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An anionic defensin from Plutella xylostella with potential activity against Bacillus thuringiensis

Published online by Cambridge University Press:  22 July 2016

X.-X. Xu ,
Y.-Q. Zhang ,
S. Freed ,
J. Yu ,
Y.-F. Gao ,
S. Wang ,
L.-N. Ouyang ,
W.-Y. Ju  and
F.-L. Jin
X.-X. Xu
Affiliation:
Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, College of Agriculture, South China Agricultural University, Guangzhou 510642, P. R. China
Y.-Q. Zhang
Affiliation:
Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, College of Agriculture, South China Agricultural University, Guangzhou 510642, P. R. China
S. Freed
Affiliation:
Department of Entomology, Faculty of Agricultural Sciences and Technology, Bahauddin Zakariya University, Multan 60800, Pakistan
J. Yu
Affiliation:
Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, College of Agriculture, South China Agricultural University, Guangzhou 510642, P. R. China
Y.-F. Gao
Affiliation:
Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, College of Agriculture, South China Agricultural University, Guangzhou 510642, P. R. China
S. Wang
Affiliation:
Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, College of Agriculture, South China Agricultural University, Guangzhou 510642, P. R. China
L.-N. Ouyang
Affiliation:
Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, College of Agriculture, South China Agricultural University, Guangzhou 510642, P. R. China
W.-Y. Ju
Affiliation:
Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, College of Agriculture, South China Agricultural University, Guangzhou 510642, P. R. China
F.-L. Jin*
Affiliation:
Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, College of Agriculture, South China Agricultural University, Guangzhou 510642, P. R. China
*
*Address for correspondence Phone: +86 2085280203 Fax: +86 20 85280293 E-mail: [email protected]
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Abstract

Insect defensins, are cationic peptides that play an important role in immunity against microbial infection. In the present study, an anionic defensin from Plutella xylostella, (designated as PxDef) was first cloned and characterized. Amino acid sequence analysis showed that the mature peptide owned characteristic six-cysteine motifs with predicted isoelectric point of 5.57, indicating an anionic defensin. Quantitative real-time polymerase chain reaction analysis showed that PxDef was significantly induced in epidermis, fat body, midgut and hemocytes after injection of heat-inactivated Bacillus thuringiensis, while such an induction was delayed by the injection of live B. thuringiensis in the 4th instar larvae of P. xylostella. Knocking down the expression of nuclear transcription factor Dorsal in P. xylostella by RNA interference significantly decreased the mRNA level of PxDef, and increased the sensitivity of P. xylostella larvae to the infection by live B. thuringiensis. The purified recombinant mature peptide (PxDef) showed higher activity against Gram-positive bacteria, with the minimum inhibition concentrations of 1.6 and 2.6 µM against B. thuringiensis and Bacillus subtilis, respectively. To our knowledge, this is the first report about an anionic PxDef, which may play an important role in the immune system of P. xylostella against B. thuringiensis.

Keywords

Antimicrobial activityBacillus thuringiensisdefensinexpressionPlutella xylostellaRNAi
Type
Research Papers
Information
Bulletin of Entomological Research , Volume 106 , Issue 6 , December 2016 , pp. 790 - 800
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Copyright
Copyright © Cambridge University Press 2016 

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