第二代测序技术用于水稻和稻瘟菌互作早期转录组的分析

doi:10.3724/SP.J.1005.2012.00102

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HEREDITAS ›› 2012, Vol. 34 ›› Issue (1): 102-112.doi: 10.3724/SP.J.1005.2012.00102

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Transcriptome analysis of early interaction between rice and Magnaporthe oryzae using next-generation sequencing technology

LI Xiang-Long^1,2, BAI Bin^1,2, WU Jun^1,3, DENG Qi-Yun^1,3, ZHOU Bo^2,4

1. Branch of Longping, Graduate School of Central South University, Changsha 410012, China 2. Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China 3. China National Hybrid Rice Research Center, Changsha 410125, China 4. Institute of Biotechniques, Zhejiang University, Hangzhou 310029, China

Received:2011-04-21 Revised:2011-05-11 Online:2012-01-20 Published:2012-01-25

Abstract

Abstract: Magnaporthe oryzae is speculated to express and translocate a set of effector proteins to interfere with rice innate immunity and to promote its invasion. In this study, we carried out a transcriptome analysis of early interaction between rice and M. oryzae using a next generation sequencing technology to identify putative effector protein genes. We totally obtained 338 942 M. oryzae derived sequence tags from around 12.5 M gross mixed tags by refering the genome sequence of reference strain 70–15. These M. oryzae sequences were finally mapped to 779 predicted genes. Of these 799 genes, 108 were predicted to be involved in the interaction between rice and M. oryzae and 42 encoded proteins predicted to be secreted. Expression of these 42 putative secreted protein genes were further assessed using a reverse transcription polymerase chain reaction (RT-PCR) method, which revealed that 12 of them were expressed during the infection. Four of these 12 expressed genes were further confirmed to be expressed specifically during the infection. This study demonstrated an attempt to isolate M. oryzae genes, especially those putative effector protein genes, which are expressed specifically at the early interaction stage using a next-generation sequencing technology.

Key words: Solexa sequencing technology, Magnaporthe oryzae, secreted protein, transcriptome analysis, expression specificity

LI Xiang-Long, BAI Bin, WU Jun, DENG Qi-Yun, ZHOU Bo. Transcriptome analysis of early interaction between rice and Magnaporthe oryzae using next-generation sequencing technology[J]. HEREDITAS, 2012, 34(1): 102-112.

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Transcriptome analysis of early interaction between rice and Magnaporthe oryzae using next-generation sequencing technology

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