白掌SSR引物筛选及12份种质资源亲缘关系分析  

Screening of SSR Primer of Spathiphyllum and Analysis of Genetic Relationship of 12 Germplasm Resources

作　　者：刘小飞 孙映波 黄丽丽 于波 Liu Xiaofei;Sun Yingbo;Huang Lili;Yu Bo(Key Laboratory of Urban Agriculture in South China(Ministry of Agriculture),Guangdong Key Lab of Ornamental Plant Germplasm Innovation and Utilization,Environmental Horticulture Institute,Guangdong Academy of Agricultural Sciences,Guangzhou,510640)

机构地区：[1]广东省农业科学院环境园艺研究所,广东省园林花卉种质创新综合利用重点实验室,农业农村部华南都市农业重点实验室,广州510640

出　　处：《分子植物育种》2022年第20期6802-6810,共9页Molecular Plant Breeding

基　　金：广州市科技计划项目(202002020025);广东省重点领域研发计划项目(2018B020202003);广东省科技计划项目(2015B070701016)共同资助。

摘　　要：本研究比较分析了白掌Spathiphyllum‘Parrish’转录组和叶绿体基因组中SSR标记的类型与分布,结果发现转录组中共有SSR标记7184个,其中,二核苷酸重复数量最多,为3454个;叶绿体基因组共有SSR标记281个,其中,单核苷酸重复数量最多,为223个。分别从转录组和叶绿体基因组的SSR标记中随机挑选32和18个,设计引物,PCR扩增后琼脂糖电泳结果显示,来自转录组的24对引物扩增条带清晰,多态性好;来自叶绿体基因组16对引物扩增条带清晰,多态性差。因此,选择来自转录组的24对引物合成M13标记的荧光引物,PCR扩增后毛细管电泳结果显示,19对引物共扩增出60个等位位点。利用60个等位位点构建了12份种质资源的系统进化树,UPGMA聚类分析将12份种质分为3组。19对引物中DN9090、24-DN9534、27-DN9108和DN101174对引物多态性较好,分别扩增出4、6、4和6个等位位点,可完全区分12份种质。本研究将为白掌种质资源鉴评提供可靠的SSR荧光标记引物。This study comparatively analyzed the types and distribution of SSR markers in the Spathiphyllum’Parrish’transcriptome and the chloroplast genome,and found that there were 7184 SSR markers in the transcriptome.Among them,the number of dinucleotide repeats was the largest,with a number of 3454.There are a total of 281SSR markers in the chloroplast genome.Among them,the number of single nucleotide repeats was the largest,with a number of 223.32 and 18 were randomly selected from the SSR markers of the transcriptome and chloroplast genome,respectively,and primers were designed.After PCR amplification,agarose electrophoresis results showed that the 24 pairs of primers from the transcriptome had clear amplified bands and good polymorphism.The amplified bands of 16 pairs of primers in the chloroplast genome were clear and had poor polymorphism.Therefore,24 pairs of primers from the transcriptome were selected to synthesize M13 labeled fluorescent primers.After PCR amplification,the capillary electrophoresis results showed that 19 pairs of primers amplified a total of 60alleles.A phylogenetic tree of 12 germplasm resources was constructed using 60 alleles.UPGMA cluster analysis divided 12 accessions into 3 groups.Among the 19 pairs of primers,DN9090,24-DN9534,27-DN9108,and DN10117 had good polymorphisms.4,6,4,and 6 alleles were amplified,and 12 germplasms could be completely distinguished.This study will provide usable SSR fluorescent markers primers for the evaluation of germplasm resources in Spathiphyllum.

关 键 词：白掌 简单重复序列 指纹图谱编码 种质资源 

分 类 号：S682.19[农业科学—观赏园艺]