摘要
为火龙果种质资源的遗传多样性评价及品种分子鉴定提供参考,利用MISA软件对获得的Unigene进行筛选,以确定火龙果转录组的SSR位点及分布,进而利用Primer 3.0软件设计SSR引物,并利用PCR扩增筛选SSR多态性引物,根据SSR扩增产物的多态性,利用UPGMA方法对15个火龙果种质材料进行聚类分析。结果表明:在碱基序列总长度为54.975Mb的79 658条Unigene中,筛选得到16 523个SSR位点,分布于13 493条Unigene序列上,SSR位点的出现频率为20.74%,平均每3.33kb出现1个SSR位点;SSR的主要类型是单核苷酸重复、二核苷酸重复和三核苷酸重复,其占比分别为59.58%、26.53%和12.93%;随机筛选获得的37对引物经PCR扩增,有23对引物可扩增出清晰条带,11对引物在15个火龙果种质材料中表现出多态性;UPGMA聚类分析将15个火龙果种质材料分为4类,其中,果肉为紫红色或粉红色的贵州紫红龙、贵州红肉-1和台湾红水晶等8个火龙果种质材料聚为Ⅰ类,果肉为红色或粉红色的贵州红龙、云南红肉-2和云南粉肉3个种质材料聚为Ⅱ类,果肉为白色的云南白肉-1、云南白肉-2和贵州野生白肉3个种质材料聚为Ⅲ类,果肉为紫红色的台湾黑龙火龙果种质材料单独为Ⅳ类。
The acquired Unigene was screened by using MISA software to determine SSR loci and distribution of pitaya transcriptome,SSR primers were designed by Primer 3.0 software and SSR polymorphism primers were screened by PCR amplification.15 pitayagermplasm materials was analyzed by UPGMA clustering method according to polymorphism of SSR amplified products to provide a reference for genetic diversity and variety molecular identification of pitayagermplasm resources.Result：16 523 SSR loci distributed on 13 493 Unigene sequences are screened from 79 658 Unigenes of base sequences with total 54.975 Mb.The occurrence frequency of SSR loci is 20.74%and there is one SSR locus within3.33 kb averagely.Mononucleotide,dinucleotide and trinucleotide are main types of repeat motif,account for 59.58%,26.53% and 12.93% respectively.23 pairs of primers in randomly screened 37 pairs of primers can amplify distinct bands and 11 pairs of primers show polymorphism in 15 pitayagermplasm materials.15 pitayagermplasm materials can be divided into 4 groups by UPGMA clustering analysis.Eight pitayagermplasm materials such as Zihonglong,Hongrou-1,Hongshuijing with purplish red or pink flesh,Honglong,Hongrou-2 and Fenrou with red or pink flesh,Bairou-1,Bairou-2 and Wild Bairou with white flesh and Heilong with purplish red flesh are divided into GroupⅠ,GroupⅡ,GroupⅢand GroupⅣrespectively.
出处
《贵州农业科学》
CAS
2018年第7期1-5,共5页
Guizhou Agricultural Sciences
基金
贵州省科学技术基金项目"基于转录组测序克隆火龙果果实甜度及色泽相关功能基因"[黔科合基础(2016)1146]
"火龙果耐低温种质收集及相关生理机制研究"[黔科合平台人才(2017)5603]
贵州省科技厅与贵州省农业科学院联合基金项目"火龙果果实糖酸积累相关酶活性研究"[黔科合LH字(2014)7705]
"钙对低温胁迫下火龙果生理生化特性的影响"[黔科合LH字(2015)7081]
贵州省重大攻关项目"火龙果产业化关键技术研究与集成示范"[黔科合重大专项字(2012)6006]
国家公益性行业(农业)科研专项(合作项目)子课题"中缅边境地区火龙果产业高效生产及产业化研究与示范"[公益性行业(农业)科研专项(2014)03036]
关键词
火龙果
转录组
微卫星标记
多态性
pitaya
tanscriptome
microsatellite marker
polymorphism
