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大豆普遍存在的脲酶编码基因座的等位基因间互补作用

Interallelic complementation at the ubiquitous urease coding locus of soybean.

作者信息

Goldraij Ariel, Beamer Lesa J, Polacco Joe C

机构信息

Department of Biochemistry, University of Missouri, Columbia, Missouri 65211, USA.

出版信息

Plant Physiol. 2003 Aug;132(4):1801-10. doi: 10.1104/pp.103.022699.

Abstract

Soybean (Glycine max [L.] Merrill) mutant aj6 carries a single recessive lesion, aj6, that eliminates ubiquitous urease activity in leaves and callus while retaining normal embryo-specific urease activity. Consistently, aj6/aj6 plants accumulated urea in leaves. In crosses of aj6/aj6 by urease mutants at the Eu1, Eu2, and Eu3 loci, F(1) individuals exhibited wild-type leaf urease activity, and the F(2) segregated urease-negative individuals, demonstrating that aj6 is not an allele at these loci. F(2) of aj6/aj6 crossed with a null mutant lacking the Eu1-encoded embryo-specific urease showed that ubiquitous urease was also inactive in seeds of aj6/aj6. The cross of aj6/aj6 to eu4/eu4, a mutant previously assigned to the ubiquitous urease structural gene (R.S. Torisky, J.D. Griffin, R.L. Yenofsky, J.C. Polacco [1994] Mol Gen Genet 242: 404-414), yielded an F(1) having 22% +/- 11% of wild-type leaf urease activity. Coding sequences for ubiquitous urease were cloned by reverse transcriptase-polymerase chain reaction from wild-type, aj6/aj6, and eu4/eu4 leaf RNA. The ubiquitous urease had an 837-amino acid open reading frame (ORF), 87% identical to the embryo-specific urease. The aj6/aj6 ORF showed an R201C change that cosegregated with the lack of leaf urease activity in a cross against a urease-positive line, whereas the eu4/eu4 ORF showed a G468E change. Heteroallelic interaction in F(2) progeny of aj6/aj6 x eu4/eu4 resulted in partially restored leaf urease activity. These results confirm that aj6/aj6 and eu4/eu4 are mutants affected in the ubiquitous urease structural gene. They also indicate that radical amino acid changes in distinct domains can be partially compensated in the urease heterotrimer.

摘要

大豆(Glycine max [L.] Merrill)突变体aj6携带一个隐性单基因损伤,即aj6,它消除了叶片和愈伤组织中普遍存在的脲酶活性,同时保留了正常的胚胎特异性脲酶活性。相应地,aj6/aj6植株的叶片中积累了尿素。在aj6/aj6与Eu1、Eu2和Eu3位点的脲酶突变体杂交时,F(1)个体表现出野生型叶片脲酶活性,F(2)出现脲酶阴性个体,表明aj6不是这些位点的等位基因。aj6/aj6与缺乏Eu1编码的胚胎特异性脲酶的无效突变体杂交的F(2)表明,aj6/aj6种子中的普遍存在的脲酶也无活性。aj6/aj6与eu4/eu4(一个先前被指定为普遍存在的脲酶结构基因的突变体,R.S. Torisky、J.D. Griffin、R.L. Yenofsky、J.C. Polacco [1994] Mol Gen Genet 242: 404 - 414)杂交,产生的F(1)具有野生型叶片脲酶活性的22%±11%。通过逆转录聚合酶链反应从野生型、aj6/aj6和eu4/eu4叶片RNA中克隆了普遍存在的脲酶的编码序列。普遍存在的脲酶有一个837个氨基酸的开放阅读框(ORF),与胚胎特异性脲酶有87%的同源性。aj6/aj6的ORF显示R201C变化,在与脲酶阳性品系杂交时,该变化与叶片脲酶活性的缺失共分离,而eu4/eu4的ORF显示G468E变化。aj6/aj6×eu4/eu4的F(2)后代中的异等位基因相互作用导致叶片脲酶活性部分恢复。这些结果证实aj6/aj6和eu4/eu4是普遍存在的脲酶结构基因受影响的突变体。它们还表明,在脲酶异源三聚体中,不同结构域的氨基酸激进变化可以得到部分补偿。

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