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.     

Pollen–pistil interaction in the inter-specific crosses of Sesamum sp

The wild species of Sesamum have long been recognized as an important source of many valuable pest and disease resistance genes and other novel traits such as male sterility and plant type for the cultivated taxon. Utilization of these species in crop improvement is marginal because of their inherent low crossability with the cultigen. An understanding of the biological nature of incompatibility systems that impede hybridization offers tools for successful introgressions into cultivated sesame. The objective of this investigation was to observe pollen germination and pollen tube growth in cross pollinated S. indicum pistils as an indicator of pre-zygotic barriers operating in wide crosses involving S. indicum. Three wild species: S. alatum, S. radiatum and S. laciniatum were used as pollen donors in hybridization with S. indicum. In the cross S. indicum × S. alatum 85.23% of pollen grains germinated and pollen tubes reached ovaries within 8 h after pollination. In other two crosses, S. indicum × S. laciniatum and S. indicum × S. radiatum the level of incompatibility was high as evidenced by a reduced pollen germination and slow rate of pollen tube growth. Measures to circumvent incompatibility in these crosses to produce interspecific hybrids are discussed.     

A reproductive calendar of Lilium longiflorum Thunb. cv. Gelria

Summary The development and timing of different processes in the floral biology and sexual reproduction of Lilium longiflorum were studied and brought together into a chronological reproductive calendar. Care must be taken when emasculating flowers later than 4 days before anthesis (DBA), because the stomium of the anther was open from 8 DBA and the pollen grains appear to be mature from 3–2 DBA. The receptivity period of the pistil lasts for a rather long period, but before wilting of an unpollinated flower as determined by a change of the colour of the perianth from white into brown, the flower had already lost its receptivity. Because L. longiflorum pollen grains can easily be stored, the pistil receptivity should thus determine the pollination schedule. The relationships between pollen tubes and pistil are discussed with respect to plant breeding.Abbreviations DAA Days After Anthesis - DAP Days After Pollination - DBA Days Before Anthesis     

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.     

麻疯树花粉管荧光染色及生长过程的研究

花粉管的生长是完成植物受精的重要生理过程,观察花粉管生长首先需要对雌蕊组织进行软化透明。本研究探索适合麻疯树雌蕊结构的透明方法,并利用苯胺蓝染色技术,在荧光显微镜下观察授粉后麻疯树花粉管的萌发和生长过程。结果表明:麻疯树雌蕊组织可采用NaOH溶液软化透明,较优的条件是2 mol·L-1的NaOH,50℃恒温箱内软化3~4 h;麻疯树授粉2 h后花粉管开始萌发,10 h后花粉管穿过花柱基部,11 h后在子房中观察到花粉管。麻疯树授粉后,花粉管进入子房至少需要11 h,完成受精则需要更长的时间。     

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.     

The breeding system of rubber (Hevea brasiliensis): an evaluation of controlled hand pollination methods

Summary Controlled hand pollinated pistils of rubber were observed using fluorescence microscopy to assess the efficiency of the universally-employed method for the production of progeny for plant breeding. The controlled hand pollination method conducted in the morning resulted in the deposition of a mean of 15.6 pollen grains on the stigma, with no stray pollination observed. Over 36% of the pistils had the potential to set fruit.Pollinations conducted in the afternoon at the normal time of anthesis, had double the fruit set potential of morning pollinations as measured by penetration of ovules by pollen tubes. Pollinator efficiency also varied, with excessive damage to stigmas resulting in reduced pollen germination and tube growth. There were differences between clones in both female and male fertility, in the proportion of pistils with more than three carpels and in the production of small abnormal stigmas. There was no difference in pollen tube growth following self- or cross-pollination, indicating that the self-sterility mechanism of rubber operates in the ovary. Pollen could be stored for 5 days at 5°C and 75% RH with a 22% loss of fertility.     

Barriers to hybridization of Solanum bulbocastanum Dun. and S. Verrucosum Schlechtd. and structural hybridity in their F1 plants

Summary Results of reciprocal crosses between Solanum verrucosum (2n=2x=24) and S. bulbocastanum (2n=2x=24) are described in terms of pollen tube behaviour in styles, of berry and seed set, of fertility and of meiotic behaviour of the F₁ hybrids. Pollen tube growth of S. verrucosum is strongly inhibited in styles of S. bulbocastanum, whereas no inhibition is observed in the reciprocal cross. Therefore S. bulbocastanum x S. verrucosum fails to set berries or seeds, whereas the reciprocal cross produces both berries (54.4% berry set) and seeds (0.3 per berry). Only 14.6% of the seeds germinate. Both the diploid and corresponding tetraploid hybrid plants are vigorous, flower abundantly, have a rather regular meiosis (mainly rod bivalents), but show a high degree of cytoplasmic-genic male sterility. Crossability of the diploid hybrid plants is nil when used as pollen parents and near to zero when used as pistillate parents. The barriers to hybridization of the parent species (unilateral inhibition of pollen tube growth, somatoplastic sterility, cytoplasmic-genic male sterility and structural differences of the parental chromosomes) are discussed and methods are suggested to overcome these barriers.     

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.     

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.     

Self-incompatibility interactions in Brassica napus

Summary The interactions of nine S-gene lines of Brassica napus were examined. Seven different S-genes were involved in the crosses, with three lines of different origin having the same S-gene. In general, the reactions on selfing the heterozygotes were as expected from the results of test crosses with the homozygous parent lines. All types of dominance relationships were found, including reversal of dominance in pollen and pistil. Weakening of activity of one or both genes was found in several cases. Background genotype was found have to some effect and reciprocal differences in reactions were also observed.     

Identification of self-incompatibility alleles in pear cultivars (Pyrus communis L.)

Self and cross-incompatibility determination by means of fruit and seed set experiments or pollen tube growth observations in the style has been frequently reported to be unclear in pear (Pyrus communis L.). Thus,in order to develop a reliable in vivo method to test pollen-pistil incompatibility in pear, pollen tube performance has been studied along the pistil following self and cross-pollinations. Results show that, while pollen tube growth in the style may be an unclear test, ovule observation at the microscope for the presence of pollen tube in the nucellus is a proper method to test incompatibility in this crop. With this analysis we could identify S-alleles of ‘Williams’ (S₁S₂) and ‘Coscia’(S₃S₄), and three of the four possible S-genotypes resulting from the ‘Williams’ × ‘Coscia’ cross, as represented by ‘Butirra Precoz Morettini’ (S₁S₃), ‘Santa Maria Morettini’ (S₂S₃)and ‘Tosca’ (S₁S₄). This result demonstrates that ‘Williams’ and ‘Coscia’ cultivars do not share any allele in common. We also established two new inter-incompatibility groups in pear. Furthermore, the presence of a common allele between ‘Williams’ and ‘Agua de Aranjuez’,and ‘Coscia’ and ‘Agua de Aranjuez’, three apparently unrelated old cultivars, may indicate a narrower genetic base than expected for European pear. This finding together with the fact that 40% of new released cultivars have direct or indirect parental relationship with the cultivars ‘Coscia’ and/or ‘Williams’, anticipates the possibility of new cases of cross-incompatibility for this crop in the future. Both the method described and the determination of the S-genotypes will facilitate the characterisation of self and cross-incompatibility relationships in this species. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.     

The yield stability of CIMMYT'S maize germplasm

Summary Relationships between whole-plant and germination NaCl tolerance and two in vitro measures of NaCl tolerance were evaluated in lucerne (Medicago sativa L.). No significant correlations were observed between relative NaCl tolerance of non-regenerating callus cultures, herbage regrowth, seed or pollen germination, and pollen tube growth. Simulated selection using indirect measures of NaCl tolerance produced small expected gains in relative NaCl tolerance during herbage regrowth and seed germination. These gains were less than 30% of those expected with direct selection for regrowth or germination tolerance. Selection based on the apparent NaCl tolerance of callus tissue or male gamephytes would appear unproductive. Direct selection for NaCl tolerance both at germination and during mature plant regrowth would appear to be necessary to rapidly develop lucerne populations with field-relevant levels of tolerance to NaCl stress.Approved by the Arizona Agric. Expt. Stn. as Journal Series no. 5015.     

Variation in pollen performance among plants of Fagopyrum esculentum

Summary In genetically-heterogeneous outcrossing species, there is the opportunity for selection based on the male gamete. Buckwheat (Fagopyrum esculentum Moench) is self-incompatible with one ovule per flower, so pollen competition at each ovule can be studied. The occurrence of selection among pollen parents was determined, as well as the relative importance of prezygotic and post-zygotic selection. Mixed pollinations from two donors produced nonrandom paternity, with one of the donors being favored about 2:1 on several females. Individual plants showed significant variation in the speed of pollen-tube growth. Therefore, prezygotic selection is likely to have occurred based on the speed of pollen tube growth. In single-donor pollinations, donors had equal success as expected in the absence of post-zygotic selection among donors. However, a significant male x female interaction was found, consistent with postzygotic selection against particular parental combinations. To test whether male fitness is reduced by increased allocation to seed filling during pollen production, large- and small-seeded lines were compared, both as pollen donor and as pollen recipient. The large-seeded line was better in both roles, thus there was no evidence that greater allocation to seeds reduced the quality of the pollen.     

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     

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.     

Pollen germination and pollen tube growth following self-pollination and intra- and interspecific pollination of Medicago species

Summary Pollen germination and pollen tube growth after self- and intra- and interspecific pollinations of annual [M. scutellata (L.) Mill., M. disciformis DC., and M. rigidula (L.) All.] and perennial (M. sativa L.) Medicago species were studied using a Carl Zeiss microscope with a fluorescence filter attachment.Self-pollination of self-fertile annual and intraspecific pollination of cross-fertile perennials were characterized by high pollen germination and rapid normal pollen tube growth. In contrast pollen germination percentages were very low and many pollen tube growth abnormalities occurred following interspecific pollination and self-pollination of self-sterile plants. The time period from pollination to fertilization for interspecific crosses was about double that following self-pollination of the annual species. However, fertilization occurred frequently following interspecific pollinations. Much of the sterility was due to factor(s) operating after fertilization.Cooperative investigations of the Kansas Agricultural Experiment Station, Dept. of Agronomy and the U.S. Department of Agriculture, Agricultural Research Service. Contribution No. 82-582-J- Mention of a trademark name or proprietary product does not constitute a guarantee or warranty by the U.S. Department of Agriculture, and does not imply its approval over other products that also may be suitable.     

Increasing resistance to Fusarium crown and root rot in asparagus by gametophyte selection

In garden asparagus, Fusarium crown and root rot is the main cause of crop decline. Since chemical treatments are inefficient, efforts should focus on the development of resistant cultivars to control the disease. Toxic culture filtrate (TCF) of F. oxysporum has affected asparagus pollen germination and tube growth. Consequently, gametophyte selection was evaluated to ascertain if the application of selective agents at this level could increase selection efficiency. Two susceptible pistillate plants and one tolerant and one susceptible staminate plants were used in controlled crosses. Before pollination, a drop of a germination vehicle with TCF or without it was applied to the stigmas. Some pollinated pistils were fixed and analyzed by fluorescence microscopy; the rest were left on the plant for seed production. Fifty to 200 seeds were obtained per treatment combination (staminate plant x pistillate plant x pollination vehicle). The derived plantlets were inoculated in vitroand evaluated for disease symptoms. The application of TCF to stigmas reduced pollen germination and tube growth compared with untreated controls,regardless of the genotypic combination. Pollen germination and tube growth was poorer for the tolerant staminate genotype than for the susceptible one. When the TCF was applied, the number of seeds per pollination in comparison with the controls diminished only when the susceptible genotype was the pollinator. The percentage of affected root area of the progenies obtained after applying the TCF was lower than in the controls only when the tolerant genotype was the pollinator. Increasing Fusarium resistance in asparagus by means of gametophyte selection seems feasible. This revised version was published online in August 2006 with corrections to the Cover Date.     

Diversity of penicillarian millets (Pennisetum glaucum and P. purpureum) as for the compatibility between their gynoecia and pollens from some other Poaceae

Summary The compatibility between gynoecia of different penicillarian millets (Pennisetum glaucum and P. purpureum) and pollens from some other Poaceae was evaluated in order to determine whether different penicillarian pistils showed the same pistil-pollen interactions with a same non-penicillarian pollen and to determine whether compatibility was dependent on the genetic divergence or the degree of sympatric evolution of the mating partners. Ten pearl millet lines (P. glaucum subsp. glaucum), six wild pearl millet accessions (P. glaucum subsp. monodii) and one P. purpureum accession were pollinated with five other Pennisetum species: P. pedicellatum, P. polystachyon, P. schweinfurthii, P. squamulatum, P. ramosum, and with three species from other genera: Cenchrus ciliaris, Panicum maximum and Zea mays. Each male species was represented by a unique accession. Pollen grain germination on the stigmata and pollen tube growth in the gynoecia were monitored by means of fluorescence microscopy after aniline blue staining. Significant compatibility differences were observed between a given pollen and gynoecia from different P. glaucum accessions. But there was no evidence of compatibility differences between the three female taxonomic groups. The eight pollen parents can be classified into three groups according to their mean degree of compatibility with the different penicillarian gynoecia. P. ramosum, P. schweinfurthii and P. squamulatum showed a high compatibility: more than 50% of penicillarian gynoecia had pollen tubes at the micropyle, six hours after pollination. Zea mays, Cenchrus ciliaris and Panicum maximum showed a low compatibility: pollen tubes were arrested in the style. P. pedicellatum and P. polystachyon showed nil compatibility: few pollen grains germinated on the stigmata and very rare tubes reached the entry of the style, where they stopped. It must be noted that such a strong gametic barrier has been observed only with pollinator species sympatric to pearl millet.