华北农学报 ›› 2025, Vol. 40 ›› Issue (3): 1-8. doi: 10.7668/hbnxb.20195519

所属专题: 小麦 生物技术

• 作物遗传育种·种质资源·生物技术 •    下一篇

小麦TaSOD-B1基因克隆及KASP标记开发与验证

侯培珂, 程宇坤, 王继庆, 孙玲, 王建鹏, 耿洪伟   

  1. 新疆农业大学 农学院,新疆农业大学优质专用麦类作物工程技术研究中心,新疆 乌鲁木齐 830052
  • 收稿日期:2024-10-23 出版日期:2025-07-04
  • 通讯作者:
    耿洪伟(1978—),男,重庆人,教授,博士,主要从事小麦遗传育种研究。
  • 作者简介:

    侯培珂(1999—),女,新疆库尔勒人,硕士,主要从事小麦遗传育种研究。

    侯培珂、程宇坤为同等贡献作者。

  • 基金资助:
    国家自然科学基金杰出青年科学基金项目(2022D01E46); 新疆小麦产业技术体系(XJARS-01)

Cloning of Wheat TaSOD-B1 Gene and Development and Verification of KASP Marker

HOU Peike, CHENG Yukun, WANG Jiqing, SUN Ling, WANG Jianpeng, GENG Hongwei   

  1. College of Agronomy,Xinjiang Agricultural University,Special High Quality Triticeae Crops Engineering and Technology Research Center,Xinjiang Agricultural University,Urumqi 830052,China
  • Received:2024-10-23 Published:2025-07-04

摘要:

克隆小麦籽粒超氧化物歧化酶(SOD)基因,开发与SOD活性相关的竞争性等位基因特异性PCR(KASP)标记,对选育高SOD活性小麦品种具有重要意义。根据基因ID,设计特异性引物克隆得到TaSOD-B1基因gDNA序列;在小麦基因组遗传变异及Ensembl Plants数据库比对得到单核苷酸多态性(SNP)位点,开发与小麦SOD活性密切相关的KASP标记,并通过287份来自国内外冬小麦品种(系)SOD活性与基因型间的相关性分析进行标记的实用性验证。利用6对特异性引物扩增中国春品种TaSOD-B1基因片段,拼接得到5B染色体上的TaSOD-B1基因,基因全长为6 491 bp,包含1 650 bp的开放阅读框(ORF),共编码549个氨基酸,预测分子质量为60.80 ku,由12个外显子及11个内含子构成,内含子符合典型的GT-AG结构。基于TaSOD-B1基因第1外显子的第44碱基处经测序验证确实存在的差异位点开发KASP标记,检测结果表明,等位变异类型为TaSOD-B1a的基因型为AA,与高SOD活性相关,用荧光基因FAM(显示为蓝色)标记;等位变异类型为TaSOD-B1b的基因型为GG,与低SOD活性相关,用荧光基因HEX标记(显示为红色)。对287份国内外冬小麦品种(系)进行检测发现,不同基因型的SOD活性差异达到显著水平。基于TaSOD-B1序列成功开发出1组与SOD活性相关,并可用于SOD活性遗传改良的KASP标记。

关键词: 普通小麦, 超氧化物歧化酶, 基因克隆, KASP标记

Abstract:

Cloning the wheat grain superoxide dismutase(SOD)gene and developing competitive allele-specific PCR(KASP)markers related to SOD activity are of great significance for breeding wheat varieties with high SOD activity.According to the gene ID,specific primers were designed to clone the gDNA sequence of TaSOD-B1 gene.The single nucleotide polymorphism(SNP)loci were obtained by comparing the wheat genome genetic variation and Ensembl Plants database,and the KASP markers closely related to the SOD activity of wheat were developed.The practicability of the markers was verified by the correlation analysis between SOD activity and genotypes of 287 winter wheat varieties(lines).The TaSOD-B1 gene fragment of Chinese spring variety was amplified by six pairs of specific primers,and the TaSOD-B1 gene on chromosome 5B was obtained by splicing.The full length of the gene was 6 491 bp,including an open reading frame(ORF)of 1 650 bp,which encoded a total of 549 amino acids.The predicted molecular weight was 60.80 ku,the gene was composed of 12 exons and 11 introns.The intron conformed to the typical GT-AG structure.The KASP marker was developed based on the 44th base of the first exon of the TaSOD-B1 gene,and was verified by sequencing.The results showed that the genotype of the allelic variation type TaSOD-B1a was AA,which was associated with high SOD activity,and was labeled with the fluorescent gene FAM(shown as blue).The genotype of TaSOD-B1b was GG,which was associated with low SOD activity and was marked with the fluorescent gene HEX(shown as red).The detection of 287 winter wheat varieties(lines)at home and abroad showed that the SOD activity of different genotypes was significantly different.Based on the TaSOD-B1 sequence,a set of KASP markers related to SOD activity was successfully developed and could be used for genetic improvement of SOD activity.

Key words: Common wheat, Superoxide dismutase, Gene cloning, KASP marker

中图分类号: 

引用本文

侯培珂, 程宇坤, 王继庆, 孙玲, 王建鹏, 耿洪伟. 小麦TaSOD-B1基因克隆及KASP标记开发与验证[J]. 华北农学报, 2025, 40(3): 1-8. doi: 10.7668/hbnxb.20195519.

HOU Peike, CHENG Yukun, WANG Jiqing, SUN Ling, WANG Jianpeng, GENG Hongwei. Cloning of Wheat TaSOD-B1 Gene and Development and Verification of KASP Marker[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(3): 1-8. doi: 10.7668/hbnxb.20195519.