Scientia Agricultura Sinica ›› 2016, Vol. 49 ›› Issue (12): 2301-2309.doi: 10.3864/j.issn.0578-1752.2016.12.006

• PLANT PROTECTION • Previous Articles     Next Articles

Cloning and Expression Analysis of Apoptosis-Related Gene FpTatD in Fusarium pseudograminearum

CHEN Lin-lin, HOU Ying, DING Sheng-li, SHI Yan, LI Hong-lian   

  1. College of Plant Protection, Henan Agricultural University/National Key Laboratory of Wheat and Maize Crop Science/ Collaborative Innovation Center of Henan Grain Crops, Zhengzhou 450002
  • Received:2016-03-15 Online:2016-06-16 Published:2016-06-16

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Abstract: 【Objective】The objective of this study is to investigate the potential biological functions of apoptosis-related genes FpTatD in Fusarium pseudograminearum, for this purpose, FpTatD genes were identified and cloned in F. pseudograminearum, and the expression of these genes were examined at mycelia, conidia and infection stages.【Method】 The known TatD proteins were obtained from GenBank, and four TatD candidates in fusariums were identified by BlastP. FpTatD genes and open reading frames (ORF) were amplified from genome DNA and cDNA by PCR, and the transcription levels in mycelia, conidia and infection were examined by qRT-PCR. The expression levels of FpTatD genes in the affinity interaction and non affinity interaction were assayed by RNA-seq.【Result】Four TatD candidates were identified in Fusariums, which divided into two significant branches by the phylogenetic tree. TatD1 and TatD2 belong to an ancient family that is conversed in almost all eukaryotic, while TatD3 and TatD4 seem to come from a unique TatD family that only exists in plants and fungi. Four FpTatD candidates, which identified in F. pseudograminearum were cloned, and the full-length sequences of FpTatD1, FpTatD2, FpTatD3 and FpTatD4 were 993, 1 331,  1 227 and 1 176 bp. FpTatD2 contains an intact open reading frame with 94 and 55 bp non-coding sequences in 5′ terminal, while FpTatD1, FpTatD3 and FpTatD4 consist continuous open reading frames. The four clones encode 36.37, 43.13, 45.59 and 44.25 kD proteins. All FpTatD contained the conserved DNase domain and most conserved amino acid residues. qRT-PCR analysis revealed that FpTatD1 and FpTatD2 were highly expressed in F. pseudograminearum, and both were highly up-regulated at early stages of infection. Especially, the expression level of FpTatD1 escalated to 8.8 and 7.6 times in 36 h and 3 d. However, FpTatD3 and FpTatD4 were poorly expressed at all stages. Thus, FpTatD1 and FpTatD2 might play important roles in F. pseudograminearum. RNA-seq analysis was consistent with qRT-PCR results that FpTatD1 and FpTatD2 were highly expressed and up-regulated at early stages of infection. In addition, Compared to the affinity interaction between F. pseudograminearum and GM301, FpTatD genes showed higher expression levels in the non affinity interaction between F. pseudograminearum and ZM24.【Conclusion】Apoptosis-related gene FpTatD might play an important role in the interaction between F. pseudograminearum and its host.

Key words: Fusarium pseudograminearum, apoptosis, TatD nuclease, gene cloning, expression analysis

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