Studies on heteromorphic self-incompatibility systems: Physiological aspects of the incompatibility system of Primula obconica

Summary In Primula obconica, a species with a heteromorphic self-incompatibility system, the distinction between compatible and incompatible pollen tubes takes place on the stigma surface in thrum flowers, self tubes growing randomly over the papillar cells. No differences were seen between self and cross tube behaviour on the pin stigma surface, but self tubes were inhibited within the stigmatic tissue with differences in tube length evident after 24 h. The stigma surface bears a proteinaceous pellicle and binds the lectin Concanavalin A. Removal of the stigma removes the incompatibility barrier in mature gynoecia. Bud pollination shows that pollen tubes cannot grow in a normal manner on immature stigmas; the random growth of tubes over the stigma surface resembles that of mature thrum selfs. Fewer compatible tubes reach the style base of young gynoecia and smaller numbers of seeds are set than in mature flowers. Pin and thrum pollen grains germinate and grow in aqueous media, thrum tubes growing longer than pin. The presence of H₃BO₄ and CaCl₂ in the growth medium promotes tube elongation and lengths equivalent to compatible styles can be obtained. The pollen grains have proteinaceous materials in their walls which diffuse out on moistening. Prolonged washing in aqueous media removes these materials but the incompatibility reaction remains unchanged. Thus the incompatibility reaction is between pollen tubes and stigmatic tissue and differs from the homomorphic, sporophytic system where pollen wall proteins elicit the incompatibility response.     

Pollen flow in a population of Primula vulgaris Huds.

A British population of Primula vulgaris was visited four times between 1971 and 1976 for study of the nature and functioning of distyly. The stainability and production of pollen from pins exceeded those of thrums. About 20% of pin pollen and 48% of thrum pollen was removed from anthers by insect visitors. Pins and thrums were present in a 1:1 ratio in 1971 but pins outnumbered thrums in 1976. Thrums produced more seeds per flower than pins though the number of ovules produced by each was similar. Two methods for collecting stigmas from open flowers and analyzing them for pollen loans produced somewhat different results. For pin stigmas, the pollen load consisted of 2–23% thrum pollen; for thrum stigmas, the pollen load consisted of 0–71% pin pollen with most stigmas having less than 50% pin pollen. In general, intermorph pollen flow is less than would be expected if pollen flow were random. It is probable that most intramorph pollen on stigmas is a result of self- or geitonogamous pollination. The extensive literature concerning the natural pollinators of the primrose is reviewed. Although Darwin's hypothesis concerning the functional significance of distyly in promoting intermorph pollination was never quantified, the pollen flow patterns observed in P. vulgaris are unexpectedly deviant and are similar to those patterns observed in several unrelated heterostylous species in other plant families.     

Phylogenetic relationships and character evolution in Primula L.: The usefulness of ITS sequence data

ABSTRACT

The main goals of this research were to reconstruct the infrageneric phylogeny of the genus Primula based on both nuclear and chloroplast DNA sequences, and to use the resulting phylogenies to elucidate the evolution of breeding systems, morphological characters, chromosome number, and biogeographic distribution in the genus. In this paper, the results of a pilot study based on the nuclear ribosomal Internal Transcribed Spacer (ITS) region are described. ITS sequences from 21 taxa produced a number of variable characters sufficient to resolve relationships among sections. The resulting phylogeny confirmed the monophyly of sections Auricula and Aleuritia. Sections Armerina, Proliferae, Crystallophlomis, Parryi, and Auricula, with a base chromosome number of x = 11, and sect. Aleuritia, with a base chromosome number of x = 9, formed two well supported clades. The ITS topology also suggested that leaves with revolute vernation, previously believed to be a derived state, might represent the ancestral condition in Primula, with later reversals to the involute condition. Finally, this initial ITS tree provides preliminary support to the proposed role of the widespread, diploid and heterostylous P. mistassinica as having given origin to the polyploid, homostylous P. incana and P. laurentiana.     

Analysis of late stage flower development in Primula vulgaris reveals novel differences in cell morphology and temporal aspects of floral heteromorphy

Heterostyly in Primula is characterized by the development of long-styled pin and short-styled thrum flowers, with anthers midway down the corolla tube in pin flowers, and at its mouth in thrum flowers. Other differences include pollen size and stigmatic papillae length. Several linked genes at the S locus control these differences.In this study we have analyzed pin and thrum flowers through the temporal development of heteromorphy.These studies indicate that the S locus linked genes that orchestrate heteromorphic flower development act in coordination, but with different temporal and spatial dynamics. Style length is differentiated by longer style cells in pin than thrum. However, our studies on cell shape and size within the corolla tube show that a different mechanism mediates the dissimilar elevation of anthers between pin and thrum types. These studies have also revealed that upper corolla tube cells in thrum flowers are wider than those in pin flowers. This results in a larger corolla tube mouth in thrum flowers and represents a new and previously undocumented heteromorphic variation between pin and thrum flowers.     

The distyly in Primula elatior (L.) Hill (Primulaceae), with a study of flowering phenology and pollen flow

Flowers of Primula elatior exhibit all the dimorphic traits typical of distyly. The difference in pollen production between pin and thrum flowers is not thought to be caused by the pollen size dimorphism. The percentage of pin pollen which is probably viable exceeds that of thrum pollen. In the population studied the morph ratio is 1:1, and plants are randomly dispersed with regard to morph type. Bumblebees are the main pollinators, and analysis of the stigmatic pollen load in naturally |xjllinated flowers shows that pin stigmas receive many more pollen grains than thrum stigmas. Intramorph pollination is considerable in pins, while thrums are subject to some differential pollination. Emasculation experiments have shown that intraflower pollination in the thrum morph is negligible. Aging of pin flowers has a reducing effect on their total pollen loads, while in thrums there is no change in total pollen load after the first days of anthesis.     

On the occurrence of unilateral incompatibility in Primula section Aleuritia Duby and the origin of Primula scotica Hook

The occurrence of unilateral incompatibility was tested for a distylous heteromorphic system, using crosses between a self-sterile and three self-fertile species in Primula L. section Aleuritia Duby. The crosses showed non-reciprocity but not in the same direction as would be predicted in the case of unilateral incompatibility. Pollen from the self-compatible (s-c) species was not always inhibited on the style of the self-incompatible (s-i) species, and cross-fertility between s-i and s-c crosses more resembled that between different s-c species, which was also non-reciprocal. Cytoplasmic-nuclear DNA interactions and the possibility of embryo-endosperm imbalance could both explain these results. In crosses between Primula farinosa L. (s-i diploid) and P. scotica Hook. (s-c hexaploid), heterostylous pin tetraploid offspring were produced. This result is discussed in relation to the genome of P. scotica and the possibility that pin morph plants may occur in wild P scotica populations.     

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。     

Comparative study of pollination biology of two closely related alpine Primula species,namely Primula beesiana and P.bulleyana (Primulaceae)

The comparative pollination biology of a population of Primula beesiana,a population of P.bulleyana,and an overlapping population of these two species in Yulong Shan,Yunnan Province,China,was studied in 2004 and 2005.The results indicate that both P.beesiana and P.bulleyana are typical heterostylous and obligate outcrossing species;the main pollinators of the two species were bees and butterflies.At the sites of all three populations,the main pollinating visitors of the two species showed preference for one...     

Comparative study of pollination biology of two closely related alpine Primula species,namely Primula beesiana and P. bulleyana (Primulaceae)

Abstract The comparative pollination biology of a population of Primula beesiana, a population of P. bulleyana, and an overlapping population of these two species in Yulong Shan, Yunnan Province, China, was studied in 2004 and 2005. The results indicate that both P. beesiana and P. bulleyana are typical heterostylous and obligate outcrossing species; the main pollinators of the two species were bees and butterflies. At the sites of all three populations, the main pollinating visitors of the two species showed preference for one of the two species, resulting in the ethological isolation of the two species by the pollinators. This ethological isolation contributes to the reproductive isolation of the two species, which supports the hypothesis that P. beesiana and P. bulleyana are two distinct species. The reproductive isolation of the two species (ethological isolation) is probably an important mechanism in maintaining species boundaries in the genus and has contributed to the species diversification of Primula in the area. In addition, gene exchanges between P. beesiana and P. bulleyana has occurred to some extent in the overlapping population, but whether natural hybridization has contributed to species diversification in Primula remains to be determined.     

Primula vallicola sp. nov. (Primulaceae) from Sichuan,China

A new species of Primula, P. vallicola Y. Xu, G. Hao & C.M. Hu, is described and illustrated from Sichuan province, China. Based on general morphology the new species is assigned to Primula sect. Aleuritia subsect. Yunnanensis. It is closely allied to P. kialensis, but can be distinguished by its larger stature, leaf shape, and the well‐differentiated petiole. The leaf shape of the new species is ovate to ovate‐elliptic and usually sub‐rounded at base. The new species also resembles P. homogama, but differs in being glabrous rather than with appressed hairs, and with longer petiole.     

滇西北玉龙雪山报春花种质资源的调查

滇西北玉龙雪山地处报春花属的现代形成与分布中心,有非常丰富的报春花种类。根据作者2003~2004年对玉龙雪山报春花属植物进行系统调查和资料整理,表明该地区有报春花40种(包括亚种与变种),占我国产报春花属植物总数的12%及云南产报春花属植物总数的29%;同时研究了该属植物在玉龙雪山的分布规律及其资源的生境类型并介绍了主要种类的生物学性状及观赏价值(包括花、叶及其开花期);而且对玉龙雪山报春花资源的保护及合理开发利用提出了建议。     

Shortened anther-stigma distance reduces compatible pollination in two distylous Primula species

Aims Distyly has been regarded as an adaptation to improve compatible pollination between two floral morphs with reciprocal herkogamy. The hypothesis that the different positions of anthers and stigmas within flowers as well as their reciprocal position between morphs, reduce the probability of self pollination raised by Darwin has been rarely tested. In this study, we measured stigmatic pollen loads in response to reduced reciprocal herkogamy in two Primula species.Methods To see whether reciprocal herkogamy can increase compatible and/or reduce incompatible pollen deposition, thus promoting compatible pollination, we shortened the distance between anthers and stigmas within the flowers by changing the position of the corolla tube, to which the anthers were fused, i.e. reduced herkogamy in natural populations of Primula secundiflora and P. poissonii and quantified stigmatic pollen loads in the field over 2 years.Important findings In both species, stigmatic pollen loads were significantly higher in the long-styled (L-morph) than in the short-styled morph (S-morph) in both control and manipulated flowers, but percentage of compatible pollen in S-morph were higher. Flowers manipulated to halve the anther–stigma distance showed a similar pattern for 2 years: total pollen grain counts on stigmas did not differ significantly but compatible pollen grains in L- and S-morphs were significantly decreased in both species. The percentage of compatible pollen loads was decreased by 68.7% in P. secundiflora and 65.3% in P. poissonii in L-morphs, while it decreased by 30.6% and 2.9% in S-morphs, respectively. Our manipulation of the relative position of anthers and stigmas in the two distylous species indicated that a lower degree of herkogamy reduced compatible but incompatible pollen transfer was likely to increase. The higher proportion of compatible pollen in the S-morph than in the L-morph in the two Primula species could be attributed to the accessibility of two-level sexual organs, floral orientations and pollinator behaviors. This is a first attempt to manipulate intraflower herkogamy for understanding adaptation of heterostyly, shedding insights into how the reciprocal herkogamy promotes compatible pollination.     

安徽羽叶报春和毛茛叶报春的微形态特征

在扫描电子显微镜和光学显微镜下,观察了安徽羽叶报春(Primula merrilliana Schltr.)与毛茛叶报春(Primula cicutariifolia Pax)的种子、花粉形态及表面纹饰与叶表皮形态结构.结果表明:2种类的种子都呈不规则七面体或多面体,多具五边形纹饰;安徽羽叶报春的种皮雕纹浅,网眼具许多小的乳头状突起;毛茛叶报春的种皮雕纹明显,网眼粗糙,具许多大的乳头状突起.2种类花粉均为球形,具散孔(孔数达10个以上),具网状雕纹.2种类的叶表皮细胞呈凹凸镶嵌状不规则排列,而在中脉处呈长方形或略带微波状长方形,上表皮细胞稍大;叶两面被腺毛,且顶端具水孔.气孔的分布属于偏叶下表面生长,在边缘处尤密,而在中脉处近无;相对而言,安徽羽叶报春的气孔略比毛茛叶报春的气孔小,且密度也低;电子显微镜下2种类的气孔外拱盖内缘光滑,保卫细胞外缘隆起明显,中间凹陷,其上着生的乳头状突起安徽羽叶报春不如毛茛叶报春明显.     

二型花柱植物海仙花报春花部性状随地理梯度的变异

植物的花部性状在异质环境中表现出不均一的适应性进化, 其自然变异可能在时空格局上呈现一定的规律性。选择同一物种的不同地理居群进行花部表型变异分析, 能揭示花部性状随地理梯度的变异模式。海仙花报春(Primula poissonii)属于典型的二型花柱植物, 依赖昆虫传粉实现严格的型间异交。该物种广布于横断山地区亚高山-高山草甸, 其分布海拔跨度大且花部性状在种内具有较高变异, 但这些变异在不同地理梯度(海拔梯度和经纬度梯度)的特定选择因子作用下的变化规律尚不清楚。本研究选择海仙花报春16个居群, 对8个花部关键性状和二型花柱繁殖器官的互补度与海拔和经纬度的关系进行研究, 探究花部性状随地理梯度变异的模式及其潜在的选择因素。研究表明, 海仙花报春两种花型的花冠管开口大小、花药高度以及短柱花柱头高度与海拔均呈正相关, 但两种花型的花冠大小, 长柱花的花瓣长度、柱头到花冠管开口的距离, 以及短柱花的花冠管长度与海拔高度间均呈负相关, 其余性状与海拔无显著相关性。除短柱花中柱头高度以外的性状均随着纬度升高而逐渐减小。长柱花中除花药和柱头间的距离以及柱头到开口的距离外, 其余性状均随着经度的增加而减小; 短柱花的花瓣长度、花药高度以及花药和柱头间的距离随着经度的增加而变大, 其余性状均随着经度的增加而减小。繁殖器官间的互补度并不随地理环境的变化而变化。花部性状的地理变异可能受访花昆虫组成的地理变化驱动。繁殖器官间互补程度的高度保守表明非选型交配在居群二态性的维持和稳定过程中起关键作用。本研究为进一步深入开展报春花属(Primula)花部性状及其选择压力的地理变异研究奠定了基础。     

Experimental insights on the function of ancillary pollen and stigma polymorphisms in plants with heteromorphic incompatibility

Most heterostylous plants possess a reciprocal arrangement of stigmas and anthers (reciprocal herkogamy), heteromorphic self‐incompatibility, and ancillary polymorphisms of pollen and stigmas. The topographical complementarity hypothesis proposes that ancillary polymorphisms function in the rejection of incompatible pollen thus promoting disassortative pollination. Here, we test this hypothesis by investigating patterns of pollen transfer and capture in populations of dimorphic Armeria maritima and A. pubigera and distylous Limonium vulgare (Plumbaginaceae), and by studying pollen adherence and germination patterns in A. maritima following controlled hand‐pollinations. Armeria lacks reciprocal herkogamy allowing the evaluation of the extent to which ancillary polymorphisms affect the composition of pollen loads. We compared the amounts of compatible and incompatible pollen on stigmas in natural populations and calculated the proficiencies of pollen transfer for each mating type. We detected disassortative pollination in each species, and mating types did not differ in compatible pollen capture, although cob stigmas captured more incompatible pollen. Controlled hand‐pollinations revealed the failure of incompatible pollen to adhere and germinate on stigmas. Our results provided evidence that, while structural in nature, pollen‐stigma dimorphisms are tightly associated with heteromorphic incompatibility and likely function to promote disassortative pollination, especially in the absence of reciprocal herkogamy.     

Importance of Queen Bumble Bees as Pollinators Facilitating Inter-morph Crossing in Primula sieboldii

Abstract Flower-visiting insects, the distribution of pollen on an insect's body, and fruit and seed sets of Primula sieboldiiE. Morren were investigated in a maritime deciduous forest habitat of the Hidaka region in Hokkaido. Queens of Bombus diversus tersatus Smith (Hymenoptera: Apidae), Bombus schrencki albidopleuralis Skorikov, and Bombus deuteronymus deuteronymus Schulz were observed to visit the flowers of the species. Bumblebee claw marks were found on the flower petals of 68% of the flowering ramets of the species. Scanning-electronmicroscopy revealed that pin and thrum pollens of P. sieboldii were deposited in different positions along the proboscis of a B. diversus tersatus queen (n=38400 grains). On the glossa, all the observed pollen was of the pin type, while more than half of the pollen grains observed on the upper part of the proboscis, galea, stipes, and maxillary palpus were of the thrum type. In the field, high fruit and seed sets were recorded for both pin and thrum morphs, indicating the occurrence of sufficient legitimate pollination during the flowering season. In addition to the effectiveness of queen bumblebees as pollinators for P. sieboldii suggested in this study, phenological and morphological matches, i.e., early spring flowering during the queen bee emergence season and the similar lengths of the corolla tube and the bee proboscis, also support the inference that queen bumble bees play an important role in legitimate pollination of P. sieboldii.