Wide mapping of a T-DNA insertion site in oilseed rape using bulk segregant analysis and comparative mapping

Doubled haploid oilseed rape lines segregating for a transgene inducing herbicide resistance (bar gene) were investigated for the wide mapping of the T-DNA insertion site. Bulk segregant analysis using presence/absence and intensity polymorphisms between the bulks, as well as comparative mapping with a linkage group deriving from another cross, led to the identification of 11 random amplified polymorphic DNA (RAPD) markers tightly or loosely linked to the bar gene. Ten RAPD loci out of 11 were located on the same side of the bar locus, strongly suggesting that the T-DNA integrated in a telomeric or subtelomeric position. The eleventh RAPD marker exhibited a strong segregation distortion, which could be the result of a heteroduplex formation. Comparison of the linkage groups obtained from the two crosses showed different recombination rates between markers, possibly reflecting differences in parental genetic backgrounds. Consequences and potential applications in transgene dispersal safety assessment studies are discussed.     

The association of genes for purple coleoptile with chromosomes of the wheat variety Mironovskaya 808

Summary The association of genes for purple pigment in the coleoptile with the chromosomes of the winter wheat variety Mironovskaya 808 was investigated using monosomic F₂ analysis. The segregation ratio for F₂ hybrids of Chinese Spring monosomics x Mironovskya 808 seems to indicate that the purple colour of the coleoptile is determined by two dominant genes, Rc₃ and Rc₄, which are located on the chromosomes 7D and 6B respectively, and which reinforce each other. Apart from these two genes, suppressors found on the chromosomes 2A, 2B, 2D, 4B and 6A also play a role in the intensity of the purple colour.With the aid of a Chinese Spring telocentric chromosome marker it was observed that the Rc₃ gene is located on the chromosome arm 7DS, at a distance of 16±4.23 crossover units from the centromere.     

Detection of molecular markers linked to the durable adult plant stripe rust resistance gene Yr18 in bread wheat (Triticum aestivum L.)

Stripe rust of wheat caused by Puccinia striiformis West. f. sp. tritici presents a serious problem for wheat production worldwide, and identification and deployment of resistance sources to it are key objectives for many wheat breeders. Here we report the detection of simple sequence repeat (SSR) markers linked to the durable adult plant resistance of cv. ‘Otane’, which has conferred this resistance since its release in New Zealand in 1984. A double haploid population from a cross between ‘Otane’ and the susceptible cv. Tiritea’ was visually assessed for adult plant infection types (IT) in the glasshouse and field, and for final disease severity in the field against stripe rust pathotype 106E139A⁺. At least three resistance loci controlled adult plant resistance to stripe rust in this population. Quantitative trait loci (QTL) mapping results revealed that two of these, one on chromosome 7DS corresponds to the durable adult plant resistance gene Yr18 and other on chromosome 5DL were contributed from ‘Otane’; while the remaining one on chromosome 7BL, was contributed from the susceptible ‘Tiritea’. Interval mapping placed the ‘Otane’‐resistant segment near the centromere of chromosome 7DS at a distance of 7 cM from the SSR marker gwm44. The stability of QTL in the two environments is discussed. SSR gwm44 is potentially a candidate marker for identifying the durable resistance gene Yr18 in breeding programmes.     

Recent advances in molecular genetics of forest trees

The use of molecular markers has greatly enhanced our understanding of the genome structure of forest trees. Conifers, in particular, have a relatively large genome, containing a very high proportion of repeated DNA, consisting of tandemly repetitive and dispersed repetitive DNA sequences. The nature of highly conserved tandemly repetitive rRNA genes has been investigated in a number of tree species, and their sites mapped on specific chromosomes by fluorescent in situ hybridization (FISH). Different families of retrotransposons (IFG, and TPE1) have been isolated and characterized from the dispersed repetitive DNA of pines. Genome maps have been constructed in a number of forest tree genera: Pinus, Picea, Pseudotsuga, Cryptomeria, Taxus, Populus, and Eucalyptus. EST databases have been established from cDNA clones of pines and poplars. The structure and maternal or paternal modes of inheritance of organelle genomes have been investigated in forest trees. Comparative mapping in conifers has shown that gene families are conserved across genera. Due to lack of polyploidy in conifers, the evolution of this group of trees may have occurred primarily by duplication and dispersal of genes, probably by retrotranspositions, to form complex gene families. The evolution of angiosperm tree species has presumably involved both gene duplication as well as genome duplication (polyploidy). Application of genetic engineering has shown that genes from phylogenetically unrelated organisms can be introduced and expressed in trees, thus offering prospects of genetic improvement of forest trees. This revised version was published online in August 2006 with corrections to the Cover Date.     

Progress in mapping agronomic genes in apple (The European Apple Genome Mapping Project)

Summary The progress of the European Apple Genome Mapping Project is described. Populations segregating for a range of agronomic genes have been established in six European countries. The need for robust methods of analysis has been identified, especially with regard to the development of molecular markers. Isozyme systems, RAPDs, RFLPs and amplified genes are being used to construct a reference genetic linkage map. Standardisation and precise definition of both genotypic and phenotypic measurements has been recognised as being essential for future exploitation of genetic markers in apple breeding. Phenotypic measurements are being replicated in different geographical locations over several years. Statistical and genetic analyses are aimed at defining components of genetic variation which account for ‘genes’, as defined by apple breeders. A relational database is being constructed which will combine disparate sources of data relating to the genetics of apple. Comparative mapping has been identified as an efficient means of expanding genetic knowledge within and between Rosaceae genomes.     

Mapping of genes involved in glutathione, carbohydrate and COR14b cold induced protein accumulation during cold hardening in wheat

Using some of the chromosome substitution lines developed from thecrosses of the donor Cheyenne to Chinese Spring we showed that theaccumulation of water soluble carbohydrates during different stages ofhardening was time dependent. Moreover there was a significantcorrelation between the rate of carbohydrate accumulation and the frosttolerance. The expression and regulation of a wheat gene homologous tothe barley cold regulated cor14b gene was compared in frost sensitiveand frost tolerant wheat genotypes at different temperatures. Studies madewith chromosome substitution lines showed that the threshold inductiontemperature polymorphism of the cor14b wheat homologous genewas controlled by loci located on chromosome 5A of wheat, while cor14b gene was mapped, in Triticum monococcum, onto the longarm of chromosome 2A^m. Our study on the effect of cold hardeningon glutathione (GSH) metabolism showed that chromosome 5A of wheathas an influence on the GSH accumulation and on the ratio of reduced andoxidised glutathione as part of a complex regulatory function during coldhardening. In addition, the level of increase in GSH content duringhardening may indicate the degree of the frost tolerance of wheat.     

Molecular mapping of a locus controlling resistance to Albugo candida in Indian mustard

Brassica juncea (L.) Czern & Coss is widely grown as an oilseed crop in the Indian subcontinent. White rust disease caused by Albugo candida (Pers.) Kuntze is a serious disease of this crop causing considerable yield loss every year. The present study was undertaken to identify molecular markers for the locus controlling white rust resistance in a mustard accession, BEC‐144, using a set of 94 recombinant inbred lines (RILs). The screening of individual RILs using an isolate highly virulent on the popular Indian cultivar ‘Varuna’ revealed the presence of a major locus for rust resistance in BEC‐144. Based on screening of 186 decamer primers employing bulked segregant analysis (BSA), 11 random amplified polymorphic DNA markers were identified, which distinguished the parental lines and the bulks. Five of these markers showed linkage with the rust resistance locus. Two markers, OPN0_l000 and OPB06₁₀₀₀, were linked in coupling and repulsion phases at 9.9 cM and 5.5 cM, respectively, on either side of the locus. The presence of only two double recombinants in a population of 94 RILs suggested that the simultaneous use of both markers would ensure efficient transfer of the target gene in mustard breeding programmes.     

对家蚕多星纹基因ms的SSR标记定位分析

对家蚕幼虫斑纹突变多星纹基因ms进行分子标记定位,有益于对多星纹性状的分子标记辅助选择和对该基因的定位克隆研究。利用家蚕雌性染色体在减数分裂过程中不发生交换的特点,采用幼虫斑纹为素斑(p)的家蚕品系C108,幼虫斑纹为多星纹(ms)的家蚕品系g02,组配正反交群体(g02×C108)F1♀×g02♂和g02♀×(g02×C108)F1♂,分别记作BC1F和BC1M,利用已经构建的家蚕SSR分子标记连锁图谱中第12连锁群上的18对SSR标记引物在亲本间进行多态性筛选。BC1F群体中的普通斑个体均表现出与(g02×C108)F1相同的杂合型带型;而所有多星纹个体的带型与亲本g02一致,为纯合型。结果筛选出S1206、S1208、S1210、C5553S3共4个与家蚕ms连锁的SSR标记。利用BC1M群体构建家蚕ms及其连锁的SSR标记遗传连锁图,连锁图的图距为34.5 cM,4个SSR标记及ms的排列次序为S1206—S1208—S1210—C5553S3—ms,ms位于34.5 cM处,与ms最近的标记为C5553S3,遗传距离为12.4 cM。依据该SSR标记遗传连锁图谱可对ms进一步精确定位。     

苹果遗传连锁图谱构建及抗早期落叶病的基因定位

以“秦冠”ד富士”F1群体的158个株系为试材,得到最佳SSR- PCR反应体系:在20 μL总的反应体系中Taq DNA聚合酶0.5U、引物0.1μmol/L、Mg2+ 1.8 mmol/L、模板DNA浓度15 ng/μL、dNTPs 0.30 mmol/L.利用SSR分子标记,采用JoinMap 3.0软件构建“秦冠”ד富士”的遗传连锁图谱.结果表明:从340对SSR引物中筛选出49对具有多态性的引物,在群体中共获得75个SSR标记,其中17个不符合孟德尔遗传规律,其余27个位点可定位到10个连锁群上.对苹果杂交F1代早期落叶病进行抗性遗传分析,最后将抗早期落叶病基因定位到了遗传连锁图谱中的第10个连锁群上.与已经发表的苹果遗传连锁框架图对比结果表明,抗早期落叶病基因被定位在已发表图谱的第8连锁群上.