杨属ITS序列的分子进化特点分析

本文通过对杨属5组代表种并以柳属中旱柳的ITS序列作为参照,用成对比较的方法对杨属21个植物样品的ITS序列比较发现,杨属ITS序列的碱基替换情况是转换数大于颠换数,ITS-1序列中碱基发生转换和颠换的数目基本相似,而ITS-2序列中碱基发生转换的数目远远大于颠换。用朱克斯和坎托一参数模型和木村二参数模型对杨属ITS序列的替换率计算,无论是ITS-1还是ITS-2序列都没有达到数理统计上的明显差异。但由于ITS-2序列碱基的变化转换大大高于颠换,所以二参数模型计算所得结果大于一参数模型;采用相对速率测验法,对杨属各组分子进化相对速率进行了检测,结果分析得知,杨属5组的ITS序列分子进化速率有差异。杨属各组的ITS-1序列比较说明,大叶杨组、青杨组、黑杨组的进化速率都小于白杨组,而胡杨组则是进化速率最快的,大叶杨组的进化速率小于青杨组、胡杨组和黑杨组,黑杨组和胡杨组大于青杨组;ITS-2序列比较发现白杨组是进化速率最慢,其次是大叶杨组、青杨组,而黑杨组则稍大于胡杨组;总ITS序列比较是大叶杨组和黑杨组相对较慢,胡杨组最快;但就总的碱基替换分析,除了黑杨组和大叶杨组在ITS-1区间及青杨组、胡杨组和白杨组在ITS-2区间进化速率有显著差异外,其余组间都无显著的差异,尤其是在整个ITS序列区段碱基替换的差异都无显著差异。     

Characterization of dinucleotide and trinucleotide EST-derived microsatellites in the wheat genome

Over the past decade microsatellites or simple sequence repeats (SSRs) have attracted a considerable amount of attention from researchers. The aim of the present paper was to analyse expressed sequence tag-derived SSR (EST-SSR) marker variability in wheat and to investigate the relationships between the number and type of repeat units and the level of microsatellite polymorphism. Two hundred and forty-one new EST-SSR markers available in a public database () were characterized in eight durum wheat cultivars (Svevo, Ciccio, Primadur, Duilio, Meridiano, Claudio, Latino, Messapia), two accessions of Triticum turgidum var. dicoccoides (MG4343, MG29896), one accession of T. turgidum var. dicoccum (MG5323) and in the common wheat cv. Chinese Spring. Of these, 201 primer pairs (83.4%) amplified PCR products successfully, while the remaining 40 (16.6%) failed to amplify any product. Of the EST-SSRs analysed, 45.2% of the primer pairs amplified one or two PCR products. Multiple discrete PCR products were observed among both di- and trinucleotide EST-SSR markers (31.2 and 40.5%, respectively). Markers based on dinucleotide microsatellites were more polymorphic than those based on trinucleotide SSRs in the 12 wheat genotypes tested (68.9 and 52.7%, respectively). An average of 2.5 alleles for dinucleotide and 2.0 alleles for trinucleotide SSRs was observed. The data reported in the present work indicate the presence of a significant relationship between motif sequence types and polymorphism. The primer set based on the AG repeat motif showed the lowest percentage of polymorphism (55.0%), while the primer set based on the AC repeat motif showed t he highest percentage (85.0%). Among trinucleotide SSRs, the AGG microsatellite markers showed the highest percentage of polymorphism (70.0%), and the ACG motif the lowest value (25.0%). The characterization of these new EST-SSR markers and the results of our studyon the effect of repeat number and type of motifs could have important applications in the genetic analysis of agronomically important traits, quantitative trait locus discovery and marker-assisted selection.     

Amplified fragment length polymorphism as a tool for molecular characterization of almond germplasm: genetic diversity among cultivated genotypes and related wild species of almond,and its relationships with agronomic traits

Amplified fragment length polymorphism (AFLP) analysis is a rapid and efficient method for producing DNA fingerprints and molecular characterization. Our objectives were to: estimate genetic similarities (GS), marker indices, and polymorphic information contents (PICs) for AFLP markers in almond cultivars; assess the genetic diversity of almond cultivars and wild species, using GS estimated from AFLP fingerprints and molecular characterization; and facilitate the use of markers in inter-specific introgression and cultivar improvement. The genetic diversity of 45 almond cultivars from Iran, Europe, and America, were studied assaying 19 primer combinations. In addition, several agronomic traits were evaluated, including flowering and maturity times, self-incompatibility, and kernel and fruit properties. Out of the 813 polymerase chain reaction fragments that were scored, 781 (96.23%) were polymorphic. GS ranged from 0.5 to 0.96, marker indices ranged from 51.37 to 78.79, and PICs ranged from 0.56 to 0.86. Results allowed the unique molecular identification of all assayed genotypes. However, the correlation between genetic similarity clustering as based on AFLP and clustering for agronomic traits was low. Cluster analysis based on AFLP data clearly differentiated the genotypes and wild species according to their origin and pedigree, whereas, cluster analysis based on agronomic data differentiated according the pomological characterization. Our results showed the great genetic diversity of the almond cultivars and their interest for almond breeding.     

水稻抗条纹叶枯病基因Stv-bi的分子标记辅助选择

水稻条纹叶枯病是我国黄淮及长江流域粳稻区重要的病害。由于水稻条纹叶枯病的发病受外界条件影响较大,人工接种抗性鉴定比较困难,利用与抗病基因紧密连锁的分子标记进行标记辅助选择对提高抗性育种效率具有重要意义。来自籼稻抗源Modan的Stv-bⁱ是水稻育种中广泛应用的条纹叶枯病抗性基因。本研究设计了与Stv-bⁱ紧密连锁的SSR及STS分子标记,用3个抗条纹叶枯病混合群体F30718(圣稻13/镇稻88)、F50701(武优34/T022//圣稻806)、F60702 (V6/T022//镇稻88)进行分子标记检测和田间条纹叶枯病抗性鉴定,其结果的符合率分别为99.3%、87.7%和91.8%。表明这些分子标记可以用于条纹叶枯病抗性基因Stv-bⁱ分子标记辅助选择。     

抗虫转基因植物鉴定方法的研究进展

农作物受害虫侵害严重,给农业生产带来巨大损失,喷施化学杀虫剂使害虫产生抗药性,且污染环境。近年来,转基因技术研究不断深入,抗虫转化研究工作进展迅速。鉴定转基因植物的方法技术有很多种,在此主要阐述了利用标记基因进行鉴定,分子标记,免役技术三大类方法,并对其原理分别进行了简单介绍。     

棉花黄萎病抗性的分子研究进展

本文综述了棉花黄萎病菌的分子鉴定，棉花黄萎病抗性的遗传基础，分子标记在棉花抗黄萎病方面的应用，棉花黄萎病抗性基因的克隆及转化等方面的研究进展。     

大白菜抗根肿病基因位点CRa和CRb的分子标记鉴定

利用大白菜抗根肿病基因CRa和CRb分子标记(SC2930和KBr H129J18R)引物组,对78份大白菜材料进行抗根肿病分子标记鉴定。结果表明,在这78份材料中,有34份材料含有SC2930-T(CRa抗病标记)标记,其中杂合抗病位点材料17份,纯合抗病位点材料17份。有37份材料含有KBr H129J18R抗病标记,其中纯合抗病位点材料15份,杂合抗病位点材料22份。有20份材料不含有CRa和CRb所对应的抗病标记,23份材料含有2个抗病标记。该研究初步明确了78份参试大白菜材料所含抗根肿病基因CRa和CRb类型,为大白菜抗根肿病育种提供材料选择依据。     

Fusarium wilt of chickpea: physiological specialization, genetics of resistance and resistance gene tagging

Chickpea wilt caused by Fusarium oxysporum f. sp. ciceris is one of the major yield limiting factors in chickpea. The disease causes 10–90% yield losses annually in chickpea. Eight physiological races of the pathogen (0, 1A, 1B/C, 2, 3, 4, 5 and 6) are reported so far whereas additional races are suspected from India. The distribution pattern of these races in different parts of the world indicates regional specificity for their occurrence leading to the perception that F. oxysporum f. sp. ciceris evolved independently in different regions. Pathogen isolates also exhibit differences in disease symptoms. Races 0 and 1B/C cause yellowing syndrome whereas 1A, 2, 3, 4, 5 and 6 lead to wilting syndrome. Genetics of resistance to two races (1B/C and 6) is yet to be determined, however, for other races resistance is governed either by monogenes or oligogenes. The individual genes of oligogenic resistance mechanism delay onset of disease symptoms, a phenomenon called as late wilting. Slow wilting, i.e., slow development of disease after onset of disease symptoms also occurs in reaction to pathogen; however, its genetics are not known. Mapping of wilt resistance genes in chickpea is difficult because of minimal polymorphism; however, it has been facilitated to great extent by the development of sequence tagged microsatellite site (STMS) markers that have revealed significant interspecific and intraspecific polymorphism. Markers linked to six genes governing resistance to six races (0, 1A, 2, 3, 4 and 5) of the pathogen have been identified and their position on chickpea linkage maps elucidated. These genes lie in two separate clusters on two different chickpea linkage groups. While the gene for resistance to race 0 is situated on LG 5 of Winter et al. (Theoretical and Applied Genetics 101:1155–1163, 2000) those governing resistance to races 1A, 2, 3, 4 and 5 spanned a region of 8.2 cM on LG 2. The cluster of five resistance genes was further subdivided into two sub clusters of 2.8 cM and 2.0 cM, respectively. Map-based cloning can be used to isolate the six genes mapped so far; however, the region containing these genes needs additional markers to facilitate their isolation. Cloning of wilt resistance genes is desirable to study their evolution, mechanisms of resistance and their exploitation in wilt resistance breeding and wilt management.     

Mitochondrial DNA Restriction Fragment Length Polymorphisms in Malus Species

Three apple (Malus×domestica) cultivars and 11 Malus accessions have been investigated by the probe hybridization method on their mitochondrial DNA (mtDNA). The gene probes used were: coxI, coxII, atpA, atp6, and atp9. Our results revealed enough variation to characterize ten mtDNA haplotypes among the Malus genotypes examined. The taxonomic and phylogenetic implications of mtDNA polymorphism are also discussed.     

北方粳稻分子育种研究进展

本文介绍了北方粳稻分子育种课题的研究情况和所取得的研究成果。目前主要进行两方面的研究，一方面为特异基因型的鉴定：主要有筛选鉴定出抗旱性材料86份，苗期抗寒性材料40份，后期抗寒性材料629份，抗稻瘟病材料28份，对筛选出的抗旱性材料进行了第二轮聚合杂交，共配组合171个。另一方面是关于遗传与生理基础研究，对穗部性状、粒形、产量和品质以及它们之间的关系进行了深入的研究。另外本文对北方粳稻分子育种材料构建过程中存在的问题进行了综合讨论，并讨论北方粳稻分子育种今后的发展方向。