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     

Segregation of 12 isozyme genes among doubled haploid lines derived from a japonica x indica cross of rice (Oryza sativa L.)

Summary The segregation of 12 heterozygous isozyme markers was analyzed among F₂ plants and 51 anther culture (AC)-derived lines obtained from the japonica × indica cross of rice, IRAT 177 × Apura. All the lines except two were homozygous products of recombination of the two parental phenotypes. Doubled haploid (DH) lines derived from plants regenerated from the same callus were identical, confirming previously obtained results in rice. Surprisingly, some lines derived from different calli were also identical, suggesting a phenomenon of early callus fragmentation. All these observations at the isozyme level were confirmed by field evaluation. Deviations of segregations from the expected 1 : 1 ratio were observed at 4 loci among the DH lines. Among these, two were also noted among the F₂ plants. The two other distortions, both in favor of the japonica allele, were observed specifically in the AC-derived materials.Although this concerns a small proportion of the genes under study, it suggests that the embryogenic microsporal population does not represent a random gametic array. On the other hand, evaluation of recombination between isozyme genes located on chromosome 6 appears consistent with F₂ data and data previously recorded on the other japonica × indica crosses. The potential use of isozymes in breeding doubled haploids derived from remote crosses in rice is discussed.Abbreviations MCPA = 2-methyl-4-chlorophenoxyacetic acid - IAA = indolacetic acid - AC plant or line = anther culture-derived plant or line - DH line = doubled haploid line     

Androgenic capability among genotypes of winter and spring barley

Variable efficiency of androgenesis remains a serious problem in many species of cereals. It is still unclear what makes certain genotypes more amenable to androgenesis than others. This study was undertaken to quantify the previously suspected advantage of winter barley genotypes over spring ones with regard to regeneration efficiency in anther culture. The material consisted of 40 barley hybrids originating from Polish breeding companies. The number of androgenic structures per 100 anthers did not differ significantly between analysed groups (119 vs. 152 non‐significant), but the average regeneration of green plants per 100 anthers was five times higher in winter genotypes (6.4 vs. 1.3). The incidence of albinism was lower for the winter than for the spring materials (70% vs. 90%), while the rate of spontaneous chromosome doubling was similar in both groups (58% vs. 56%). The results strongly support the notion that winter genotypes are more amenable to androgenesis and this may be a consequence of their better adaptation to stress conditions.     

Factors affecting oat haploid production following oat × maize hybridization

Doubled haploids (DHs) are becoming increasingly important in crop breeding programmes but methods for producing oat DHs remain inefficient. In this study haploid and DH oat plants were produced using the oat × maize hybridization method. Factors influencing the rate of caryopsis and haploid embryo production including genotype, post‐pollination plant growth regulator application and temperature were investigated. The four growth regulators tested showed significant differences in their capacity to induce caryopsis formation with dicamba producing the highest numbers of caryopses, followed by picloram, 2,4‐dichlorophenoxyacetic acid (2,4‐D) and gibberellic acid (GA₃). No significant differences were observed between these growth regulators for their effect on embryo production. The concentration of dicamba was also important and was found to influence caryopsis but not embryo production, with 50 and 100 mg/l dicamba producing significantly more caryopses than 25 or 5 mg/l. Temperature had a significant impact on both caryopsis and embryo production with the magnitude and direction of response depending on genotype. Rates of haploid embryo production observed were between 0.8% and 6.7% of the pollinated florets. The proportion of haploids, which survived and were successfully doubled with colchicine following transfer to soil was between 72% and 81%.     

Effect of genotype,environment and carbohydrate on anther culture response in head cabbage (Brassica oleracea L. convar. capitata (L.) Alef.)

Summary The effect of genotype, growing conditions for donor plants and type and concentration of carbohydrate in the culture medium was investigated for anther culture of head cabbage (white cabbage, savoy cabbage, pointed-headed cabbage). Strong genotypic effects on embryo formation from the cultured anthers were shown as well as superior embryo formation from anthers of field grown donor plants compared to plants grown in the greenhouse. When comparing 7, 10 and 13% sucrose in the medium, embryo response increased with increasing sucrose concentration. With maltose, which was generally inferior to sucrose as carbohydrate source for anther culture, the embryo response did not increase with maltose concentration above 10 per cent.     

Pleiotropic effects on the ml-o mildew resistance gene in barley in different genetical backgrounds

Summary The effects on quantitative characters of the ml-o5 allele has been investigated by means of doubled haploid lines produced by the Hordeum bulbosum method. In one series of experiments the genetic background in each cross was found significantly to modify the negative pleiotropic effects. Doubled haploids were also produced from a F1 cross and tested in the field during two years, one with strong and one with weak mildew infections. In the first year only an insignificant reduction in grain yield was detected in the mlo lines, in the second it was significantly reduced with 5.9%. Thousand grain weights were, however, higher in the resistant lines. In both series of experiments the possibility to extract very high yielding mlo genotypes was demonstrated. The nature of the genetic background effect is discussed.     

QTL analysis of seed dormancy in rice (Oryza sativa L.)

Effective cumulative temperature (ECT) after heading would be a more reasonable parameter for seed sampling of pre-harvest sprouting/seed dormancy (SD) tests in segregating populations than the days after flowering. SD is an important agronomic trait associated with grain yielding, eating quality and seed quality. To identify genomic regions affecting SD at different grain-filling temperatures, and to further examine the association between SD and ECT during grain-filling, 127 double haploid (DH) lines derived from a cross between ZYQ8 (indica)/JX17 (japonica) by anther culture were analyzed. The quantitative trait loci (QTLs) and their digenic epistasis for SD were identified using a molecular linkage map of this population. A total of four putative QTLs for SD (qSD-3, qSD-5, 6 and 11) were detected on chromosomes 3, 5, 6 and 11, together explaining 41.4% of the phenotypic variation. Nine pairs of digenic epistatic loci were associated with SD on all but chromosome 9, and their contributions to phenotypic variation varied from 2.87%–8.73%. The SD QTL on chromosome 3 was identical to the QTLs found in other mapping populations with different genetic backgrounds, which could be a desirable candidate for gene cloning and marker-assisted selection in rice breeding.     

Genetic and environmental effects on glucosinolate content and chemoprotective potency of broccoli

Broccoli is well recognized as a source of glucosinolates and their isothiocyanate breakdown products. Glucoraphanin is one of the most abundant glucosinolates present in broccoli and its cognate isothiocyanate is sulphoraphane, a potent inducer of mammalian detoxication (phase 2) enzyme activity and anti‐cancer agent. This study was designed to measure: glucosinolate levels in broccoli florets from an array of genotypes grown in several environments; the elevation of a key phase 2 enzyme, quinone reductase, in mammalian cells exposed to floret extracts; and total broccoli head content. There were significant environmental and genotype‐by‐environment effects on levels of glucoraphanin and quinone reductase induction potential of broccoli heads; however, the effect of genotype was greater than that of environmental factors. The relative rankings among genotypes for glucoraphanin and quinone reductase induction potential changed, when expressed on a per head basis, rather than on a concentration basis. Correlations of trait means in one environment vs. means from a second were stronger for glucoraphanin and quinone reductase induction potential on a per head basis than on a fresh weight concentration basis. Results of this study indicate that development of a broccoli phenotype with a dense head and a high concentration of glucoraphanin to deliver maximum chemoprotective potential (high enzyme induction potential/glucoraphanin content) is a feasible goal.     

Segregation distortion in doubled haploid lines of barley (Hordeum vulgare L.) detected by simple sequence repeat (SSR) markers

A total of 147 simple sequence repeat (SSR) markers (including86 barley and 61 wheat microsatellite markers) were tested for their segregation in a doubled haploid (DH) and an F₂ population of barley. The DH population consisted of 71 doubled haploid lines, developed from F₁ plants of a cross between Tadmor and WI2291using isolated microspore culture technique. A genetic linkage map consisting of 43 microsatellite markers was constructed using the DH population. Particularly on chromosome 4H microsatellite markers showed distorted segregation ratios. Segregation of DH lines based on molecular markers were compared with segregation of 92 F₂ lines from the same cross. The proportion of loci deviating from the expected monogenic segregation ratios in the DH population was significantly higher (19/43loci, 44%) than in the F₂ population (7/43 loci, 16%). The deviation was biased towards the WI2291 parent alleles. In line with this observation, WI2291 was found to perform better than Tadmor in regenerating green plantlets with the isolated microspore-culture technique. This revised version was published online in July 2006 with corrections to the Cover Date.     

QTL mapping and associated marker selection for the efficacy of green plant regeneration in anther culture of rice

Anther culturability of rice is a quantitative trait controlled by nuclear‐encoded genes. The identification of quantitative trait loci (QTL) and associated marker selection for anther culturability is important for increasing the efficiency of green plant regeneration from microspores. QTL associated with the capacity for green plant regeneration in anther culture of rice were mapped on chromosomes 3 and 10 using 164 recombinant inbred (RI) lines from a cross between ‘Milyang 23’ and ‘Gihobyeo’. The quantitative trait locus located on chromosome 10 was detected repeatedly when three anther culture methods were applied and was tightly linked to the markers, RG323, RG241 and RZ400. Associations between these markers and the efficacy of green plant regeneration in 43 rice cultivars and two F₂ populations, ‘MG RI036’/‘Milyang 23’, and ‘MG RI036’;/‘IR 36’ were analysed. One of these markers, RZ400, was able to identify effectively genotypes with good (> 10.0%) and poor (< 3.0%) regenerability, based on the marker genotypes in the cultivars and two F₂ populations. This marker enables the screening of rice germplasm for anther culturability and introgression into elite lines in breeding programmes.