Impact of Humidity on Temperature-Induced Grain Sterility in Rice (Oryza sativa L)

High temperature‐induced grain sterility in rice is becoming a serious problem in tropical rice‐growing ecosystems. We studied the mechanism of high temperature‐induced grain sterility of different rice (Oryza sativa L) cultivars at two relative humidity (RH) levels. Four varieties of Indica and Japonica rice were exposed to over 85 % RH and 60 % RH at 36/30 °C, 34/30 °C, 32/24 °C and 30/24 °C day/night air temperatures from late booting to maturity inside sunlit phytotrons. Increasing both air temperature and RH significantly increased spikelet sterility while high temperature‐induced sterility decreased significantly with decreasing RH. Neither Indica nor Japonica rice types were superior to the other in the response of their spikelets to increased air temperature and RH. Increased spikelet sterility was due to increased pollen grain sterility which reduced deposition of viable pollen grains on stigma. Reduction in sterility with decreased RH was more due to decreased spikelet temperature than to air temperature. Thus the impact of RH should be considered when interpreting the effect of high temperature on grain sterility. Spikelet fertility was curvilinearly related to spikelet temperature. Grain sterility increased when spikelet temperature increased over 30 °C while it became completely sterile at 36 °C. The ability of a variety to decrease its spikelet temperature with decreasing RH could be considered as avoidance while the variability in spikelet sterility among varieties at a given spikelet temperature could be considered as true tolerance.     

The effects of high-temperature stress on the germination of pollen grains of upland cotton during square development

The reproductive stage of flowering plants is sensitive to high-temperature stresses. High temperature is a major factor influencing pollen grain viability in upland cotton (Gossypium hirsutum). The objective of this study was to identify the relationship between cotton pollen germination percentage and temperature by assaying the pollen germination of four upland cotton cultivars in vitro at different temperatures during the blooming period. The results showed that in vitro pollen germination percentage was related to the culture temperature of pollen germination and the temperature of the square development process. High temperature affected pollen development and germination, and high-temperature tolerance differed among the cotton cultivars. The pollen germination percentage decreased rapidly with changes in the culture temperature from 30 to 39 °C. A culture temperature of 35 °C might be a critical temperature for the pollen viability transition and could be used to screen cotton cultivars that have pollen grains with high-temperature resistance. Before the high-temperature stage, cultivars with rates of decrease in the percentage of pollen germination of less than 41 % at 35 °C relative to the rates at 30 °C might be considered as high-temperature tolerance cultivars, and cultivars with rates of decrease in the percentage of pollen germination greater than 41 % might be considered as susceptible cultivars. The high-temperature stress for pollen grain germination in vitro was greater than 30 °C, and the high-temperature stress for square development might be greater than 33 °C. Boll retention was significant; it was positively correlated with the pollen germination percentage and negatively correlated with temperature during the high-temperature stage. This study provided a method for rapidly screening cultivars (lines) with high-temperature tolerance pollen in upland cotton breeding.     

Establishment of reliable methods of in vitro pollen germination and pollen preservation of Brassica rapa (syn. B. campestris)

A simple and reliable method for evaluating the viability of Brassica pollen was established in which the in vitro germination rate of pollen was adopted as the index of the viability of pollen grains. Pollen grains were preincubated in an atmosphere in which the relative humidity (RH) was fixed to 52% or 66% at 20 °C for 5 hours. They were cultured for 16 hours at 25 °C in a liquid Kwack's medium (1964) supplemented with 20% sucrose, and the pH was adjusted to 8.0. They were then observed under a microscope and the number of germinating and unchanged pollen grains were counted. The germination rate of pollen was improved and stabilized by preincubation and the use of a high pH medium. More than 90% of the freshly harvested pollen grains of Brassica rapa (syn. B. campestris) germinated constantly in these conditions Undehisced anthers were collected from flowers at anthesis and dehydrated by incubation at 20 °C for 16–24 hours in an atmosphere where the RH was fixed to 15% or 32%. They were put into a plastic vial and preserved in a freezer at -20 °C. The germination percentage of the preserved pollen was scored at intervals during preservation. The germination rate of the pollen grains preserved at -20°C for 1 year was higher than 50% and the pollen proved to be efficient for seed set. Most of the seeds germinated normally. This revised version was published online in August 2006 with corrections to the Cover Date.     

Viability and storage of bromeliad pollen

Several bromeliad species from two different subfamilies, were used to develop a reliable method to evaluate pollen viability. Pollen germination on a medium containing 20% sucrose, 0.001%H₃BO₃ and 0.5% agar was comparable to germination on a compatible stigma. Maximum germination was reached within 2 to 10 hours depending on the species. Based on this test, six species were considered as being good pollen donors with germination percentages between 49%and 83%. Furthermore, pollen from these species and cultivars could be stored in liquid nitrogen (–196 ^°C) without a considerable loss of viability. For all species, a dehydration period of 4 hours prior to cryopreservation and a rehydration period of 1 hour after cryostorage were essential. Greenhouse humidity influenced anther moisture content and cryostorability. This revised version was published online in July 2006 with corrections to the Cover Date.     

Hybrid Tea-rose pollen. I. Germination and storage

Summary Germination and storage trials were carried out with pollen of several rose varieties. The pollen grains germinated well in a 15% sucrose solution with 40 ppm boric acid. Staining the pollen with a 0.1% tetrazolium solution and standardizing the degree of colour at which the pollen grains are counted as viable, provided a good viability estimate, simpler to carry out than in vitro germination. Germination capacity and staining ability of the pollen were greatly impeded-about halved-by dehydration during storage in desiccators at low humidity. This effect could be corrected by humidifying the pollen beforehand for about one hour, though this pre-treatment increased the percentage of germinated pollen grains more than the percentage stained. There was no difference between the two percentages in fresh or in deep-frozen pollen.Pollen stored at 1°C and high relative humidity soon lost its germination capacity: between 0 and 20% humidity a considerable proportion of the pollen remained viable for 9 months and longer. Storage for the same period in vacuum-sealed glass tubes at –24°C maintained viability as well or better and would probably prolong it further. Some of the cold-stored pollen induced a reasonable seed set after one year, a low seed set was obtained even after two years of storage at 1°C and low humidity.     

Effects of desiccation and a change in temperature on germination of immature grains of wheat (Triticum aestivum L.)

A study was made of the effects of desiccation and a change in temperature on the germination of wheat grains harvested 20 days after anthesis. When the germination test was performed immediately after harvesting, germination percentages ranged from 5.2% to 10.7%. Germination percentages increased to 48.2% to 90.3% after grain had been desiccated at 20 °C and then hydrated at 10 °C. This increase occurred even if grains had been desiccated in an atmosphere of high relative humidity. The germination percentage of non-desiccated grains depended on the germination temperature. When the pericarp and testa were removed from embryos, the germination percentage of grains incubated at 20 °C and then at 10 °C was higher than that of grains incubated at 10 °C. In general, a low germination temperature is believed to be effective in breaking dormancy of wheat grains. However, a change in temperature stimulated germination to a greater extent than a constant low temperature. The germination percentage of 5-day cycle alternating temperature was greater than that with 1-day, 2-day and 10-day cycles. Although the germination of immature wheat grains was stimulated by both high and low germination temperatures, it is likely that cycles shorter and, also, longer than a critical period induce limited germination. Loss of dormancy commonly occurs when development of wheat grains proceeds at a high temperature, with imbibition at a low temperature. However, germination ability of non-desiccated immature wheat grain was enhanced by a change in temperature during germination. This revised version was published online in August 2006 with corrections to the Cover Date.     

In vitro germination and viability of buckwheat (Fagopyrum esculentum Moench) pollen

An in vitro method for the germination of common buckwheat pollen was developed. Pollen grains were successfully germinated in an artificial medium consisting of 0.2 g each of MnSO₄, Ca(NO₃)2.4H₂O and KNO₃, 0.04 g H₃BO₃, 15 g sucrose and 30 g polyethylene glycol (molecular weight approximately 20,000) dissolved in 100 ml of double distilled water. The viability of pollen was assessed by in vivo and in vitro germination tests at 20 °C and 25 °C over a 38 h time period. Pollen grains were collected and germinated at 4 h intervals from freshly harvested flowers grown under 16 h day length and a constant temperature. Maximum pollen viability was found 2 h and 6 h after first light when plants were maintained at 25 °C and 20 °C, respectively. Viability, as measured by germination percentage, was similar at both temperature regimes. Some pollen remained viable for approximately 34 to 38 h in intact flowers, but all pollen lost viability in less than an hour when stored at room temperature without humidity control. This revised version was published online in July 2006 with corrections to the Cover Date.     

Pollen storage at low temperature as a procedure for the improvement of cold tolerance in spring rape, Brassica napus L.

The possibility of selecting spring rape for cold tolerance at the mature pollen grain stage was studied by investigating the effects of pollen storage at low temperatures on the quality of pollen grains and on the cold tolerance of the plants generated from them. Pollen treatments of F¹ hybrids affected fertilization ability much more than viability and even after 10 days storage at 3 or 10°C the pollen germination percentage was reasonably high. Pollen storage for 7 or 10 days at 3 or 10°C significantly increased the cold tolerance of F² seed germination, with 3°C being more effective. Pollen storage for a shorter time had no effect upon the number of resulting genotypes tolerant to low temperature. This approach may be successfully applied in plant breeding to enrich segregating plant populations with cold-tolerant genotypes.     

Cryopreservation of Delphinium pollen at –30 °C

Pollen of garden varieties and species of Delphinium that was cryopreserved at –30 ^°C after 3 hours of air drying and storage on silica gel, had high germination and pollen tube growth in vitroeven after 180-day storage. On the other hand, pollen stored at 25 ^°C showed a marked decrease in the germination rate within 10 days. The best in vitro germination of Delphinium pollen was on a 1% water agar containing 15% sucrose and 0.005 to 0.01% boric acid and at 15 to 20 ^°C. Field pollination with the cryopreserved pollen showed higher fruit and seed set than pollination with pollen stored at 25 ^°C, and was not significantly different from fresh pollen. This revised version was published online in August 2006 with corrections to the Cover Date.     

Inter-varietal pollen tube growth and ovule penetration in the avocado

Summary Results are presented showing number of pollen tubes in the pistil, and ovule penetrations in crosses involving the avocado varieties Edranol, Ryan, Hass, Reed, Talbot, Jalna, Fuerte, Bacon and Sharwil. Between 5% and 100% of the ovules in each cross were penetrated by a pollen tube. The female parent exerted more control than the male on numbers of tubes and ovule penetrations and the results for type A varieties were more consistent than those for type B varieties. Penetration of an ovule by two tubes was observed in all varieties except Reed and Fuerte and penetration of an ovule by three tubes was observed in the variety Edranol only. With the possible exception of the variety Bacon as female parent there was no difference between pollination with pollen from type A or B varieties.     

水稻开花期花粉活力和结实率对高温的响应特征

采用远红外增温设施在水稻开花第1 d分别对4个水稻品种（南粳41、武香粳14、扬粳6号和汕优559）进行5 h（10:00~15:00）高温处理（40℃）,然后转入常温,观测当天及其后4 d开花颖花的花粉活力及同期开花颖花成熟期的结实对高温的响应特征。结果表明,高温处理后,处理后各天颖花的花粉活力均显著下降（P< 0.05）,但随开花日序的后移其下降幅度逐步减小。其中,下降最大为处理当天或其后1 d,花粉在柱头和培养基上的萌发率以及花粉I-KI溶液的可染率分别平均降低了16.00、25.85和11.74个百分点,而处理后4 d的降幅仅为8.49、6.63和6.02个百分点。相同的趋势也表现在同期开花颖花的结实特征上,高温处理的当天和其后4 d开花颖花的结实率分别平均下降了14.04和5.95个百分点;而高温处理当天的空粒率和秕粒率平均分别提高了10.06和3.98个百分点,处理后4 d分别提高了3.98和1.97个百分点。另外还发现,高温处理下品种间存在一定差异,其中,汕优559的花粉活力和籽粒结实受高温的影响最小。相关分析发现结实率、空粒率和秕粒率与柱头花粉的萌发率、花粉在培养基上的萌发率均呈显著直线相关,而花粉I-KI溶液的可染率仅与空粒率存在显著相关（P< 0.05）。可见,水稻遭遇短期高温后,随着开花日序的后移高温对颖花形成的潜在热害逐渐降低,在生产实践中花粉在柱头和培养基上的萌发率可作为品种选育和高温热害发生的几率、热害程度预测的可靠指标。     

Assessment of Cold and Heat Tolerance of Winter-grown Canola (Brassica napus L.) Cultivars by Pollen-based Parameters

Winter‐grown canola (Brassica napus L.) production is limited mostly by frost and winter kill in the southern canola‐growing regions of the United States. Tolerance to cold and heat were assessed by studying percentage of pollen viability (PV), in vitro pollen germination (PG) and pollen tube length (PTL) for 12 field‐grown cultivars. Freshly collected pollen from all cultivars were incubated on artificial solid growth media at a constant temperature ranging from 10 to 35 °C at 5 °C interval for 30 h to determine PG and PTL. A modified bilinear model best described the temperature response functions of PG and PTL. Canola cultivars showed significant variability (P < 0.001) for PV (61.3 % to 89.7 %), PG (29.0 % to 48.2 %) and PTL (463 to 931 μm). The average cardinal temperatures, T_min, T_opt and T_max, for PG and PTL were 6.4, 24.3 and 33.7 °C, respectively. Principal component analysis revealed that maximum PG, PTL, T_min and T_opt of both PG and PTL were the most important factors in determining cold tolerance, whereas T_max of PG and PTL, and maximum PG and PTL were more responsible in separating the cultivars for heat tolerance. The canola cultivar, KS3077, was the most cold tolerant with the lowest T_min and the widest temperature adaptability range, and the cultivar Kadore was the most heat tolerant with the highest T_max for the PG. The identified cold‐ and heat‐tolerant cultivars may be useful in canola‐breeding programmes to develop cultivars suitable for a niche environment.     

Storage of avocado pollen

Summary Avocado pollen was stored at a range of temperatures and relative humidities (RH) for up to one year and the pollen was tested for viability in vivo.Pollen stored for one month was capable of germination on the stigma and penetrating the ovule when stored at 4°C with <1,23,55 and 75% r.h. and at -196°C with 0% r.h. Most pollen samples stored at 25 and -15°C at a range of r.h. would germinate on the stigma but none would penetrate the ovule.After one year of storage, pollen at 4°C and <1 and 23% r.h. would germinate on the stigma but would not penetrate the ovule. There was no germination of pollen stored at 4°C and 55 and 75% r.h. Only pollen stored at -196°C and 0% r.h. would penetrate the ovule, but thawing and refreezing once during the year destroyed the viability.     

Quantitative Genetic Analysis of Traits in Avocado Cultivars *

This research is aimed at quantitative genetic analysis of several avocado traits and cultivars. The experimental material consists of our avocado breeding project in which data were collected from several crosses as well as selfings of cultivars. Parenthood was determined by isozymes and the seedling progenies were assessed for eight traits. A biometrical genetic approach for analysis of this breeding project is presented. Genetic profiles of the traits and cultivars were detailed by several characteristics: level of heterozygosity and dominance deviation, dominance of alleles, dominance direction, general evaluation of additive and non-additive genetic variance, maternal inheritance, and allelic differences among the various cultivars in major genes controlling the same quantitative trait. Five avocado cultivars: ‘Fuerte’, ‘Hass’, ‘Ettinger’, ‘Tova’, and ‘Rosh-Hanikra’, were characterized separately for each trait. Practical conclusions for the breeder regarding economically important quantitative traits are discussed.     

In vitro interspecific fertilization,embryo development and formation of hybrid seedlings between Gossypium hirsutum and G. arboreum

Summary In vitro hybridization between Gossypium hirsutum, and G. arboreum was carried out. Hybrid seedlings were obtained after successive use of the following five kinds of media: 1) pollen grain germination medium, 2) double-fertilization medium, 3) embryo development medium, 4) seedlings formation medium, 5) green seedlings growth medium. The factors affecting in vitro interspecific fertilization, embryogenesis and seedling formation were studied. The key factors were temperature and relative humidity (RH). The optimum RH for in vitro interspecific fertilization was 65%, and a suitable temperature range was 26–30°C. Pollen grain germination ratio decreased rapidly at a RH of 80%. When the temperature and the RH were higher than 32°C and 80%, respectively, the fertilization rate decreased to zero.Effects of petals and calyces of the maternal flowers on in vitro interspecific fertilization and ovule growth were identified. Even though the petals or calyces were excised, hybrid plantlets were also obtained after media were improved and a shake culture was used. In addition, the process of double-fertilization and embryo development were studied cytologically. The developmental characteristics of the hybrid embryos derived from in vitro interspecific fertilization were a later occurrence of the double-fertilization process and a much lesser division of endosperm cells. But the embryo development was not affected by these characteristics, and young hybrid embryos can develop to their cotyledon stage finally on artificial media.     

Effects of short-term storage on germinability of pistachio pollen

The effects of short‐term storage under room conditions (constant 25°C and 35% relative humidity), refrigeration (4°C) and freezing (‐20°C) on the germinability of pollen of pistachio, Pistacia vera L., were studied and the effects of desiccation and pollen age on its germinability were reassessed. It was found that: (1) weight loss as a result of desiccation was positively correlated with germinability; (2) pollen grains stored in room conditions only germinated after prehydration, which restored germinability even after 7 days of exposure; (3) refrigerated (4°C) pollen grains retained their germinability for at least a week; (4) frozen pollen grains irreversibly lost most of their germinability; (5) 2‐day‐old pollen grains, within the anthers, retained their germinability under room conditions. These findings will contribute to the improvement of pistachio pollen preservation and help to find a simple and inexpensive method for its short‐term storage, while retaining its germinability.     

Production of polyhaploids of hexaploid wheat using stored pearl millet pollen

The effects of drying and freezing on viability of pearl millet pollen were examined with the aim of using stored pollen in polyhaploid production of hexaploid wheat. Freshly collected pollen of pearl millet line NEC 7006 with 55% water content, germinated at a frequency of 80%. Pollen that was dried for two hours to 6% water content showed 50% germination frequency and maintained similar frequencies after the freezing process. In crosses of hexaploid wheat variety Norin 61 with fresh pearl millet pollen, embryos were obtained at a frequency of 27.6%. In crosses with pollen stored at -196 °C, -80 °C and -20 °C for one month, embryo formation frequencies ranged from 27.5 to 17.4%. After five and twelve months of storage, the frequencies ranged from 29.7 to 14.6% at storage temperatures of -196 °C and -80 °C, and from 8.0 to 3.2% at -20 °C, indicating significant differences among storage temperatures. However, no significant frequency difference was found among pollen water contents at the time of collection. All plants regenerated from crosses with pearl millet pollen stored for five months were wheat polyhaploids. These results suggest that stored pearl millet pollen is an efficient medium for producing polyhaploids in hexaploid wheat. This revised version was published online in August 2006 with corrections to the Cover Date.     

Relationships between Yield and Quality Parameters of Malting Barley (Hordeum vulgare L.) and Phenological and Meteorological Data

In this study, phenological and meteorological data have been used to interpret variations in a time series of regional average yields and quality parameters of malting barley (Hordeum vulgare L). The analyses were focused mainly on the grain filling period. Duration and occurrence of this development stage showed remarkable differences from year to year, as heading varied over more than four weeks and yellow ripeness over two weeks in the investigation period from 1974 to 1996. Yields above average were achieved only in years when grain filling duration exceeded 42 d. Protein concentrations below 10.5 % and grading percentages over 90 % required at least 44 d of grain filling. Temperature had the strongest influence on the length of grain filling, even though the calculated Growing Degree Days (base temperature 3 °C) were not absolutely constant. Mean daily temperature and relative air humidity were the best estimators with respect to grain yield. An optimum temperature range was found between 14 and 18 °C. Assuming a linear relationship, yield reductions between 4.1 and 5.7 % have been calculated for every 1 °C increase of the mean daily temperature. Relative air humidity was the best single estimator for grain protein concentration. The results of this study suggest that relative humidity during grain filling can be a more suitable parameter to describe drought stress effects than precipitation amounts from heading to yellow ripeness or from January 1 to yellow ripeness.     

Techniques for collecting,handling germinating,and storing of pollen of the filbert (Corylus spp.)

Summary Temperature and humidity immediately before dehiscence were important in maintaining viability and germination of filbert pollen. Air temperatures at or above 23°C during dehiscence and pollen storage resulted in nonviability after 8–24 h. Cut branches bearing elongating catkins when exposed to temperatures of 18°±2°C during dehiscence yielded viable pollen. Manganese and boron added to the medium did not significantly increase germination, while 10–100 p.p.m. boron appeared to inhibit it. Storage tests indicated that filbert pollen viability may be maintained at 92% relative humidity and –18°C for at least 12 months.Also published as Tech. Paper No. 2305, Oregon Agricultural Experiment Station.     

Evaluation of Seed Vigor Tests for Predicting Seedling Establishment at Low Temperature in Rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

This study was carried out to develop an evaluation method to predict rice seedling establishment (SE) under low-temperature conditions. Two Korean-bred japonica cultivars, Shindongjin and Hopum were used in the experiment. Fresh seeds were treated with an accelerated aging (AA) at 40°C and 100% RH for 1-15 days. The SEs of the fresh and AA seeds were evaluated in nursery beds at 17°C, and their correlation coefficients with seed vigor values measured by 9 test methods including standard germination test (SGT), cool germination test (CLT), cold germination test (CDT), seedling growth rate test (SGRT), ascorbate peroxidase (APX), peroxidase (POX), superoxide dismutase (SOD), catalase (CAT), and α ‐amylase (AMY) activities. The percentage of SE decreased slowly from 75 to 0% with an increasing of AA period from 0 to 15 days. The result of nine vigor tests showed different correlations with the SE. SGT, CLT, SGRT, and POX were significantly correlated with the SE. In the correlation analysis with only short-term aging seeds (1-7 days), the SE was very highly correlated with SGT, CLT, CDT, SGRT, POX, and CAT. These results suggest that seed vigor values measured by several methods including SGT and POX could be used as a reference value to secure SE at low temperatures in nursery bed rice seedling culture.