摘要
粗山羊草(Aegilops tauschii,2n=2x=14,DD)是六倍体普通小麦的祖先之一,其高分子量谷蛋白亚基(HMW-GS)变异类型丰富,是小麦品质改良的重要基因资源。利用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分析了黄河中游地区161份粗山羊草的HMW-GS,发现3种编码序列未知的y-型亚基,即Dy10.5t、Dy10.4t和Dy10.5*t亚基。通过AS-PCR扩增、克隆、测序和氨基酸序列推导,发现3种未知序列均具有典型HMW-GS的序列结构特征且较为相似,仅Dy10.4t与Dy10.5t亚基存在一个氨基酸重复单元的缺失,Dy10.5t与Dy10.5*t亚基在信号肽部位有一个氨基酸的替换(L?F)。通过对这3种HMW-GS与32个已知氨基酸序列的HMW-GS多序列比对和系统进化关系分析,证实Dy10.5t、Dy10.4t和Dy10.5*t亚基是D基因组编码的高分子量谷蛋白y-型亚基家族的新成员。
Aegilops tauschii （2n = 2x = 14, DD）, an ancestral specie of Tiriticum aestivum （2n = 6x = 42, AABBDD）, possesses extensive allelic variation in high molecular weight glutenin subunits （HMW-GS）. The elite genetic recourses are of great value in quality improvement of T. aestivum. Ae. tauschii is distributed in the middle reaches of Yellow River, and its HMW-GS genes have not been investigated in large scales. To detect HMW-GS genes in Ae. tauschii resources in this area and to understand its structure and function by sequencing, 161 Ae. tauschii accessions collected in July 2006 were used in this study with Yumai 49 as a control. Through sodium dodecyl sulphate polyacrylamide gel electrophoresis （SDS-PAGE） technique, three y-type subunits were observed, named as Dy10.5^t, Dy10.4^t , and Dy10.5^＊t. Genes of the three subunits were amplified, cloned and sequenced by allele-specific PCR. The results revealed that the deduced amino acid sequence had typical characteristics of HMW-GS and a general resemblance to each other, but Dy 10.4^t had a repeated amino acid motif deletion to Dy10.5^t, and Dy10.5^＊t had an amino acid substitution sites in signal peptide domain to Dy10.5t （L-F）. From multiple sequence alignment and phylogenetic analysis of storage protein gene family, it is concluded that the subunits Dy10.5^t, Dy10.4^t, and Dy10.5^＊t from Ae. tauschii are similar to the Glu-1 locus found in wheat and related species, and are new members of y-type HWM glutenin family.
出处
《作物学报》
CAS
CSCD
北大核心
2009年第7期1244-1252,共9页
Acta Agronomica Sinica
基金
河南省高校杰出人才创新工程项目(HAIPURT
2007KYCX009)资助
关键词
粗山羊草
高分子量谷蛋白
分子克隆
系统进化
Aegilops tauschii
HWM-GS
Molecular cloning
Phylogenetic relationships
