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The alcohol dehydrogenase with a broad range of substrate specificity regulates vitality and reproduction of the plant-parasitic nematode Bursaphelenchus xylophilus

Published online by Cambridge University Press:  15 October 2018

Linsong Wang Open the ORCID record for Linsong Wang [Opens in a new window] ,
Tingting Zhang ,
Zhengsong Pan ,
Lulu Lin ,
Guoqing Dong ,
Min Wang  and
Ronggui Li
Linsong Wang
Affiliation:
College of Life Sciences, Qingdao University, Qingdao 266071, PR, China
Tingting Zhang
Affiliation:
College of Life Sciences, Qingdao University, Qingdao 266071, PR, China
Zhengsong Pan
Affiliation:
School of Life Sciences and Technology, Shanghai Jiaotong University, Shanghai 200240, PR, China
Lulu Lin
Affiliation:
College of Life Sciences, Qingdao University, Qingdao 266071, PR, China
Guoqing Dong
Affiliation:
College of Life Sciences, Qingdao University, Qingdao 266071, PR, China
Min Wang
Affiliation:
College of Life Sciences, Qingdao University, Qingdao 266071, PR, China
Ronggui Li*
Affiliation:
College of Life Sciences, Qingdao University, Qingdao 266071, PR, China
*
Author for correspondence: Ronggui Li, E-mail: lrg@qdu.edu.cn
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Abstract

Pine wilt disease, which is caused by the pine wood nematode (PWN), Bursaphelenchus xylophilus, has caused huge damage to pine forests around the world. In this study, we analysed the PWN transcriptome to investigate the expression of genes related to the associated bacterial species Pseudomonas fluorescens and found that the gene adh-1 encoding alcohol dehydrogenase (ADH) was upregulated. The open reading frame of adh-1, which encoded a protein of 352 amino acid residues, was cloned from B. xylophilus. Recombinant ADH with a relative molecular weight of 39 kDa, was present mainly in inclusion bodies and was overexpressed in Escherichia coli BL21 (DE3) and purified after refolding. The biochemical assay revealed that recombinant ADH could catalyse the dehydrogen reaction of eight tested alcohols including ethanol in the presence of NAD+. Quantitative real-time RT-PCR analysis indicated that ethanol upregulated adh-1 expression in PWN. Results of RNA interference and inhibition of ADH treatment indicated that downregulating expression of adh-1 or inhibition of ADH could reduce ethanol tolerance and the vitality and reproduction ability of B. xylophilus, suggesting that adh-1 is involved in pathogenicity of PWN.

Keywords

Alcohol dehydrogenaseBursaphelenchus xylophilusgene cloningreproductionsubstrate specificityvitality
Type
Research Article
Information
Parasitology , Volume 146 , Issue 4 , April 2019 , pp. 497 - 505
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Copyright
Copyright © Cambridge University Press 2018 

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