Molecular characterization, haplotype analysis and development of markers specific to dzs18 gene regulating methionine accumulation in kernels of subtropical maize.

IF 2.6 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY 3 Biotech Pub Date : 2024-10-01 Epub Date: 2024-09-21 DOI:10.1007/s13205-024-04088-2
Hriipulou Duo, Rashmi Chhabra, Vignesh Muthusamy, Subhra J Mishra, Ikkurti Gopinath, Gaurav Sharma, Jayanthi Madhavan, Chirravuri N Neeraja, Rajkumar U Zunjare, Firoz Hossain
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Abstract

Maize kernel protein is deficient in sulfur-containing essential amino acid such as methionine. The dzs18 gene encodes methionine-rich 18-kDa δ-zein in maize kernels. In this study, we sequenced full-length of dzs18 gene (820 bp) among 10 maize inbreds, revealing 43 SNPs and 22 InDels (average length-7.58 bp). Three InDels (4 bp at 113th, 15 bp at 463rd and 3 bp at 615th position) distinguished the wild-type (functional) from the mutant (non-functional) allele of dzs18. The 4 bp (TTAT) insertion caused a frameshift mutation, resulting in truncated DZS18 protein. The 15 bp insertion (ATG-TCT-TCG-ATG-ATA) added methionine-serine-serine-methionine-isoleucine, while the 3 bp deletion (CAA) led to loss of a glutamine residue in the mutant allele. Three gene-based PCR markers were developed for diversity analysis of dzs18 gene among 48 inbreds, which had an average methionine content of 0.136 %. (range: 0.031-0.340 %). Eight haplotypes were identified with methionine content varying from 0.066 % (Hap7) to 0.262 % (Hap3). Haplotypes with 4 bp deletion accumulated more methionine (0.174 %) than haplotypes with 4 bp insertion (0.082 %). The average methionine in 15 bp deletion and insertion haplotypes was 0.106 % and 0.150 %, respectively. The 3 bp insertion had 0.140 % methionine, while the deletion possessed 0.117 % methionine. Protein-protein association analysis predicted that DZS18 protein interacts with 19-kDa α-zein, 27- and 16-kDa γ-zeins, WAXY and O2 protein. A paralogue of dzs18 gene with 74 % sequence identity was identified. The functional markers reported here could facilitate the development of high methionine maize cultivars, which holds great significance to combat malnutrition, especially in developing countries.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04088-2.

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亚热带玉米籽粒中调节蛋氨酸积累的 dzs18 基因的分子特征、单体型分析和特异性标记的开发。
玉米籽粒蛋白缺乏含硫必需氨基酸,如蛋氨酸。dzs18 基因编码玉米籽粒中富含蛋氨酸的 18 kDa δ-玉米蛋白。本研究对 10 个玉米近交系的 dzs18 基因全长(820 bp)进行了测序,发现了 43 个 SNPs 和 22 个 InDels(平均长度为 7.58 bp)。三个 InDels(第 113 位 4 bp、第 463 位 15 bp 和第 615 位 3 bp)区分了 dzs18 的野生型(功能性)和突变型(非功能性)等位基因。4 bp (TTAT)插入导致了框架移位突变,从而产生了截短的 DZS18 蛋白质。15 bp 的插入(ATG-TCT-TCG-ATG-ATA)增加了蛋氨酸-丝氨酸-丝氨酸-蛋氨酸-异亮氨酸,而 3 bp 的缺失(CAA)导致突变等位基因中谷氨酰胺残基的缺失。为分析 48 个近交系中 dzs18 基因的多样性,开发了三种基于基因的 PCR 标记,其蛋氨酸平均含量为 0.136%(范围:0.031-0.340%)。鉴定出 8 个单倍型,蛋氨酸含量从 0.066 %(Hap7)到 0.262 %(Hap3)不等。缺失 4 bp 的单倍型比插入 4 bp 的单倍型(0.082 %)积累了更多的蛋氨酸(0.174 %)。15 bp缺失单倍型和插入单倍型的平均蛋氨酸含量分别为 0.106 % 和 0.150 %。3 bp 插入型的蛋氨酸含量为 0.140%,而缺失型的蛋氨酸含量为 0.117%。蛋白质-蛋白质关联分析表明,DZS18 蛋白与 19 kDa α-玉米素、27 kDa 和 16 kDa γ-玉米素、WAXY 和 O2 蛋白相互作用。发现了一个具有 74 % 序列同一性的 dzs18 基因旁系。本文报告的功能标记可促进高蛋氨酸玉米品种的发展,这对消除营养不良,尤其是发展中国家的营养不良具有重要意义:在线版本包含补充材料,可查阅 10.1007/s13205-024-04088-2。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
3 Biotech
3 Biotech Agricultural and Biological Sciences-Agricultural and Biological Sciences (miscellaneous)
CiteScore
6.00
自引率
0.00%
发文量
314
期刊介绍: 3 Biotech publishes the results of the latest research related to the study and application of biotechnology to: - Medicine and Biomedical Sciences - Agriculture - The Environment The focus on these three technology sectors recognizes that complete Biotechnology applications often require a combination of techniques. 3 Biotech not only presents the latest developments in biotechnology but also addresses the problems and benefits of integrating a variety of techniques for a particular application. 3 Biotech will appeal to scientists and engineers in both academia and industry focused on the safe and efficient application of Biotechnology to Medicine, Agriculture and the Environment.
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