作物学报
作物學報
작물학보
ACTA AGRONOMICA SINICA
2010年
4期
580-589
,共10页
朱西平%李鑫%李雅轩%晏月明
硃西平%李鑫%李雅軒%晏月明
주서평%리흠%리아헌%안월명
α-醇溶蛋白%普通小麦%粗山羊草%SNP%系统进化
α-醇溶蛋白%普通小麥%粗山羊草%SNP%繫統進化
α-순용단백%보통소맥%조산양초%SNP%계통진화
α-gliadins%Common wheat%Aegilops tauschii%SNP (single nucleotide polymorism)%Phylogenetics and evolution
通过特异PCR引物设计,从普通小麦品种(豫麦34和娴农19)和粗山羊草(T9、T197、T48、T176和T17)中扩增、克隆了7个新的α-醇溶蛋白基因,分别命名为Gli-YM34、Gli-YN19、Gli-T9、Gli-T197、Gli-T48、Gli-T176和Gli-T17,基因序列长度为846～891 bp,编码282～297个氨基酸残基,都具有α-醇溶蛋白的典型结构特点.其中Gli-YM34和Gli-YN19基因推导的醇溶蛋白都含有一个额外的半胱氨酸残基,可能对面筋品质有正向作用.根据α-醇溶蛋白氨基酸序列所具有的4种T细胞抗原表位和多聚谷氨酰胺重复区的平均长度以及中国春缺体四体分析,将来自普通小麦品种的Gli-YM34和Gli-YN19基因定位在6D染色体上的Gli-D2位点,而且Gli-YM34和Gli-YN19与来自粗山羊草的α-醇溶蛋白基因具有很高的序列相似性,进一步证明粗山羊草是普通小麦D基因组的供体.在克隆的4个典型α-醇溶蛋白基因中检测到21个SNP和1个9 bp的缺失.系统进化分析表明,α-醇溶蛋白基因与低分子量谷蛋白亚基基因关系较近,在大约43.69百万年时分化,与ω-醇溶蛋白和HMW-GS基因亲缘关系较远,它们的分化时间大约为79.39百万年.
通過特異PCR引物設計,從普通小麥品種(豫麥34和嫻農19)和粗山羊草(T9、T197、T48、T176和T17)中擴增、剋隆瞭7箇新的α-醇溶蛋白基因,分彆命名為Gli-YM34、Gli-YN19、Gli-T9、Gli-T197、Gli-T48、Gli-T176和Gli-T17,基因序列長度為846～891 bp,編碼282～297箇氨基痠殘基,都具有α-醇溶蛋白的典型結構特點.其中Gli-YM34和Gli-YN19基因推導的醇溶蛋白都含有一箇額外的半胱氨痠殘基,可能對麵觔品質有正嚮作用.根據α-醇溶蛋白氨基痠序列所具有的4種T細胞抗原錶位和多聚穀氨酰胺重複區的平均長度以及中國春缺體四體分析,將來自普通小麥品種的Gli-YM34和Gli-YN19基因定位在6D染色體上的Gli-D2位點,而且Gli-YM34和Gli-YN19與來自粗山羊草的α-醇溶蛋白基因具有很高的序列相似性,進一步證明粗山羊草是普通小麥D基因組的供體.在剋隆的4箇典型α-醇溶蛋白基因中檢測到21箇SNP和1箇9 bp的缺失.繫統進化分析錶明,α-醇溶蛋白基因與低分子量穀蛋白亞基基因關繫較近,在大約43.69百萬年時分化,與ω-醇溶蛋白和HMW-GS基因親緣關繫較遠,它們的分化時間大約為79.39百萬年.
통과특이PCR인물설계,종보통소맥품충(예맥34화한농19)화조산양초(T9、T197、T48、T176화T17)중확증、극륭료7개신적α-순용단백기인,분별명명위Gli-YM34、Gli-YN19、Gli-T9、Gli-T197、Gli-T48、Gli-T176화Gli-T17,기인서렬장도위846～891 bp,편마282～297개안기산잔기,도구유α-순용단백적전형결구특점.기중Gli-YM34화Gli-YN19기인추도적순용단백도함유일개액외적반광안산잔기,가능대면근품질유정향작용.근거α-순용단백안기산서렬소구유적4충T세포항원표위화다취곡안선알중복구적평균장도이급중국춘결체사체분석,장래자보통소맥품충적Gli-YM34화Gli-YN19기인정위재6D염색체상적Gli-D2위점,이차Gli-YM34화Gli-YN19여래자조산양초적α-순용단백기인구유흔고적서렬상사성,진일보증명조산양초시보통소맥D기인조적공체.재극륭적4개전형α-순용단백기인중검측도21개SNP화1개9 bp적결실.계통진화분석표명,α-순용단백기인여저분자량곡단백아기기인관계교근,재대약43.69백만년시분화,여ω-순용단백화HMW-GS기인친연관계교원,타문적분화시간대약위79.39백만년.
Seven novel a-gliadin genes from common wheat cultivars (Yumai 34 and Yannong 19) and Aegilops tauschii accessions (T9, T197, T48, T176, and T17) were amplified and cloned by using a PCR-hased strategy. They were designated as Gli-YM34, Gli-YN19, Gli-T9, Gli-T197, Gli-T48, Gli-T176, and Gli-T17, respectively. Their length of the open reading frame (ORF) ranged from 846 to 891 bp, encoding the putative proteins of 282-297 amino acid residues. Comparative analysis showed that all genes isolated had typical structural characters of a-gliadin genes reported previously. Particularly, Gli-YM34 and GIi-YN19 α-gliadins from common wheat possessed an additional cysteine residue, suggesting a possible positive effect on dough quality. Both genes were assigned to Gli-D2 locus on the chromosome 6D by the analysis of four celiac disease toxic epitopes and glutamine residues in the polyglutamine domain as well as nullisomic-tetrasomic lines of Chinese Spring. A total of twenty-one single nucleotide polymorphisms (SNPs) and a 9 bp deletion among the four typical a-gliadin genes were identified. Phylogenetic and evolutionary analysis revealed that the a-gliadin genes are closely related to LMW-GS genes and their divergence occurred 43.69 million year ago. Less homology was found between α-gliadin genes and HMW-GS and ω-gliadin genes, and they diverged about 79.39 million year.
