Effect of flowering stage and storage conditions on pollen quality of six male date palm genotypes

Availability of efficient male genotypes is critical for successful artificial pollination and regular bearing of female date palms. The effect of flowering stage and storage conditions on pollen quality of six male date palm genotypes encoded ‘ABD1′, ‘P4′, ‘P3′, ‘P8′, ‘P7′ and ‘P13′were evaluated. Pollen collected from spathes developed at the middle of flowering stage exhibited the best viability (90%) and germinability (85%) compared to other stages. Pollen viability was greater than 90%, except for ‘P8′ that exhibited 80%, while, germinability greatly varied among the genotypes. Pollen quality decreased during 4 months of storage upon genotype and temperature, with a minimum reduction at ?30 °C followed by 4 °C. Heat shock exposure (33 ± 2 °C) following storage revealed that pollen stored at ?30 °C or 4 °C should be used for pollination on the same day of take out to avoid dramatic quality loss. The ‘ABD1′, an early flowering genotype, proved highest pollen quality both at fresh stage and after storage. While, the ‘P3′, a late flowering genotype, retained its pollen quality during storage. However, the ‘P13′ genotype exhibited excellent pollen quality when fresh, but greatly loses germinability during storage.     

Deterioration of ovary plays a key role in heat stress-induced spikelet sterility in sorghum

In sorghum (Sorghum bicolor [L.] Moench), the impact of heat stress during flowering on seed set is known, but mechanisms that lead to tolerance are not known. A diverse set of sorghum genotypes was tested under controlled environment and field conditions to ascertain the impact of heat stress on time-of-day of flowering, pollen viability, and ovarian tissue. A highly conserved early morning flowering was observed, wherein >90% of spikelets completed flowering within 30 min after dawn, both in inbreds and hybrids. A strong quantitative impact of heat stress was recorded before pollination (reduced pollen viability) and post pollination (reduced pollen tube growth and linear decline in fertility). Although viable pollen tube did reach the micropylar region, 100% spikelet sterility was recorded under 40/22°C (day/night temperatures), even in the tolerant genotype Macia. Heat stress induced significant damage to the ovarian tissue near the micropylar region, leading to highly condensed cytoplasmic contents and disintegrated nucleolus and nucleus in the susceptible genotype RTx430. Whereas, relatively less damages to ovarian cell organelles were observed in the tolerant genotype Macia under heat stress. Integrating higher tolerance in female reproductive organ will help in effective utilization of the early morning flowering mechanism to enhance sorghum productivity under current and future hotter climate.     

Pollen,ovules, and pollination in pea: Success,failure, and resilience in heat

Field pea (Pisum sativum), a major grain legume crop, is autogamous and adapted to temperate climates. The objectives of this study were to investigate effects of high temperature stress on stamen chemical composition, anther dehiscence, pollen viability, pollen interactions with pistil and ovules, and ovule growth and viability. Two cultivars (“CDC Golden” and “CDC Sage”) were exposed to 24/18°C (day/night) continually or to 35/18°C for 4 or 7 days. Heat stress altered stamen chemical composition, with lipid composition of “CDC Sage” being more stable compared with “CDC Golden.” Heat stress reduced pollen viability and the proportion of ovules that received a pollen tube. After 4 days at 35°C, pollen viability in flower buds decreased in “CDC Golden,” but not in “CDC Sage.” After 7 days, partial to full failure of anthers to dehisce resulted in subnormal pollen loads on stigmas. Although growth (ovule size) of fertilized ovules was stimulated by 35°C, heat stress tended to decrease ovule viability. Pollen appears susceptible to stress, but not many grains are needed for successful fertilization. Ovule fertilization and embryos are less susceptible to heat, but further research is warranted to link the exact degree of resilience to stress intensity.     

Effects of high-temperature stress on microsporogenesis in heat-sensitive and heat-tolerant genotypes of Phaseolus vulgaris

High ambient temperature (32/27 °C, day/night, 12 h photoperiod) applied prior to anthesis to Phaseolus vulgaris plants results in abnormal pollen and anther development during microsporogenesis. Scanning and transmission electron microscopy were used to examine anther and pollen morphology and pollen wall architecture after heat stress was applied to two genotypes that differ with respect to yield potential under high‐temperature field conditions: one, a heat‐sensitive, Mesoamerican genotype, A55, the second, a heat‐tolerant, Andean genotype, G122. High‐temperature treatment of both genotypes was applied 1–13 d before anthesis. Under heat stress, the heat‐tolerant genotype showed anther and pollen characteristics that were generally similar to the low temperature controls. In contrast, after 9 d of heat treatment before anthesis, the anthers of the heat‐sensitive genotype were indehiscent and contained abnormal pollen. Pollen wall architecture was also affected in the 12 and 13 d treatments. In addition to the morphological changes, the heat‐sensitive genotype also experienced reduced pollen viability and reduced yield in high‐temperature experiments conducted in both the greenhouse and field.     

Pollinator preference and pollen viability mediated by flower color synergistically determine seed set in an Alpine annual herb

Gentiana leucomelaena manifests dramatic flower color polymorphism, with both blue‐ and white‐flowered individuals (pollinated by flies and bees) both within a population and on an individual plant. Previous studies of this species have shown that pollinator preference and flower temperature change as a function of flower color throughout the flowering season. However, few if any studies have explored the effects of flower color on both pollen viability (mediated by anther temperature) and pollinator preference on reproductive success (seed set) in a population or on individual plants over the course of the entire flowering season. Based on prior observations, we hypothesized that flower color affects both pollen viability (as a function of anther temperature) and pollen deposition (as a function of pollinator preference) to synergistically determine reproductive success during the peak of the flowering season. This hypothesis was tested by field observations and hand pollination experiments in a Tibetan alpine meadow. Generalized linear model and path analyses showed that pollen viability was determined by flower color, flowering season, and anther temperature. Anther temperature correlated positively with pollen viability during the peak of the early flowering season, but negatively affected pollen viability during the peak of the mid‐ to late flowering season. Pollen deposition was determined by flower color, flowering season (early, or mid‐ to late season), and pollen viability. Pollen viability and pollen deposition were affected by flower color that in turn affected seed set across the peak of the flowering season (i.e., when the greatest number of flowers were being pollinated). Hand pollination experiments showed that pollen viability and pollen deposition directly influenced seed set. These data collectively indicate that the preference of pollinators for flower color and pollen viability changed during the flowering season in a manner that optimizes successful reproduction in G. leucomelaena. This study is one of a few that have simultaneously considered the effects of both pollen viability and pollen deposition on reproductive success in the same population and on individual plants.     

Influence of Temperature and Ageing of Ovules and Pollen on Reproductive Success in Trifolium repens L.

Four clones of Trifolium repens L., originating from Iceland,Denmark and Israel, were cultivated in controlled environmentchambers at 10, 14, 18 and 22°C. Seed set at 10°C was,on average, 45% lower than at 18°C. Generally, seed frequencieswere higher in the stylar ovule positions than in the basal.This polarization became more pronounced at low temperatures.At least 30% of the decrease in seed set was caused by inadequatepollen tube growth. This was further supported by in vitro studieswhich showed that pollen developed significantly shorter pollentubes at low temperatures and a strong positive correlationbetween the capacity of male parents to induce seed set in basalovule positions and the length of pollen tubes in vitro. Femaleage, determined by days relative to anthesis, was also foundto have a significant influence on seed set, i.e. when pollinationwas delayed until the fifth day of flowering, seed set decreasedby an average of 60% when compared to florets pollinated onthe first day of anthesis. Pollen age did not influence seedset. Ageing of pollen decreased in vitro germinability, tubeelongation and the percentage of pollen tubes with sperm cells.Seed set, pollen germinability, pollen tube lengths, and abilityto form sperm cells in vitro varied significantly among clones.Copyright1994, 1999 Academic Press Trifolium repens, white clover, temperature, pollen tube, sperm cells, ovule fertility, seed set     

束花石斛的繁育生物学特性

为了探究束花石斛（Dendrobium chrysanthum）的繁育生物学特性,对其开花习性、花粉活力、柱头分泌特性、人工授粉结实率、自然结实种子育性和贮藏特性进行研究,结果表明：（1）束花石斛花芽萌动期为19d,现蕾期为23 d,群体花期为28 d,单花寿命10～15 d;(2)开花1～3 d花粉活力保持在90%左右,开花至第9天时花粉活力仅为23.7%,初始3 d开花的花粉活力与其他时间取样的活力差异均达极显著水平（P<0.01）,花粉活力随开花时间延长呈由强到弱的趋势;（3）束花石斛的柱头腔从开花第2天开始有分泌物,第6 d时分泌物最多,柱头过氧化氢染色结果表明在开花5～6 d时柱头可授性最强,柱头可授性随开花时间延长呈由弱到强再到弱的趋势;（4）选取花朵开放4 d的花粉和柱头人工授粉,同株异花和同花授粉未见结实。野外结实果质量2.02～4.83 g,种子长0.628～0.845 mm,宽0.082～0.110 mm,长宽比为7.798,成胚率达97.35%,种子活力为90.80%,室温条件保存6个月的种子活力维持在73.20%。上述结果说明,束花石斛单花花期不长,开花初...     

Pollen dispersion, pollen viability and pistil receptivity in Leymus chinensis

BACKGROUND AND AIMS: Leymus chinensis is an economically and ecologically important grass that is widely distributed across eastern areas of the Eurasian steppe. A major problem facing its propagation by man is its low sexual reproductivity. The causes of low fecundity are uncertain, largely because many aspects of the reproductive biology of this species remained unknown or incomplete. This study aims to address some of these issues. METHODS: Pollen dispersion, pollen viability, pollen longevity and pistil receptivity were studied in a representative, natural population of L. chinensis growing in Inner Mongolia. KEY RESULTS: Flowering of L. chinensis occurred at the end of June and lasted for 5 d. Pollination peaked between 1600 h and 1700 h, and about 56.1 % of the total pollen grains were released at this time. Pollen density was highest towards the middle of flowering spikes and lowest at the bottom over the 5 d measurement period. Pollen viability (62.4 %) assessed using TTC was more accurate than using IKI (85.6 %); 50 % of pollen arriving on stigmas germinated. Pollen remained viable for only 3 h and the pollen : ovule ratio was 79 333 : 1. Pistil receptivity lasted for only 3 h and, overall, 86.7 % of pistils were pollinated. Within the spike, the relative fecundity of different positions was middle > lower > upper throughout the period of pollination; daily variation of fecundity was similar to that of the pollen flow. The spikes that opened on the day of highest pollen density exhibited the highest fecundity (36.0 %). No seeds were produced by self-pollination. CONCLUSIONS: The data suggest that low pollen viability, short pollen longevity and short pistil receptivity all appear to contribute to the low seed production typical of this important forage crop.     

The effects of time,temperature and plant variety on pollen viability and its implications for gene flow risk

• Pollen viability affects the probability that a pollen grain deposited on a plant's stigma will produce a viable seed. Because a mature seed is needed before a gene flow event can occur, pollen viability will influence the risk of escape for genetically engineered (GE) crops.
• Pollen viability was measured at intervals for up to 2 h following removal of the pollen from the anthers. It was quantified at three temperatures and for different alfalfa varieties, including both conventional and Roundup Ready (RR) varieties. Pollen viability was assessed using in vitro germination.
• Time since removal from the anthers was the most prevalent factor affecting pollen viability in alfalfa. Pollen viability declined with increasing time at all three temperatures and for all varieties tested. Pollen viability was not affected by temperatures ranging between 25 and 37 °C and did not vary among plant varieties, including conventional and RR varieties.
• Bee foraging behaviour suggested pollen viability within the first 10 min following pollen removal from a flower to most affect seed production. Pollen longevity was predicted to have little impact on seed set and gene flow. Linking pollinator behaviour to pollen viability improved our understanding of its impact on gene flow risk.

     

濒危植物蒙古扁桃花粉活力和柱头可授性研究

用过氧化氢酶法测定蒙古扁桃花粉的活力和寿命,用联苯胺-过氧化氢法测定柱头可授性。结果表明,蒙古扁桃花粉活力在散粉后第2天最高,3d后活力迅速下降,三种类型的植株的花粉活力下降速度存在差异,其活力可持续30d左右。短柱头植株柱头在开花后1d可分泌黏液,长柱头和中柱头花植株的柱头在开花后2d才分泌黏液,短柱头分泌黏液的持续时间较后者长。柱头可授性持续时间同种之间也存在差异,长柱头和中柱头植株可达6d,而短柱头花植株可持续10d左右。     

High temperature stress of Brassica napus during flowering reduces micro- and megagametophyte fertility, induces fruit abortion, and disrupts seed production

High temperature stress (HTS), during flowering, decreases seed production in many plants. To determine the effect of a moderate HTS on flowering, fruit and seed set in Brassica napus, plants were exposed to a HTS (8/16 h dark/light, 18 degrees C night, ramped at 2 degrees C h-1, over 6 h, to 35 degrees C for 4 h, ramped at 2 degrees C h-1 back to 23 degrees C for 6 h) for 1 or 2 weeks after the initiation of flowering. Although flowering on the HTS-treated plants, during both the 1 week and 2 week HTS treatments, was equal to that of control-grown plants, fruit and seed development, as well as seed weight, were significantly reduced. Under HTS, flowers either developed into seedless, parthenocarpic fruit or aborted on the stem. At the cessation of the HTS, plants compensated for the lack of fruit and seed production by increasing the number of lateral inflorescences produced. During the HTS, pollen viability and germinability were slightly reduced. In vitro pollen tube growth at 35 degrees C, from both control pollen and pollen developed under a HTS, appeared abnormal, however, in vivo tube growth to the micropyle appeared normal. Reciprocal pollination of HTS or control pistils with HTS or control pollen indicated that the combined effects of HTS on both micro- and megagametophytes was required to knock out fruit and seed development. Expression profiles for a subset of HEAT SHOCK PROTEINs (HSP101, HSP70, HSP17.6) showed that both micro- and megagametophytes were thermosensitive despite HTS-induced expression from these genes.     

Seed set,pollen morphology and pollen surface composition response to heat stress in field pea

Pea (Pisum sativum L.) is a major legume crop grown in a semi‐arid climate in Western Canada, where heat stress affects pollination, seed set and yield. Seed set and pod growth characteristics, along with in vitro percentage pollen germination, pollen tube growth and pollen surface composition, were measured in two pea cultivars (CDC Golden and CDC Sage) subjected to five maximum temperature regimes ranging from 24 to 36 °C. Heat stress reduced percentage pollen germination, pollen tube length, pod length, seed number per pod, and the seed–ovule ratio. Percentage pollen germination of CDC Sage was greater than CDC Golden at 36 °C. No visible morphological differences in pollen grains or the pollen surface were observed between the heat and control‐treated pea. However, pollen wall (intine) thickness increased due to heat stress. Mid‐infrared attenuated total reflectance (MIR‐ATR) spectra revealed that the chemical composition (lipid, proteins and carbohydrates) of each cultivar's pollen grains responded differently to heat stress. The lipid region of the pollen coat and exine of CDC Sage was more stable compared with CDC Golden at 36 °C. Secondary derivatives of ATR spectra indicated the presence of two lipid types, with different amounts present in pollen grains from each cultivar.     

High‐temperature‐induced reductions in cropping in everbearing strawberries (Fragaria × ananassa) are associated with reduced pollen performance

Heat‐induced short‐term reductions in cropping (often referred to as ‘thermodormancy’ by UK strawberry growers) occur frequently in extended‐season everbearing strawberry production in the UK. A 2‐year study evaluated the effects of high temperatures on the performance of pollen in two everbearing strawberry cultivars (cvs ‘Everest’ and ‘Diamante’). Pollen from both cultivars lost its ability to germinate following exposure to a period of high temperature (30/20°C day/night). However, after termination of the high‐temperature episode, germination capacity recovered to levels similar to control plants that were not exposed to high temperature. High temperature also resulted in slower rates of pollen tube growth. As a result, only a small proportion of high‐temperature‐affected flowers developed into fruits. These results suggest that heat‐induced reductions in cropping in everbearing strawberries are associated with reduced pollen performance.     

Direct analysis of pollen fitness by flow cytometry: implications for pollen response to stress

Sexual reproduction in flowering plants depends on the fitness of the male gametophyte during fertilization. Because pollen development is highly sensitive to hot and cold temperature extremes, reliable methods to evaluate pollen viability are important for research into improving reproductive heat stress (HS) tolerance. Here, we describe an approach to rapidly evaluate pollen viability using a reactive oxygen species (ROS) probe dichlorodihydrofluorescein diacetate (i.e. H₂DCFDA‐staining) coupled with flow cytometry. In using flow cytometry to analyze mature pollen harvested from Arabidopsis and tomato flowers, we discovered that pollen distributed bimodally into ‘low‐ROS’ and ‘high‐ROS’ subpopulations. Pollen germination assays following fluorescence‐activated cell sorting revealed that the high‐ROS pollen germinated with a frequency that was 35‐fold higher than the low‐ROS pollen, supporting a model in which a significant fraction of a flower's pollen remains in a low metabolic or dormant state even after hydration. The ability to use flow cytometry to quantify ROS dynamics within a large pollen population was shown by dose‐dependent alterations in DCF‐fluorescence in response to oxidative stress or antioxidant treatments. HS treatments (35°C) increased ROS levels, which correlated with a ~60% reduction in pollen germination. These results demonstrate the potential of using flow cytometry‐based approaches to investigate metabolic changes during stress responses in pollen.     

珍稀濒危植物细果秤锤树开花生物学特性和繁育系统

为了解珍稀濒危植物细果秤锤树（Sinojackia microcarpa）开花特征和有性繁殖,对其花部形态特征、开花动态、花粉胚珠比（P/O）、柱头可授性、花粉活力、套袋试验、访花昆虫及访花频率进行观测。结果表明：（1）细果秤锤树的杂交指数（OCI）为4,单花期5~7 d,种群花期可持续20 d左右。花粉与胚珠比为4 093.21±498.56。开花后第3天的花粉活力最高（76.21%）,而开花后第7天时花粉活力较低（18.37%）。细果秤锤树柱头最适可授期在开花后第2天。（2）套袋试验表明,细果秤锤树存在部分自交不亲和性,同时不存在无融合生殖,传粉昆虫是其完成生殖过程所必需的,且异株授粉能够提高其坐果率和结籽率。细果秤锤树的访花昆虫有3目5科7种,主要访花昆虫有黄胸木蜂（Xylocopa appendiculata）、熊蜂（Bombus sp.）、胡蜂（Vespa sp.）、中华蜜蜂（Apis cerana）、黑带食蚜蝇（Episyrphus balteatus）、其中熊蜂平均访花频率最高,达（8.67±0.21） 次·h^-1。对该物种开花生物学特征与繁育系统进行深入研究,有利于进一步探究其濒危机制,为后续珍稀濒危植物的研究提供理论依据和参考。     

Physiological factors limit fruit set of tomato (Lycopersicon esculentum Mill.) under chronic, mild heat stress

The effects of chronic, mild heat stress on fruit set, fruit production, release of pollen grains, photosynthesis, night respiration and anther dehiscence were examined in tomatoes (Lycopersicon esculentum Mill.) differing in high‐temperature sensitivity. Plants were grown under three temperature regimes: (1) 28/22 or 26/22 °C (optimal temperature); (2) 32/26 °C (high temperature); and (3) 32/26 °C day/night temperatures relieved at 28/22 °C for 10 d before anthesis, then returned to 32/26 °C (relieving treatment). FLA 7156 was the only cultivar with fruit set at 32/26 °C. All five cultivars, however, had fruit set under the relieving treatment (RT). The longer the relief, the higher the percentage of fruit set. Longer periods of relief also increased the number of pollen grains released, and linear regression analysis showed a significant relationship between the number of pollen grains released and the percentage of fruit set. Germination of pollen grains was also lowered in high‐temperature‐grown plants. The number of pollen grains produced, photosynthesis and night respiration did not limit fruit set under chronic, mild heat stress, however. This suggested that cultivar differences in pollen release and germination under heat stress are the most important factors determining their ability to set fruit.     

Comparing heat stress effects on male-fertile and male-sterile tomatoes

To separate the effects of heat stress on male and female reproductive tissues, male-sterile (MSs) and male-fertile tomatoes (MFs) were placed in growth chambers at 12 h day/12 h night temperatures of 28/22, 30/24 or 32/26 °C from flower appearance to seed maturation (daily mean temperatures of 25, 27 or 29 °C). Pollen from MFs was applied individually to MS flowers. As MFs were self-pollinated, heat stress was experienced by both male and female tissues. At growth temperatures of 29 °C fruit number, fruit weight per plant, and seed number per fruit were only 10%, 6·4% and 16·4%, respectively, compared with those at 25 °C. Heat stress also adversely affected fruitset in MSs, especially when experienced by donor pollen. No fruit at all developed on MSs receiving pollen produced at 29 °C, even when ovule development, pollen germination and subsequent embryo development all took place at 25 °C. Effects on fruitset in MSs were reduced if donor pollen had not experienced heat stress. MSs grown at 29 °C but receiving pollen developing at 25 °C produced 73% as much fruit (both on number and weight basis), had 40% as high fruitset and produced 87% of the seed per fruit as MSs grown at 25 °C. This use of male-sterile and male-fertile lines of tomato provides new evidence that impairment of pollen and anther development by elevated temperature will be an important contributing factor to decreased fruit set in tomato, and possibly other crops, with global warming.     

Evaluation of Viability,Shedding Pattern,and Longevity of Pollen from Genetically Modified (GM) Herbicide-tolerant and Wild-type Zoysiagrass (<Emphasis Type="Italic">Zoysia japonica</Emphasis> Steud.)

Knowledge about pollen viability is important when evaluating the risk of genetically modified (GM) plants. Here, staining via iodine potassium iodide (IKI) or triphenyl tetrazolium chloride (TTC) could not distinguish between live and dead pollen from Zoysia japonica. Therefore, to obtain a reliable assessment of such viability and longevity, we developed an optimum germination medium containing 20% sucrose and 50 ppm H₃BO₃. Pollen grains transferred to the germination medium at about 1000 hours had a germination rate of >90%. Pollen was most predominantly shed at approximately 1000 hours, with viability declining to nearly 0% at 1200 hours. All germinability was lost within 150 min when stored at 25°C. No significant difference was found between GM and non-GM plants in their pollen viability or longevity.     

The size and germinability of Scots pine pollen in different temperatures in vitro

The effect of genotype, the origin of genotype, and germination temperature on Scots pine pollen grain size, hydration rate, germinability, and tube growth was studied in vitro. The mean sizes of dry and germinated pollen grains varied among pollen genotypes in different ways, thus the hydration rate varied among genotypes. Pollen from Scots pine that originates in northern Finland hydrated more than pollen from a population in southern Finland. Germination temperature had no effect on the hydration rate. Germinability and tube growth rate of northern genotypes were higher at 20 °C than at 15 °C. Differences among southern genotypes were not significant. At 15 °C, the germinability and pollen tube growth rate of northern genotypes were lower than southern genotypes. At 20 °C, the differences were not significant. It appears that germination and growth of pollen from northern populations are enhanced at higher temperatures whereas pollen from southern populations is unaffected.     

野菊与菊花杂交中花粉活力和柱头可授性及胚胎发育研究

应用石蜡制片、活体压片、光学显微镜及扫描电子显微镜观察等方法,研究了四倍体河南云台山野菊(Dendranthema indicum)与栽培菊花'钟山金山'(D.grandiflorum 'Zhongshanjinshan')种间杂交中父本花粉活力、花粉在柱头萌发、花粉管生长及胚胎发育情况等.结果发现,父本云台山野菊的花粉活力在授粉时为12%左右.人工授粉后的不同时间,在柱头上都观察到正常萌发的花粉粒,且花粉管都能进入柱头,其中,在授粉后0.5 h时,平均每柱头有5.9粒花粉萌发;12 h时,为59.9粒;而24和48 h时,则分别降为47.1和35.7粒.此外,在授粉后8、10、12和15 d时,分别在49.1%、40.8%、39.7%和38.5%子房内观察到正常发育的胚胎,最终杂交结实率为44.8%,而母本自然开放结实率为52.3%.研究表明,授粉前其多数母本雌蕊发育良好、授粉后多数花粉能在柱头正常萌发和花粉管正常生长,在受精后大部分胚胎发育正常是野菊与栽培菊种间杂交较高结实率的重要保证,而授粉前父本较低的花粉活力对杂交结实率影响不大.