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分子特征分析表明,玉米基因sugary2编码淀粉合酶同工型SSIIa。

Molecular characterization demonstrates that the Zea mays gene sugary2 codes for the starch synthase isoform SSIIa.

作者信息

Zhang Xiaoli, Colleoni Christophe, Ratushna Vlada, Sirghie-Colleoni Mirella, James Martha G, Myers Alan M

机构信息

Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, 1210 Molecular Biology Building, IA, USA.

出版信息

Plant Mol Biol. 2004 Apr;54(6):865-79. doi: 10.1007/s11103-004-0312-1.

Abstract

Mutations in the maize gene sugary2 ( su2 ) affect starch structure and its resultant physiochemical properties in useful ways, although the gene has not been characterized previously at the molecular level. This study tested the hypothesis that su2 codes for starch synthase IIa (SSIIa). Two independent mutations of the su2 locus, su2-2279 and su2-5178 , were identified in a Mutator -active maize population. The nucleotide sequence of the genomic locus that codes for SSIIa was compared between wild type plants and those homozygous for either novel mutation. Plants bearing su2-2279 invariably contained a Mutator transposon in exon 3 of the SSIIa gene, and su2-5178 mutants always contained a small retrotransposon-like insertion in exon 10. Six allelic su2 (-) mutations conditioned loss or reduction in abundance of the SSIIa protein detected by immunoblot. These data indicate that su2 codes for SSIIa and that deficiency in this isoform is ultimately responsible for the altered physiochemical properties of su2 (-) mutant starches. A specific starch synthase isoform among several identified in soluble endosperm extracts was absent in su2-2279 or su2-5178 mutants, indicating that SSIIa is active in the soluble phase during kernel development. The immediate structural effect of the su2 (-) mutations was shown to be increased abundance of short glucan chains in amylopectin and a proportional decrease in intermediate length chains, similar to the effects of SSII deficiency in other species.

摘要

玉米基因sugary2(su2)的突变以有益的方式影响淀粉结构及其产生的物理化学性质,尽管该基因此前尚未在分子水平上得到表征。本研究检验了su2编码淀粉合酶IIa(SSIIa)的假设。在一个Mutator活性玉米群体中鉴定出了su2位点的两个独立突变,即su2 - 2279和su2 - 5178。比较了野生型植株与任一新型突变纯合植株中编码SSIIa的基因组位点的核苷酸序列。携带su2 - 2279的植株在SSIIa基因的外显子3中总是含有一个Mutator转座子,而su2 - 5178突变体在外显子10中总是含有一个类似小反转录转座子的插入。通过免疫印迹检测到六个等位的su2(-)突变导致SSIIa蛋白丰度丧失或降低。这些数据表明su2编码SSIIa,并且这种同工型的缺乏最终导致了su2(-)突变体淀粉物理化学性质的改变。在su2 - 2279或su2 - 5178突变体中,可溶性胚乳提取物中鉴定出的几种特定淀粉合酶同工型之一不存在,这表明SSIIa在籽粒发育过程中在可溶性阶段具有活性。su2(-)突变的直接结构效应表现为支链淀粉中短葡聚糖链的丰度增加,中间长度链成比例减少,这与其他物种中SSII缺乏的效应相似。

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