Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (20): 3982-3991.doi: 10.3864/j.issn.0578-1752.2014.20.006

• PLANT PROTECTION • Previous Articles     Next Articles

EST-SSR Information Analysis and Markers Development in Fusarium oxysporum

LI Xin-feng1, ZHANG Zuo-gang1, WANG Jian-ming1, GAO Jun-ming1, TIAN Hong-xian2   

  1. 1College of Agriculture, Shanxi Agricultural University, Taigu 030801, Shanxi
    2Crops Research Institute for Alpine, Shanxi Academy of Agricultural Sciences, Datong 037000, Shanxi
  • Received:2014-04-01 Revised:2014-05-30 Online:2014-10-16 Published:2014-10-16

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Abstract: 【Objective】 The objective of this study is to investigate the distribution of Fusarium oxysporum SSRs (simple sequence repeat) derived from ESTs (expressed sequence tag) and to develop EST-SSR markers. These researches could provide technical supports for the genetic diversity analysis of Fusarium. 【Method】 All the 9 304 EST sequences of F. oxysporum were obtained from NCBI. SSRIT (simple sequence repeat identification tool) was used to search SSRs in these sequences and Primer Premier 5.0 was used to design primers. Fourteen pairs of primers were used to analyze the genetic diversity of 23 F. oxysporum strains from different hosts and areas, and the PCR products of these primers were detected by PAGE.【Result】A total of 92 SSRs were identified from 90 F. oxysporum EST sequences, with the frequency of 1.0%. The dinucleotide repeats (59.78%) were most abundant followed by tri-(17.39%) and penta-nucleotide repeats (11.96%). AC/GT (0.51%) occurred more often than any other repeat motif. Among the 30 synthesized primers, 20 (66.7%) amplified distinct bands, and 14 (46.7%) amplified expected polymorphic products. The genetic diversity analysis based on the SSR data showed that a total of 62 bands were amplified by 14 selected SSR primers, 59 (95.2%) of which were polymorphic, with an average of 4.2 bands per primer. The genetic similarity ranged from 0.487 to 0.933, with an average of 0.686. The genetic variation of the strains was obvious. The cluster analysis based on the SSR data showed that except No.14 strain, F. oxysporum strains were distinctly clustered into different groups according to their hosts at 0.781.【Conclusion】Exploitation of SSR markers from F. oxysporum ESTs is convenient and practicable. Fourteen EST-SSR markers identified in this study would be useful for genetic variation study of F. oxysporum.

Key words: Fusarium oxysporum, expressed sequence tag (EST), simple sequence repeat (SSR), genetic diversity

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