一种鉴别栽培柿品种SNP标记的新方法  

A Novel SNP Marker for the Identification of Persimmon(Diospyros kaki)Cultivars

作　　者：梁晋军 朱溯远 张宇琴 张鹏飞[1] 温鹏飞[1] 杨运良 LIANG Jin-jun;ZHU Su-yuan;ZHANG Yu-qin;ZHANG Peng-fei;WEN Peng-fei;YANG Yun-iang(College of Horticulture,Shanxi Agricultural University,Taigu 030801;Institute of Cotton Research,Shanxi Agricultural University,Yuncheng 044000)

机构地区：[1]山西农业大学园艺学院,太谷030801 [2]山西农业大学棉花研究所,运城044000

出　　处：《生物技术通报》2024年第1期160-167,共8页Biotechnology Bulletin

基　　金：山西农业大学博士科研启动项目(2021BQ32);山西农业大学生物育种工程项目(YZGC113);山西省青年科学研究项目(20210302124067);运城市科学技术局现代农业项目(YCKJ-2021021,YCKJ-2022048);运城市科学技术局基础项目(YCKJ-2021043)。

摘　　要：【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nrDNAITS序列SNP分子标记的新方法能将18份栽培柿品种分成11类。【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.

关 键 词：栽培柿 nrDNAITS SNP 杂合位点 遗传差异 

分 类 号：S665.2[农业科学—果树学]