Pollen germination,pollen tube growth and fertilization following self and interspecific pollination of Paspalum species

Summary Crossability between most Paspalum species is very low. This study was undertaken to identify the impediments to hybridization. Accessions of P. intermedium Munro. ex Morong, P. jurgensii Hackel and P. dilatatum Poir were self-pollinated and crossed with one anther. Paspalum intermedium is essentially self-sterile but P. jurgensii and P. dilatatum are highly self-fertile. Following pollination, pollen germination and tube growth were studied by observing the pollinated pistils with fluorescent microscopy. Examination of self-pollinated pistils revealed that the pollen germinated shortly after contacting the stigmas. Germination was over 80% for the P. intermedium and P. dilatatum accessions but only 57% for P. jurgensii. Pollen tubes grew to the micropyle within 45 minutes after pollination in P. dilatatum and 1 hour and 15 minutes in P. jurgensii. However, in the P. intermedium accessions most tubes did not grow beyond the stigma and very few penetrated the style and ovary. Apparently stylar-incompatibility is the reason for the low selfed seed set. In the cross-pollinations, pollen germinated shortly after pollination and germination ranged from 57 to 88% for the different crosses. In all crosses the pollen tubes grew to the micropyle within 30 minutes to 2 hours after pollination indicating that a cross-incompatibility system is not the cause for low crossability among these species. By examining embryo sacs from P. intermedium × P. dilatatum, its reciprocal and P. intermedium × P. urvillei crosses, it was determined that gametes failed to unite in some crosses and this is a major reason for low crossability.     

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.     

The transfer of triazine resistance from Brassica napus L. to B. oleracea L. I. Production of F1 hybrids through embryo rescue

Summary Triazine resistant Brassica napus ssp. oleifera and ssp. rapifera were hybridized to cultivars of B. oleracea ssp. italica, ssp. botrytis, ssp. capitata and ssp. fimbriata. The interspecific embryos did not survive in vivo but could be rescued in vitro using a culture medium developed by Monnier (1973). The embryos did not grow directly into normal plants but were successfully regenerated using the protocol developed by Keller (1984). Hybridization efficiency ranged from 0 to 2.64 hybrids per pollination. Interspecific embryo abortion may be related to abnormal endosperm development.     

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.     

Relationships between African species of the genus Cucumis L. Estimated by the production,vigour and fertility of F1 hybrids

Summary The production, vigour, and fertility of F₁ hybrids between nine African species of the genus Cucumis L. were studied as a measure of the relationships between the species. Hybrid plants were obtained from 29 out of the 72 possible cross combinations. Two F₁ hybrids died as seedlings, and 27 hybrids were raised to maturity. Pollen production and stainability varied greatly amongst these hybrids, as did fruit and seed set following self pollination and backcrossing with either parental species. The fruit shape of the hybrids was always intermediate between that of both parental species.Two species appeared to be closely related: C. prophetarum L. and C. anguria var. longipes A. Meeuse. Most other species produced highly to moderately fertile F₁ hybrids with at least one other species. C. metuliferus Naud. produced only sterile hybrids with C. zeyheri Sond. 2x. The results sustain the recent taxonomic classification of the genus (Jeffrey, 1980).     

Interspecific hybrids of Zinnia elegans Jacq. and Z. angustifolia HBK: embryology,morphology and powdery mildew resistance

Summary Reciprocal crosses between 13 cultivars of Zinnia elegans Jacq. and one source of Z. angustifolia HBK resulted in interspecific hybrids. Florets examined 14 days after pollination showed that all 13 cross combinations of Z. angustifolia x Z. elegans yielded embryos in 47.2% of the florets pollinated. Six cross combinations of Z. elegans x Z. angustifolia produced embryos in 4.6% of the total Z. elegans florets pollinated. Interspecific crosses resulted in seed-propagated hybrids when Z. angustifolia was the maternal parent. Hybrids were obtained from reciprocal crosses between the two species when embryos were excised and cultured. Morphologically, most hybrids resembled Z. angustifolia but were sterile. Colchicine treatments of selected sterile hybrids resulted in shoots producing fertile inflorescences. Preliminary tests showed that hybrids were resistant to powdery mildew (Erysiphe cichoracearum DC ex Mecat).Scientific Article No. A 3108, Contribution No. 6173 of the Maryland Agricultural Experiment Station, Department of Horticulture.     

Effect of mentor pollen on pistil-pollen incongruities among species of Cucumis L.

Summary The merits of mentor pollen in certain interspecific hybridizations in the genus Cucumis L. were evaluated. African wild cucumber species Cucumis metuliferus Naud. and C. africanus L. and Asian species C. sativus var. hardwickii Alef and var. sikkimensis Hook, were reciprocally crossed. Pollen tube growth was arrested halfway down the style in all combinations except in C. sativus × C. africanus. Mentor pollen irradiated with 100 and 200 krad grew through the style and into the ovules in all species. Fruitset was obtained in all cases when mentor pollen was used either alone or in mixtures. Fruits set after self pollination with mentor pollen yielded only seeds without embryos.In several crosses, aided by mentor pollen, a number of relatively large seeds (with enlarged embryosac) were obtained. A sample of these embryosacs contained a globular structure like an embryo. Embryosacs with embryo-like structures were explanted on several different media but no development was obtained.     

Use of Vaccinium octoploids to facilitate 4x-6x gene transfers

The use of an 8x Vaccinium produced through doubling a tetraploid hybrid with colchicine was studied as a means of facilitating gene exchange between V. corymbosum L. and V. ashei Reade. Analysis included meiotic observations and crossability studies of the 8x plant, as well as attempting to develop 6x breeding lines. Meiotic analysis revealed the presence of micronuclei in Telphase II products and polyspory in sporads. In the crossability studies five 4x highbush plants were pollinated with 8x o2 4x pollen. Data gathered included: % fruit set, average weight/fruit, average number of seeds/fruit, average number of seedlings/pollination, and number of 6x seedlings. Octoploid pollinations were significantly lower than 4x pollinations in all parameters. One 6x plant was produced from the 4x-8x crosses but was found to be mitotically unstable, having somatic cells with chromosome numbers ranging from 48 to 168.     

Methods of overcoming interspecific barriers in Trifolium

Summary Barriers to interspecific hybridization in Trifolium were investigated by manipulation of mentor pollen treatments, ploidy levels, and compatibility and male sterility systems. Crosses involving the addition of mentor pollen produced fewer seeds and hybrids than crosses involving normal pollination. Lower seed set with mentor pollen was deduced to result from the use of less viable pollen, approximately half the pollen having been killed by alcohol. Pollinations at the diploid level resulted in more hybrids than at the tetraploid level, perhaps because genes for male sterility produced higher female sterility in the tetraploids. The self-compatible stock produced more seeds, mostly selfs, than the self-incompatible stock, but produced more hybrids only in one cross, T. pratense L. × T. diffusum Ehrh. The use of male-sterile female parents reduced selfing but produced fewer hybrids than male-fertile female parents. Techniques of this study were designed to affect prefertilization barriers, but the lack of effect may indicate that postfertilization barriers in Trifolium are of greater importance.Journal Article No. 98-3-208 of Kentucky Agricultural Experiment Station. Published with approval of the Director.     

Natural hybridization between Solanum acaule Bitt. and S. megistacrolobum Bitt. in the province of jujuy,Argentina

Summary Evidence is provided that hybridization of the tetraploid (2n=4x=48), self-fertile tuber-bearing species Solanum acaule Bitt. with the diploid (2n=2x=24), self-incompatible, tuber-bearing S. megistacrolobum Bitt. takes place in several localities of the province of Jujuy in the high, cold plateaux of the Argentine Puna. The triploid hybrids (2n=3x=36) closely resemble S. acaule in growth habit, leaf morphology and floral structures and for this reason they can be easily overlooked for that species in the field. Experimental data show that S. acaule can be crossed with S. megistacrolobum though the crossability is rather low and variable according to the particular cross considered. The artificial hybrid obtained compares well with the natural hybrid in morphology and chromosome number. The hybrids, though almost completely male sterile, are successful colonizers of disturbed areas around farmers' dwellings, cattle enclosures and other areas where the soil is rich in organic matter.There is some evidence that the Tilcara material of S. acaule subsp. aemulans and the hybrids of S. acaule x S. megistacrolobum have some characters in common, which can be interpreted as having a similar origin.It is postulated that S. acaule subsp. aemulans, in Jujuy at any rate, is not a primitive form of S. acaule as thought by Hawkes and Hjerting, but rather a fertile hybrid derivative of S. acaule x S. megistacrolobum through the functioning of 2n gametes.We also provide evidence that S. bruecheri Correll should not be considered a hybrid of S. acaulle x S. megistacrolobum but a synonym for S. gourlayi Hawkes. The new name, S. x indunii Okada et Clausen, is proposed to designate this hybrid.     

Resistance to Heterodera schachtii in patellares section of the genus Beta

Summary Fifty-two accessions of the section Patellares wild beet (including 26 Beta patellaris Moq., 13 B. procumbens Chr. Sm and 13 B. webbiana Moq.) and 14 progeny families were selected and tested against sugarbeet cyst nematode, Heterodera schachtii Schm. All Patellares species tested were highly resistant, but not immune, to the development of H. schachtii, after infection. This is the first report that mature female nematodes developed in the roots of B. webbiana plants. The occasional development of nematode cysts in roots of juvenile wild beets was, however, not a heritable genetic factor.Mention of a trademark, vendor, or proprietary product does not constitute a guarantee or warranty of the product by the USDA and does not imply its approval to the exclusion of other products or vendors that may also be suitable.     

The reproductive biology of three perennial Sesbania species (Leguminosae)

Summary The reproductive biology of Sesbania sesban, S. goetzei and S. keniensis (Leguminosae; Papilionoideae) was studied. Fifty percent flowering was observed at 102 to 153 days after germination for S. sesban accessions; 96 to 146 days for S. goetzei and 131 to 176 days for S. keniensis accessions. Flowers opened in the afternoon and remained fresh for 2–3 days. Bee species including Xylocopa sp., Apis mellifera, Megachile bituberculata and Chalicodoma sp. visited the flowers. Hand pollination experiments showed that all three species were self and cross compatible. The percentage of fully developed seeds was greater in pods formed after cross pollination compared to self pollination. No evidence was found for stigmatic or stylar self incompatibility. Outcrossing is probably the common method of reproduction under natural conditions, although in isolated trees substantial seed set by selfing might occur. Pod production under natural conditions was 34% for S. sesban; 49% for S. goetzei and 39% for S. keniensis. Considerable variation was found in pod production under open pollination between accessions of the same species. Selective abortion was observed within pods, with more mature seeds formed at the distal end of the pod.     

A possible gene source for early,day-length neutral pigeon peas,Cajanus cajan (L.) Millsp.

Summary Atylosia scarabaeoides Benth. and A. platycarpa Benth., close relatives of the species Cajanus cajan (L.) Millsp., were screened for photoperiodic response. Four photoperiods ranging from 12 h 45 min to 19 h were studied in three environments. A. scarabaeoides flowered freely only in the first photoperiod. A. platycarpa not only flowered early (39 to 63 days after planting) in all four photoperiods, but also exhibited a relatively constant vegetative phase up to 16 h of illumination. Cajanus cajan, in which most cultivars-if not all-exhibit a response to photoperiod, was successfully hybridized with the two Atylosia species. It is suggested that genes for earliness and insensitivity to day-length could be transferred from A. platycarpa to C. cajan.     

The nature of the Bolivian weed potato species Solanum sucrense Hawkes

Summary Two theories for the origin of the Bolivian weed potato species Solanum sucrense Hawkes have been tested. The first was that it had an allopolyploid origin as a hybrid of the cultivated tetraploid S. tuberosum ssp. andigena (Juz. et Buk.) Hawkes with the wild/weed tetraploid cytotype of S. oplocense Hawkes. The second hypothesis postulated that it was a cross of the weed diploid species S. sparsipilum (Bitt.) Juz. et Buk. with S. oplocense.Synthetic hybrids of S. tuberosum ssp. andigena x S. oplocense and of S. sparsipilum x S. oplocense have been compared morphologically with S. sucrense accessions. Their crossability has also been investigated.The crossability and morphological studies strongly suggested that the former hypothesis was the more likely. This was borne out by observations made on the 1974 University of Birmingham Expedition when segregating populations of S. sucrense and S. oplocense were collected growing with feral S. tuberosum ssp. andigena.The evidence from this study indicates that the S. tuberosum ssp. andigena x S. oplocense hybrid has formed the basis of the S. sucrense gene pool. It is suggested that this hybridogenic taxon be maintained under the name Solanum x sucrense.The clarification of the taxonomic relationships of these three species will be of interest to Globodera resistance breeders, in view of the fact that these taxa are resistant to various pathotypes of the golden nematode.     

Optimal pollination conditions for seed set after a self-pollination,an intraspecific cross and an interspecific cross of marrow-stem kale (Brassica oleracea var. acephala)

Summary The effects of different pollination techniques, with and without emasculation and delayed pollination, were investigated to find the conditions for maximum seed set after self-pollination and intraspecific and interspecific crosses of Brassica oleracea var. acephala. The results indicated that the pollination conditions achieving maximum seed set vary with the type of pollination. After controlled self-pollination, the best seed set occurs in bud 3 to bud 10. For the intraspecific cross, the youngest flower and the oldest bud produced the largest number of developed ovules but bud pollination was productive to bud 8. The yields from these two pollination types were best when the female parent was not emasculated. In the interspecific cross with B. campestris cv. Marco the best results came from the youngest flowers and the oldest buds subjected to the standard practice of pollinating directly after emasculation. Possible reasons for these effects are discussed.     

Cytoplasmic homology between Aegilops mutica Boiss. and Ae. ovata L.

Summary Triticum aestivum L. em Thell. (2n=42; AABBDD), and T. durum Desf. (2n=28; AABB) genomes were substituted into the cytoplasms of Aegilops mutica Boiss. (2n=14; M_tM_t), Ae. heldreichii Holzm. (2n=14; MM), Ae. uniaristata Vis. (2n=14; M^uM^u), and Ae. ovata L. (2n=28; C^uC^uM^oM^o), to identify the M-genome diploid cytoplasm donor of Ae. ovata. Substitution of the T. durum genome into Ae. uniaristata cytoplasm resulted in a large proportion of shriveled inviable seeds. A few plump viable seeds were obtained all of which produced male-sterile plants having one univalent or telocentric chromosome from Ae. uniaristata. The T. aestivum plants having Ae. uniaristata or Ae. mutica cytoplasms were fertile. However, Ae. mutica was similar to Ae. ovata in the induction of delayed maturity and tall robust growth habit to the T. durum and T. aestivum plants. Cytoplasms of the other C- and M-genome diploids Ae. umbellulata Zhuk. (2n=14; C^uC^u) and, Ae. heldreichii (2n=14; MM) earlier had been shown to differ from that of Ae. ovata. Therefore, Ae. mutica is the most likely cytoplasm and M-genome donor to Ae. ovata.     

Cytology and leafspot resistance in Arachis hypogaea x wild species hybrids

Summary Introgression of germplasm from diploid wild Arachis species to A. hypogaea has great potential for improving pest resistance in cultivated peanuts. This investigation evaluated methods for incorporating exotic germplasm into cultivated peanuts, especially for Cercospora arachidicola Hori resistance. Interspecific hybrids between A. hypogaea (cvs. NC 2 and NC 5) and the wild species A. cardenasii Krap. et Greg. nom. nud. and A. chacoense Krap. et Greg. nom. nud. were analyzed cytologically and for leafspot resistance. All F₁ hybrids were sterile, had irregular meiosis, and very few multivalents. They were highly resistant to C. arachidicola in field tests and had a 10-fold reduction of conidia per lesion in the greenhouse as compared to A. hypogaea cultivars. After colchicine treatments of F₁ hybrids, hexaploids (2n=60) and aneuploids (2n=54, 56, 63) were observed. The hexaploids had up to 18 univalents per pollen mother cell and very few multivalents, indicating a low frequency of intergenomic chromosome pairing. For C. arachidicola resistance, significant differences were not found among wild species parents, F₁ hybrids and two generations of hexaploids. Most hexaploids were stable at 2n=60 and embryos aborted when backcrosses with the respective wild species were attempted. However, when hexaploids were backcrossed to A. hypogaea, several fertile pentaploid (2n=50) offspring were obtained. Use of self-pollinating pentaploids is believed to be the quickest method to recover 40-chromosome hybrid derivatives in these hybrids.     

Results of multiple pollination in blueberry (Vaccinium elliottii Chapm.)

Blueberry (Vaccinium section Cyanococcus, family Ericaceae) is a shrub that produces multiple-seeded berries in which only a fraction of the ovules develop into viable seeds. A recessive yellowleaf marker gene was used to evaluate the efectiveness of a single pollination versus multiple pollinations in producing seeds in Vaccinium elliottii Chapm. A multiple-pollination experiment was conducted to see if the first pollen applied or the second produced more progeny, and to see if multiple pollinations increased the number of seedlings produced. Flowers of diploid yellowleaf Vaccinium elliottii were pollinated from one to four times at daily intervals. Pollen from redleaf and yellowleaf plants was used to produce two types of seedlings that could be distinguished visually. Generally, percent berry set, seeds per berry, and seed germination were not increased by multiple pollination. After multiple pollination, the pollen applied first always produced the most seedlings. Prior pollination greatly reduced seed set from subsequent pollination, but subsequent pollination caused little or no decrease in the number of seeds set by the first pollination. This revised version was published online in July 2006 with corrections to the Cover Date.     

The mildew resistance gene Sph 2in relation to gall mite resistance in the black currant

Summary In black currant progenies derived from parents heterozygous for resistance to American gooseberry mildew. Sphaerotheca mors-uvae (Schw.) Berk., and to gall mite, Cecidophyopsis ribis Westw. (genes Sph ₂from the Swedish cultivar Öjebyn and Ce from Ribes grossularia L., respectively), there is invariably a defieit of mildew resistant plants. Linkage in repulsion between Sph ₂and Ce would account for the significantly greater deficit of mildew resistants in the gall mite resistant class. In contrast with published data for the mildew resistance gene Sph ₃, there is no evidence of linkage of Sph ₂with the leafing out gene Lf ₁, suggesting that the gene order is Sph ₂-Ce-Lf ₁-Sph ₃.     

Natural triploid hybrids between Solanum acauleBitter and S. infundibuliformePhilippi in the province of Jujuy,Argentina

Summary The occurrence of natural triploid hybrids between tetraploid Solanum acauleBitter subsp. acaule and diploid S. infundibuliformePhilippi both tuber-bearing Solanums, in the Puna region of Jujuy, is reported. The natural hybrids, which are morphologically intermediate between their putative parents and completely male sterile, have a somatic chromosome number of 36. The artificial hybrids, obtained by crossing both parental species, closely resemble the natural ones thereby confirming their parentage. It is proposed to designate the natural hybrids Solanum x viirsooi hybr. nov.