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Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (01): 62-70.doi: 10.3724/SP.J.1006.2012.00062

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

cDNA-AFLP Analysis of Differentially Expressed Genes in Tobacco Infected by TMV

CHEN Rong-Ping1,2,LIU Lie3,WAN Xiu-Qing2,QIU En-Jian2,WANG Chun-Jun2,SONG Bao-Gang2,YAN Pei-Qiang2,YANG Tie-Zhao1,*   

  1. 1 College of Tobacco Science, Henan Agricultural University, Zhengzhou 450002, China; 2 Mudanjiang Tobacco Science and Research Institute of Heilongjiang Province Tobacco Company, Mudanjiang 157011, China; 3 Centers for Disease Control and Prevention in Mudanjiang, Mudanjiang 157022, China
  • Received:2011-05-10 Revised:2011-09-12 Online:2012-01-12 Published:2011-11-07
  • Contact: 杨铁钊, E-mail: yangtiezhao@126.com, Tel: 0371-63358030

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Abstract: Flue-cured tobacco variety Longjiang 925 with high resistance to tobacco mosaic virus (TMV) was used as the tested material, and we selected 240 pairs of primers and amplified cDNA from the tobacco leaf inoculated by TMV. The results showed that approximately 9500 gene transcript fragments were obtained, and twelve inducible expressed gene fragments were selected out by cloning and sequencing. The inducible fragments functions, involve in the nucleic acid metabolism, protein synthesis and modulation, energy metabolism, stress responding, intracellular transport, metabolism of carbohydrates. To validate the functions of these differentially expressed gene sequences obtained, we analyzed TIF2 fragment by real-time PCR with the samples collected at 0, 12, 24, 48, and 72 hours after inoculation, and the mRNAs samples were isolated. The result of real-time PCR indicated that the genes isolated were related to TMV-resistance. Then, both 5′- and 3′-rapid amplification of cDNA ends (RACE) were preformed. A full length cDNA sequence of TIF2 contained 875 bp, with a coding zone of 101 bp to 613 bp conjecturably, encoding 170 amino acids. Analyses of Blastn and Blastp showed that the gene was not homologous to tobacco, and probably was the TMV-resistance related novel gene. The results will be helpful for the future research on tobacco resistant-breeding to TMV in molecule level and so on.

Key words: Tobacco, TMV, Differentially Expressed Genes, cDNA-AFLP

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