【作者】 董长海；

【导师】 张彩英； 马峙英；

【作者基本信息】 河北农业大学 ， 作物遗传育种， 2011， 硕士

【摘要】 颜色性状是决定小麦外观品质的重要指标,对面条及其它相关制品品质有重要影响。克隆小麦色泽相关基因和开发与之有关的分子标记对小麦品质改良具有重要意义。本文在前人研究的基础上,对黄色素合成相关基因做了进一步研究,应用同源基因克隆技术结合PCR验证的方法,克隆与黄色素含量紧密相关的基因TaPds-A1、TaPds-B1、TaZds-A1、TaLcye-B1和TaLcye-D1,并初步探讨它们的功能;分析其在普通小麦中的等位变异,针对TaPds-B1、TaZds-A1和TaLcye-B1的等位变异开发用于快速鉴别育种材料和品种相应基因的功能标记,为以黄色素改良为目标的小麦分子育种提供材料和选择工具。主要结果如下:1.克隆了普通小麦4A和4B染色体上的TaPds-A1与TaPds-B1基因的全长编码序列,两者都含有14个外显子和13个内含子以及一个1728 bp的开放读码框。针对TaPds-B1位点的2个等位变异TaPds-B1a和TaPds-B1b,开发了相应的显性标记YP4B-1和YP4B-2,并检测了217份中国冬小麦品种,确定了它们在TaPds-B1位点的基因型。2.克隆了普通小麦2A染色体上的TaZds-A1基因的全长编码序列,其与TaPds-A1、TaPds-B1基因结构相似,都含有14个外显子和13个内含子以及一个1707 bp的开放读码框,编码一个包含568个氨基酸残基的蛋白质前体。针对TaZds-A1位点的等位变异TaZds-A1a和TaZds-A1b开发了共显性标记YP2A-1。在中优9507/CA9632的DH群体中检测到一个与TaZds-A1共分离的QTL,可解释11.3%的表型变异。通过对217份中国冬小麦品种的检测,发现开发的功能标记可以在TaZds-A1a基因型中扩增得到183 bp片段,在TaZds-A1b基因型中扩增得到179bp的片段;且TaZds-A1a基因型与低黄色素含量相关,而TaZds-A1b基因型与高黄色素含量相关。3.克隆了普通小麦3B和3D染色体上的LCYE基因TaLcye-B1和TaLcye-D1的全长,两者均含有10个外显子和9个内含子以及一个1611 bp的开放读码框,编码一个含有536个氨基酸残基的蛋白质前体。针对TaLcye-B1位点的等位变异TaLcye-B1a和TaLcye-B1b开发了一个显性标记YP3B-1,确定了217份中国冬小麦品种的基因型。更多还原

【Abstract】 Flour colour is an important quality trait, exerting significant influences on the apparence of wheat-based products, and thus it is of great importance to investigate the genes related to colour for wheat products. In the present study, TaPds-A1, TaPds-B1, TaZds-A1, TaLcye-B1 and TaLcye-D1 were cloned by the method of in silico cloning in combination with PCR amplification, and functional markers were developed for the genes TaPds-B1, TaZds-A1 and TaLcye-B1 according to their allelic varants in different wheat cultivars. The main results are summarized below.1. Full-length genomic DNA sequences of TaPds-A1 and TaPds-B1 on chromosomes 4A and 4B, respectively, were cloned from common wheat. Both have 14 exons, 13 introns and an open reading frame (ORF) of 1,728 bp, encoding 568 amino acid residues. For the two allelic variants TaPds-B1a and TaPds-B1b at TaPds-B1 locus, dominant functional markers YP4B-1 and YP4B-2 were developed, respectively. The two functional markers were used to test the 217 Chinese winter wheats to determine the genotype of TaPds-B1 in the cultivars.2. The full-length DNA sequence of a ZDS gene on wheat chromosome 2A, designated TaZds-A1, was cloned, with 14 exons and 13 introns, and it has an ORF of 1,707 bp, encoding 568 amino acid residues. A co-dominant functional marker, YP2A-1, was designed based on the polymorphisms of two alleles at the locus, TaZds-A1a and TaZds-A1b, yielding 183-bp and 179-bp fragments in TaZds-A1a and TaZds-A1b genotypes, respectively. A new QTL for YP content was detected on chromosome 2A, co-segregating with the functional marker YP2A-1 and TaZds-A1; it explained 11.3% of the phenotypic variance for YP content in a doubled haploid (DH) population from Zhongyou 9507/CA9632. Among 217 Chinese wheat cultivars and advanced lines, the averaged grain YP content of 126 cultivars with TaZds-A1b allele was 7.8% higher than that of 91 cultivars with TaZds-A1a allele.3. Full-length genomic DNA sequences of TaLcye-B1 and TaLcye-D1 on chromosomes 3B and 3D, respectively, were cloned, with 10 exons and 9 introns, and it has an ORF of 1,611 bp, encoding 536 amino acid residues. A dominant functional marker, YP3B-1, was designed based on the polymorphisms of two alleles at TaLcye-B1 locus, TaLcye-B1a and TaLcye-B1b. Using this functional marker, the genotypes of the 217 Chinese winter wheats were identified.更多还原