基于SNP标记的4个品种复烤烟叶鉴别研究  

Discrimination of Redried Leaves of Four Tobacco Varieties Based on SNP Markers

作　　者：杨金初 童治军[2] 李萌 赵旭 徐永明 冯颖杰 杨宗灿 曲鹏 李悦 孙九喆 张轲 YANG Jinchu;TONG Zhijun;LI Meng;ZHAO Xu;XU Yongming;FENG Yingjie;YANG Zongcan;QU Peng;LI Yue;SUN Jiuzhe;ZHANG Ke(Technology Center,China Tobacco Henan Industrial Co.,Ltd.,Zhengzhou 450000,China;Yunnan Academy of Tobacco Agricultural Sciences/Key Laboratory of Tobacco Biotechnological Breeding/National Tobacco Genetic Engineering Research Center,Kunming 650021,China;College of Food and Bioengineering,Zhengzhou University of Light Industry,Zhengzhou 450001,China;Yunnan Tobacco Quality Inspection&Supervision Station,Kunming 650106,China)

机构地区：[1]河南中烟工业有限责任公司技术中心,河南郑州450000 [2]云南省烟草农业科学研究院/烟草行业烟草生物技术育种重点实验室/国家烟草基因工程研究中心,云南昆明650021 [3]郑州轻工业大学食品与生物工程学院,河南郑州450001 [4]云南省烟草质量监督检测站,云南昆明650106

出　　处：《河南农业科学》2023年第11期174-180,共7页Journal of Henan Agricultural Sciences

基　　金：中国烟草总公司重点研发计划项目(110202202038);中国烟草总公司云南省公司科技项目(2020530000241034,2023530000241027)。

摘　　要：为探究单核苷酸多态性(SNP)标记在复烤烟叶品种鉴别上的可行性,开发基于SNP标记的复烤烟叶高效鉴别技术,对4个主栽烟草品种云烟87、红花大金元、K326及NC102的复烤烟叶进行了全基因组测序,应用全基因组数据发掘SNP位点,依据SNP位点设计了PCR引物,并对不同烟草品种进行了基因分型和区分。结果显示,4个品种复烤烟叶全基因组测序共获得21.5 Gb数据,共检测出27137个SNP位点。挑选其中53个SNP位点设计不同错配类型等位基因引物570条,扩增结果均与预期一致。SNP位点35、45和51在品种间具有多态性,红花大金元在35、45、51位点基因型为T/T、C/C、T/T;K326为C/T、C/T、C/C;NC102为C/T、C/T、C/T;云烟87烟叶在35位点无扩增,在45、51位点的基因型为C/T、C/T。依据3个SNP位点的基因型构建不同品种的分子ID,可准确区分4个品种的复烤烟叶。To efficiently discriminate varieties of redried tobacco leaves,single nucleotide polymorphism(SNP)molecular marker was explored and used to construct DNA fingerprinting.The whole genome sequencing technology was used to discover the SNP sites of four varieties of redried tobacco leaves including Yunyan 87,Honghuadajinyuan,K326 and NC102,and the allele specific primers for PCR were designed and used for allele typing.The results showed that 21.5 Gb data were obtained from the whole genome sequencing of four tobacco leaf samples,and a total of 27137 SNP sites were detected.Based on 53 SNP sites,570 allele specific primers of different mismatch types were designed,and multiple polymorphic sites were successfully screened to achieve allelic typing of four varieties.SNP loci 35,45 and 51 were found to be polymorphic among different varieties.For loci 35,45 and 51,SNP genotypes were T/T,C/C and T/T for Honghuadajinyuan,C/T,C/T and C/C for K326,and C/T,C/T and C/T for NC 102,respectively.For Yunyan 87,there was no amplification in loci 35,and the SNP genotypes in loci 45 and 51 were C/T and C/T.The molecular ID of each tobacco variety was constructed based on the genotype of three SNP loci,and the four varieties of redried tobacco leaves could be accurately discriminated.

关 键 词：复烤烟叶 品种 鉴别 全基因组测序 SNP位点 等位基因 多态性 

分 类 号：S57[农业科学—作物学] TS45[农业科学—烟草工业]