Aspects of sexual failure in the reproductive processes of a rare bulbiliferous plant,<Emphasis Type="Italic"> Titanotrichum oldhamii</Emphasis> (Gesneriaceae), in subtropical Asia

Titanotrichum oldhamii produces both flowers and asexual bulbils on its inflorescences. However, field observations and herbarium collections indicate that seed set is infrequent and that most reproduction is from vegetative bulbils. We have investigated the failure of sexual reproduction and identified four major causes: (1) in the wild, the seed:ovule ratio for open pollination was only 1.9%, in contrast to 10.1% for artificial cross-pollination, implying poor pollinator services. (2) The overall reproductive success was 5–10 times greater in glasshouse pollination treatments compared to field treatments. This suggests suboptimal environmental conditions for seed set in natural habitats, which may eliminate natural seed set completely. (3) Pollination experiments showed that outcrossed populations set significantly more seed and had higher germination rates than intra-populational and self crosses, in both field and glasshouse experiments. T. oldhamii thus appears to benefit from wide outcrossing. Pollen transfer between populations, however, seems to be infrequent because of the rarity and scattered distribution of Titanotrichum populations. (4) Flowers near the apex of the inflorescence are less likely to set seed, especially late in the season when inflorescences convert to bulbil production. In these late flowers, pollen tubes showed poor guidance as they approach the micropyle of the ovules. Even under optimal glasshouse conditions, the average outcrossing success was only 0.229. Almost one-half of the ovules remained undeveloped and 13.5% of ovules aborted after pollination, indicating a strong shift of resource allocation toward vegetative bulbils and rhizome development. Efficient reproduction from asexual bulbils may thus have released Titanotrichum from strong selection for efficient sexual reproduction. However, occasional seed set observed in the wild may be very important for maintaining some genetic diversity in populations, and promoting overall fitness.     

蛛网萼开花物候、花部特征及繁育系统研究

为探讨蛛网萼(Platycrater arguta)传粉生物学特性,该研究以江西上饶广丰铜钹山国家级自然保护区、江西阳际峰国家级自然保护区蛛网萼为材料,采用野外观察和人工授粉等方法,对蛛网萼开花物候、花部形态及繁育系统进行了研究。结果表明:(1)蛛网萼为伞形花序,种群花期一般在6月中下旬至8月中旬,单花花期为2~3 d,花序花期持续10 d。两个保护区由于年均温度与光照的不同,花期进程有显著不同。(2)蛛网萼可孕花与不孕花从绿色变为白色时对访花昆虫有吸引作用,不孕花能够增加昆虫的访花频率。(3)蛛网萼花粉胚珠比(P/O)为2 520~3 100,杂交指数即是3或4。(4)繁育体系属于以异交为主,部分自交亲和,需要传粉者;熊蜂(Bombus sp.)、四条蜂(Apis florea)和油茶地蜂(Andrena cammelia)是蛛网萼的有效传粉昆虫。(5)套袋和人工授粉结果显示,蛛网萼为兼性自交和异交授粉,异株异花授粉能显著提高坐果率及结籽率,这主要与生境中有效传粉昆虫种类、数量少,花粉传递效率低,种群间基因流交流降低以及柱头自花及同株异花传粉等有关,从而表明在蛛网萼现有生境中植株的早期生殖成功受到影响。蛛网萼种群分布范围扩张以有性繁殖为主,无性繁殖作为重要补充及繁殖保障。     

Breeding system of the threatened endemic Primula cusickiana var. maguirei (Primulaceae)

Maguire primrose (Primula cusickiana var. maguirei) is a threatened, narrow endemic found only along a 20‐km section within a single canyon in northern Utah, USA. Here we describe the breeding strategy of this perennial, distylous, primarily cliff‐dwelling species to test for reproductive constraints on the plants. Intermorph outcrossing treatments (i.e. between pin and thrum flowers) were the most successful hand pollinations performed, but those were usually inferior to natural pollination. Intramorph out‐crossing also was somewhat successful, but was consistently inferior to intermorph outcrossing. Seed‐set resulting from autogamy and geitonogamy treatments was least successful. Morph‐specific differences in seed set were observed for our hand pollinations, but not in natural pollination conditions. Our results suggest that pollinators play a crucial role by providing the pollen transfer necessary for maximal seed set in this rare and narrowly distributed distylous plant.     

Variation in breeding system among populations of the common woodland herbAnemone nemorosa (Ranunculaceae)

Recent studies of germination in natural habitats, of genetic variation within populations and of the relative proportion of vegetative and sexual reproduction in the clonal plant speciesAnemone nemorosa suggest that sexual recruitment by seeds from outcrossed flowers is important for the maintenance of this species' populations. Because published reports on its breeding system are controversial, pollination experiments were performed in five natural populations ofA. nemorosa. Differences in ovule number per flower were recorded among populations, but they were not related to obvious habitat differences. Seed/ovule-ratios were significantly higher after open pollination and artificial crossing than after either artificial or spontaneous selfing. Populations had no effect on seed/ovuleratios. Different breeding indices indicated thatA. nemorosa is mainly self-incompatible. Nevertheless, some seed set also occurred after selfing, and both artificial and spontaneous selfing exhibited higher variation in seed/ovule-ratios than open pollination and artificial crossing. Continuous variation in seed/ovule-ratios after selfing suggested that the expression and effectiveness of the self-incompatibility system ofA. nemorosa is influenced by both genetic variation and phenotypic plasticity.     

Reproductive ecology of a perennial outcrosser with a naturally dissected distribution

We surveyed four populations of contrasting size (two isolated and two large stands) of a woody outcrossing species, Discaria trinervis (Rhamnaceae), which has a naturally dissected distribution. Our main goal was to investigate the floral biology, breeding system and pollination mechanism of the species, which in turn may help to explain the mechanisms underlying the survival of the isolated populations. Discaria trinervis is both wind- and insect-pollinated, showing interactions with a large fraction of the available flower visitors at both stand sizes. In the larger populations, stigmatic loads were larger, more pollen tubes reached the ovules, and fruit and seed set were higher; however, the reproductive output in the smaller populations (over 2000 seeds per plant per year) seemed adequate for population persistence. Human disturbance (e.g. destruction of trees for wood extraction) may put these smaller populations at risk through loss of incompatibility alleles. The results suggest that plant species with naturally dissected distributions may provide clues about which reproductive mechanisms ensure survival under fragmentation.     

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.     

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。     

All in an afternoon: mixed breeding system in one-day lasting flowers of Hypericum elodes L. (Hypericaceae)

Knowledge about mixed mating systems can improve our understanding of the evolutionary dynamics of reproductive systems. Here we report a study of the floral and reproductive biology of Hypericum elodes, an Atlantic-European soft-water pools specialist which shows a floral architecture consistent with both self- and cross-pollination. Controlled pollination experiments were performed in a natural population during three consecutive years. Marked flowers were monitored until fruit production, and laboratory germination experiments were conducted with the seeds produced. Plants were self-compatible (SCI>0.75), however, compared with selfing, cross-pollination enhanced fruit-set, seed-set and seedling growth, but not seed germination. Inbreeding depression (δ) was mild in the pre-dispersal stages (δ = 0.22 for fruit set, 0.18 for seed set and 0.13 for seed mass), low for germination percentage (δ = 0.003) and mild for seedling growth (δ = 0.23). The breeding system of H. elodes promotes outcrossing and assures reproductive success by means of competitive autogamy. Our results suggest a mixed mating strategy for the studied population, characterized by mild inbreeding depression (cumulative δ = 0.57), highlighting the benefit of this reproductive mode in unpredictable habitat, as the typical shallow-water meadows where H. elodes grows.     

REPRODUCTIVE BIOLOGY OF THREE SPECIES OF EUPHORBIA SUBGENUS CHAMAESYCE (EUPHORBIACEAE)

The breeding system, pollen tube development, and floral biology of three species of Euphorbia subgenus Chamaesyce from southeastern Arizona have been examined in field and greenhouse studies. E. albomarginata is entirely dependent on insect vectors for pollination and seed production, E. capitellata requires insect visitors for maximum seed set but is capable of limited self-pollination, and E. hyssopifolia is self-pollinating and entirely independent of insect vectors. All three species are self-compatible. The differences in reliance on insect vectors, diversity of insects and probable amount of outcrossing are related to differences in cyathial size and arrangement. The reproductive biology of the three species is similar to that described for other weedy plant taxa.     

POLLINATOR LIMITATION IN THE FACULTATIVELY AUTOGAMOUS ANNUAL,LUPINUS NANUS (LEGUMINOSAE)

The importance of pollinator visitation for determining both maternal reproductive success and outcrossing rates was investigated in the facultatively autogamous annual, Lupinus nanus. Sixty plants in each of two adjacent sites were assigned to either a pollinator-exclusion, pollen-augmentation, or open-pollinated (control) treatment. Flower, fruit, and seed production were recorded for each plant, and outcrossing rates were determined for plants in all three treatments at each site. Pollinator-exclusion reduced the reproductive success of plants at Site 1, but had no effect on reproduction for plants at Site 2. Pollen-augmentation increased fruit production by 22% and seed production by 45% at Site 2, but had no effect on reproduction for plants at Site 1. Plants in the open-pollinated, control treatment outcrossed at intermediate rates at both sites, with plants at Site 2 (outcrossing rate, t = 0.66) outcrossing at a significantly higher rate than plants at Site 1 (t = 0.40). The pollen-augmentation treatment increased the outcrossing rate for plants at both sites (Site 1, t = 0.72; Site 2, t = 0.78). These results indicate that pollinator availability can limit maternal reproductive success for this facultatively autogamous plant species. Additionally, the intermediate outcrossing rates observed in nature for this species are not a consequence of plant characters alone, but instead result from plant characters combined with the local pollination ecology.     

Pollinator activity and pollination success of Medicago sativa L. in a natural and a managed population

Medicago sativa L. is an important cash crop in the arid region of northwest China. Pollinator activity is an essential aspect of pollination success, but the relationships between pollinator visitation rate and seed set still need further study of M. sativa. We investigated the following characteristics of M. sativa in natural and managed populations: floral traits, pollinator activity, and breeding system. Our results indicated the management could affect the number of flowers produced; however, there was no detectable effect on the seed set per flower. We found the percentage of seeds among pollinated flowers in the managed population was significantly higher than that in the natural population. Moreover, the increase in the proportion of pollinated flowers could significantly increase seed set per flower, and pollinator visitation rate was the important limiting factor for seed set in both populations. Andrena lebedevi Popov was found to be the most frequent pollinator in both populations. Outcrossing was dominant in the breeding system and insect pollination played an important role in outcrossing. Our study suggested that proper management (artificial selection) could promote pollination success of M. sativa.     

Reproductive biology of a rare cactus, Opuntia spinosissima (Cactaceae), in the Florida Keys: why is seed set very low?

 Opuntia spinosissima (Martyn) Mill. (Cactaceae) is an extremely rare taxon with a single remaining wild population of 13 plants located on Little Torch Key, Florida. The plants rarely set viable seeds and most ovaries abscise without initiation of fruits. Pollination and breeding system were examined in Little Torch Key and in plant accessions located at Fairchild Tropical Garden. Four pollination treatments were carried out to test for apomixis, self-pollination, outcrossing, and autogamy; a control treatment was also monitored. Additionally, pollen viability, pollinators, and seed set and viability were investigated. On selfed, crossed, and open (control) stigmas pollen grains germinated, but the tubes usually did not reach the base of the style. This suggests that O. spinosissima has pre-zygotic self-incompatibility. None of the pairwise crosses set seed, so the extant plants were apparently not inter-compatible. Out of 173 manipulated and control flowers, only one set fruit. Although this flower was outcrossed, no pollen tubes germinated on the stigma; this suggests agamospermy, a process common in the Cactaceae. In those flowers where pollen tubes did reach the ovary they failed to penetrate ovules, suggesting ovarian inhibition or that this taxon has lost the ability to be fertilized. Most field-collected seeds were viable, but there is no seedling recruitment under natural conditions, and vegetative reproduction is common. Based on these findings, I hypothesize that O. spinosissima is a sterile polyploid and that the 13 extant plants are asexually derived from a single lineage. Received: 17 April 1998 / Revision accepted: 3 July 1998     

THE LEVEL OF AGAMOSPERMY IN A NEBRASKA POPULATION OF SPIRANTHES CERNUA (ORCHIDACEAE)

This study investigated reproduction in a prairie population of Spiranthes cernua in eastern Nebraska. Spiranthes cernua reproduces both sexually and asexually, through adventitious embryony. Agamospermous seeds are monoembryonic, polyembryonic, have extruded or free embryos, or lack embryos. These kinds of seeds are extremely rare in Spiranthes vernalis, a close relative that relies exclusively on sexual reproduction. Sexually derived seeds are monoembryonic or lack embryos. One objective was to quantify the amount of sexual reproduction in a natural population. In 1985 and 1986, maximum estimates of sexual seed production per capsule were 19.2% and 33.6%, respectively. Thus, every capsule examined contained a high proportion of agamospermous seeds. In addition, attempts were made to manipulate the reproductive system with experimental crosses performed in the field. The treatments examined agamospermy, autogamy, and outcrossing. In all treatments the majority of seeds in a capsule were agamospermous.     

Wind pollination and reproductive assurance in Linanthus parviflorus (Polemoniaceae), a self-incompatible annual

Wind pollination was experimentally demonstrated in Linanthus parviflorus (Polemoniaceae), a predominantly beefly-pollinated, self-incompatible annual. Seed set in plants enclosed in mesh tents that excluded pollinators but allowed airborne pollen flow provided evidence for wind pollination, and the extent of seed set due to wind pollination was compared to that in open-pollinated controls and pollen-supplemented treatments. Additional controls were included to test for possible confounding effects of the mesh tent. Mean seed number in open-pollinated plants was 72.8–81.1% of that in pollen-supplemented plants, while wind pollination alone produced 49.5–52.2%, a smaller but substantial proportion of seed set with pollen supplementation. Further evidence for wind pollination was found in a comparison of sites differing in the extent of wind exposure in two populations of L. parviflorus. Airborne pollen counts were higher in exposed sites than in protected sites, and the difference was marginally significant. Seed set was significantly pollen limited in protected sites, but not in exposed sites. Taken together, the data suggest that wind pollination provides some reproductive assurance in this obligately outcrossing species. Wind pollination is hypothesized to represent an alternative to selfing as an evolutionary solution to the problem of temporal or spatial variation in pollination visitation.     

高山物种栗色鼠尾草(Salvia castanea Diels)访花昆虫多样性与传粉行为变化

唇形科鼠尾草属是世界性分布、物种多样性丰富的大类群,具有独特的传粉模式和多样化的生态类型及繁殖策略,以往对鼠尾草属的花结构和雄蕊杠杆的生态功能、雄蕊杠杆机制对传粉者空间变异的进化响应和表型选择等方面已有较深入的研究,但至今对分布于不同区系代表性物种的传粉者组成、访花行为和繁育系统特征仍知之甚少。对分布于喜马拉雅东段至横断山高海拔地区栗色鼠尾草的传粉生态、交配系统和繁殖特性进行了研究,分析了2014年至2016年的年际间、2014年晴天和阴雨天的传粉者组成和传粉行为变化,探讨了这些变化对繁殖成功的影响。研究结果表明：3年共发现栗色鼠尾草有9种访花昆虫,传粉者组成及其行为变化较为明显。2014年有桔尾熊蜂Bombus friseanus、灰熊蜂B.grahami、圣熊蜂B.religiosus和中华蜜蜂Apis cerana 4种有效传昆虫,2015年仅有桔尾熊蜂是有效传粉者,而2016年的有效传粉昆虫改变为灰熊蜂和圣熊蜂。栗色鼠尾草的盗蜜现象较为普遍,主要盗蜜昆虫为灰熊蜂和桔尾熊蜂。桔尾熊蜂访花行为的年际变化较大,受天气条件的影响明显,同时其有效传粉行为可能显著影响了灰熊蜂的盗蜜行为。栗色鼠尾草的繁育系统为专性异交,主要依靠传粉者进行授粉,缺少传粉者时低程度的主动自花授粉可为其提供繁殖保障,且没有花粉限制和近交衰退。研究揭示了在高海拔地区,多变的天气条件可能显著影响着鼠尾草属植物的传粉者种类组成、访花行为和传粉效率,进而影响植物的繁殖成效和种群稳定。有效传粉频率是保证栗色鼠尾草较高自然结实率的主要因素,在一定程度上,盗蜜强度对繁殖成功具有中性的影响。本研究结果为阐明高山鼠尾草物种的繁殖如何受气候环境变化的影响,以及保证繁殖成功的可能策略奠定基础。     

NONFRUITING HERMAPHRODITIC FLOWERS OF CALOCHORTUS LEICHTLINII (LILIACEAE): POTENTIAL REPRODUCTIVE FUNCTIONS

The breeding system, flowering display, and fruiting pattern of Calochortus leichtlinii Hook. (Liliaceae) was studied in the eastern Sierra Nevada. This species is capable of both autogamy and insect-mediated outcrossing. Either of these two means of pollination alone could affect full seed set; the populations studied were not pollination limited in 1983. Most individuals produced two flowers but set only one fruit. Flowers are not selectively matured on the basis of the quantity or purported quality of the pollen they receive; the first flower to open was matured in all experiments. Nonfruiting flowers have at least three potentially important reproductive functions: disseminating pollen, allowing plants to produce more seeds in response to an abundance of soil moisture, and setting seeds if the first flower fails to develop fruit.     

BREEDING SYSTEM EVOLUTION IN LEAVENWORTHIA: BREEDING SYSTEM VARIATION AND REPRODUCTIVE SUCCESS IN NATURAL POPULATIONS OF LEAVENWORTHIA CRASSA (CRUCIFERAE)

Populations of Leavenworthia crassa (Cruciferae) studied for 3 years exhibited among- and within-population genetic variation for a suite of floral and reproductive traits (flower width, petal length, anther position, ability to set seeds in the absence of pollinators, time to first flowering) associated with breeding system. We used electrophoretic markers to show that a population with small, monomorphically colored flowers with introrse anthers had a significantly lower outcrossing rate (t = 0.03) than did a population with larger, polymorphically colored flowers with extrorse anthers (t = 0.33). In the more-outcrossing population the correlation between higher maternal plant outcrossing rate and the suite of six traits approached significance (P < 0.067), with greater petal size, greater flower width, and reduced ability to set seeds in the absence of pollinators contributing significantly. Plants in selfing populations had a generally higher reproductive success, with a higher number of flowers per plant, a smaller proportion of unfertilized ovules, a smaller proportion of fertilized ovules aborted, a higher rate of fruit set, and overall a larger number of seeds matured than did plants from the more outcrossing populations. Pollen limitation did not appear to account for lower reproductive success in outcrossing populations. Resource limitation did not differ substantially between populations. However, within-ovary patterns of fertilization, abortion, and seed weight were significantly less random in outcrossing populations than in selfing populations, suggesting that differential gamete and embryo success may be responsible for lower reproductive success in outcrossing populations.     

Relatively weak inbreeding depression in selfing but also in outcrossing populations of North American Arabidopsis lyrata

Hermaphroditic plants can potentially self‐fertilize, but most possess adaptations that promote outcrossing. However, evolutionary transitions to higher selfing rates are frequent. Selfing comes with a transmission advantage over outcrossing, but self‐progeny may suffer from inbreeding depression, which forms the main barrier to the evolution of higher selfing rates. Here, we assessed inbreeding depression in the North American herb Arabidopsis lyrata, which is normally self‐incompatible, with a low frequency of self‐compatible plants. However, a few populations have become fixed for self‐compatibility and have high selfing rates. Under greenhouse conditions, we estimated mean inbreeding depression per seed (based on cumulative vegetative performance calculated as the product of germination, survival and aboveground biomass) to be 0.34 for six outcrossing populations, and 0.26 for five selfing populations. Exposing plants to drought and inducing defences with jasmonic acid did not magnify these estimates. For outcrossing populations, however, inbreeding depression per seed may underestimate true levels of inbreeding depression, because self‐incompatible plants showed strong reductions in seed set after (enforced) selfing. Inbreeding‐depression estimates incorporating seed set averaged 0.63 for outcrossing populations (compared to 0.30 for selfing populations). However, this is likely an overestimate because exposing plants to 5% CO₂ to circumvent self‐incompatibility to produce selfed seed might leave residual effects of self‐incompatibility that contribute to reduced seed set. Nevertheless, our estimates of inbreeding depression were clearly lower than previous estimates based on the same performance traits in outcrossing European populations of A. lyrata, which may help explain why selfing could evolve in North American A. lyrata.     

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.     

Pollination biology of Eriocaulon parkeri in Connecticut

Eriocaulon parkeri B. L. Robinson is a monoecious, pioneer species of tidal mudflats that displays characteristics that suggest outcrossing as a preferred breeding system. Analyses of breeding system dynamics, fruit set, and pollen and seed viabilities were undertaken in Connecticut and Wisconsin to test the hypothesis of entomophyly and outcrossing as a preferred breeding strategy. Potential pollinators included syrphid and long-legged flies. Seed viability was estimated at 94 ± 16% (n = 133); pollen viability at 88 ± 13%. Pollen production averaged ∼500 grains per flower. Pollen–ovule ratios within inflorescences averaged 196:1, suggesting facultative autogamy. Seed set in natural populations averaged 74%, not significantly different between early and late season plants or between greenhouse controls and hand pollinations. Emasculated plants produced small amounts of seed under controlled greenhouse conditions. Pollen tube growth through the styles of E. aquaticum, a related species, was much more prevalent than that of E. parkeri. Results suggest that E. parkeri relies heavily on geitonogamy for seed production. Some seed may be produced by agamospermy.