Observation of Flowering Process of Grape (<i>Vitis vinifera</i> L.)—Insight into the Starting of Pollination of Flower Hiding in Calyptras (Cap)

It is generally agreed that many Vitis vinifera L. cultivars are self-fertile, where self-pollination often occurs before capfall in a process called cleistogamy. Therefore, it is difficult to identify the right time to remove stamens before self-pollination during the cross-breeding of grape. For this paper, we observed the process of grape flowering and measured the pollen viability and stigma receptivity of grape flowers of ‘Shine Muscat’ in order to identify the starting time of self-pollination before capfall and to provide useful information for improving the efficiency of cross-breeding. The results demonstrate that the anther is not cracked during the visible clusters and separated clusters stages. Meanwhile, in the separated floral buds, flowering begins, and full bloom stages, the pollen viability is 60.7%, 73.2% and 80.3%, respectively; however, at the berry set stage, pollen viability drops to zero. The top of the mature stigma is composed of a layer of nearly cylindrical papillary cells, and the stigma receptivity for pollen changes with the development of flowers: in particular, no reaction was observed in the visible clusters stage; weak positive reaction at the separated clusters stage; strong positive reaction at the separated floral buds, flowering begins, and full bloom stages; and no reaction at the berry set stage. In the separated floral buds stage, pollen tubes were seen germinating in the style. In the flowering begins stage, more pollen tubes were observed at the entry of the ovary. During the full bloom stage, most pollen tubes elongated into the ovary base and some entered the pearl hole. At the berry set stage, newborn endosperm nucleus could be seen in the ovule. From the above, we can conclude that the initiation time of closed fertilization for ‘Shine Muscat’ grape can be judged as the separated floral buds stage, and it is best to discard the stamen before the separated floral buds stage when conducting cross-breeding.     

七叶树花粉活力和柱头可授性变化的研究

采用花粉离体萌发法研究不同蔗糖浓度、硼酸浓度和不同温度对七叶树花粉萌发的影响及花粉活力测定,用联苯胺-过氧化氢法测定柱头可授性.结果表明:七叶树花粉萌发的最佳培养基是12％蔗糖+30 mg/L硼酸,花粉萌发的最适温度为25℃.雄花在开花当天花粉活力最高达75.69％,并在开花当天的上午10:00时,花粉活力最强,10:00～16:00花粉活力保持较高活力,是最佳授粉时段.两性花的柱头可授期持续时间较长,为8～9 d,开花3d达到最强,开花1～4 d柱头可授性保持较高水平,为授粉的最佳时间段.因此,从七叶树的花部特征、花粉活力与柱头可授性及花粉萌发的条件看,在长期的自然选择下七叶树在花部结构和开花生理上都是相配合的,以保障生殖成功.     

Nectar Sugar Production across Floral Phases in the Gynodioecious Protandrous Plant Geranium sylvaticum

Many zoophilous plants attract their pollinators by offering nectar as a reward. In gynodioecious plants (i.e. populations are composed of female and hermaphrodite individuals) nectar production has been repeatedly reported to be larger in hermaphrodite compared to female flowers even though nectar production across the different floral phases in dichogamous plants (i.e. plants with time separation of pollen dispersal and stigma receptivity) has rarely been examined. In this study, sugar production in nectar standing crop and secretion rate were investigated in Geranium sylvaticum, a gynodioecious plant species with protandry (i.e. with hermaphrodite flowers releasing their pollen before the stigma is receptive). We found that flowers from hermaphrodites produced more nectar than female flowers in terms of total nectar sugar content. In addition, differences in nectar production among floral phases were found in hermaphrodite flowers but not in female flowers. In hermaphrodite flowers, maximum sugar content coincided with pollen presentation and declined slightly towards the female phase, indicating nectar reabsorption, whereas in female flowers sugar content did not differ between the floral phases. These differences in floral reward are discussed in relation to visitation patterns by pollinators and seed production in this species.     

Delayed stigma receptivity in Collinsia heterophylla (Plantaginaceae): genetic variation and adaptive significance in relation to pollen competition, delayed self-pollination, and mating-system evolution

To increase our knowledge about mating-system evolution, we need to understand the relationship between specific floral traits and mating system. Species of Collinsia (Plantaginaceae) vary extensively in mating system; this variation is associated with variation in floral morphology and development and with the timing of self-pollination. Counterintuitively, large-flowered, more outcrossing species tend to have delayed stigma receptivity, reducing the amount of time that the stigma is receptive to cross-pollination before autonomous self-pollination. To understand how the timing of stigma receptivity is related to mating-system evolution, we studied in detail the timing of both stigma receptivity and self-pollination (anther-stigma contact) in two greenhouse-grown populations of large-flowered Collinsia heterophylla. Crosses on emasculated flowers at different stages of floral development always produced seeds, suggesting that cross-fertilization can be effected by pollen arriving prior to physiological receptivity. Phenotypic and genetic variation within populations in the timing of stigma receptivity and anther-stigma contact was substantial, although slightly less for the contact. Despite strong interspecific and interpopulation correlations, we did not find an among-genet phenotypic correlation between the traits. This indicates that each trait may respond independently to selection, and the trait association may be the result of correlational selection.     

Pollen size strongly correlates with stigma depth among Pedicularis species

Darwin proposed that pollen size should be positively correlated with stigma depth rather than style length among species given that pollen tubes first enter the stigma autotrophically, then grow through the style heterotrophically. However, studies often show a positive relationship between pollen size and style length.Five floral traits were observed to be correlated among42 bumblebee-pollinated Pedicularis species(Orobanchaceae) in which stigmas are distinct from styles. The phylogenetic independent contrast analysis revealed that pollen grain volume was more strongly correlated with stigma depth than with style length, consistent with Darwin's functional hypothesis between pollen size and stigma depth.     

异型花柱植物喀什补血草的传粉者功能群与花粉转移效率

异型花柱是受遗传控制的花柱多态现象, 被达尔文认为是植物通过在传粉者体表不同部位滞落花粉以促进型间花粉准确传递的一种适应。该现象虽已受到广泛关注, 但在一些花型变异较大且不稳定的传粉系统中, 不同传粉者对各花型繁殖所产生的影响仍知之甚少。该研究以分布于新疆天山南坡的一个有同长花柱共存的异型花柱植物喀什补血草(Limonium kaschgaricum)种群为研究对象, 对其花型构成及频率、传粉者及花粉转移效率等进行了调查分析。结果表明: 1)种群中除了存在雌/雄蕊长度交互对应的长(L)/短(S)花柱型花外, 还有雌/雄蕊同长的花(H型), 且各花型花的花冠口直径、花冠筒长及花粉量等参数间无差异, 但花粉纹饰和柱头乳突细胞形态具二型性。其中, H型花的花粉和柱头形态与L型花(或S型花)的一致。2)花型内和自花授粉均不亲和; 型间授粉时, 花粉和柱头形态不同的花型间亲和, 反之不亲和。3)种群内存在长/短吻两类传粉昆虫。在以短吻传粉者为主的盛花初、中期, L和H型花柱头上的异型花粉数均显著高于S型花的, 且L和S型花高位性器官间的异型花粉传递效率高于低位性器官间的; 而在以长吻传粉者为主的盛花后期, L和S型花的柱头间异型花粉数无显著差异, 且高/低位性器官间具有相同的异型花粉转移效率; 与传粉者出现时期相对应的、在花期不同阶段开放花的结实率也明显不同。4)长/短吻昆虫具明显不同的传粉功能, 短吻昆虫只能对L和H型花进行有效传粉, 且访花频率和型间花粉转移效率较低, 为低效传粉者; 而长吻昆虫对各花型均能有效传粉, 具高的访花频率和型间花粉转移效率, 为高效传粉者。因为长吻昆虫的阶段性出现所形成的不稳定传粉系统, 使低效的短吻昆虫可能会成为种群中花型变异的驱动力, 并使S型花受到更大的选择压力。H型花克服了柱头缩入的弊端, 可能会成为不稳定传粉系统下的一个替代花型而持续存在。     

Pollen dynamics of bumble-bee visitation on Echium vulgare

1. We quantified pollen deposition on the stigma, pollen removal from the anthers and pollen losses in Echium vulgare , visited by workers of Bombus terrestris under controlled conditions. We used dye as a pollen analogue. Bumble-bees were trained to visit a sequence of non-emasculated flowers to estimate pollen carryover and to visit individual flowers to estimate pollen loss.
2. Carryover of pollen grains and dye particles between flowers was similar, which justifies using dye as a pollen analogue. On average 93·8% of the dye particles on the bee were carried over to the next flower. Only a small fraction of the pollen grains was deposited on the stigma (0·15%). A much larger fraction (6·1%) was lost in another way: passively during flight, through grooming or on floral parts other than the stigma. The bees removed 44% of the pollen grains from a fresh flower and 50·3% of this removed pollen adhered to the bee.
3. We predict that, using the parameters mentioned above, during a single visit to a newly opened flower, a bee collects an amount of pollen grains which will bring about 60% geitonogamous self-pollination in the next flower visited. The expected percentage of self-pollination is considerably less if bees visit flowers that have been visited before.     

Pollination mechanisms in sixCampanula species (Campanulaceae)

Pollination mechanisms within the genusCampanula were studied. Tests were undertaken to examine whether in vitro culture of pollen grains can serve as a useful tool for understanding the self- and cross-pollination mechanisms among species. Characteristics of pollen germination were interpreted in relation to mating system and floral biology. Four annual species (Campanula kremeri, C. dichotoma, C. afra, C. lusitanica), and two perennial species (C. rotundifolia andC. persicifolia) were investigated. In the annual species pollen germinability is controlled by (1) the age of pollen and/or (2) in what position pollen is deposited around the style. Correlations were found between pollen germinability and mating system in three of the four annual species. No correlations were found either between germinability and age of pollen or position on the style in the perennial species. Pollen germinability reached its maximum in the middle of the male phase in all species, except forC. dichotoma, which had a decreasing germinability throughout anthesis. The germinability was lowest at the time of stigma receptivity for all species except forC. persicifolia, where the stigma did not develop as long as pollen remained on the style. The pollen collecting hairs and pollen removal have been found to play an important role controlling the stigma development, thus affecting self-pollination. This was especially pronounced inC. persicifolia. Further studies will be undertaken to elucidate factors influencing pollination within the genusCampanula.     

海滨锦葵花内五柱头裂片弯曲的独自调节(英文)

海滨锦葵花柱有5柱头裂片,若传粉失败,柱头裂片向下弯曲使柱头与自身花粉接触,发生延迟自交。传粉不足条件下发生的延迟自交可能提供繁殖保障。该研究定量分析了花粉搁置和授精胚珠数对花内未授粉柱头裂片运动的影响,并测定了花粉—胚珠比及柱头可授性和花粉生活力。结果表明,花内未授粉柱头裂片的弯曲不受其他柱头接受花粉量及授精胚珠数的影响,仅响应于其自身是否接受到花粉,花粉-胚珠比值显示海滨锦葵交配系统属兼性异交;未授粉柱头经弯曲与自身花粉授触时的强柱头可授粉和高花粉生活力为自交授粉的发生提供了可能。包括柱头裂片运动在内的多个花性状有机地展示了一种新花内混合交配系统,且花内柱头裂片弯曲的独自调节为从花水平验证被广泛接受的自交进化解释——繁殖保障假说提供了可能。     

Delayed autonomous self-pollination in the colonizer Crotalaria micans (Fabaceae: Papilionoideae): structural and functional aspects

We investigated the reproductive biology of Crotalaria micans, a colonizing species that occurs in disturbed sites. The flowers have two whorls of stamens with differential growth and dimorphic anthers. The inner anthers serve two functions: 1) to cooperate with the stylar brush in the pollen presentation, and 2) to push the remaining pollen within the keel onto the stigma at the end of the receptivity period. Flowers pass through three phases: a) male phase, when outer anthers release their pollen, but stigmas are not receptive; b) female phase, when stigmas are receptive, but still separated from flowers' own pollen, and c) an autogamous phase, when the round anthers grow towards the stigma, leading to delayed autonomous self-pollination. Because C. micans is completely self-compatible, delayed self-pollination and geitonogamy result in approximately 76% of seeds being self-fertilized.     

Breeding system in a population of Trigonella balansae (Leguminosae)

BACKGROUND AND AIMS: Although some taxonomic studies in the genus Trigonella have been conducted, there has been no concerted effort to study the breeding system. This paper examines the floral structure and pollination system in a population of T. balansae, an annual pasture legume. METHODS: Floral morphology, hand and vector pollination, stigma receptivity, pollen tube growth, using scanning electron and fluorescence microscopy, were conducted. KEY RESULTS: Measurements of floral structure from before to after anthesis indicates an inability for T. balansae to self-pollinate and a requirement for an external vector to effectively transfer pollen from the anthers onto the stigmas of this species. Seed set can be obtained by hand or honeybee manipulation of T. balansae flowers. CONCLUSIONS: Trigonella balansae is a self-compatible species, but which requires vectors such as honeybees to bring about pollination.     

多伞阿魏的繁育系统及其生态适应性

多伞阿魏（Ferula ferulaeoides）是新疆特有的类短命植物,具有巨大的生态作用和潜在的经济价值。实验通过研究多伞阿魏的花部形态特征、繁殖系统及传粉昆虫,来阐明其对生态环境的响应。结果表明：（1）多伞阿魏花序花期一般持续10~15 d,单花花期为7~10 d,两性花呈现雌雄异位和雌雄异熟的现象;（2）其繁育系统类型为兼性异交,存在自交部分亲和,自然结果率较高,为72.24%;（3）传粉昆虫多种多样,其中以双翅目和膜翅目昆虫居多;（4）开花当天的花粉活力呈单峰曲线,在12：00时活力最高,柱头可授性在花药完全散粉后第2天最强。     

Breeding system in the dichogamous hermaphrodite Silene acutifolia (Caryophyllaceae)

The breeding system of the dichogamous hermaphrodite species Silene acutifolia, endemic to north-west Spain and north and central Portugal, is examined. Pollen germinability and style-stigma receptivity were analysed to determine whether protandry is a barrier to self-fertilization. By 48 h after anthesis, pollen germinability had declined to approx. 10 %. The short straight styles are not receptive when flowers first open. They gradually elongate and curve outwards, develop stigma papillae and become receptive. There is no clear separation between stigma and style: the stigma papillae appear in a line along the length of the style. Fruit set is high regardless of pollen source; however, seed set is significantly reduced after both spontaneous and facilitated autogamy. Seed set following spontaneous autogamy was 30 % (86 % in controls) in 1998 and 33 % (87 % in controls) in 1999. Seed set following facilitated autogamy was 62 % (86 % in controls) in 1998 and 67 % (89 % in controls) in 1999. Thus, separation of the male and female phases does not prevent production of seeds by self-pollination, although it does reduce the likelihood of this. Furthermore, results of the present experiments indicate that this species has no self-incompatibility mechanisms (self-compatibility index = 0.98). The selfing rate in the study population was 0.41, which is supported by the lack of self-incompatibility systems and by the incomplete protandry.     

The avoidance of self-interference in the Tuscan endemic spring geophyte Crocus etruscus Parl. (Iridaceae)

Spatial separation between sexes within hermaphrodite flowers (herkogamy) is widespread among angiosperms and is traditionally viewed as an adaptation that reduces the likelihood of self-pollination. While different degrees of herkogamy have been reported for Crocus, the relationship between sexual organ positions and reproductive fitness has not been explored yet, and studies of the breeding system within natural populations remain scarce within the genus. We analyzed the effects of different breeding systems (spontaneous self-pollination, facilitated self-pollination and cross-pollination) and of herkogamy degree on female reproductive success of the Tuscan endemic Crocus etruscus. Results showed that C. etruscus is monomorphic for style length and self-incompatible (SCI = 0.29). We also detected a significant negative effect of herkogamy on seed set of open-pollinated flowers. Thus, we conclude that the species avoids self-fertilization by self-incompatibility, but interestingly shows also a certain degree of spatial separation of male and female reproductive structures within the same flower. This floral trait could be useful to avoid self-interference thus reducing ovule discounting and pollen loss on its own stigma.     

高山植物扁蕾的延迟自交机制

扁蕾(Gentianopsis barbata)具有鲜艳的花和显著的腺体,并且花开放的前5 d柱头和花药始终处于不同的位置(雌雄异位),这些花综合征表明该植物应为异花传粉。为检验这一假设,我们对青藏高原植物扁蕾的海北站种群进行了3年的传粉生物学研究实验。与花综合征所表明的繁育系统相反,两年的野外观察发现昆虫的访花频率十分低,不去雄并隔离昆虫处理也能产生大量种子,说明这一种群的繁殖主要是依赖于自花传粉。尽管利用种子结实评价的柱头可授性从花开放4 d后开始下降,但随着花的发育进程,雄蕊的伸长能使得花药与柱头完全接触。实验也证明,柱头可授性和花粉活力都超过5 d,说明花药和柱头的接触能够发生自花授粉。扁蕾的这种自花传粉机制应属于典型的延迟自交类型。自花授粉发生在单花花期快要结束前,自交之前仍然保持异交传粉机制,这种延迟自交避免了自交与异交竞争造成的花粉或者种子折损,并为扁蕾在青藏高原极端环境下由于访花昆虫缺乏造成的异交失败提供了繁殖保障。     

Stylar trimorphism in four functionally andromonoeciousProsopis species (Mimosaceae)

The reproductive system ofProsopis chilensis, P. pugionata, P. flexuosa, andP. torquata is described. Observations were made of floral morphology, stigmatic receptivity, pollen grain viability, and the nature of pollen reserves. Scanning electrone microscopy and statistical analysis were carried out. The results allow us to conclude that: 1) The studied species have three distinct floral morphs characterized by significantly different style lengths and, to a lesser extent, different stamen lengths; 2) style precocity found in all species does not indicate protogyny; 3) flowers with shortest styles are not receptive; 4) the pollen grains of all floral morphs showed high viability; 5) peroxidase is present in both the pollen and pistil; 6) viable pollen grains invariably have both starch and pollenkitt; 7) the studied species are andromonoecious.     

黑河中游和烟台海滨中国柽柳的传粉生态学研究

中国柽柳(Tamarix chinensis)是干旱及半干旱地区的重要建群物种.为便于其有效利用和引种栽培,对黑河中游和烟台海滨的中国柽柳的开花动态、花粉活力、柱头可授性、访花者种类、花粉-胚珠比(P/O)、杂交指数(OCI)及座果率进行观测,结果表明:黑河中游和烟台海滨柽柳的花期和柱头可授性相似,花序花期为2-5天,开花当天柱头可授活性最大,之后逐渐下降,第4天柱头无可授性;花粉活力和访花者种类差别较大,黑河柽柳开花第1天花粉活力为(62±4.31)％,最高为(82.12±7.23)％,4天后花粉完全失活.烟台柽柳的花粉活力在开花后第1天最高为(89.13±7.23)％,第4天花粉活力下降为(11±1.12)％.黑河中游和烟台海滨柽柳的访花者均以膜翅目昆虫为主,黑河柽柳以黑带斑沙蜂(Bembix melanura)、卡切叶蜂[Megechile (Amegachile) kagiana]和黑带食蚜蝇(Episyrphus balteatus)等为主要访花者;烟台柽柳以姬蜂(Ichneumon sp.)、大灰食蚜蝇(Metasyrphus corollae)和灰地种蝇(Delia platura)等为主要访花者.二者P/O值、OCI及座果率差异不显著,黑河和烟台柽柳的P/O值分别为373.6±71.9和382.1±86.2;最高座果率分别为(89.72±7.76)％和(93.17±8.21)％;OCI均为3.上述研究结果显示,黑河中游和烟台海滨柽柳均为兼性异交的繁育系统.     

Changes in Florets’ Vertical Direction within Inflorescence Affects Pollinator Behavior,and Fitness in <i>Trifolium repens</i>

Ecological interactions between flowers and pollinators greatly affect the reproductive success. To facilitate these interactions, many flowers are known to display their attractive qualities, such as scent emission, flower rewards and floral vertical direction, in a rhythmic fashion. However, less is known about how plants regulate the relationship between these flower traits to adapt to pollinator visiting behavior and increase reproduction success. Here we investigated the adaptive significance of the flower bending from erect to downward in Trifolium repens. We observed the flowering dynamic characteristics (changes of vertical direction of florets, flowering number, pollen grain numbers, pollen viability and stigma receptivity over time after blossom) and the factors affecting the rate of flower bending in T. repens. Then we altered the vertical direction of florets in inflorescence of different types (upright and downward), and compared the pollinator behaviors and female reproductive success. Our results showed that florets opened sequentially in inflorescence, and then bend downwards slowly after flowering. The bending speed of florets was mainly influenced by pollination, and bending angle increased with the prolongation of flowering time, while the pollen germination rate, stigma receptivity and nectar secretion has a rhythm of “low-high-low” during the whole period with the time going. The visiting frequency of all the four species of pollinators on upward flowers was significantly higher than that of downward flowers, and they especially prefer to visit flowers with a bending angle of 30°–60°, when the flowers was exactly of the highest flower rewards (nectar secretion and number of pollen grains), stigma receptivity and pollen germination rate. The seed set ratio and fruit set ratio of upward flowers were significantly higher than downward flowers, but significantly lower than unmanipulated flowers. Our results indicated that the T. repens could increase female and male fitness by accurate pollination. The most suitable flower angle saves pollinators’ visiting energy and enables them to obtain the highest nectar rewards. This coordination between plants and pollinators maximizes the interests of them, which is a crucial factor in initiating specialized plant-pollinator relationships.     

An experimental evaluation of self-interference in Narcissus assoanus: functional and evolutionary implications

Floral traits that reduce self-pollination in hermaphroditic plants have usually been interpreted as mechanisms that limit the genetic consequences of self-fertilization. However, the avoidance of sexual conflict between female and male function (self-interference) may also represent an important selection pressure for the evolution of floral traits, particularly in self-incompatible species. Here, we use experimental manipulations to investigate self-interference in Narcissus assoanus, a self-incompatible species with a stigma-height dimorphism in which the degree of spatial separation between sex organs (herkogamy) differs strikingly between the long- and short-styled morphs (hereafter L- and S-morphs). We predicted that weak herkogamy in the L-morph would cause greater self-pollination and hence self-interference. Experimental self-pollination reduced seed set when it occurred prior to, or simultaneously with, cross-pollination in the L-morph, but only if it occurred prior to cross-pollination in the S-morph. In the field, autonomous self-pollination was greater in the L-morph than the S-morph, but we found no evidence that self-interference reduced maternal or paternal fitness in either morph. One-day-old flowers of the L-morph have reduced stigma receptivity and hence exhibit protandry, whereas stigma receptivity and anther dehiscence are concurrent in the S-morph. This suggests that the two style morphs have alternative strategies for reducing self-interference: dichogamy in the L-morph and herkogamy in the S-morph. These results provide insight into the mechanisms that reduce sexual conflict in hermaphrodite plants and are of significance for understanding the evolution and maintenance of sexual polymorphisms.