Pollen limitation meets resource allocation: towards a comprehensive methodology

The standard method of measuring pollen limitation is to add pollen to a number of flowers, preferably to a whole plant, and to compare fruit and seed set with that of naturally pollinated flowers on other plants. In 25 yr of research, this method has yielded valuable data, but it is difficult to use in large plants. This has caused a bias in the available data towards smaller, herbaceous plants with relatively few flowers. I argue that, in order to widen our knowledge of how pollen limitation affects plants, we should go beyond whole-plant pollen addition and change our concept of how a flowering plant functions. The traditional method does not take into account the variation in and dynamics of resource allocation and pollen availability. The concept of integrated physiological units (IPUs) does, but, although it has been applied to pollination biology, it has not received the attention it deserves. I use this article to present its merits again, to propose a step-by-step methodology for studying pollen limitation, and to examine factors influencing possible plant strategies.     

A Top-Down Hierarchy in Fruit Set on Inflorescences in Iris fulva (Iridaceae)

Abstract: In Iris fulva, the apical flower on an inflorescence opens first, and the flowering sequence then proceeds from the most basal flower upward to the apex (acropetally). Flowering order and flower position are thus partially decoupled, and this provides an opportunity to separately investigate the effects of these two factors upon fruit formation. We recorded natural patterns of fruit set and seed production, and found that fruit set patterns were determined by what appears to be a form of apical dominance. The first, apical flower had the highest fruit set and seed production. Fruit set decreased towards the base of the inflorescence, with later-opening flowers having a higher fruit set. This is contrary to the pattern usually observed in other acropetally flowering plants, which is a higher fruit set for basal, early flowers. By performing additional hand pollinations and counting pollen grains on naturally pollinated flowers, we found that pollen deposition was not a major factor limiting fruit set, and that it could not explain the large difference in fruit set between the apical and basal flowers. Removing the first flower after it had wilted increased fruit set in the remaining flowers, but mainly in the more apical flowers. Only by removing the two topmost flowers could we obtain an increase in fruit set for the basal flower. The fact that the basal flower rarely sets fruit, despite being closer to resources, suggests that the apical meristem is either a strong sink for resources or produces hormones that form a gradient along the inflorescence, which is comparable with apical dominance.     

Effects of salt stress on the reproductive biology of the halophyte <Emphasis Type="Italic">Plantago crassifolia</Emphasis>

Floral phenology, pollen quality and seed set of Plantago crassifolia plants, grown in the presence of increasing NaCl concentrations, were studied to test how this Mediterranean halophyte responded to salt stress during the reproductive phase of its life cycle. Reproductive success was maximal in plants grown in non-saline conditions, or in the presence of 100 mM NaCl, but it was negatively affected by higher salinities, due to a progressive reduction of pollen fertility, seed set, and seed viability.     

Spatio-temporal variation in pollen limitation and reproductive success of two scape morphs in Primula farinosa

In plant populations where reproductive output is limited by pollinator visitation, plants with attractive floral displays should have a selective advantage. We examined the effect of inflorescence height on pollination success in Primula farinosa, which is dimorphic for scape length. To test the hypothesis that fruit and seed initiation are more strongly pollen-limited in the short-scaped than in the long-scaped morph, and that this difference is affected by spatio-temporal variation in pollen limitation, we conducted a hand-pollination experiment in four populations over 2 yr. Pollen limitation of fruit initiation varied among populations and years, and was stronger in the short-scaped than in the long-scaped morph. The results suggest that interactions with pollinators will need to be considered for a full understanding of the maintenance of this striking polymorphism. The study also shows that, although pollen limitation is likely to vary in space and time in many plant species, such variation is not necessarily associated with variation in selection on floral characters.     

Pollinator‐mediated phenotypic selection does not always modulate flower size and number in the large‐flowered Mediterranean shrub Cistus ladanifer (Cistaceae)

Flower size and number usually evolve under pollinator‐mediated selection. However, hot, dry environments can also modulate display, counteracting pollinator attraction. Increased pollen deposition on larger flower displays may not involve higher female fitness. Consequently, stressful conditions may constrain flower size, favouring smaller‐sized flowers. The large‐flowered, self‐incompatible Mediterranean shrub Cistus ladanifer was used to test that: (1) this species suffers pollen limitation; (2) pollinators are spatially–temporally variable and differentially visit plants with more/larger flowers; (3) increased visits enhance reproduction under pollen limitation; (4) stressful conditions reduce female fitness of larger displays; and (5) phenotypic selection on floral display is not just pollinator‐mediated. We evaluated pollen limitation, related floral display to pollinator visits and fruit and seed production and estimated phenotypic selection. Flower size was 7.2–10.5 cm and varied spatially–temporally. Visitation rates (total visits/50 min) ranged from 0.26 to 0.43 and increased with flower size. Fruit set averaged 80% and seed number averaged 855, but only fruit set varied between populations and years. Selection towards larger flowers was detected under conditions of pollen limitation. Otherwise, we detected stabilizing selection on flower size and negative selection on flower number. Our results suggest that selection on floral display is not only pollinator‐dependent through female fitness in C. ladanifer. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 176 , 540–555.     

Fitness components of the hybrid Phlomis x margaritae Aparicio & Silvestre (Lamiaceae)

Studies of microsporogenesis, pollen germination, pollen stainability, fruit and seed set and ovary predation have been undertaken to evaluate the fitness of the hybrid genotype of Phlomis x margaritae relative to its parents (P. purpurea and P. x composita) and allied taxa (P. x composita nothom. composita, P. x composita nothom. trulknqueiz , and P. lychnitis). Phlomis x margaritae shows irregular meiosis with very low pollen germination rate and very high pollen sterility, no seed set, and very high ovary predation. It thus has very low reproductive success and fitness. Phlomis x composita nothom. trullenquei also shows low pollen germination and seed set, but these follow regular microsporogenesis. The existence of cryptic structural sterility is suggested for this taxon. The rest of the studied taxa (P. purpurea, P. lychnitis and P. x composita nothom. composita) were fertile and can support the specific or subspecific taxonomic status of P. x composita. A fixed ovule abortion pattern in the taxa with yellow flowers was found, since 'brood size' consisted just of one seed per fruit. On the contrary, P. purpurea was found to produce from one to four seeds per fruit. Some conservation implications are suggested.     

Floral sex ratios, disease and seed set in dioecious Silene dioica

1 In the dioecious, perennial herb Silene dioica , the density of pollen donors in a population is determined by overall plant density, the sex ratio and the proportion of plants infected with the anther-smut fungus Microbotryum violaceum , which results in permanent sterility of both male and female plants.
2 Pollinators ( Bombus spp.) were found to prefer male flowers and to avoid diseased flowers. This may result in an overall lower visitation frequency and increased risk for pollen limitation in populations with a low density of males or a high incidence of disease.
3 Compared with open-pollinated flowers, hand pollination resulted in a significant increase in the number of seeds produced per fruit in populations with an experimentally reduced proportion of males (25% and 50% male flowers) but not in a naturally male-dominated population (75% male flowers). Seed production per plant was increased by hand pollination only in the most female-dominated population. Because the floral sex ratio is often male-biased, resources rather than pollen availability are likely to set the upper limit for total seed production per individual in most healthy populations of S. dioica.
4 There was a negative relationship between seed set and incidence of disease across 22 populations in both years of a field study. However, there was no consistent difference between the responses of highly diseased populations (incidence 30–56%) and populations with a low disease incidence (incidence 0–8%) to hand pollination.
5 In a greenhouse experiment with cloned hand-pollinated females, the presence of spores on healthy flowers was found to reduce seed set significantly. In highly diseased populations, therefore, the frequent deposition of spores by flower visitors onto remaining healthy plants may decrease seed production below the potential level determined by resources or pollen availability.     

西藏高原高寒植物长鞭红景天生殖特征成功及影响因子

为了探索高寒植物长鞭红景天(Rhodiola fastigiata)濒危的生殖生态学机理,寻找保护和恢复该物种的有效途径,对其繁育系统、开花物候特征、生殖过程(开花、结实、种子产量和发育)及其影响生殖成功的主要因子进行调查分析。结果表明:长鞭红景天包括两性植株和雌性植株两种类型,其中两性植株仅执行雄性功能,种群繁殖为异交,属风媒传粉方式。种群花期出现在1a中降雨和气温较高的季节,6月下旬开始开花,7月下旬结束,时间30d左右。单株开花动态呈现单峰和双峰曲线两种模式,两性植株花期(9.29d)大于雌性植株花期(7.06d),并先于雌性植株开放,但雌性植株开花的高峰期比两性花早。从花蕾到开花,雌性单株有16%—25%的花蕾损失;开花到结实,单株36%—57%的花败育;至种子成熟,共有84%—94%的胚珠损失,仅6%—16%的胚珠发育成种子。雌性单株平均种子产量估计值约为285粒,说明长鞭红景天生殖潜力较大。人工条件下种子发芽率为93.6%,可见种子质量不是导致其濒危的原因,花粉限制和种子产量可能是影响生殖成功的主要因素。     

Proximate Limitations to Fruit and Seed Set in Phellodendron amurense var. sachalinense

Abstract Fruit and seed set in a dioecious tree, Phellodendron amurense var. sachalinense grown in isolation, were studied for three years in Hokkaido, northern Japan. Hand pollination did not affect fruit set during the three years, but did affect seed-set by increasing the proportion of fruits with higher seed numbers. Artificial defoliation resulted in decreased mean seed mass, but caused no significant differences in mean seed number per fruit. Total seed mass per infructescence increased with inflorescence size except with artificial defoliation where it reached an upper limit. Although the results of the artificial defoliation treatment indicate that resources are most important for fruit and seed set, it is concluded that fruit and seed set of the tree are not limited by resources, since the number of fruits increased with inflorescence size and an artificial defoliation treatment did not decrease seed-number.     

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.     

花粉和资源限制对青阳参坐果率的影响

以生长于自然种群的青阳参植株为材料进行补充授粉试验,以人工种植的青阳参植株为材料进行资源限制试验,探讨花粉和资源限制对青阳参坐果率的影响.结果表明:自花授粉、天然授粉和异株异花授粉植株的结实率分别为1.7％、2.25 ％和24％,花粉来源对青阳参坐果存在显著影响;补充施肥和不补充施肥植株的坐果率分别为5.12％和3.1...