Haploid plant regeneration from unpollinated ovule cultures of niger (<Emphasis Type="Italic">Guizotia abyssinica</Emphasis> (L. f.) Cass.)

Haploid plants were regenerated in vitro from unpollinated ovules of niger (Guizotia abyssinica (L. f.) (Cass.) on Murashige and Skoog nutrient medium (MS) supplemented with 10 μM naphthaleneacetic acid or 10 μM NAA + 1.5 μM kinetin and 30 g/l sucrose. Gamborg (B5) medium was the best for plant regeneration (in comparison with MS, Nitsch and Nitsch (NN), and Chu (N6) media) from cultured ovules, and 6.66 and 7.33 ovules of JNC-6 and Ootacamund cultivars were involved in direct plant regeneration on this medium. Matured ovules (ovules collected one day before anthesis or on the day of anthesis) only responded to cultural regimes and involved in direct plantlet development. Cytological preparation of root tips and chloroplast counts in the guard cells of leaf stomata of regenerated plants confirmed their haploid nature. This text was submitted by the authors in English.     

Ribosomal DNA as a convenient probe to follow segregation and possible divergency from expected homozygosity after haploidization of an androgenetic process

Summary Restriction fragment length polymorphism of the wheat nuclear ribosomal DNA has been studied in several steps of a breeding scheme, including parental genotypes, F1 hybrid, F9 generation, and anther-derived doubled haploid lines obtained from F9. Ribosomal DNA represents a suitable molecular marker in following segregation and possible divergency from expected homozygosity after haploidization of an androgenetic process. It has been shown to undergo variations among the first cycle-doubled haploid lines in the relative amount of two different sizes of ribosomal DNA repeat units. The specificity and peculiar properties of the plant system used allowed us to assign an intrachromosomal location (short arm of the chromosomes 1B, 1R or 6B) to several ribosomal DNA repeat units that differ by the length of their nontranscribed spacer region.     

A comparison of androgenetic doubled-haploid,and single seed descent lines in Triticale

Summary Sixty single seed descent (SSD) lines and about 25 anther-derived doubled-haploid (DH) lines were obtained from two triticale crosses. The frequency distributions of 10 quantitative agronomic traits were compared using parametric and non-parametric tests. A multivariate discriminant analysis was subsequently carried out. Gliadin patterns obtained from each line by polyacrylamide gel electrophoresis were used to calculate intra- and inter-population diversities from relative dissimilarity indices. It was found that DH and SSD lines show significant differences in frequency distributions of 1000 grain weight in both crosses, of heading date for one cross, and of lodging susceptibility for the other cross. The results of intra- and inter-population gliadin diversity indicate that although the SSD method theoretically provides more opportunity for recombination to occur than the DH method, it did not produce a greater range of recombinants. Since there is no significant difference between SSD- and DH-line distributions for grain yield, anther culture appears to be an efficient method for producing high yielding homozygous lines from F₁ hybrids of triticale in a relatively short time.     

Genetical investigations into β-glucan content in barley

Summary Random inbred lines produced by doubled haploidy (DH) and single seed descent (SSD) have been used to investigate the genetics of -glucan (gum) content in barley (Hordeum vulgare). Genetical analyses indicated that gum content is controlled by a simple additive genetic system. Significant negative genetic correlations were observed between -glucan content, thousand grain weight and height in the DH samples. These correlations were much reduced in the SSD samples and would suggest linkage of the genes controlling these characters. The presence of repulsion linkages could be exploited in a barley breeding programme by producing F1 derived DH to generate recombinants with high thousand grain weight and low -glucan content. Genetical parameters estimated from DH and F3 samples have successfully been used to predict the number of inbred lines transgressing the parental range for -glucan content and bivariate combinations involving -glucan.     

The genetic characterisation of novel multi-addition doubled haploid lines derived from triticale x wheat hybrids

Two novel 46-chromosome doubled haploid lines, W66 and M17, derived from separate hexaploid triticale x bread wheat crosses, were characterised using cytological and biochemical markers. Both lines were shown to be relatively stable cytologically, over 11 and 8 generations of selfing, respectively. By examining mitotic and meiotic chromosomes, the stabilities of the two lines were shown to be similar with frequencies of 2n=46 in 74.2–85.5% of cells. However, over selfed generations, the rye chromosomes were shown to have lost some of their heterochromatin, which made it difficult to establish their continued presence using cytological techniques, such as C-banding alone. Cytological evidence from pairing studies, C-banding, and fluorescence in-situ hybridization, showed that both M17 and W66 are wheat/rye multi-addition lines with rye chromosome constitutions of 1R+6R, and 1R+4R, respectively. These conclusions were confirmed by isozyme and storage-protein analysis.     

Plant regeneration through culture of isolated microspores of Triticum aestivum L.

Wheat microspores mechanically isolated from the anthers before culture and isolated from the anthers during the hole culture period in a chemically defined medium resulted in proembryos, embryos and finally plants. Of the four genotypes included, all responded with proembryos, and the two spring wheats Ciano and Walter gave rise to macroscopic embryos and plants. The frequency of embryo regeneration and the frequency of albino plants in both Ciano and Walter was in accordance with previously obtained results with anther culture derived material.Abbreviations 2,4-d 2,4-dichlorophenoxy acetic acid - NAA 1-naphthaleneacetic acid     

Barley anther culture using membrane rafts

When compared to agarose solidified media in small petri dishes, membrane rafts used in conjunction with liquid induction media significantly improved anther culture response in the Australian, malting-quality, spring barley cultivar Clipper. In contrast, the German cultivar Gimpel did not show an increased response on rafts.Abbreviations BA 6-benzylaminopurine - IAA indoleacetic acid - DH doubled haploid     

Methods for estimating gene numbers for quantitative characters using doubled haploid lines

Summary Three methods of estimating the numbers of genes segregating for quantitative characters using doubled haploid lines are presented. The first uses estimates of the range and genetical variance of an F₁ or F₂ derived population. The second adapts the genotype assay method of Jinks and Towey (1976) to F₂ derived lines. The third uses the variances of an F₂ derived population. Statistical problems of obtaining meaningful estimates using these methods are discussed and it is concluded that genotype assay is the best method for distinguishing between few and many genes. These methods are illustrated using data from an experiment containing doubled haploid lines of barley developed using the H. bulbosum system.     

Glycoalkaloid Profile in Potato Haploids Derived from Solanum tuberosum–S. bulbocastanum Somatic Hybrids

Cultivated and wild potato species synthesize a wide variety of steroidal glycoalkaloids (GA) that may affect either human health or biotic stress resistance. Therefore, GA composition must be a major criterion in the evaluation of breeding products when species genomes are merged and/or manipulated. This work reports the results of GA analysis performed on unique haploid (2n=2x=24) plants obtained from tetraploid (2n=4x=48) Solanum bulbocastanum–S. tuberosum hybrids through in vitro anther culture. Glycoalkaloids were extracted from tubers and analyzed by HPLC. Haploids generally showed the occurrence of parental GA. However, in several cases loss of parental GA and gain of new GA lacking in the parents was observed. It may be hypothesized that new GA profiles of our haploids is the result of either genetic recombination or combinatorial biochemistry events. To highlight differences between haploids and parents, soluble proteins and antioxidant activities were also determined. Both were always higher in haploids compared to their parents. The nature of the newly formed GAs will be further investigated, because they may represent new metabolites that can be used against pest and diseases, or are useful for human health.     

抗疫病加工型辣椒细胞质雄性不育恢复系的创制

为加快抗疫病加工型辣椒细胞质雄性不育(cytoplasmic male sterility,CMS)恢复系的创制,该试验以单生、长果自交系481 4 10为母本,以抗疫病的恢复系939 1和1021(1) 1为父本配制杂交组合,采用花药培养技术将抗疫病恢复系的Rf基因和抗疫病基因导入单生、长果自交系中,并利用分子标记辅助选择(molecular marker assisted selection, MAS)技术鉴定DH系(Double Haploid line)的Rf基因以及抗病性,进一步用室内苗期抗性鉴定的方法验证MAS筛选的含Rf DH系对疫病的抗性。结果表明：(1)花药培养的供体亲本(481 4 10×939 1)F₁诱导出22个胚状体,成苗后经倍性鉴定获得11个花培DH系;而由供体亲本［481 4 10×1021(1) 1］F₁获得 9个DH系。(2)分子标记CRF SCAR对花培DH系进行分子标记辅助选择验证结果表明,来自供体(481 4 10×939 1)F₁的DH系中有7个可扩增出870 bp的特异条带,占63.6%;而供体［481 4 10×1021(1) 1］F₁获得的DH系中则有8个能扩增出870 bp的特异条带,占88.9%。(3)分子标记FQ01/RQ01对DH系进行分子标记辅助选择筛选结果发现,来自供体(481 4 10×939 1)F₁的DH系有5个能扩增出717 bp的特异条带,占45.5%;而来自供体［481 4 10×1021(1) 1］F₁的DH系有4个可扩增出717 bp的特异条带,占44.4%。(4)MAS技术初步筛选到7个携带Rf的抗疫病DH系,分别命名为‘渝辣选3 2/3 3/3 5/7 1/7 5/7 6/7 9’;苗期抗性鉴定结果显示,7个DH系中5个DH系抗疫病,2个中抗疫病;农艺性状调查和测交实验表明,‘渝辣选3 2’和‘渝辣选7 1’是单生朝天椒、果实较长,味辣,均为CMS恢复系。该研究创制的2个DH系为利用辣椒CMS三系配套选育抗病新品种奠定了基础。