基于TILLING技术筛选大豆GmAGL15基因突变体  

Screening AGL15 Mutant in Soybean by TILLING

作　　者：孙彦波 李忠峰 王俊[1] 邱丽娟 SUN Yan-bo;LI Zhong-feng;WANG Jun;QIU Li-juan(College of Agriculture,Yangtze University,Jingzhou 434025,China;Key Laboratory of Crop Germplasm Utilization,Ministry of Agriculture,Institute of Crop Sciences,Chinese Academy of Agricultural Science/National Key Facility for Gene Resources and Genetic Improvement,Beijing 100081,China;College of Agriculture,Henan Agricultural University,Zhengzhou 450002,China)

机构地区：[1]长江大学农学院,湖北荆州434025 [2]中国农业科学院作物科学研究所/国家农作物基因资源与遗传改良重大科学工程/农业部种质资源利用重点实验室,北京100081 [3]河南农业大学农学院,河南郑州450002

出　　处：《大豆科学》2019年第6期906-913,共8页Soybean Science

基　　金：大豆种质资源保护与利用(2019NWB036);国家农作物资源共享平台(NICGR2019)

摘　　要：AGL15是MADS(MCM1-AGAMOUS-DEFICIENS-SRF)基因家族中的一员,MADS蛋白通过与下游基因启动子的顺式作用元件特异性结合调控植物种子中贮藏物质的积累。为验证大豆中AGL15基因的功能,利用TILLING技术在EMS诱变的中品661的M5中筛选出8个GmAGL15突变体。共检测到10个突变位点,其中6个突变位点发生在基因的编码区导致氨基酸发生改变,4个突变位点发生在基因的内含子。1个SNP导致该基因第2个外显子的第54个碱基由C变为T,并使该基因编码的第79位氨基酸由丙氨酸突变为缬氨酸;5个SNP发生在该基因的第6个外显子的112位碱基上,使得碱基由T变为A,导致该基因编码的第568位氨基酸由亮氨酸突变为甲硫氨酸。大豆GmAGL15的突变分布频率为1/805 kb。考种发现5个非同义突变体和1个内含子突变体的蛋白质或脂肪含量与野生型ZP661相比差异显著。本研究发现的等位变异有助于阐明GmAGL15在大豆种子中的作用机制,并为大豆的遗传改良提供宝贵的种质资源。AGL15 is a member of the MADS(MCM1-AGAMOUS-DEFICIENS-SRF)gene family.MADS proteins regulate the accumulation of storage substances in plant seeds by binding specifically to cis-acting elements of downstream gene promoters.To verify the function of AGL15 gene in soybean,eight GmAGL15 mutants were screened from M5 of ZP661 mutant induced by EMS with TILLING.Ten mutation sites were detected,six of which occurred in the coding region of the gene,resulting in changes in amino acids,and four occurred in the intron of the gene.One SNP changed from C to T at the 54 th base of the second exon,and mutated the 79 th amino acid from alanine to valine.Five SNPs occurred on the 112 base of the sixth exon,which changed the base from T to A,resulting in the mutation of the 568 th amino acid from leucine to methionine.The mutation frequency of soybean GmAGL15 was 1/805 kb.The protein or fat contents of five non-synonymous mutants and one intron mutant were significantly different from those of wild type ZP661.Allelic variation found in this study is helpful to elucidate the mechanism of GmAGL15 in soybean seeds and provide valuable germplasm resources for soybean genetic improvement.

关 键 词：大豆 转录因子 AGL15 EMS TILLING 

分 类 号：S51[农业科学—作物学]