The division of the generative nucleus and the formation of callose plugs in pollen tubes of Aechmea fasciata (Bromeliaceae) cultured in vitro

The effect of different external factors on pollen germination and pollen tube growth is well documented for several species. On the other hand the consequences of these factors on the division of the generative nucleus and the formation of callose plugs are less known. In this study we report the effect of medium pH, 2-[N-morpholino]ethanesulfonic acid (MES) buffer, sucrose concentration, partial substitution of sucrose by polyethyleneglycol (PEG) 6000, arginine (Arg), and pollen density on the following parameters: pollen germination, pollen tube length, division of the generative nucleus, and the formation of callose plugs. We also studied the different developmental processes in relation to time. The optimal pH for all parameters tested was 6.7. In particular, the division of the generative nucleus and callose plug deposition were inhibited at lower pH values. MES buffer had a toxic effect; both pollen germination and pollen tube length were lowered. MES buffer also influenced migration of the male germ unit (MGU), the second mitotic division, and the formation of callose plugs. A sucrose concentration of 10% was optimal for pollen germination, pollen tube growth rate and final pollen tube length, as well as for division of the generative nucleus and the production of callose plugs. Partial substitution of sucrose by PEG 6000 had no influence on pollen germination and pollen tube length. However, in these pollen tubes the MGU often did not migrate and no callose plugs were observed. Pollen tube growth was independent of the migration of the MGU and the deposition of callose plugs. In previous experiments Arg proved to be positive for the division of the generative nucleus in pollen tubes cultured in vitro. Here, we found that more pollen tubes had callose plugs and more callose plugs per pollen tube were produced on medium with Arg. After the MGU migrated into the pollen tube (1 h after cultivation), callose plugs were deposited (3 h). After 8 h the first sperm cells were produced. The MGU moved away from the active pollen tube tip until the second pollen mitosis occurred, thereafter the distance from the MGU to the pollen tube tip diminished. Callose plug deposition never started prior to MGU migration into the pollen tube. Pollen tubes without a MGU also lack callose plugs (±30% of the total number of pollen tubes). Furthermore, we found a correlation between the occurrence of sperm cells in pollen tubes and the synthesis of callose plugs.     

Branch direction in Carnegiea gigantea (Cactaceae): Regional patterns and the effect of nurse plants

Abstract. Branch direction in Carnegiea gigantea (saguaro) cacti affects PAR interception and CO₂ uptake, and may influence reception of photomorphogenic cues. Branch direction data were collected in 29 saguaro populations over Arizona to determine if the southerly bias observed in previous studies at two locales can be generalized to the American range of the saguaro, and to determine the effect of shading by nurse plants on branch direction. Locally observed S bias from previous studies can be extended and generalized to include the American saguaro range. Branches were statistically uniform across environmental variability regarding temperature and precipitation. The S bias was not found in saguaros shaded by large nurse plants, supporting the assertion that branch direction occurs, in part, to maximize receipt of insolation.     

Variation of pollen and ovule parameters among different ploidy levels ofCorydalis (Fumariaceae)

The size and number of pollen grains and ovules are compared between 20 populations of different ploidy levels in two self-incompatible species ofCorydalis (Fumariaceae), to test the presence of ploidy-related variation in these reproductive characters. In both species, higher ploidy levels are associated with larger pollen grains, ovules and corolla, but the number of pollen grains and ovules are not different between ploidy levels. The investment per flower is consequently larger at higher ploidy levels, but the flower number per individual is lower, suggesting that the mode of partition of the investment for sexual reproduction varies between different ploidy levels. InC. orthoceras, sex allocation estimated by pollen:ovule ratios in number and volume is more female-biased in polyploids than in diploids. In spite of these variations, the characters studied can not be used as indicators of ploidy level due to the large overlaps between the ploidy levels.     

桔梗花粉萌发与花粉管生长研究

以2年生桔梗植株为材料,采用液体培养法研究了培养基种类、PEG、蔗糖、pH以及培养温度、培养时间对桔梗花粉离体萌发生长的影响,结果表明:(1)浓度为100~150 g.L-1的PEG可显著促进桔梗花粉萌发和花粉管的生长;200~250 g.L-1PEG显著促进花粉萌发,但对花粉管生长的作用不显著。(2)100 g.L-1的蔗糖有利于花粉萌发和花粉管生长,高浓度蔗糖(200 g.L-1)有明显抑制作用;(3)桔梗花粉离体萌发和花粉管生长的适宜培养基为ME3+BK+10%蔗糖+150 g.L-1PEG(pH5.8);(4)25~40℃条件下桔梗花粉均可较好萌发,以30℃培养1.5 h为最佳培养条件。     

Variation in plant functional traits across and within four species of Western Australian Banksia (Proteaceae) along a natural climate gradient

Plant traits are fundamental components of the ecological strategies of plants, relating to how plants acquire and use resources. Their study provides insight into the dynamics of species geographical ranges in changing environments. Here, we assessed the variation in trait values at contrasting points along an environmental gradient to provide insight into the flexibility of species response to environmental heterogeneity. Firstly, we identified how commonly measured functional traits of four congeneric species (Banksia baxteri, B. coccinea, B. media and B. quercifolia) varied along a longitudinal gradient in the South Western Australian Floristic Region. This regional gradient provides significant variation in moisture, temperature and soil nutrients: soil nitrogen content decreases with declining rainfall and increasing temperature. We hypothesized that (i) the regional pattern in trait–environment associations across the species would match those observed on a global scale and (ii) that the direction and slopes of the within‐species relationships would be similar to those across species for each of the measured traits. Along the regional gradient we observed strong shifts in trait values, and cross‐species relationships followed the expected trend: specific leaf area was significantly lower, and leaf N_area and seed dry mass significantly higher, at the drier end of the rainfall gradient. However, traits within species were generally not well correlated with habitat factors: we found weak patterns among populations, either due to the small ecological gradient or perhaps because fine‐scale structuring among populations (at a micro‐evolutionary scale) was low due to high gene flow within species. Understanding how species traits shift as a result of climatic influences, both at the regional (across species) and local (within species) scale, provides insight into plant adaptation to the environment. Such studies have important applications for conservation biology and population management in the face of global change.     

Differential self‐fertilization rates in response to variation in floral traits within inflorescences of Aquilegia buergeriana var. oxysepala (Ranunculaceae)

To assess variation in the proportion of self‐fertilized seeds among flowers within inflorescences and the relationship between floral traits and the rate of self‐fertilization, the proportion of self‐fertilized seeds among individual flowers was estimated using ten microsatellite markers in self‐compatible plants of Aquilegia buergeriana var. oxysepala. Within‐inflorescence variation in floral traits, such as the duration of the male and female phases, flower size, herkogamy and the number of pollen grains and ovules in two natural populations, were investigated. The first flower in an inflorescence produced more seeds and a higher proportion of self‐fertilized seeds than the second flower. The higher proportion of self‐fertilized seeds in the first flowers was accompanied by a higher number of pollen grains and ovules in the bud stage and the female phase. These results indicate that the high proportion of self‐fertilized seeds in the first flowers in an inflorescence may be due to the high number of remaining pollen grains in the female phase. This suggests that variation in floral traits within inflorescences affects seed quality and quantity among flowers within inflorescences.     

Effect of male age on sperm traits and sperm competition success in the guppy (Poecilia reticulata)

Deleterious mutations can accumulate in the germline with age, decreasing the genetic quality of sperm and imposing a cost on female fitness. If these mutations also affect sperm competition ability or sperm production, then females will benefit from polyandry as it incites sperm competition and, consequently, minimizes the mutational load in the offspring. We tested this hypothesis in the guppy (Poecilia reticulata), a species characterized by polyandry and intense sperm competition, by investigating whether age affects post‐copulatory male traits and sperm competition success. Females did not discriminate between old and young males in a mate choice experiment. While old males produced longer and slower sperm with larger reserves of strippable sperm, compared to young males, artificial insemination did not reveal any effect of age on sperm competition success. Altogether, these results do not support the hypothesis that polyandry evolved in response to costs associated with mating with old males in the guppy.     

The effect of temperature on the duration of the effective pollination period in ‘Oblica’ olive (Olea europaea) cultivar

The effective pollination period (EPP) was determined for ‘Oblica’ olive trees in response to sequential pollinations at anthesis and at increasing days after. The role played by each of three components of the EPP, namely the stigmatic receptivity, pollen tube growth and ovule longevity was examined and the duration of the EPP under different environmental conditions was calculated. An increase in temperature in the field was caused by enclosing bearing trees within polyethylene cages. The duration of the EPP varied between years and environmental conditions. Initial fruit set (IFS) declined gradually in response to sequential cross‐pollination. The significant decrease with respect to the maximum levels of IFS occurred 5 days after anthesis (DAA) in 2007 and between 4 and 5 DAA in 2010 at both environmental conditions. However, the fruit set decrease with sequential pollinations was sharper for the trees inside the cages than for trees growing at the open air, suggesting that high temperatures induce earlier and more pronounced decrease in flower fertility. The stigma receptivity was quite extended decreasing slowly with time. In 2010, the stigma receptivity was prolonged beyond sampling date (7 DAA) in both environmental conditions. Stigmas first lost the capacity to support pollen germination and later their capacity to adhere pollen grains. Pollen tube growth occurred rapidly and fertilization took place 1 or 2 days after pollination. Cross‐pollination resulted with higher fertilization levels, whereas fertilization was reduced and delayed under self‐pollination. Enclosing the trees within the cages had small effect on cross‐pollination but impaired the self‐pollination which resulted with reduced levels of fertilization. The ovule longevity was prolonged beyond the end of sampling in both years and conditions (beyond 9 DAA in 2009 or 7 DAA in 2010). Discrepancy between the estimates of the duration of the EPP based on analyses of its components and those indicated by the declines in fruit set from delayed pollination suggests that a different component of the pistil, the style, could have the critical role in determining the duration of the EPP.     

Evolution of mating system and the genetic covariance between male and female investment in Clarkia (onagraceae): selfing opposes the evolution of trade-offs

Sex allocation theory has assumed that hermaphroditic species exhibit strong genetically based trade-offs between investment in male and female function. The potential effects of mating system on the evolution of this genetic covariance, however, have not been explored. We have challenged the assumption of a ubiquitous trade-off between male and female investment by arguing that in highly self-fertilizing species, stabilizing natural selection should favor highly efficient ratios of male to female gametes. In flowering plants, the result of such selection would be similar pollen:ovule (P:O) ratios across selfing genotypes, precluding a negative genetic correlation (r(g)) between pollen and ovule production per flower. Moreover, if selfing genotypes with similar P:O ratios differ in total gametic investment per flower, a positive r(g) between pollen and ovule production would be observed. In outcrossers, by contrast, male- and female-biased flowers and genotypes may have equal fitness and coexist at evolutionary equilibrium. In the absence of strong stabilizing selection on the P:O ratio, selection on this trait will be relaxed, resulting in independence or resource-based trade-offs between male and female investment. To test this prediction, we conducted artificial selection on pollen and ovule production per flower in two sister species with contrasting mating systems. The predominantly self-fertilizing species (Clarkia exilis) consistently exhibited a significant positive r(g) between pollen and ovule production while the outcrossing species (C. unguiculata) exhibited either a trade-off or independence between these traits. Clarkia exilis also exhibited much more highly canalized gender expression than C. unguiculata. Selection on pollen and ovule production resulted in little correlated change in the P:O ratio in the selfing exilis, while dramatic changes in the P:O ratio were observed in unguiculata. To test the common prediction that floral attractiveness should be positively genetically correlated with investment in male function, we examined the response of petal area to selection on pollen and ovule production and found that petal area was not consistently genetically correlated with gender expression in either species. Our results suggest that the joint evolutionary trajectory of primary sexual traits in hermaphroditic species will be affected by their mating systems; this should be taken into account in future theoretical and comparative empirical investigations.     

叶幕PAR光能截留和分配对葡萄群体光合同化物源库关系的调控

 田间自然条件下在葡萄园群体水平上多年的研究证明：叶幕PAR光能截留率与葡萄群体净光合速率、叶幕和单叶PAR光能截留率与果实干物质总量占地上部生物量干重的百分比、与果实总糖产量占果实本身干物质总量的百分比、以及与果皮色素产量占果实本身干物质总量的千分比之间呈现显著或极显著的线性正相关关系。说明利用叶幕结构变异调节叶幕PAR光能截留和分配,可以对光合同化物源库关系和果实中物质代谢方向进行有效的调控。PAR光能截留率较高和分配合理的叶幕,不但通过较高的群体光合速率为产量和品质形成提供了丰富的同化物“源”,而且通过调节器官间“库”关系使同化物以较高的比例流向果实,同时使果实中物质代谢过程有利于合成构成品质的要素。     

梅花个体内花柱长度的变异及其对繁殖成功的影响

植物个体内花型的变异影响繁殖成功, 雌性繁殖性状的变异可能影响雌性的繁殖成功, 也可能作为花粉的受体影响雄性的繁殖成功。然而, 植物个体内不同花柱长度的花产生的花粉是否影响植物的繁殖成功却少有研究。梅(Armeniaca mume)是原产我国的重要木本花卉和经济果树, 我们的野外观察发现在同一植株内, 不同花的花柱长度有变异, 存在长柱型、短柱型和雄花型(雌蕊败育)三种花型, 是比较雌性繁殖性状的变异对两性繁殖成功的影响的理想材料。本文主要测量了不同花型的花部特征, 统计花期, 并开展体外花粉萌发以及人工控制授粉实验。结果表明: 长柱型的花冠直径、雌蕊长、单花花粉数、花粉体积显著大于短柱型和雄花型。长柱型的单花期以及雌花期显著长于短柱型。长柱型、短柱型以及雄花型花粉在活体柱头上的萌发率没有显著性差异, 雄花型的花粉管长度显著高于长柱型和短柱型。长柱型为母本的花粉萌发率以及花粉管的长度要显著高于短柱型。长柱型、短柱型、雄花型花粉授粉与自然对照处理的坐果率没有显著性差异, 而长柱型为母本的坐果率要显著高于短柱型为母本的坐果率。这些结果表明野生梅花的长柱型为母本有利于花粉的萌发和花粉管的伸长, 有高的坐果率; 但其作为花粉供体的雄性功能与其他花型没有差异。     

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.     

Differential pollination success in the course of individual flower development and flowering time in Gentiana pneumonanthe L. (Gentianaceae)

The effect of flowering time and anthesis stage on fecundity of Gentiana pneumonanthe L. (Gentianaceae) in a greenhouse experiment was studied. The species is self-compatible, not apomictic, and rarely self-pollinates spontaneously. On average, flowers lived for 11.4 days, starting with a male stage of 3.6 and continuing with a female stage of 7.9 days. Pollen viability was highest during the first two days of the male stage, but never exceeded 50%. Seed-set from cross-pollination was higher earlier in the flowering period (77%) than later (63Vo). For self-pollinations, no significant difference was observed. The success of hand-pollinations declined dramatically after the fourth day of the female stage. Self-pollination yielded lower seed-set on the first day of the female stage than on the following three days. Also, there was a significant difference in seed-set between self- and cross-pollinations (25% vs 55%, respectively) only on the first day. This suggests that selection against self pollen was stronger during the first stage of the female phase and weakened as the stylar tissue aged. In conclusion, two different ways to enhance outcrossing were observed: (1) within the population's flowering period, and (2) within an individual flower's lifespan. Both may have important effects on the genetic variability of G. pneumonanthe offspring under unfavourable conditions. Wese include small population size (associated with reduced pollinator service), general pollinator scarcity, or rainy summers.     

Phenotypic selection on floral traits in the arctic plant Parrya nudicaulis (Brassicaceae)

The evolution of floral traits is often attributed to pollinator‐mediated selection; however, the importance of pollinators as selective agents in arctic environments is poorly resolved. In arctic and subarctic regions that are thought to be pollen limited, selection is expected to either favor floral traits that increase pollinator attraction or promote reproductive assurance through selfing. We quantified phenotypic selection on floral traits in two arctic and two subarctic populations of the self‐compatible, but largely pollinator‐dependent, Parrya nudicaulis. Additionally, we measured selection in plants in both open pollination and pollen augmentation treatments to estimate selection imposed by pollinators in one population. Seed production was found to be limited by pollen availability and strong directional selection on flower number was observed. We did not detect consistently greater magnitudes of selection on floral traits in the arctic relative to the subarctic populations. Directional selection for more pigmented flowers in one arctic population was observed, however. In some populations, selection on flower color was found to interact with other traits. We did not detect consistently stronger selection gradients across all traits for plants exposed to pollinator selection relative to those in the pollen augmentation treatment; however, directional selection tended to be higher for some floral traits in open‐pollinated plants.     

不同脱水处理对桔梗花粉离体萌发及贮藏性能的影响

对桔梗花粉离体萌发条件进行了筛选,并采用花粉离体萌发法测定了不同干燥方式和贮藏时间处理对桔梗花粉生活力的影响;同时采用人工授粉的方法研究了不同处理后桔梗花粉授粉结实能力的变化。结果表明：（1）30℃培养1.5h是桔梗花粉离体萌发和花粉管生长的最佳培养条件。（2）不同干燥方式、不同干燥时间处理后桔梗花粉萌发率和花粉管长度均有所下降,但下降幅度不同,其中以干燥箱45℃烘干处理1.5h~2h下降最少,其花粉耐藏性也有明显提高。（3）不同干燥方式处理桔梗花粉后其授粉结籽率的变化明显不同,阴置2h处理的花粉结籽率最低,而干燥器内放置6h、日晒2h和烘干1.5h处理后花粉结籽率略有变化;干燥处理后随贮藏时间的增加,其授粉结籽率均呈下降趋势,但45℃烘干1.5h处理的花粉结籽率下降速度最慢,其授粉结籽能力可保持5d左右。     

Intra‐annual variation of the spatiotemporal distribution and abundance of Talitridae and Oniscidea (Crustacea,Peracarida) at Bizerte Lagoon (northern Tunisia)

During 1 year from spring 2007 to winter 2008, we conducted four seasonal samplings along a transect in the supralittoral zone of Bizerte Lagoon at Menzel Jmil (Tunisia) to study the intra‐annual variation of Peracarida diversity, Talitridae and Oniscidea. For each season, one transect was studied and quadrates were placed successively from the shoreline to the road backing the shore. Talitridae and Oniscidea were the most abundant taxa in our samples reaching highest densities in summer, when fourteen species were found. The minimum and maximum values of species richness of Talitridae were observed in winter (four species) and summer (eight species), respectively, while for Oniscidea, species richness ranged from one species in winter and five species in spring. Both Talitridae and Oniscidea exhibited intra‐annual variation in their spatial distribution; in that, they moved away from the shoreline in winter and were found near the waterline in the other seasons. ANOVA test shows a high significant correlation between both Talitridae with Suaeda maritima and Oniscidea with the plants S. maritima,Salicornia arabica and Obione portulacoides.     

In vitro asymbiotic germination and seedling development of Limodorum abortivum (Orchidaceae)

Abstract

Germination tests were carried out using immature seeds of Limodorum abortivum and applying in vitro techniques. The results proved that BM₁ culture medium is suitable to promote both germination and further growth stages. Details of the developmental pattern, and some micromorphological features, are described.