Geographical distribution of allozyme patterns in shallot (Allium cepa var. ascalonicum Backer) and wakegi onion (A. × wakegi Araki)

Summary Shallot and wakegi were collected in the main islands of Indonesia, and in Japan, Korea, Taiwan, Malaysia, Thailand and Bangladesh. Five isozyme resolutions, phosphoglucomutase (PGM), glutamate oxaloacetate (GOT), glutamate dehydrogenase (GDH), esterase (EST) and peroxidase (POX) were employed for demonstrating inter-and intraspecific differences. A dendrogram separated 189 collected accessions into 25 types of wakegi onion and 18 types of shallot. All accessions of Japan, Korea and Taiwan were determined to be wakegi onion, whereas those of Bangladesh, Malaysia and Thailand were shallot. Twenty-six out of 165 Indonesian accessions indicated wakegi onion distribution in Sumatra, West Java province and in Sulawesi Island. This confirmed that there is mixed-cultivation of the two Allium species with no distinction made between them. Japan and Indonesia had respectively 12 and eight unique types of wakegi onion, while Korea had only one type. West Java showed the most various type of wakegi onion, whereas East Java had many types of shallot. Shallots collected from Bangladesh were distinetly different from those of South East Asian types.     

Seed productivity test of CMS lines of Japanese bunching onion (Allium fistulosum L.) possessing the cytoplasm of a wild species, A. galanthum Kar. et Kir.

In a previous study, we developed male sterile lines of Japanese bunching onion (Allium fistulosum L.) possessing the cytoplasm of a wild species, A. galanthum Kar. et Kir., by backcrossing. To evaluate seed productivity of the male sterile lines in practise, they were crossed with the male fertile line, cultivar 'Kujyo', using honeybees as pollinators under field conditions. The number of florets and seeds per inflorescence, seed set and seed germination of the material were investigated. Although variation was observed among the male sterile lines, there were several lines having seed productivity equal to cultivar 'Kujyo'. Our data demonstrate that the male sterile lines of A. fistulosum possessing the cytoplasm of A. galanthum are useful as seed parents for the commercial F seed production of A. fistulosum.     

Variation in pollen viability in the onion (Allium cepa L.)

Summary The pollen viability of onions in a glasshouse was recorded from May to October 1975, using the fluorescein test. The average viability was 60–95% for most of this period but fell to less than 1% during the last two weeks of August. There was great variation in pollen viability between anthers within a flower and between flowers within a head. Attempts to induce pollen inviability by low temperature treatments at various stages of inflorescence development were unsuccessful. Low levels of pollen inviability appear to be a characteristic feature of onions, but the high level of inviability which was found both in this and in a previous season was associated particularly with the August period.     

Culture of unpollinated ovules,ovaries, and flower buds in some species of the genus Allium and haploid induction via gynogenesis in onion (Allium cepa L.)

Summary Induction of haploid plants is of great importance for breeding purposes because of the possibility to obtain from haploids homozygous material by artificial chromosome doubling in relatively short times. The present study reports the first evidence of successfull haploid induction in onion. Isolated ovules, ovaries, or whole flower buds of different Allium species were cultured on BDS agar medium. Testa browning in the ovules and an extensive growth of the latter were observed. In cultures of ovaries and flower buds, development of callus and subsequent regeneration of plantlets from the region of the nectaries were observed. In leek, sometimes supernumerary flower organs like ovules were formed in this callus. In onion (Allium cepa L.), plantlets developed from the ovules in all culture methods. Chromosome numbers of these plantlets were counted in root tip squash preparations. They were found to be haploid. Haploid plants were significantly smaller than diploid ones. They were transferred to soil and developed until bulb formation. Because of their importance for breeding, haploid plants obtained by gynogenesis are further stored in vitro.     

Gibberellic acid as a gametocide for the common onion (Allium cepa L.). II. The effect of GA 4/7

Summary The gametocidal effect of Berelex GA 4/7 on onions was investigated. Frequent spraying during the bolting period with relatively high concentrations (0.1 and 0.3%) resulted in a very high percentage of completely male sterile plants in the beginning of the flowering period. The gametocidal effect was accompanied by a considerable reduction in seed production.Differences in reaction to GA 4/7 between the three varieties involved were small.     

Identification of Allium cepa L. cultivars by means of statistical analysis of C-banded chromosomes

Summary The C-banded karyotypes of six Allium cepa cultivars, including five dry onions and one shallot, were investigated. Chromosome length and banding patterns were statistically analysed. The value of C-banding for distinguishing these Allium cepa cultivars was tested by the application of a discriminant analysis. It appeared that chromosomal differences between cultivars exist, but are too small to be clearly demonstrated with moderate efforts. Large variation within plants and between plants within cultivars, presumably for the greater part caused by the applied method, obscured the variation between the cultivars.     

Linkage of downy mildew resistance genes Pd 1 and Pd 2from Allium roylei Stearn in progeny of its interspecific hybrid with onion (A. cepa L.)

Summary Male meiosis of the interspecific hybrid between Allium roylei and A. cepa is undisturbed relative to its parents. Based on meiotic data, A. roylei is concluded to be a closer relative of A. cepa than A. fistulosum. Segregation ratios for downy mildew resistance among BC₁ and F₂ progenies from the F₁ between A. roylei and A. cepa indicate the presence of two dominantly inherited, weakly linked nuclear resistance genes, Pd ₁ and Pd ₂, in A. roylei (recombination frequency 0.32±0.03). Presumably Pd ₁ and Pd ₂are the first genes described in Allium residing in one linkage group. The prospects of exploiting A. roylei in onion breeding seem very promising.     

Chromosome and cytoplasm analyses of somatic hybrids between onion (Allium cepa L.) and garlic (A. sativum L.)

Chromosomes and cytoplasms were analyzed in two lines of a somatic hybrid between onion (Allium cepa L.) and garlic (A. sativum L.). One line of the somatic hybrid had 40 chromosomes and the other 41chromosomes. Genomic in situhybridization successfully revealed the chromosome constitution of the two lines. One line had 20 chromosomes from onion and17 chromosomes from garlic, and the other had 21 chromosomes from onion and 17chromosomes from garlic. Interestingly, both lines had three chimeric chromosomes. PCR-RFLP analyses of chloroplast and mitochondrial DNAs of both lines showed that these were identical to the onion parent. This revised version was published online in July 2006 with corrections to the Cover Date.     

Pre- and post-fertilization barriers to backcrossing the interspecific hybrid between Allium fistulosum L. and A. cepa L. with A. cepa

Summary To facilitate the introgression of desirable traits of Allium fistulosum into the genome of A. cepa, several accessions of the hybrid between these species were pollinated with A. cepa as the recurrent parent, and in vitro ovary and ovule culture were performed to obtain an increase in the recovery of backcross progeny. Compared to the results obtained from seed development in planta, the increase in the number of backcross progeny was generally very limited, and in some cases even a decrease was found. Raising the sucrose concentration in the ovary culture medium resulted in a higher frequency of ovules developing back seed coats but this was not followed by an increase in the number of backcross progeny obtained. Pollen tube growth of A. cepa was disturbed in the styles of the interspecific hybrids. Per ovule, frequencies of micropylar penetration exceeded frequencies of backcross progeny only to a limited extent. Hence, it was concluded that in the tested interspecific hybrid accessions the attainable gain in viable backcross progeny by the application of in vitro culture techniques is limited by strong pre-fertilization barriers acting at the level of stylar incongruity.     

Measuring onion cultivars with image analysis using inflection points

Summary The suitability of image analysis was studied to measure bulb characteristics for varietal testing of onions (Allium cepa L.). Eighteen genotypes were used, which covered a whole range of onion shapes, including some quite identical ones. The characteristic height and diameter were measured both by image analysis and by hand. The methods gave comparable results with regard to precision and discriminating power. The discriminating power of the image analysis method could be increased by adding a new characteristic: the relative position of the diameter.For determining the height of the bulb with image analysis two methods are described to locate the top of a bulb. One method is to use the location of the inflection point as the top of the bulb. For most bulb shapes, this point coincides with the top of the bulb as indicated by a crop expert. For ovate shaped onions, however, the inflection point is below the top of the bulb as indicated by the crop expert. The other method is to use the location of the intercept of the tangent at the inflection point with the length axis of the bulb as the top of the bulb. This point is always above the top as indicated by the crop expert. Moreover it seems to perform slightly worse for the discrimination of cultivars. Therefore the location of the inflection point as the top of the bulb is preferred.     

Heritability estimates and response to selection for the pungency and single center traits in onion

Summary Heritabilities of the pungency and single center traits were estimated in onion breeding populations using selection response and half-sib family analyses. Pungency was determined indirectly by measuring enzymatically produced pyruvic acid in individual bulbs. After one generation of selection, pungency was lowered by 8.1% and 8.9% in the populations 90-61-1 and 89-69-8, respectively, and realized heritabilities of 0.21 and 0.51 were estimated.Selection had no effect in lowering the pungency of population 90–62. Heritability estimates calculated through half-sib progeny analysis were 0.53, 0.48, and 0.25 for pungency in the populations 90-61-1, 90–62 and 89-69-8, respectively. The number of single centered onions was increased by 19% and 22% in populations 90–62 and 89-69-8, respectively, after one generation of selection, and the realized heritability estimates were 0.37 and 0.34, respectively.     

Introgression of leaf blight resistance from Allium roylei Stearn into onion (A. cepa L.)

Summary Segregation analysis in BC₁ and F₂ progenies from interspecific hybrids between Allium roylei and A. cepa demonstrated that leaf blight resistance is conditioned by one single dominant gene from A. roylei. This gene, designated Bs ₁ , was unlinked to the genes Pd ₁ and Pd ₂ determining downy mildew resistance. The prospects of exploiting A. roylei as a source for disease resistances in onion breeding are very promising.     

Isolation of 1,796 SSR clones from SSR-enriched DNA libraries of bunching onion (Allium fistulosum)

Bunching onion (Allium fistulosum L.) is one of the most important vegetables in Japan. To establish a genetic basis for molecular breeding of bunching onion, we isolated 1,796 simple sequence repeat (SSR) clones by large-scale sequencing of SSR-enriched genomic DNA libraries. Of these, 1,331 (74.1%) contained (GT) _n repeats (n > 5), while 314 (17.5%) were (GA) _n -containing clones. The average number of SSR repeats was 10.5 and 10.4 in the (GT) _n - and (GA) _n -containing clones, respectively. In a sample of five bunching onion inbred lines, an average of 3.2 alleles were detected in the 100 SSR loci investigated, with the polymorphic information content averaging 0.55. These results indicate that bunching onion SSRs are very rich sources of highly informative genetic markers.     

Callus induction and plant regeneration from anther and inflorescence culture of Sorghum

Summary Twenty-five inbred lines, including grain and forage types from the USA and China, two hybrids, one Sorghum almum, and one Parasorghum (S. versicolor) were tested for their response to anther culture. Three nutrient media were effective in inducing anther calli from six cultivars (Xin White, TX 403-TSB, DDY Sommer Milo, TX 2779, Brawley, and Spur Federal) and one was effective for plant regeneration for one cultivar, Xin White. Averaged over media, callus induction frequency (number of calli per 100 anthers) was highest in cultivars Xin White and TX 403-TSB (6.7 and 3.9%, respectively). The means of cultivars for media C17-2 and Ms-t-z-2, 4.3 and 3.2%, respectively, were superior to that for medium 85D3-2 (0.1%). Expressed as an average of the six cultivars and three media the mean calli induction frequency was 2.6%; however, differential responses of genotype and medium were noted. Among the 10 regeneration media tested, medium MS-d-4 containing Murashige and Skoog basal components plus 2.0 mg/l indole-3-acetic acid (IAA) and 2.5 mg/l kinetin was the most effective for plant regeneration. Numbers of albino plants and calli developing only roots increased directly with callus-induction time, whereas the frequency of plant regeneration decreased. Regenerated plants had varied numbers of chromosomes in root tip cells: 10, 15, 20, 40, and 60. The 29 regenerated plants that reached maturity, however, were highly fertile and contained only 10 bivalents in pollen mother cells. Normal chromosome number and behavior for the regenerated plants suggest that induced calli originated from cells other than microspores. However, spontaneous chromosome doubling in microspore-derived haploids may occur. The appearance of albinos also implies that haploids may have been produced from anther culture.Joint contribution of the Dept. of Agronomy and USDA-ARS, Kansas Agricultural Experiment Station, Manhattan, KS 66506-5501, USA. Contribution no. 88-566-J.     

Onion germplasm interacts with sulfur fertility for plant sulfur utilization and bulb pungency

Summary Sixty-two onion (Allium cepa L.) entries grown under high (4.0 mequiv.liter^-1) and low (0.1 mequiv.liter^–1) S fertility were evaluated for foliar and bulb S utilization, and pungency. Significant differences were detected among entries at both S fertility levels for foliar and bulb %S, and enzymatically developed pyruvate (pungency) in bulb tissue. Correlations between foliar %S and enzymatically developed pyruvate and between bulb %S and enzymatically developed pyruvate were poor suggesting that S is differentially partitioned into flavor and non-flavor compounds in the bulbs of onion germplasm. Significant S by entry interactions were found for all variables tested indicating that selection for S utilization and bulb pungency in onions be conducted at S fertility levels indicative of targeted production areas.     

Production of interspecific hybrids between Glycine max and G. tomentella through embryo culture

Summary Interspecific hybrids have been obtained in an incompatible cross between Glycine max and G. tomentella through the in vitro culture of hybrid embryos. The percentage of successful pod setting in the crosses averaged 12.8% but there were marked differences depending on the soybean cultivar used as the female parent. Hybrid embryos at globular to heart stages were extracted from the embryo sac 15–25 days after pollination and cultured in vitro. Hybrid plants were successfully obtained by culturing the embryos on B5 medium supplemented with 0.1 mg/l IBA followed by culture on B5 medium supplemented with 0.1 mg/l TBA plus 0.25 mg/l 2-iP. The F1 plants resembled the wild male parent in growth form, but had an intermediate leaf shape between that of the parents.     

Production of interspecific hybrids between Alstroemeria pelegrina L. var. rosea and A. magenta Bayer by ovule culture

Interspecific hybrids were efficiently produced in the cross-incompatible combination between Alstroemeria pelegrina L. var. rosea and A. magenta Bayer by culturing immature ovules with placenta 7–14 days after pollination on 2 g/l Gelrite-solidified MS medium containing 3% (w/v)sucrose. The plants showed intermediate characteristics between the parents and their hybridity was confirmed by karyotype and DNA analyses. The mean number of chromosome association per PMC at metaphase I was 2.60I+6.70II, pollen stainability was20.8%, and they produced viable seeds after self-pollination. Furthermore, mature plants were obtained when the hybrids were backcrossed as male parents with both the parents. The backcross-progeny from A. pelegrina var. rosea × hybrids exhibited 3.8 to 79.7% pollen stainability and that from A. magenta × hybrids 78.8 to 98.3%. Almost all of these plants produced viable seeds after self-pollination, which implies that they can beutilized for breeding of novel cultivars of Alstroemeria. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparison of fertility between the F1, F2 and anther derived lines in the crosses of indica/japonica and japonica/indica in rice (Oryza sativa L.)

Summary Response of anthers in in vitro culture was examined in the indica-japonica hybrids of rice (Oryza sativa L.). Significant genotypic differences were observed for callus induction and regeneration among the different interracial hybrids of indica-japonica races. Induction frequency of haploids ranged from 57.7 to 72.9 per cent and doubled haploid androgenic lines ranged from 27.1 to 42.3 per cent in the anther culture of the different hybrids. The indica-japonica hybrids recorded partial pollen grain and spikelet fertility in F1 (29.9 to 41.5% and 19.4 to 48.7% respectively) as well as in F2 (42.7 to 50.6% and 37.1 to 54.4% respectively). In contrast, the androgenic doubled haploid lines recorded significant increase and the pollen grain and spikelet fertility was 76.3 and 78.6 per cent respecitively. The results suggested that the sterility barriers for realising genetic recombinants and fixation of fertile homozygous lines in indica-japonica hybridization programme could be overcome through F1 anther culture technique.Abbreviations BAP Benzyl Amino Purine - 2,4-D 2,4-Dichlorophenoxy acetic acid - MS Murashige & Skoog medium - IAA Indole Acetic Acid     

Analysis of the yield potential and yield components of F1 and F2 hybrids of crosses between wheat (Triticum aestivum L.) and spelt (Triticum spelta L.)

Summary Crosses of wild barley (Hordeum vulgare ssp. spontaneum and Hordeum vulgare ssp. agriocrithon) with Hordeum vulgare ssp. vulgare were used to select high yielding grain types under dryland Mediterranean conditions. No special difficulties were faced in making the crosses, in eliminating the brittle rachis genes from the grain types or in selecting 6-rowed types in crosses between 2-rowed wild barley and 6-rowed ssp. vulgare varieties. Brittle rachis genotypes, present in the segregating populations were used in developing self-reseeding permanent pastures for dry areas. The best selections were tested in seven trials during 1989–92 and some of them outyielded their parents and also the best improved check variety by 13–22%. Indications for transgressive segregation were obtained for grain yield, straw yield, total biological yield, harvest index and volume weight. The crude protein content of some of the selections was significantly higher than that of the checks. For breeding programs aiming at large seeds, special ssp. spontaneum lines should be used as parents. High grain yield was positively correlated with high straw yield, total biological yield, earliness in heading date, high harvest index and negatively with volume weight. It was concluded that unexploited useful genes, even when not directly observed in wild barley, could be transfered easily into high yielding genotypes by breeding.     

Interspecific hybrids of Cyclamen persicum Mill. and C. purpurascens Mill. produced by ovule culture

Summary Interspecific crosses were made to introduce the scent of flowers of C. purpurascens into C. persicum cultivars and ovule culture was used to rescue the abortive hybrid embryos. Cultivars of C. persicum diploid (CPD, 2n=2×=48) and C. persicum tetraploid (CPT, 2n=4×=96) were the pistillate parents and wild species of C. purpurascens (CP, 2n=34) were staminate parents. After pollination, crossed ovaries were collected periodically and examined using paraffin sections. Histological observations suggested that both hybrid ovules of CPD x CP and CPT x CP should be transferred to culture medium 35 days after pollination. Based up on this observation, crossed ovaries were collected 28 days after pollination and ovules with placenta were transferred to MS (1962) medium containing 3% sucrose. These ovules were cultured in the dark at 25° C. The hybrids (2n=41) derived from CPD x CP had the scent of C. purpurascens, whereas the hybrids (2n=65) derived from CPT x CP had the scent of C. persicum. Although both hybrids had complete genomes from the parents and produced a few viable pollen grains, they failed to yield viable seeds by self- and cross-pollination with fertile pollen grains of C. persicum cultivars.Abbreviations CPD C. persicum diploid - CPT C. persicum tetraploid - CP C. purpurascens