Use of irradiated pollen as mentor pollen to induce self-fertilization of two self-incompatible Upper Amazon cacao clones

Summary Two self-incompatible Upper Amazon cacao clones, T85/799 and T79/501, were pollinated with compatible Amelonado pollen subjected to varying doses of gamma irradiation (10–100 Gy). The proportion of flat non-viable beans to fully formed, viable beans in the pods increased with an increase in dosage of gamma rays. At 60 Gy all the beans produced were flat and non-viable, beyond this dosage fruit set was zero. Pollinating the self-incompatible cacao clones with a 1 : 1 mixture of compatible mentor pollen irradiated at 60 Gy and normal self pollen produced a mixture of flat, non-viable beans and fully-formed viable beans. Similar experiments using irradiated pollen with a marker gene suggested that the fully-formed viable beans resulted from selfing. Increasing the proportion of the radiation-treated compatible pollen in the mixture increased the number of fully-formed beans. However, when compatible pollen which had been treated either at 80 Gy or with temperatures of 35° C, 40° C and 45° C for periods of five, ten and fifteen minutes in factorial combination were mixed with self pollen, no successful pollinations were achieved. Pollen viability tests indicated that, whilst pollen treated at 60 Gy were about 50% viable, those treated at either 80 Gy or with temperatures of 35–45° C were mostly not viable. This suggests that, to overcome the incompatibility in cacao, the tubes of the mentor pollen grains used should at least grow into the style. The possible causes for overcoming the self-incompatibility in cacao are discussed.     

Incompatible pollen tube growth and protein composition in styles of Japanese pear following high temperature treatments

Summary Immersing the excised-prepollinated styles of self-incompatible Japanese pear, Pyrus serotina Rehd., into distilled water at 45°C for 1.5–2.5 min promoted the growth of self-pollen tubes; both the number of styles penetrating the pollen tubes through styles to cut ends and of pollen tubes protruding from cut ends were increased. The subsequent growth of the protruded pollen tubes on agar media was also enhanced by the treatments. Treatments at 40°C did not promote the growth of self-pollen tubes and those at 50°C led the styles to necrosis. In the stylar proteins analyzed by isoelectricfocusing polyacrylamide gel electrophoresis, several changes in banding pattern were found following the heat treatment; one of the bands was a glycoprotein that increases in quantity in developing styles as self-incompatibility of the style becomes stronger (Hiratsuka et al., 1986). This protein band was stained more weakly by Coomassie Brilliant Blue-G250 dye. Antigenic substances in the styles examined by immunodiffusion seemed not to be denatured by high temperature treatments.Abbreviations IEF-PAGE isoelectricfocusing polyacrylamide gel electrophoresis - CBB Coomassie Brilliant Blue     

The progamic phase in Cichorium intybus L. Pollen tube growth in the style,incompatibility reaction and gametophytic competition

Summary Pollen germination and tube growth were studied following compatible, incompatible and pseudo-compatible pollinations in chicory. Pollen germination begins 3 minutes after compatible pollinations. The earliest pollen tubes reach the ovary 17 minutes later. Many of the later germinating pollen tubes are arrested and burst at the stigma papillae. In the transmitting tissue inhibitional effects due to negative interactions between pollen tubes are frequently observed. Complete self-incompatibility results in total inhibition of germination. In case of pseudo-self-compatibility, some pollen germinate but germination and stigma penetration are delayed and often result in pollen bursting. There is no self-incompatibility reaction in the transmitting tract but if the pollen tubes reaching this tissue are relatively numerous, negative interactions between them occur as after compatible pollinations. An hypothesis is presented which attributes the negative interactions between pollen tubes to the diffusion of a substance from the growing pollen tubes. This substance would also provoke pollen bursting on the stigma.     

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.     

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.     

Pollen grain germination and pollen tube growth following in vivo and in vitro self and interspecific pollinations in annual Cicer species

Summary Germination of pollen grains and growth of pollen tubes were studied to determine the cause of barreness in crosses among annual Cicer species. In vivo and in vitro time-course studies and fluorescent microscopy revealed no pollination incompatibility among the selfs, crosses and reciprocals of C. arietinum L., C. reticulatum Lad. and C. cuneatum Rich. In general, Cicer pollen grains germinated and grew on styles of Cicer species. Pollen tube growth was characterized by irregularly spaced and intermittent callose deposits. Failure of seed formation in interspecific pollinations may be attributed to the slowness of pollen tube growth or collapse of fertilized ovules. In addition to these causes, shortness of stamens and sparsity of pollen grains were responsible for flower drop in natural selfs. Although the number of pollen tubes entering the micropyle in interspecific pollinations was low, it may be possible to grow the fertilized ovules on an artificial medium to obtain F₁ plants.     

Overcoming the self-incompatibility of Lilium longiflorum Thunb. by application of flower-organ extract or temperature treatment of pollen

Summary The application of flower organ extracts to stigmas and the temperature treatment of pollen were tried to overcome self-incompatibility of Lilium longiflorum cv. Georgia.Substances in stigma, style, ovary and anther were extracted with ethanol and fractionated with ethylacetate into the acidic, basic and aqueous fractions. The extracs melted in a small volume of distilled water were applied to stigmas prior to self-pollination. Hinomoto stigma extract, self-pollinated and cross-pollinated Georgia stigma extracts of high concentrations and Georgia anther extract of high concentration were effective in overcoming the self-incompatibility and resulted in a high percentage of fruit set and many normal seeds. Extracts from Hinomoto ovary, style and anther were ineffective, except a basic fraction, which was very slightly effective.Pollen was treated with 40°C for 60 or 90 minutes and 50°C for 30 or 60 minutes, and a half of each lot was followed by –20°C for 24 h, prior to self-pollination. All treatments were effective, especially at 40°C for 60 minutes or 50°C for 30 minutes, and 40°C for 90 minutes or 50°C for 60 minutes followed by –20°C for 24 h.     

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。     

Incompatibility problems in species crosses of tulips

Summary An investigation into the conditions that lead to higher seed yields from species crosses of tulip has revealed that the temperature has an effect on seed production. Under constant temperatures, the best results in the phytotron were obtained at 14 °C.Various problems associated with such crosses have been explained on the basis of microscopical observations. One of the factors associated with a poor seed yield from interspecific tulip crosses is the abnormal growth of the pollen tubes in the embryo-sac cavities of numerous ovules, with the result that fertilization does not take place.The failure of the cross between Red Pitt, a Darwin tulip of the species T. gesneriana and the T. fosteriana tulips Mad. Lefeber and Princeps appears to be due to an inhibition of the pollen tube growth in the stigma.     

甘蓝自交不亲和系柱头和花粉粒不亲 和互作的电镜扫描研究

扫描电镜观察到甘蓝(Brassica oleracea L.)自交不亲和系02-E1柱头上不能萌发的花粉在授粉后18小时内就会萎缩变小直至干瘪。能够萌发的花粉从一个萌发沟萌发后，另外两个萌发沟就自动萎缩，其花粉管有的扭曲不能伸入乳突细胞，仅少量花粉管能伸长进入花柱。经过5%NaCl溶液处理后的甘蓝自交不亲和系的柱头表膜乳突细胞或皱缩或破裂，花粉粒很容易萌发并长出花粉管，然后花粉管穿破乳突细胞壁而伸入花柱完成受精作用。     

Differential cold sensitivity of pollen grain germination in two Prunus species

Summary Pollen grains from selected cutivars of almond [Prunus dulcis (Mill.) D. A. Webb] and peach (Prunus persica Batsch L.) were exposed to a range of temperatures between 1°C and 34°C on a thermogradient plate. Pollen germination at temperatures below 9°C was conspicuously greater in almond than peach. Miximal germination percentages were attained at about 16°C (almond) and 23°C (peach). The two species did not differ in their capacity for pollen tube elongation over a broad range of temperatures. Maximal pollen tube elongation occurred at temperatures 5°C to 8°C higher than maximal pollen germination. Species affiliation appeared to be of much greater consequence than chilling requirement or bloom date of the sporophyte in predicting gametophytic response to temperature.     

Self-incompatibility inRibes

Summary Seven species ofRibes were studied; all of them proved to be self-incompatible. They were found to have hollow styles and incompatible pollen tubes were not inhibited until they reached the ovary. The extent to which self-pollen tubes penetrated the ovary before being arrested varied with different species.     

A procedure suitable for in vitro pollen selection inBrassica oleracea

Summary Experiments to determine whetherBrassica oleracea pollen could effect fertilisation following incubation in liquid culture medium are reported. Pollen was incubated in vitro, collected by centrifugation and used to pollinate compatible pistils. While retaining viability and the capacity to germinate such pollen was unable to penetrate the stigma papillae. However, incubated pollen produced functional tubes following pollination of styles from which the stigmas had been removed. These tubes grew through the style to the ovary and viable seed was obtained. The potential application of this procedure in pollen selection is discussed.     

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.     

Unusual behaviour of growing pollen tubes in the styles and ovules of Spinacia oleracea L.

Summary Pollen tubes of spinach, while growing in the styles, produce branches. In the ovules and the surrounding tissue, at later stages, they grow like fungus mycelium and ramify in all directions. Haustoria-like structures are produced from the pollen tubes near the micropyle of the ovule. These structures penetrate into integuments, nucellus and other tissues and produce branches. Several pollen tubes enter a single ovule and appear to bring about polyspermy. Though the functional significance of some of these phenomena is still speculative, it is likely that the persisting pollen tubes and haustoria may be responsible for the nourishment of the growing embryo.     

The performance of double or mixed pollinations with compatible and self-incompatible or incongruous pollen of pear and apple

Summary Six pear and five apple trials were carried out to ascertain the outcome of combinations of compatible pollen (C) with self (S) or incongruous pollen (I) as to the pollination index (PI=seeds/pollinated flower). The PI of the mixture C+I (1:5) was consistently depressed as compared to that of the control C. The results of the double pollinations S/C and I/C were affected by the temperature at pollination; their PI's at <15°C were twice as high as those at >15°C, being well above and below the PI of C in the former and latter case respectively. The opposite was true for the C/S combination, the PI of which increased with the pollination temperature; the PI of C/I did not differ much from the PI of C, irrespective of temperature. The conclusion was reached that the interaction previously and presently found between compatible and self-incompatible pollen also exists to a fair extent between compatible and incongruous pollen. However, in pear neither the mentor nor the pioneer pollen technique proved to aid its hybridization with apple, the formation of self seed was not observed either. In apple the production of apple × pear hybrids was likewise doubtful, but the double pollinations S/C and C/S formed 4–10% self seed.     

Inheritance of sexual incompatibility in apricot

The study was carried out with 3- to 6-year-old apricot progenies from the breeding programmes of ARS in Fresno, California, USA and of CEBAS in Murcia, Spain. Progenies from 19 different crosses were studied to determine the inheritance of the genetic incompatibility trait. The 19 crosses included those where both parents were self-compatible or both self-incompatible, and also those where a self-compatible parent and another self-incompatible were crossed. Segregating progenies were studied either by bagging a sufficiently high number of flowers in balloon stage and determining the percentage of fruit set several weeks later or by following pollen tube growth with fluorescence microscopy. Results indicated that a monofactorial system with a multiallelic series appears to control the trait. Alleles for self-compatibility would allow the pollen tubes to grow in any style and reach the ovules. Alleles for self-incompatibility would stop the pollen tube growth if the same allele is present in the pollen grain and in the pistil.     

小麦花粉管生长途径及受精过程经历时间的研究

应用常规石蜡切片和荧光显微镜技术对小麦花粉管生长途径及受精过程相应时间进行研究。结果表明，授粉后，花粉随即萌发，2个精子进入花粉管，营养核留在花粉粒内，不久解体。花粉管进入羽毛状柱头分支结构的细胞间隙，继续生长于花柱基部至子房顶部的引导组织的细胞间隙内。随后穿过子房壁，在子房壁与外珠被之间的缝隙中向珠孔方向生长。自花粉萌发至花粉管长入珠孔大约需要1 h。自花粉萌发并到达柱头分支结构中，花粉管均具明显的绿色荧光；但在引导组织区以及子房壁与外珠被之间生长的花粉管几乎看不到荧光。授粉后1 h花粉管经珠孔及珠心表皮细胞间隙进入1个助细胞，并释放精子。授粉后2～6 h精卵融合，授粉后16 h合子第一次分裂。授粉后2～3 h精子与极核融合，授粉后4 h初生胚乳核第一次分裂。授粉后2～16 h，即精卵融合至合子分裂前为花粉管通道法转化合适的时间，采用花柱横切滴加法转入外源DNA。     

Pollen tube growth and fertilization after different in vitro pollination techniques of Aechmea fasciata

The effect of cut style and placental pollination on fertilization efficiency was studied for compatible Aechmea fasciata plants. These alternative in vitro pollination techniques resulted in lower penetration rates of the ovules by pollen tubes in comparison to pollination on the stigma. An explanation was found in the intervention of the normal pollination process,through which less pollen-pistil interactions were built up. After cut style pollination the percentage of ovule penetration by pollen tubes increased when a longer style part was left at the ovary. Probably fewer factors that control pollen tube growth are present in the lower style part. Pollen germinated on the ovules after placental pollination but only rarely penetration of the micropyle by a pollen tube occurred. Activation of the ovary, induced by in vivo prepollination for 6 hours, and pollination two days after anthesis did not increase the fertilization percentage. Grafting a style with active growing pollen tubes to ovules on the placenta (placental grafted style pollination) resulted in a higher fertilization percentage. Pollen tube growth through the style was essential for pollen tube guidance to the ovules and penetration of the micropyle. This revised version was published online in August 2006 with corrections to the Cover Date.     

Studies on the self-incompatibility of Petunia hybrida in excised-style culture. An attempt at improving a technique in excised-style culture

Summary A technique of excised-style culture was developed. In twelve compatible combinations of pollen and style, a correlation was found between the number of pollen tubes protruding from the style and the number of mature seeds formed in vivo. This suggests that the technique of improving excised-style culture is effective not only for studying the interactions between style and pollen tubes during tube growth, but also for predicting the production of seeds in vivo.