量比例为1∶1.两型花具有精确的交互式雌雄异位特征,并且该特征与花冠长度相关性显著.长/短花柱型花的柱头裂片长度、花粉大小及淀粉含量等具有二型性.花粉体外培养时花粉萌发率及花粉管生长速率无显著性差异.人工授粉后,艳丽耳草长/短花柱型花型间异交花粉管生长形态正常,授粉24 h后花粉管均已进入子房.而长/短花柱型花在自交及型内异交下均表现为不亲和,花粉管生长停止于柱头,花粉管顶端累积胼胝质并膨大.艳丽耳草没有无融合生殖现象,型间人工辅助异交授粉结实率为100％,显著高于自然结实率.本研究结果表明,艳丽耳草是典型的二型花柱植物,并具有异型自交不亲和系统.     

Heteromorphic incompatibility and efficiency of pollination in two distylous Pentanisia species (Rubiaceae)

BACKGROUND AND AIMS: Distyly has been hypothesized to promote cross-pollination by reducing intrafloral and geitonogamous self-pollination, and enhancing intermorph pollination. Distylous plants typically display both reciprocal herkogamy and a heteromorphic incompatibility system, which allows mating only between morphs. Distyly and its pollination consequences were examined in two Pentanisia species with long-tubed flowers which are pollinated almost exclusively by butterflies. METHODS: Anther and stigma heights were measured to quantify reciprocal herkogamy. The type of incompatibility system was determined by observing pollen tubes and seed production following controlled hand pollination. Pollen loads on pollinators and stigmas were also examined to assess the efficiency of intermorph pollen flow. KEY RESULTS: Pentanisia prunelloides and P. angustifolia exhibit reciprocal herkogamy and a host of ancillary dimorphisms, including pollen colour, exine sculpturing, stigmatic papilla shape and floral-tube pubescence. Controlled hand-pollinations revealed the presence of a strong heteromorphic incompatibility system in both species. The site of incompatibility differed between the morphs; intramorph pollen tubes were blocked in the style of the short-styled morph and on the stigmatic surface of the long-styled morph. Butterflies carried pollen from the short- and long-styled morphs primarily on their head and proboscis, respectively. Natural pollination resulted in a higher proportion of pollen transfer from long- to short-styled plants than vice versa. Nevertheless, fruit set did not differ between morphs. CONCLUSIONS: Both Pentanisia species are fully distylous. Reciprocal herkogamy results in pollen from the two morphs being carried on different locations on pollinators' bodies, which in turn promotes intermorph pollination. Intramorph pollination does not result in fertilization, because of an effective heteromorphic incompatibility system.     

The function and adaptive significance of tristyly in Pontederia cordata L. (Pontederiaceae)

Darwin proposed that the adaptive significance of tristyly is to promote insect-mediated pollination among floral morphs with anthers and stigmas at equivalent levels (legitimate pollination). Strong pollen trimorphism in Pontederia cordata enables an evaluation of this hypothesis. In an investigation of pollen flow patterns in different parts of the North American range of the species, legitimate pollination of some morphs was observed in all but one population investigated. However at only one site (Ft. McCoy, Florida) was Darwin's hypothesis confirmed for the three floral morphs. The long-styled morph most frequently exhibited legitimate pollination, the mid-styled morph was intermediate, and the short-styled morph rarely experienced legitimate pollination.
Temporal variation in pollen flow was observed during the flowering season of P. cordata . Total stigmatic pollen loads decreased as the density of inflorescences and activity of pollinators declined. In spite of variable total loads the legitimate component was relatively constant in the long- and mid-styled forms, while in the short-styled form this measure increased as the season progressed. Little variation was detected between stigmatic samples collected at different times during a single day.
Legitimate pollination may be inhibited by local foraging of pollinators and the spatial segregation of floral morphs. At Paugh Lake (Ontario) over 75% of all bumble bee flights are among the five nearest neighbours of a particular inflorescence. In the same population, there is a probability of over 70% that the nearest three neighbours of an inflorescence are of the same floral morph. Despite these influences, legitimate pollination occurs in P. cordata , suggesting substantial pollen carryover.     

Pollination biology and the impact of floral display, pollen donors, and distyly on seed production in Arcytophyllum lavarum (Rubiaceae)

In animal-pollinated plants, two factors affecting pollen flow and seed production are changes in floral display and the availability of compatible mates. Changes in floral display may affect the number of pollinator visits and the availability of compatible mates will affect the probability of legitimate pollination and seed production. Distyly is a floral polymorphism where long-styled (pin) and short-styled (thrum) floral morphs occur among different individuals. Distylous plants frequently exhibit self and intra-morph incompatibility. Therefore changes in morph abundance directly affect the arrival of compatible pollen to the stigmas. Floral morph by itself may also affect female reproductive success because floral morphs may display differences in seed production. We explored the effects of floral display, availability of neighboring compatible mates, and floral morph on seed production in the distylous herb ARCYTOPHYLLUM LAVARUM. We found that floral display does not affect the mean number of seeds produced per flower. There is also no effect of the proportion of neighboring legitimate pollen donors on seed production in pin or thrum flowers. However, floral morphs differed in their female reproductive success and the thrum morph produced more seeds. Hand pollination experiments suggest that differences in seed production between morphs are the result of pollen limitation. Future research will elucidate if the higher seed production in thrum flowers is a consequence of higher availability of pollen donors in the population, or higher efficiency of the pin morph as pollen donor.