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八倍体草莓DNA指纹图谱的构建与初步遗传分析
刘国霞1,张全芳1,杨雪1,谭晴晴1,胡悦1,范阳阳1,姚川1,崔刚2,步迅1,陈雪燕1*,刘炳福3*
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(1.山东省农业科学院 农作物种质资源研究所, 济南 250100;2.山东普朗特农业技术有限公司, 济南 250100;3.山东省农业科学院 休闲农业研究所, 济南 250100)
摘要:
为对草莓品种进行分子鉴定,从59对SSR引物中筛选出18对多态性好的引物,采用毛细管电泳检测方法对84份八倍体草莓品种进行标记分析,检测每个标记等位变异大小,并进行初步遗传分析。将每个引物对84份草莓扩增的带型按照一定原则进行数字标注,为了简化编码,引物P1~P18分别编号为字母A~R,将引物编号与相应的带型编码进行组合得到该样品在特定引物的扩增产物编码,按照18对引物顺序串联这些编码形成该品种的DNA指纹图谱。结果表明:18对SSR引物共扩增得到多态性条带150个,不同引物扩增到多态性条带数4~17个,共检测到377个带型;各引物的多态性信息含量(PIC)值为0.494~0.908,平均值为 0.738;利用18对SSR引物共构建了84个草莓品种的DNA指纹图谱。聚类分析结果表明:在遗传相似系数为0.99时能将所有品种区分开,一些来源相同的品种被优先聚在一起。综上,本研究成功构建84个草莓品种的DNA指纹图谱,这些品种遗传关系较近。
关键词:  草莓  SSR  DNA 指纹图谱  遗传分析
DOI:10.11841/j.issn.1007-4333.2023.02.04
投稿时间:2022-04-15
基金项目:山东省农业科学院揭榜科技难题项目(SHJB2021-47)
Construction of DNA fingerprinting of octoploid strawberry and preliminary genetic analysis
LIU Guoxia1,ZHANG Quanfang1,YANG Xue1,TAN Qingqing1,HU Yue1,FAN Yangyang1,YAO Chuan1,CUI Gang2,BU Xun1,CHEN Xueyan1*,LIU Bingfu3*
(1.Institute of Crop Germplasm Resources, Shandong Academy of Agricultural Sciences, Jinan 250100, China;2.Shandong Plant Agricultural Technology Co., Ltd., Jinan 250100, China;3.Institute of Leisure Agriculture, Shandong Academy of Agricultural Sciences, Jinan 250100, China)
Abstract:
In order to screen strawberry cultivars at molecular level, 18 SSR primer pairs with high polymorphism were identified from 59 primer pairs, and 84 octoploid cultivated strawberry samples were screened. By using capillary electrophoresis method, the size of different alleles were detected, and the molecular fingerprinting and genetic analysis for all samples were established. Different alleles of every prime were digitally labelled on 84 strawberry variety according to certain principles. For simplicity, the primers P1 to P18 were labelled as A to R, then the code for one strawberry variety on specific primer was got through combining the primer label and the corresponding allele label. Combination of these codes of 18 primers according to the primer sequence to form one DNA fingerprint for one particular variety. The results showed that: A total of 150 polymorphic bands were amplified, and the number of polymorphic bands amplified by different primers ranged from 4 to 17. Finally, 377 band combinations were obtained in this study. The value of polymorphism information content ranged from 0. 494 to 0. 908, and the average value was 0. 738. DNA fingerprint of all 84 strawberry variety was constructed following the above method. The result of clustering analysis showed that all varieties were completely differed from each other by using these 18 primer pairs based on the threshold of genetic similarity coefficient at 0. 99. Some cultivars from same origin were clustered together obviously. In conclusion, the DNA fingerprint of all 84 strawberry variety was constructed, and the relationship among them was close.
Key words:  strawberry  SSR  DNA fingerprinting  genetic analysis