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与玉米(Zea mays L.)低植酸籽粒相关的肌醇(3)磷酸1合酶基因的表达及核苷酸序列

Expression and nucleotide sequence of an INS (3) P1 synthase gene associated with low-phytate kernels in maize (Zea mays L.).

作者信息

Shukla Soni, VanToai Tara T, Pratt Richard C

机构信息

Department of Horticulture and Crop Science, The Ohio State University Columbus, OH 43210, USA.

出版信息

J Agric Food Chem. 2004 Jul 14;52(14):4565-70. doi: 10.1021/jf049976b.

Abstract

Most of the phosphorus (P) in maize (Zea mays L.) kernels is in the form of phytic acid. A low phytic acid (lpa) maize mutant, lpa1-1, displays levels reduced by 66%. A goal of genetic breeding is development of low phytic acid feedstocks to improve P nutrition of nonruminant animals and reduce the adverse environmental impacts of excess P in manure. The genetic basis of the lpa1-1 mutation is not known, but previous genetic mapping has shown both the mutant phenotype and the Ins (3) P(1) synthase (MIPS) gene, which encodes the first enzyme, myo-inositol phosphate synthase, in the phytic acid biosynthetic pathway, map to the same chromosomal region in maize. Research was conducted to determine the expression of the MIPS gene in lpa1-1 and wild-type kernels with similar genetic backgrounds and to ascertain if variation in the MIPS coding sequence could be inferred to be the basis of the mutation. MIPS enzyme activity determined by TLC was reduced 2-3-fold in mutant kernels. RT-PCR-based experiments using primers specific for the 1S-MIPS sequence indicated gene expression was reduced 50-60% in the mutant. Sequence analysis of the MIPS genomic sequence revealed 10 exons and 9 introns that are identical in both mutant and wild-type developing kernels. These findings support an association between reduced MIPS gene activity and low phytic acid content, but additional research should examine the promoter, the 5'UTR, or transcriptional controlling elements of the MIPS gene to ascertain the possible presence of a genetic lesion in those regions.

摘要

玉米(Zea mays L.)籽粒中的大部分磷(P)以植酸的形式存在。一种低植酸(lpa)玉米突变体lpa1-1,其植酸水平降低了66%。遗传育种的一个目标是培育低植酸原料,以改善非反刍动物的磷营养,并减少粪便中过量磷对环境的不利影响。lpa1-1突变的遗传基础尚不清楚,但先前的遗传图谱显示,突变体表型和肌醇-1-磷酸合酶(MIPS)基因(该基因编码植酸生物合成途径中的第一种酶——肌醇磷酸合酶)都定位于玉米的同一染色体区域。开展研究以确定MIPS基因在具有相似遗传背景的lpa1-1和野生型籽粒中的表达情况,并确定是否可以推断MIPS编码序列的变异是突变的基础所在。通过薄层层析法测定的MIPS酶活性在突变体籽粒中降低了2至3倍。使用针对1S-MIPS序列的特异性引物进行的基于逆转录聚合酶链反应(RT-PCR)的实验表明,该基因在突变体中的表达降低了50%至60%。对MIPS基因组序列的分析揭示了10个外显子和9个内含子,它们在突变体和野生型发育籽粒中是相同的。这些发现支持了MIPS基因活性降低与低植酸含量之间的关联,但进一步的研究应检查MIPS基因的启动子、5'非翻译区(5'UTR)或转录控制元件,以确定这些区域是否可能存在遗传损伤。

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