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Scientia Silvae Sinicae ›› 2015, Vol. 51 ›› Issue (4): 110-115.doi: 10.11707/j.1001-7488.20150414

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Response of Transcription Factors of Populus tomentosa to Inoculation with Botryosphaeria dothidea

Zhangxiuying1, Songruiqing2, Zhangxingyao3   

  1. 1. Changchun University of Technology Changchun 130000;
    2. College of Forestry, Northeast Forestry University Harbin 150040;
    3. Research Institute of Forest Ecology, Environment and Protection, CAF Beijing 100091
  • Received:2013-12-20 Revised:2014-11-24 Online:2015-04-25 Published:2015-05-20

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Abstract:

【Objective】This study aims at investigating the response of transcription-related genes of Populus tomentosa induced by Botryosphaeria dothidea infectation.【Method】The suppression subtractive hybridization (SSH) technique was used to construct a cDNA library, and the bioinformatics were used to analyze the expression of transcription related gene of Populus tomentosa infected with Botryosphaeria dothidea. The bark mRNA was purified and transcribed into cDNA. A poplar-pathogen interaction cDNA library of inoculated poplar bark 72 h post-inoculation (hpi) was constructed. Expressed sequence tags (ESTs) were obtained, which were assembled into unisequences prior to their annotation. The unisequences related to transcription were selected, and were further analyzed by quantitative real-time PCR (qRT-PCR). The randomly selected 199 positive clones were sequenced, and compared with the sequences of NCBI gene pool, among which 172 EST sequences were found homologous. The homologous analysis for these EST was conducted in dbEST database retrieval, 【Result】The result showed that 10 sequences were associated with transcription, and accounted for 5.8% of the total sequences. They were divided into four categories, i.e. Zinc finger protein (Cys/His-rich), containing the EAR transcriptional repressor; CONSTANS-like protein, Encoding a nuclear protein; ordinary transcription factor, regulating transcription and WRKY21 transcription factor, participating in the plant defense. Through further RT-qPCR analyses on three kinds of transcription factors, the result showed that the expression amount of Zinc finger protein (Cys/His-rich) after inoculation was 50 times of the non-inoculated; The WRKY21 expression amount was 0.94 times of the non-inoculated; The CONSTANS-like protein expression amount was 2 times of the non-inoculated.【Conclusion】 The expression of these transcription factor genes was significantly increased in Populus tomentosa infected by Botryosphaeria dothidea.

Key words: Botryosphaeria dothidea, Populus tomentosa, suppression subtractive hybridization, real time PCR (qRT-PCR), transcription factors

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