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根据33个低分子量麦谷蛋白亚基(LMW-GS)基因5′侧翼序列的相似性进行聚类分析,可将其划分成8个类群,这与基于N末端推导氨基酸序列进行的类群划分结果完全一致.序列比对发现,各类群基因5′侧翼保守序列间存在DNA多态性,共发现34个多态性位点,其中18个为潜在单核苷酸多态性位点(SNPs,singlenucleotidepolymorphisms).除1个LMW-GS类群之外,其余7个类群的5′侧翼序列均具有类群特异性DNA变异位点.根据类群间的DNA多态性对这7个类群设计了特异引物,利用普通小麦(TriticumaestivumL.)品种中国春及其第1同源群双端体系对其进行染色体定位分析,揭示了1AS,1BS和1DS上分别有第2,1和4类群.对PCR产物的克隆测序进一步验证了不同染色体组上的LMW-GS基因类群间5′侧翼序列具有特异性.这些结果表明,LMW-GS基因的编码区及其5′侧翼保守序列可能是协调进化的.本文报道的7对引物可对7类LMW-GS基因的完整编码区进行特异扩增,因而能在小麦复杂的遗传背景下有目的地对某一类LMW-GS基因进行分离克隆,这有助于弄清单个LMW-GS对小麦品质的贡献.同时,在小麦育种中,这些标记对于有效地选择与品质密切相关的LMW-GS组分有一定应用价值.
Cluster analysis based on the similarity of 5 ’flanking sequences of 33 low molecular weight glutenin subunits (LMW-GS) genes can be divided into 8 groups, which is completely consistent with the grouping based on N-terminal deduced amino acid sequence The results of sequence alignment showed that there were 34 polymorphic sites among the 5 ’flanking conserved sequences in each group of genes, of which 18 were potential single nucleotide polymorphisms (SNPs, single nucleotide polymorphism. Except for one LMW-GS group, the 5 ’flanking sequences of the other seven groups all had group-specific DNA variation sites. According to the DNA polymorphism among the groups, specific primers were designed for these seven groups, Chromosome mapping of Triticum aestivum L. var. Chinensis and its first homologue double-end system revealed that there were 2, 1 and 4 groups on 1AS, 1BS and 1DS, respectively. The cloning of PCR products Sequencing further confirmed the specificity of the 5 ’flanking sequences between the LMW-GS gene groups on different chromosome sets.These results indicate that the coding region of LMW-GS gene and its 5’ flanking conserved sequences may be coordinated evolution.In this paper Seven pairs of primers were available for class 7 LMW-GS Gene specific amplification of the complete coding region, which can be purposely in the wheat genetic background of a particular type of LMW-GS gene cloning, which helps to understand the single LMW-GS on wheat quality contribution. At the same time, these markers have some value in the effective selection of LMW-GS components closely related to quality in wheat breeding.