Effect of enhanced UV-B radiation on pollen quantity,quality, and seed yield in Brassica rapa (Brassicaceae)

We conducted three experiments to examine the influence of ultraviolet-B radiation (UV-B; 280–320 nm) exposure on reproduction in Brassica rapa (Brassicaceae). Plants were grown in a greenhouse under three biologically effective UV-B levels that simulated either an ambient stratospheric ozone level (control), 16% (“low enhanced”), or 32% (“high enhanced”) ozone depletion levels at Morgantown, WV, USA in mid-March. In the first experiment, we examined whether UV-B level during plant growth influenced in vivo pollen production and viability, and flower production. Pollen production and viability per flower were reduced by ≈50% under both enhanced UV-B levels relative to ambient controls. While plants under high-enhanced UV-B produced over 40% more flowers than plants under the two lower UV-B treatments, whole-plant production of viable pollen was reduced under high-enhanced UV-B to 17% of that of ambient controls. Whole-plant production of viable pollen was reduced under low-enhanced UV-B to 34% of ambient controls. In the second experiment, we collected pollen from plants under the three UV-B levels and examined whether source-plant UV-B exposure influenced in vitro pollen germination and viability. Pollen from plants under both enhanced-UV-B treatments had initially lower germination and viability than pollen from the ambient level. After in vitro exposure to the high-enhanced UV-B levels for 6 h, viability of the pollen from plants grown under ambient UV-B was reduced from 65 to 18%. In contrast, viability of the pollen from plants grown under both enhanced UV-B treatments was reduced to a much lesser extent: only from ≈43 to 22%. Thus, ambient source-plant pollen was more sensitive to enhanced UV-B exposure. In the third experiment, we used pollen collected from source plants under the three UV-B levels to fertilize plants growing under ambient-UV-B levels, and assessed subsequent seed production and germination. Seed abortion rates were higher in plants pollinated with pollen from the enhanced UV-B treatments, than from ambient UV-B. Despite this, seed yield (number and mass) per plant was similar, regardless of the UV-B exposure of their pollen source. Our findings demonstrate that enhanced UV-B levels associated with springtime ozone depletion events have the capacity to substantially reduce viable pollen production, and could ultimately reduce reproductive success of B. rapa.     

Polyester and nylon powders used as pollen diluents preserve pollen germination and tube growth in controlled pollinations

Pollen acquisition for seed production, breeding programs and supplemental pollination can be costly and difficult. The identification of dry particulates for use as pollen diluents would facilitate the use of limited amounts of pollen and aid in accurate pollen application and dispersion. Four powders - Rilsan ES, polyester, wheat flour, and Lycopodium spores - were evaluated as pollen diluents using petunia as a model system. Diluents were combined with petunia pollen at a 5:1 (v/v) ratio. Two types of studies were conducted: (1) storage studies evaluated the viability of pollen combined and held with diluent for different durations; and (2) in vivo studies evaluated pollen tube growth in the styles of flowers pollinated with pollen-diluent mixtures. Pollen germination was not affected when stored as pollen-diluent mixtures for 4 days. Slight detrimental effects on pollen germination were observed after 6 days storage with Rilsan ES powders. Pollination with all the pollen-diluent mixtures resulted in fewer pollen tubes growing in the style compared to controls diluted with heat-killed pollen instead of diluent powders. Lycopodium-pollen mixtures were the most inhibitory, providing only 8% of the tube numbers observed in controls. Pollen mixed with polyester powders, Rilsan ES powders or wheat flour had tube numbers ranging from 47 to 61% of the control, but still had 175 or more pollen tubes per style, which would be sufficient for high rates of seed set in petunia. Wheat flour-pollen mixtures tended to clump and degrade pollen flow. Rilsan ES and polyester were identified as two promising pollen diluent powders that can facilitate accurate metering and distribution of pollen, produce large numbers of pollen tubes, and maintain pollen viability under storage.     

Pollen viability and limitation of seed production in a population of the circumpolar cushion plant, Silene acaulis (Caryophyllaceae)

Pollen viability among genders and limitation of female seed production in a natural trioecious population of the circumpolar cushion plant Silene acaulis was examined. Pollen viability was estimated by an in vitro pollen germination experiment. Both male and hermaphrodite flowers displayed large variation in pollen viability (0–53% in hermaphrodite and 0–54% in male flowers). There was a significant difference between genders in pollen viability: male plants had on average higher pollen viability than hermaphrodite plants. Resource and pollen limitation of seed production was studied by an experiment consisting of three treatments; (I) hand-pollination and removal of all other flowers on the cushion, (II) hand-pollination without removal of other flowers, and (III) open pollination without removal of flowers. Hand-pollination increased seed production, whereas removal of flowers had no effect on seed production. Abortion of pollinated ovules during seed development and seed mass did not differ among treatments. To control for effect of fruit number on seed production, data from naturally pollinated individuals was used. There was a positive correlation between both total number of seeds and fruit number, mean seed number per fruit and fruit number, respectively. These results indicate that seed production of 5. acaulis is mainly limited by pollen availability whereas resource competition between fruits is not important as a limiting factor. The possible role of male quality differences between genders and pollen limitation of seed production for maintenance of trioecious reproductive systems is discussed.     

Ripe pollen carbohydrate changes in Trachycarpus fortunei: the effect of relative humidity

Pollen of the palm Trachycarpus fortunei was kept at 25°C and relative humidities (RH) of 20, 55 and 98%. Changes in viability, water content and carbohydrates were measured over 2–17 days. Water content remained almost constant at 20 and 50% RH and increased dramatically at 98%. Pollen viability and germination rate remained almost constant over 14 days at 20% RH and decreased to about 2% after 7–9 days at 55% and to even less at 98% RH. Although the three experimental conditions were constant, qualitative and quantitative variations in pollen carbohydrates were recorded, even after pollen had lost its viability. The quantities of mono-, di- and polysaccharides varied with the period of pollen storage at the various RH. The greatest changes in glucose, fructose and sucrose content were recorded at 55 and 98% RH. At these relative humidities, maximum glucose and fructose content and minimum sucrose content occurred at maximum water content. Starch was not present in mature pollen but appeared and peaked after 7–9 days of pollen storage at 55 and 98%. Appearance of starch coincided with an increase in pectin content. PAS-positive cytoplasmic polysaccharides showed an increasing trend at 20% RH. A relation was found between pollen viability, water content and monosaccharide content. Pollen viability and germination capacity remained high at 20% RH for 14 days. At this relative humidity, pollen water, glucose and fructose contents remained almost constant, while sucrose reached its maximum value. The fluctuations of more complex carbohydrates (starch, pectins and PAS-positive cytoplasmic polysaccharides) were less easy to interpret. Changes observed under experimental conditions could simulate processes occurring in nature during pollen presentation and dispersal.     

Pollen germination in vitro at low temperature in European and Andean tetraploid potatoes

Summary Confined design combined with use of tolerance ratio was used to compare pollen germination capacity at low and high temperature in Andean and European potato material. Four clones of Solanum tuberosum from the European gene pool were compared with four Andean potato clones derived from the breeding program for frost resistance at the International Potato Center (CIP), Lima, Peru. For each clone, the same pollen lot was used throughout each replication. Pollen were germinated at 9 °C and 21 °C. Fortification of media with potato starch and 14 min preincubation at 25 °C were used as variables. The Andean material maintained its germination capacity better than the European material when temperature was decreased. It was possible significantly to distinguish potato clones with low temperature requirement for pollen germination if incubation proceeded germination at 21 °C, but not at 9 °C. Fortification with starch had no significant effect.     

水分胁迫对澳洲坚果花粉生活力和贮藏性的影响

采集经不同水分胁迫处理的澳洲坚果植株的花粉进行生活力测定和贮藏性研究,结果表明,新鲜花粉生活力随贮藏时间增加而下降。常温下正常发育花粉可保存6d左右,此后萌发率逐渐下降;而处于水分胁迫下的花粉,6d后生活力迅速降低;水分胁迫程度越严重,花粉生活力下降越快。几种处理的花粉有效贮藏期分别是:饱和灌溉处理6d,正常灌溉处理30d,水分亏缺处理10d,严重干旱处理(CK)4d。研究还表明,品种间花粉贮藏性表现为Kau >Pahala >O.C.。澳洲坚果花粉生活力的最佳染色鉴定方法为联苯胺染色法。     

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.

     

Viability and longevity of pollen from transgenic and nontransgenic tall fescue (Festuca arundinacea) (Poaceae) plants

Pollen is an important vector of gene flow in plants, particularly for outcrossing species like tall fescue. Several aspects of pollination biology were investigated using pollen from transgenic and nontransgenic plants of tall fescue (Festuca arundinacea Schreb.), the most important forage species worldwide of the Festuca genus. To effectively assess in vitro pollen viability in tall fescue, an optimized germination medium (0.8 mol/L sucrose, 1.28 mmol/L boric acid and 1.27 mmol/L calcium nitrate) was developed. Treatment with relatively high temperatures (36° and 40°C) and high doses of UV-B irradiation (900-1500 μW/cm(2)) reduced pollen viability, while relative humidity did not significantly influence pollen viability. Viability of pollen from transgenic progenies (T1 and T2) was similar to that from seed-derived control plants. Pollen from primary transgenics (T0) and primary regenerants (R0) had various levels of viability. Hand pollination using the primary regenerants and transgenics revealed that no seed set could be obtained when pollen viability was lower than 5%. Pollen from transgenic progenies and nontransgenic control plants could survive up to 22 h under controlled conditions in growth chamber. However, under sunny atmospheric conditions, viability of transgenic and nontransgenic pollen reduced to 5% in 30 min, with a complete loss of viability in 90 min. Under cloudy atmospheric conditions, pollen remained viable up to 240 min, with about 5% viability after 150 min. This report is the first on pollen viability and longevity in transgenic forage grasses and could be useful for risk assessment of transgenic plants.     

Storage of Pollen Grains in Organic Solvents: Effect of Organic Solvents on Leaching of Phospholipids and its Relationship to Pollen Viability

In vitro germinability and membrane integrity (as revealed bythe fluorochromatic reaction (FCR) test) of pollen grains ofCrotalaria retusa L. stored in various organic solvents forsix months at –20±2 °C were studied and correlatedwith leaching of lipids, phospholipids, sugars and free aminoacids from pollen grains into organic solvents during storage.Pollen grains stored in organic solvents with low dielectricconstants (a measure of their non-polar nature), such as hexane,cyclohexane and diethyl ether, showed high scores for germinationand FCR and very little leaching of phospholipids, sugars andamino acids. Pollen grains stored in solvents with high dielectricconstants (a measure of their polar nature) such as isopropanoland methanol did not show germination or positive FCR scores,but showed extensive leaching of phospholipids, sugars and freeamino acids. The viability of pollen grains stored in organicsolvents seems to be determined largely by the effect of theorganic solvents on pollen phospholipid composition, which inturn affects membrane integrity and consequently pollen viability. Crotalaria retusa, organic solvents, pollen storage, viability, phospholipids     

马占相思花粉离体萌发研究

马占相思(Acacia mangium)是含羞草科(Mimosaceae)金合欢属(Acacia Mill.)树种,在我国的广东、广西、福建以及海南等省(区)大面积种植推广。马占相思用途广泛,其杂交育种工作近年来逐渐受到重视,马占相思花粉研究对其杂交育种成败具有重要影响。该研究以新鲜的马占相思花粉为供试材料,开展不同蔗糖浓度(50、100、150、200、250和300 g·L~(-1))、不同硼酸浓度(100、200和300 mg·L~(-1))以及不同培养温度(26、28和30℃)对马占相思花粉离体萌发影响的研究,同时使用筛选出的马占相思花粉离体萌发的最佳处理,开展对另外3株与初试的马占相思生长发育状况接近的马占相思的花粉活力检测研究,并观察记录马占相思花粉离体萌发特征。结果表明:将刚开放的马占相思花序采回室内阴干,于次日10:00后,用毛笔刷法成功收集大量马占相思花粉,经显微镜观察检测,花粉纯度较高,可以保证后续研究备用;马占相思花粉离体萌发的最佳处理为200 g·L~(-1)蔗糖、100 mg·L~(-1)硼酸、28℃培养温度;马占相思花粉在培养24 h之后,花粉萌发率最高(为94.28%),花粉管最长(为6.0 D);检测3株马占相思的花粉萌发率分别为90.11%、82.31%、85.67%,具有显著差异。该研究结果为今后进一步开展人工控制授粉、选育优良的相思杂交新品种提供了基础。     

Effects of γ-ray treatment onCannabis saliva pollen viability

The viability and thein vitro germination capability of hemp pollen (cv. Carmagnola) were studied. Viability tests were based on the microscopic observation of the fluorescence of loaded fluorescein diacetate (FDA), while, for germinability tests, five different media were tested. The effects of irradiation with γ-rays on pollen viability and germination and on seed set were also studied, at three different irradiation doses (20, 60 and 100krad). The results show that in one of the media tested, about 85–90% of the pollen grains are viable and able to germinate in control samples, and that while viability measured by FDA test is not affected by increased γ-ray doses, the pollenin vitro germinability drops to about one-half of the controls at the maximum γ-ray dose employed, 100krad. Seed set of hemp plants pollinated with the irradiated pollen dropped to less than 1% of that of plants pollinated by untreated pollen for the higher dose used. The different media suitable forin vitro germination of hemp pollen, and the observed lack of correspondence between viability and germination capacity tests are discussed.     

Responses of tobacco pollen to high humidity and heat stress: viability and germinability in vitro and in vivo

Summary Responses of pollen grains of Nicotiana tabacum to high humidity (95% RH, 4 h) and temperature (38°/45° C, 4 h) stresses were investigated. Pollen grains were subjected to only RH or only temperature, or to both of these stresses. Their viability was assessed on the basis of the fluorochromatic reaction (FCR) test, and vigour was assessed on the basis of the time taken for in vitro germination as well as on the emergence of pollen tubes through the cut end of semi-vivo implanted styles. None of the stress conditions affected pollen viability and high RH or high temperature stress did not individually affect pollen vigour. However, pollen vigour was markedly affected when both the stresses were given together. Pollen grains subjected to high RH at 38° C took a longer time to germinate in vitro and the pollen tubes emerged later from the cut end of the semi-vivo styles; division of the generative cell was also delayed. Pollen grains subjected to high RH at 45° C failed to germinate in vitro, but did germinate on the stigma. Many pollen tubes subjected to this treatment showed abnormalities, and the growth of pollen tubes in the pistil was much slower than that observed in other treatments. Pollen samples subjected to all of the stress conditions were able to induce fruit and seed set. The implications of these results on the relationship between the FCR test and viability, and between viability and vigour, especially in stressed pollen, are discussed.     

花粉的保存及其生活力测定

花粉是种子植物的雄配子体，在有性繁殖中发挥着重要作用。采集的花粉和贮存的花粉，在使用之前必须作生活力的鉴定，以估价花粉是否有授精能力，并掌握花粉的形态和生理特征。本文对花粉的采集与保存、花粉生活力的概念及花粉生活力的测定方法作了详细的介绍，并对各种方法的优缺点进行了讨论。     

花粉的保存及其生活力测定

花粉是种子植物的雄配子体,在有性繁殖中发挥着重要作用,采集的花粉和贮存的花粉,在使用之前必须作生活力的鉴定,以估价花粉是否有授精能力,并掌握花粉的形态和生理特征,本文对花粉的采集与保存,花粉生活力的概念及花粉生活力的测定方法作了详细的介绍,并对各种方法的优缺点进行了讨论。     

小果短柱茶花粉离体培养系统的研究

山茶的短柱茶组是优良种质资源,有必要对小果短柱茶(Camellia confusa Chang 1941)的花粉萌发和花粉管生长的生理特性进行研究.本文研究了花粉生活力、培养温度及pH对小果短柱茶花粉萌发和花粉管生长的影响.结果表明:最适离体萌发培养基为5%蔗糖、0.003%的硼酸,0.005%的氯化钙和12%的PEG...     

Loss of viability of tomato pollen during long-term dry storage is associated with reduced capacity for translating polyamine biosynthetic enzyme genes after rehydration

The possibility that a loss of pollen viability during dry storage in a freezer is caused by the reduced pollen capacity to enhance polyamine biosynthetic enzyme activity after rehydration was investigated using pollen grains of tomato (Solanum lycopersicum=Lycopersicon esculentum) stored at -30 degrees C under dry conditions for up to 42 months. Pollen grains showed normal germinability for at least 12 months in storage, but those stored for longer than 24 months exhibited a significant reduction in germinability and fruit-setting ability. This age-dependent reduction in pollen viability coincided with the extent to which the pollen lost the capacity to increase arginine decarboxylase (ADC) and S-adenosylmethionine decarboxylase (SAMDC) activities and polyamine contents upon rehydration. Immunoblot analysis indicated that the capacity of pollen to translate ADC and SAMDC mRNAs was impaired in accordance with the loss of viability. Also, the capacity to synthesize proteins in general decreased with the increase in storage duration. The addition of 1 mM putrescine, spermidine, or spermine to incubation medium promoted germination, impregnation of pollen grains with 1 mM spermidine restored fertilization ability, and the addition of 1 mM spermidine to incubation medium promoted protein synthesis exclusively in pollen grains which had been stored for a long time. These results indicate that the reduction in viability of tomato pollen during long-term dry storage in a freezer involves a decline in the capacity to enhance gene translation for polyamine biosynthetic enzymes upon rehydration.     

培养条件及贮藏温度和时间对木麻黄花粉萌发率的影响

用离体培养的方法研究了不同蔗糖、硼酸浓度,以及不同贮藏温度和贮藏时间对木麻黄(Casuarina)花粉萌发的影响.结果表明:15%的蔗糖是木麻黄花粉萌发的最佳浓度;在15%蔗糖培养基上添加硼酸显著促进木麻黄花粉萌发,250 mg kg-1硼酸是木麻黄花粉萌发的最佳浓度;添加了琼脂的固体培养基更有利于木麻黄花粉萌发;在常温下木麻黄生活力丧失很快,但低温下花粉的萌发力可保持较长时间.     

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.     

苏铁属花粉萌发及保存条件研究

以不同浓度梯度的蔗糖与硼酸组合在不同pH条件下用悬浮培养法测定德保苏铁、叉叶苏铁、元江苏铁和越南篦齿苏铁花粉的活力;将元江苏铁和越南篦齿苏铁花粉分别保存在不同低温、不同湿度的环境中,研究温度和湿度对保存花粉的影响。结果表明:(1)最适合苏铁属植物花粉萌发的培养液配方为蔗糖(1%～2.5%)+硼酸(100～500 mg/L),pH6.0～7.0;(2)在室温下,将苏铁花粉密封保存在有干燥剂的容器中,可存活30 d以上;(3)在0℃条件下,不加干燥剂,花粉可保存4个月以上;(4)用液氮保存后的越南篦齿苏铁花粉进行人工授粉,结实率高达90.3%,与用新鲜花粉人工授粉的结实率无明显差异;(5)将花粉含水率降低到15.5%～13.2%后,能在液氮中进行长期保存,表明花粉液氮保存可以作为苏铁花粉长期和超长期保存的方法。     

Histological and Morphological Studies of Pollen Grains from Elongata, Reduced Elongata and Staminate Flowers in Carica papaya L.

The quality and quantity of pollen from three types of flowers; elongata, reduced elongata and staminate from the commercialized Thai papaya cultivar ‘Khak Nual’ were determined using pollen morphology, pollen physical characters and pollen development processes. Pollen development progressed at the same pace in the three types of pollen-producing flowers and was consistent with pollen development in many angiosperms. Pollen morphology showed that papaya pollen grains are tricolporate, with three apertures, and there is no significant difference in diameter (25.18–25.72 μm) and weight (11.76–15.45 ng) among pollen sources. The staminate flower shows the lowest amount of pollen, with 12,368 pollen per anther, but higher viability and germination rates of 95.53 % and 53.64 %, respectively. In contrast, the elongata type shows the highest amount of pollen grains with 14,884 pollen per anther and the lowest viability and germination rates, 93.06 % and 46.33 % respectively. The physical characteristics of pollen grains from reduced elongate and elongate flowers are similar. Reduced elongate flower type can donate pollen without self-pollinating.