Association between sporophytic reaction to Alternaria helianthi and gametophytic tolerance to pathogen culture filtrate in sunflower (Helianthus annuus L.)

Seven sunflower genotypes comprising of populations and hybrids showing differential sporophytic reaction to Alternaria leaf and stem blight were studied for their gametophytic reaction to pathogen culture filtrate. The sunflower pollen grains germinate well in the liquid medium and give good pollen tube growth in the absence of the culture filtrate. The addition of increasing concentrations of culture filtrate to medium significantly reduced the pollen germination and tube growth in all the genotypes. The reduction in pollen germination and tube growth in vitro due to culture filtrate was more in highly susceptible genotypes L-101 and Morden than the moderately resistant genotypes Acc. nos. 1229, 180 and ISFH-306. Pretreating the stigma and style with the culture filtrate before pollination reduced the number of pollen grains germinating compared to untreated control suggesting toxin stress can be created on the stigmatic surface before pollination. There was correspondence between pollen germination on stressed stigma (in vitro) and sporophytic reaction of the genotype suggesting pollen grain having resistance would germinate on the stressed stigma and fertilise the ovule achieving selective fertilisation. The correlation analysis indicated that there is a negative relation between sporophytic per cent disease index value and gametophytic parameters such as in vitro and in vivo pollen germination, culture filtrate required to inhibit 50% pollen germination and pollen tube growth. The association between pollen and the sporophytic reaction to the disease indicate the possibility of rapid screening of a large number of genotypes by means of pollen assay as an alternate technique with regard to sporophytic disease index in sunflower. The study also indicate the possibility of pollen selection before fertilisation to achieve rapid improvement in disease resistance. This revised version was published online in August 2006 with corrections to the Cover Date.     

Fertility and pollen tube growth in polyploid melons (Cucumis melo L.)

The high sterility of tetraploid melon (Cucumis melo L.) plants is a serious inconvenient to obtain economically useful polyploid lines. In order to study the alterations on diploid pollen tube growth and their influence on sterility, in vitro and in vivo pollen tube growth from three commercial varieties and its tetraploids was observed. Polyploid pollen grains show a lower germination percentage in vitro and a slower germination rate than haploid pollen. Furthermore, diploid pollen grains producing two or three tubes and tube branchings were observed on diploid pollen; this seems to indicate a decrease in its functionality. In vivo and in vitro pollen behaviours are quite similar, but an interaction between the pollen and the pistillar tissues was detected in vivo. In the diploid × tetraploid cross, where fruits were not obtained, pollen tubes showed a slow growth rate at the base of the pistil. The possible effects of these alterations in sterility are discussed in terms of the factors that could affect fecundation. This revised version was published online in July 2006 with corrections to the Cover Date.     

珍稀濒危物种大别山五针松花粉离体萌发的研究

为了研究大别山五针松濒危的原因,并为人工杂交育种和保护该物种提供理论依据。本试验采用花粉离体萌发法,设置正交试验,研究了蔗糖、硼、钙对其萌发率及花粉管生长的影响。结果表明：蔗糖是影响大别山五针松花粉萌发和花粉管生长的关键因子,其次是硼酸,氯化钙对其影响最小;蔗糖的添加,明显促进花粉萌发,但高浓度蔗糖花粉管的生长有很大的抑制作用;适当添加硼酸可以促进花粉萌发,但不利于花粉管的生长;氯化钙对花粉萌发影响最小,不同浓度的氯化钙均可以获得较高的萌发率。综合萌发率、花粉管生长,大别山五针松花粉萌发的最适培养基为：4%蔗糖+0.05%硼酸+0.025%氯化钙。     

In vitro pollen germination and tube growth of tomato (Lycopersicon esculentum Mill.) and its relation with plant growth

Summary In vitro pollen germination at 10°C, 14°C and 22°C of four groups of two pure line tomato varieties was compared with their plant growth at 19°C D/10°C N under controlled environmental conditions. Generally, pollen germination was slow at 10°C but after 6 h the percentage of germination was similar to that at 22°C. Maximum germination was obtained at 14°C already after 4 h. The longest pollen tubes occurred at 22°C. The two varieties within each group differed significantly from each other for percentage of pollen germination. For one group, this difference was greater at 10°C than at 22°C. The varieties in two groups also differed significantly for pollen tube growth. These differences in pollen tube growth rate were greater at 22°C than at 10°C. There was no clear relationship between pollen germination, pollen tube growth and plant growth in any of the four pairs of varieties. The results are discussed with regard to the possibility of pollen selection at low temperature in order to improve the efficiency of breeding for growth at low temperature.     

Effects of gamma irradiation on in vitro pollen germination of different Cucumis species

Summary Pollen grains of 14 Cucumis accessions were irradiated with 0, 1, 2, 3 or 4 kGy acute gamma rays and germinated in vitro directly afterwards. Pollen germination was significantly reduced by increasing irradiation dose for all species, except C. melo var. agrestis. Pollen tube growth was generally reduced likewise. Pollen of two C. anguria subspecies was most sensitive to irradiation. Sensitivity of the pollen with respect to pollen tube growth in relation to irradiation dose was inversely related to total DNA amount per nucleus. In vitro germination was not related to DNA amount per nucleus. Results show that the examined Cucumis species, especially C. melo var. agrestis, are sufficiently resistant to irradiation to be used as donor species for in vivo egg cell transformation of the cucumber.     

Comparison of different pollen viability assays to evaluate pollen fertility of potato dihaploids

Summary The main obstacle in breeding potato (Solanum tuberosum L.) dihaploids is the severe limitation of male fertility. To determine pollen viability assays that correlate well to fertility in crosses, results of five different pollen viability assays were compared by correlation analysis with fruit and seed set characters in test crosses, and to pollen tube growth in situ (PL-test). The methods used were: staining the pollen cells with carmino acetic acid (CAA-test); in vitro pollen germination (PG-test); and detection of pollen staining rates after incubation with fluoresceine diacetate (FDA-test), 2-3-5-triphenyle tetrazolium chloride (TTC-test), and 5-bromo-4-chloro-3-indolyle--galactoside (X-Gal-test).The results of test crosses and pollen tube growth in situ correlated with the results of all other assays with the following ranking, from highest to lowest: enzyme activity assays (X-Gal-test, FDA-test, TTC-test), in vitro pollen germination (PG-test), and pollen staining by CAA. The newly developed X-Gal-test for monitoring -galactosidase activity showed the least variation of all assays investigated. Thus, this highly reproducible simple procedure is recommended for male fertility screening.Abbreviations B/F Berries obtained per 100 flowers - CAA Carmino acetic acid - FDA Fluoresceine diacetate - PG Pollen germination rate in vitro - PL Pollen tube growth in situ - S/B Seeds per berry - S/F Seeds per pollinated flower - TTC 2-3-5-triphenyle tetrazolium chloride - X-Gal 5-bromo-4-chloro-3-indolyle--galactoside     

油松花粉离体萌发及萌发特征研究

为了加强油松有性杂交,为种子园育种研究奠定基础,对油松花粉离体萌发的适宜条件及萌发特征进行了系统研究。结果表明,油松花粉在无蔗糖培养基中仍具一定萌发能力,蔗糖的添加可显著促进花粉萌发,但高浓度蔗糖可抑制花粉管的生长;油松花粉管易形成分枝,蔗糖浓度与分枝花粉频率呈正相关;硼酸的添加在培养初期可有效加速花粉萌发,但过高浓度的硼酸对花粉萌发具抑制作用;Ca2+对花粉萌发的影响较小,不同浓度Ca2+处理均可获得较高的萌发率;培养温度对花粉萌发的促进作用明显,但温度的持续升高将抑制花粉萌发。综合萌发率、花粉管生长及分枝花粉管频率,油松花粉萌发的最适液体培养基组成为：2.00%蔗糖+0.075%硼酸+0.005% Ca2+,26℃下光照培养,72 h时萌发率可达(96.10±0.47)%。     

桃花粉活性检测和离体萌发特性的研究

果树花粉活力直接影响授粉、受精乃至座果。为了解果树品种的花粉生活力,给引种栽培和杂交育种工作创造有利条件,以桃花粉为试验材料,采用染色方法检测花粉的活性。试验结果表明,含有蓝墨水的花粉培养基,花粉萌发率可以达到64.67%,与对照相比差异不明显,因此该方法比较适合于快速检测花粉活性;固体培养基培养2个品种桃花粉,萌发率比液体培养基培养分别高25%和20%。另外温度、花粉密度、培养基中蔗糖及硼酸浓度都对花粉萌发率和花粉管长度有影响,在温度为24℃、1粒花药/100 μL、10%的蔗糖或100 mg/L的硼酸的条件下,均有利于花粉萌发和花粉管生长。     

The effects of cis-trans ABA on embryo germination and seed dormancy in wheat

Pre-harvest sprouting of wheat grain can cause economic losses especially in cultivars with low levels of seed dormancy. The aim of this study was to determine genotype differences in embryo sensitivity to germination in response to exogenous (+/–) cis-trans ABA treatments at different concentrations. Six white and four red seed-colored bread wheat genotypes that differed in dormancy were grown in a field near Swift Current, Saskatchewan in 2000 as a randomized complete block design with four replicates. The seed samples from this experiment were germinated in a controlled environment at 20 °C without light. The exogenous ABA treatments were 0 μM – whole seed (control), 0 μM-embryos, 25 μM – embryos and 50 μM – embryos. The ABA experiment was a factorial design with four randomized complete blocks with four ABA treatments in all combinations with the ten genotypes. A weighted (by day) germination index (WGI) was calculated for each genotype in each ABA treatment. Genotypes differed in response to ABA. The genotypes, ABA concentration and genotype by ABA concentration interaction effects were significant (p ≤ 0.05). Excised embryos showed significantly decreased dormancy in most of the experimental genotypes. The addition of exogenous ABA enhanced embryo dormancy of most genotypes. This revised version was published online in August 2006 with corrections to the Cover Date.     

阔叶杨桐花粉离体萌发和花粉管生长特性的研究

研究了不同浓度的蔗糖和硼酸,在不同温度和培养时间内对阔叶杨桐花粉萌发率和花粉管生长的影响,以期为阔叶杨桐的繁殖育种研究提供依据.结果表明:1)不同浓度的蔗糖和硼酸处理组之间存在显著性差异,蔗糖和硼酸都能促进杨桐花粉萌发和花粉管的生长,但超过一定浓度则会起到抑制作用;2)随着培养温度的升高,花粉萌发率和花粉管长度呈先升后降的趋势;3)随着培养时间的延长,花粉萌发率和花粉管长度逐渐增加,但最初的前12h内花粉萌发快,在培养的前9h内花粉管生长较快.4)实验显示:阔叶杨桐花粉培养的最适培养基是15％蔗糖和0.02％硼酸,最适培养温度为25℃,培养24 h后花粉萌发率为84.32％,花粉管长达2 003.51 μm.     

Factors affecting haploid production in wheat using the Hordeum bulbosum system. 1. Genotypic and environmental effects on pollen grain germination,pollen tube growth and the frequency of fertilization

Summary A quantitative examination of pollen grain germination and pollen tube growth within the overy wall was made in selected wheat × tetraploid H. bulbosum hybridizations, to investigate the effect of the wheat and the H. bulbosum genotype on these characters. As expected, variation at the known crossability loci had no effect on pollen grain germination. The frequency of pollen tube penetration of the ovary wall was, however, severely reduced when the dominant alleles were present. Pollen tube penetration was nevertheless observed in 3 of the 110 ovaries examined of the non-crossable cultivar Highbury. The H. bulbosum genotype had a much smaller effect on these characters, but significant differences between the clones were observed in the numbers of pollen tubes initially penetrating the ovary wall. Although two H. bulbosum genotypes showed no significant differences in the number of ovaries with pollen tubes at the base of the ovule, significant differences in the frequency of fertilization were observed. The possible cause of this discrepancy is discussed.The frequency of fertilization in crossable wheat × H. bulbosum hybridizations was improved by the application of gibberellic acid within 10 minutes of pollination, and reduced by an increase in the ambient temperature from 20°C to 26°C. Fertilization following the pollination of non-crossable wheat genotypes was not affected by either of these factors.     

植物生长调节剂对‘秦冠’苹果花粉管生长和花粉萌发的影响

研究吲哚乙酸、萘乙酸、赤霉素、2,4-D、6-BA5种植物生长调节剂对苹果花粉管生长和花粉萌发的影响。以贮藏6个月的‘秦冠’花粉为材料,配制不同浓度梯度植物生长调节剂的固体培养基培养花粉,研究不同浓度植物激素对花粉管生长和花粉萌发的影响。结果表明,当吲哚乙酸≤1 mg/L、萘乙酸≤1 mg/L、6-BA≤5 mg/L、赤霉素≤10 mg/L时对花粉萌发有轻微促进作用,当浓度增大时抑制作用较突出;2,4-D对花粉萌发存在明显抑制作用;当赤霉素、2,4-D、吲哚乙酸、萘乙酸、6-BA浓度分别达到50、30、30、15、25 mg/L时,花粉遭受浓度迫害出现畸形扭曲。吲哚乙酸、萘乙酸、赤霉素、2,4-D、6-BA对‘秦冠’花粉管生长和花粉萌发均无明显促进作用,并且随浓度增大抑制作用较明显,人工辅助授粉时用于添加剂效果不理想。     

Effects of γ-radiation on vitality and competitive ability of Cucumis pollen

Summary Pollen of 4 Cucumis sativus genotypes and of the related species C. metuliferus and C. zeyheri 2x was irradiated with four doses of -rays: 1, 2, 4 and 5 kGy. Higher radiation doses were found to inhibit germination and pollen tube growth in vitro with C. sativus, the most sensitive species.Irradiated pollen of cucumber never performed normal fertilization. However, after radiation with 1 kGy, cucumber pollen was able to occupy all available ovules, of which about 50% developed into empty seeds without embryo. This makes 1 kGy-irradiated maternal pollen less suitable to serve as mentor pollen in interspecific hybridizations. Doses of 2 kGy and more appeared to eliminate the ability to compete for ovules, while fruit set after a 2 kGy radiation was still high.     

The site of action of the crossability genes (Kr 1, Kr 2) between Triticum and Secale. I. Pollen germination,pollen tube growth,and number of pollen tubes

Summary Seven genotypes of common wheat (Triticum aestivum L.) were crossed with rye (Secale cereale L.) in order to find the site or sites of action of the crossability genes, Kr ₁ and Kr ₂, of wheat. The data obtained, by fluorescence microscopy, were compared to the controls (wheat x wheat). The results indicate that the crossability genes have little effect on pollen germination and on the time taken for the pollen tubes to reach the microphyle, irrespective of their crossabilities with rye. The number of pollen tubes reaching the microphyle is, however, affected by the Kr-genes, as high crossable genotypes have more pollen tubes than the low crossable ones. There was a high correlation between the mean number of pollen tubes at the micropyle with seed set, which also reflects the crossability. The Kr-genes seem to manifest themselves in the retardation and inhibition of pollen tube growth between the style base and the top of the embryo sac, where the effect is most distinct in the low crossable genotypes.     

中国石蒜花粉活力测定研究

为了解中国石蒜的花粉活力及其最佳体外萌发条件,分别采用TTC染色法和离体萌发法对中国石蒜的新鲜花粉的活力进行了测定。pH 7.0,5%TTC在35℃条件下染色结果显示99%的中国石蒜花粉具有活力。对影响中国石蒜花粉萌发和花粉管伸长的3个因素（蔗糖、硼酸和氯化钙）进行单因素实验,发现三者的最适浓度分别为蔗糖150 g/L、硼酸60 mg/L、氯化钙50 mg/L。3种浓度的培养液中,中国石蒜的花粉萌发率分别为36.7%、77.6%和44.2%。在中国石蒜花粉萌发最适蔗糖浓度(150 g/L)的基础上,正交设计硼酸和氯化钙浓度,实验结果表明,其最适浓度与单因素实验相同,但花粉萌发率达87%,花粉管伸长量达897 μm,远远高于单因素实验的结果,表明三者协同作用,促进中国石蒜花粉的萌发和花粉管的伸长。上述实验证实中国石蒜花粉具有较高的活力。为中国石蒜与其他石蒜属植物的种间杂交可行性提供了实验依据。     

Effects of high temperatures on mature pollen grains in wild and cultivated maize accessions

Summary The effects of high temperature on mature pollen of various maize lines were investigated. Genotypic differences in pollen reaction to high temperature were revealed. Pollen grains resistant to high temperature (35°C, 26°C) were characterized by higher germination capacity and better ability to develop normal pollen tubes. The studies are of interest to evaluate reproductive system tolerance and conduct gamete selection at the mature pollen grain stage in maize.     

Breeding tomato for pollen tolerance to low temperatures by gametophytic selection

Haploid selection for traits related to pollen cold tolerance in tomato was performed in segregating populations derived from a Lycopersicon esculentum × L. pennellii hybrid. BC₁ populations were obtained by combining normal and low temperature treatments on two stages of pollen development: pollen formation, and germination and pollen tube growth. F₁ hybrids were cultivated under low and normal temperatures and their pollen was used to pollinate L. esculentum plants at low and normal temperatures. The four BC₁ populations obtained were tested for the quality and quantity of pollen produced at low temperatures. The population obtained by cold treatment at both stages had a significantly improved pollen germination ability at low temperatures. The two other coldselected BC₁ populations showed no differences compared with the unselected BC population. A second cycle of pollen selection, corresponding to BC₂, was applied in order to test its persistence in the subsequent generations and the possibility to further improve the character. This second cycle showed no improvement although some plants retained the high pollen germination ability at low temperatures that was observed in the first cycle. Hence, gametophytic selection of some characters related with tomato pollen performance may be feasible, at least for the first cycle of selection.     

同源四倍体西瓜自交花粉萌发生长与内源激素的关系

对花粉离体培养和花粉在雌蕊柱头上萌发、生长的研究发现,同源四倍体西瓜自交,花粉萌发率低,花粉管在花柱中生长缓慢,在花粉萌发和生长过程中伴随出现结节状、花粉管尖端膨大呈球形花粉管及双花粉管等异常现象。经过内源激素测定分析表明,上述现象与四倍体西瓜雌芯和花粉中细胞分裂素－－玉米素核苷（ＺＲ）含量过高、雌蕊的ＩＡＡ和ＧＡ３与ＡＢＡ的比值低、及花粉和雌蕊共同作用有关。     

万寿菊花粉活力及柱头可授性研究

为探明造成万寿菊雄性不育系结实率低的原因,对万寿菊花粉活力及柱头可授性进行了研究。采用花粉离体萌发法研究万寿菊自交系V-01花粉萌发适宜温度、花粉活力日变化和适宜贮存条件;用联苯胺-过氧化氢法检测万寿菊雄性不育系S-261、S-17-06-29、S-0191不育株柱头的可授性;水溶性苯胺蓝染色法检测花粉在柱头的萌发情况。结果表明：（1）万寿菊V-01适宜萌发温度介于25~30℃之间;V-01花粉活力日变化趋势为先升高后降低,11-13时采集的花粉萌发率最高;4℃干燥贮存是最适宜的花粉贮存条件。（2）万寿菊柱头形态呈‘γ’状时有可授性,可授性可持续3天。（3）万寿菊花粉授粉到柱头上1 h内即可萌发,授粉后2 h,花粉细胞达到花粉管内。     

Investigations into the Barrier(s) to Interspecific Hybridization between Cicer arietinum L. and eight other Annual Cicer species

Barrier(s) to interspecific hybridization between the cultivated chickpea, Cicer arietinum L., and eight other annual wild species, i.e. C. reticulatum Lad., C. echinospermum Dav., C. pinnatifidum J. and S., C. judaicum Boiss., C. bijugum Rech., C. chorassanicum (Bge) M. Pop., C. yamashitae Kit. and C. cuneatum Rich., were investigated. In general, good pollen germination and pollen tube growth were observed in all eight crosses and their reciprocals. En spite of a few pollen cube growth abnormalities in most crosses, pollen tube penetration into the ovule and, thus, fertilization was observed in all cross combinations. However, differences were observed in the time from pollination to fertilization, not only between different interspecific crosses but also between reciprocals of a particular interspecific cross. The crossability barrier is, therefore, believed to be due to factor(s) operating after fertilization.