中国农业科学 ›› 2015, Vol. 48 ›› Issue (11): 2108-2117.doi: 10.3864/j.issn.0578-1752.2015.11.003

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

普通烟草及其祖先种基因组SSR位点分析

童治军,焦芳婵,肖炳光   

  1. 云南省烟草农业科学研究院/烟草行业烟草生物技术育种重点实验室/国家烟草基因工程研究中心,昆明 650021
  • 收稿日期:2014-10-13 出版日期:2015-06-01 发布日期:2015-06-01
  • 通讯作者: 肖炳光,Tel:0871-65179532;E-mail:xiaobg@263.net
  • 作者简介:童治军,E-mail:tzj861@163.com
  • 基金资助:
    中国烟草总公司项目(110201301006(JY-06))、中国烟草总公司云南省公司项目(2013YN01)

Analysis of SSR Loci in Nicotina tabacum Genome and Its Two Ancestral Species Genome

TONG Zhi-jun, JIAO Fang-chan, XIAO Bing-guang   

  1. Yunnan Academy of Tobacco Agricultural Sciences/Key Laboratory of Tobacco Biotechnological Breeding/National Tobacco Genetic Engineering Research Center, Kunming 650021
  • Received:2014-10-13 Online:2015-06-01 Published:2015-06-01

摘要: 【目的】普通烟草(N. tabacum ,2n=24Ⅱ=48 TTSS)及其2个祖先种-绒毛状烟草(N. tomentosoformis,2n=12Ⅱ=24 TT)和林烟草(N.sylvestris,2n=12Ⅱ=24 SS)基因组SSR位点信息的统计分析,有助于烟草属植物的遗传分析。【方法】从公共数据库NCBI(National Center for Biotechnology Information)中下载上述3个烟草基因组数据,应用SSRIT和TRF软件分析其各自SSR位点分布特征,每个基因组随机合成50对SSR引物扩增多态性。【结果】在绒毛状烟草基因组、林烟草基因组和普通烟草基因组中分别获得218 081、263 478和397 432个SSR总位点,其间的平均距离分别为7.52、7.78和9.06 kb。绝大部分的SSR位点分布在内含子和UTR(尤其是5′-UTR)区域;以2核苷酸和3核苷酸类型为主,占基因组内SSR位点总数目的2/3以上,其中,2核苷酸类型丰度最高;含有A(T)n基序结构的频率及数量最高;除单核苷酸类型外,重复次数多在3—10。150对合成的引物对8个烟草种DNAs进行PCR反应,所有材料均能扩增出清晰稳定的目标片段,其中36对引物显示多态性。【结论】绒毛状烟草、林烟草和普通烟草基因组内SSR呈现一定的分布特征,表明SSR位点在亲缘关系相对较近的烟草种间具有高度保守性。

关键词: 绒毛状烟草, 林烟草, 普通烟草, 基因组, SSR位点

Abstract: 【Objective】 The statistical analysis of SSR loci genomic information in Nicotina tabacum (2n = 24Ⅱ= 48 TTSS) and its two ancestral species (N. tomentosiformis 2n = 12Ⅱ= 24 TT and N. sylvestris 2n = 12Ⅱ= 24 SS) are carried out for the genetic analysis of plants of the Nicotiana genus. 【Method】 The data in the above-mentioned three tobacco genomes were downloaded from a public database NCBI (National Center for Biotechnology Information), 50 pairs of random amplified polymorphic SSR were synthesized for each genome, and analyzed by the SSRIT and TRF soft-wares for their SSR-loci distribution. 【Result】 A total of 218 081, 263 478 and 397 432 SSR loci, with their average distance 7.52, 7.78 and 9.06 kb were found in the genomes of N. tomentosiformis, N. sylvestris and N. tabacum, respectively. Almost all the SSR loci are distributed in the intron domains and especially the 5′-UTR domains. The SSR motifs within the genome are dominated by the types of Di-nucleotide and Tri-nucleotide, with more than 2/3 in the total number of SSR motifs, and the type of Di-nucleotide motifs has a most abundant peak, and contains the highest frequency and quantity of A (T)n motif structure. The most of SSR motifs types have 3-10 repetitions, except mono-nucleotide type. There are 150 pairs of primers synthesized are subjected to PCR for the DNAs from eight tobacco species, all of which can amplify the clearly stable target fragments, with 36 pairs of primers shows polymorphisms. 【Conclusion】 Some distribution characteristcs of SSRs in the genomes of N. tomentosiformis, N. sylvestris and N. tabacum were observed, which indicated a high conservation for the SSR loci among the relatively closed tobacco species.

Key words: N. tomentosiformis, N. sylvestris, N. tabacum, genome, SSR loci