首页|一种鉴别栽培柿品种SNP标记的新方法

一种鉴别栽培柿品种SNP标记的新方法

扫码查看
[目的]为了鉴别栽培柿品种间的遗传差异,开发一种基于核糖体DNA内转录间隔区(nrDNA internally transcribed spacer)序列简单、高效的SNP分子标记新方法,为种质的收集、利用及推广应用提供参考.[方法]以 18 种六倍体栽培柿品种叶片为试材,对其ITS区进行扩增测序,并分析序列差异,最后用Sau96 I限制性内切酶对 151 处杂合位点进行酶切验证.[结果]18 份柿品种ITS长度均为 730 bp,共存在 6 个杂合位点,分别在 151、168、205、278、279 和 622 bp处.六倍体柿杂合位点处碱基峰图面积比例存在一定的规律,即C∶T=2∶1、C∶T=1∶1、C∶T=1∶2 和A∶G=1∶1,利用出现的杂合位点及其峰图面积比例规律差异将 18 份栽培柿品种分成 11 类.[结论]151 处位点的酶切结果验证了酶切产物浓度与峰图面积比例一致.这种基于nrDNA ITS序列SNP分子标记的新方法能将 18 份栽培柿品种分成 11 类.
A Novel SNP Marker for the Identification of Persimmon(Diospyros kaki)Cultivars
[Objective]In order to identify the genetic differences among persimmon(Diospyros kaki)cultivars,a simple and efficient SNP molecular marker based on nrDNA internally transcribed spacer sequence was developed,which may provide reference for the collection,utilization and promotion of germplasm.[Method]The leaves of 18 hexaploid cultivated persimmon cultivars were used as experimental materials,and the ITS regions of 18 cultivars were amplified and sequenced,and sequence differences were analyzed.Finally,Sau96 I restriction endonuclides were used to verify 151 heterozygous sites.[Result]Sequencing results showed that ITS length of 18 persimmon varieties was 730 bp.Six heterozygous loci were found in the sequencing peak map,which were 151,168,205,278,279 and 622 bp.The proportion of base peak area of hexaploid persimmon heterozygous sites had special pattern,that is,C∶T= 2∶1,C∶T= 1∶1,C∶T= 1∶2 and A∶G= 1∶1.The 18 cultivated persimmon varieties could be divided into 11 categories according to the differences of heterozygous sites and the proportion of peak area.[Conclusion]The results of enzyme digestion at 151 sites verified that the concentration of enzyme digestion products was consistent with the proportion of its peak map area.The results showed that the SNP molecular labeling method based on nrDNA ITS sequence could divide 18 cultivated persimmon varieties into 11 groups.

cultivated persimmonnrDNA ITSSNPheterozygous sitegenetic difference

梁晋军、朱溯远、张宇琴、张鹏飞、温鹏飞、杨运良

展开 >

山西农业大学园艺学院,太谷 030801

山西农业大学棉花研究所,运城 044000

栽培柿 nrDNA ITS SNP 杂合位点 遗传差异

山西农业大学博士科研启动项目山西农业大学生物育种工程项目山西省青年科学研究项目

2021BQ32YZGC113

2024

生物技术通报
中国农业科学院农业信息研究所

生物技术通报

CSTPCD北大核心
影响因子:0.505
ISSN:1002-5464
年,卷(期):2024.40(1)
  • 16