摘要
以秋石斛‘水芙蓉’为试材,采用生物信息学手段,研究了秋石斛转录组SSR位点的出现频率、基元种类、重复次数、长度分布等信息,并对设计挑选的50对SSR引物进行评价分析,以期为筛选适用于石斛种质资源评价和杂交后代鉴定的分子标记引物提供参考依据。结果表明:从20 782条秋石斛Unigene(>1 000 bp)中搜索到8 458个SSR位点,SSR位点发生频率和出现频率分别为31.53%和40.70%。进一步分析,发现SSR位点以单核苷酸重复为主(60.90%),共检测到106种SSR重复基元种类,单核苷酸中A/T重复基序最多,二核苷酸中AG/CT、GA/TC重复基序较多;SSR基元的重复次数主要集中在5~16次,占总SSR位点数量的95.78%;序列长度在10~20 bp的SSR位点有7 493个(88.59%),六核苷酸重复的平均长度最大(36 bp),单核苷酸重复的平均长度最小(11.41 bp)。通过琼脂糖凝胶电泳,从50对引物中筛选出42对有效扩增引物,其中14对引物具有多态性。该研究结果说明了秋石斛转录组SSR位点出现频率较高,具有开发多态性引物的潜力,可为后续石斛遗传多样性分析、分子辅助育种等研究提供候选标记资源。
Taking Dendrobium‘Rainbow-compactum’as the test material,bioinformatics methods were used to study the frequency of occurrence,types of motifs,repetition times,length distribution,and other information of SSR loci in the transcriptome of autumn dendrobium.50 pairs of SSR primers were designed and selected for evaluation and analysis,in order to provide reference for screening molecular marker primers suitable for the evaluation of Dendrobium germplasm resources and identification of hybrid offspring.The results showed that 8458 SSR loci were identified from 20782 Unigenes,the occurrence frequency and distribution frequency of SSR loci were 31.53%and 40.70%,respectively.Further analysis showed that the mononucleotide repeat was the main type(60.90%).A total of 106 kinds of SSR repeats were detected.The A/T repeats were the most common in the mononucleotide repeat,and the AG/CT and GA/TC repeats were the most common in the dinucleotide repeat.The number of repeats of SSR primitives mainly concentrated in 5-16 times,accounting for 95.78%of the total loci.There were 7493 SSR loci(88.59%)with sequence length between 10 bp to 20 bp,the average length of hexanucleotide repeats was the largest(36 bp),and the average length of mononucleotide repeats was the smallest(11.41 bp).By agarose gel electrophoresis,42 effective primers were selected from 50 primers,among which 14 were polymorphic.The results indicated that SSR loci of the Dendrobium transcriptomes were more frequent,which had the potential to develop polymorphism primers and provide candidate marker resources for genetic diversity analysis and molecular assisted breeding.
作者
林榕燕
孔兰
陈艺荃
钟淮钦
叶秀仙
LIN Rongyan;KONG Lan;CHEN Yiquan;ZHONG Huaiqin;YE Xiuxian(Crop Research Institute,Fujian Academy of Agricultural Sciences(Fujian Germplasm Resource Center)/Fujian Engineering Research Center for Characteristic Floriculture,Fuzhou,Fujian 350013)
出处
《北方园艺》
北大核心
2025年第15期42-48,共7页
Northern Horticulture
基金
福建省省属公益类科研院所资助项目(2022R1031009)
福建省农业科学院自由探索科技创新资助项目(ZYTS2023003)
福建省农业科学院科技创新团队建设资助项目(CXTD2021010-2)
福建省人民政府、中国农业科学院‘5511’协同创新资助项目(XTCXGC2021016)。
关键词
秋石斛
转录组
SSR位点
多态性
Dendrobium spp.
transcriptome
SSR locus
polymorphism
