Phylogenetic Analysis and Development of Molecular Tool for Detection of Diaporthe citri Causing Melanose Disease of Citrus

Plants
Chingchai ChaisiriChao-Xi Luo

Abstract

Melanose disease caused by Diaporthe citri is considered as one of the most important and destructive diseases of citrus worldwide. In this study, isolates from melanose samples were obtained and analyzed. Firstly, the internal transcribed spacer (ITS) sequences were used to measure Diaporthe-like boundary species. Then, a subset of thirty-eight representatives were selected to perform the phylogenetic analysis with combined sequences of ITS, beta-tubulin gene (TUB), translation elongation factor 1-α gene (TEF), calmodulin gene (CAL), and histone-3 gene (HIS). As a result, these representative isolates were identified belonging to D. citri, D. citriasiana, D. discoidispora, D. eres, D. sojae, and D. unshiuensis. Among these species, the D. citri was the predominant species that could be isolated at highest rate from different melanose diseased tissues. The morphological characteristics of representative isolates of D. citri were investigated on different media. Finally, a molecular tool based on the novel species-specific primer pair TUBDcitri-F1/TUBD-R1, which was designed from TUB gene, was developed to detect D. citri efficiently. A polymerase chain reaction (PCR) amplicon of 217 bp could be specifically amplified with the d...Continue Reading

References

Apr 1, 1995·Applied and Environmental Microbiology·N L Glass, G C Donaldson
May 21, 2008·Studies in Mycology·Pedro W CrousKevin D Hyde
Feb 24, 2012·Systematic Biology·Fredrik RonquistJohn P Huelsenbeck
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Dec 16, 2017·IMA Fungus·Vladimiro Guarnaccia, Pedro W Crous
Feb 8, 2018·Nature·Guohong Albert WuManuel Talon
Jul 13, 2018·Studies in Mycology·Y Marin-FelixP W Crous
Sep 1, 2014·Plant Disease·S MahadevakumarG R Janardhana

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Methods Mentioned

BETA
PCR
electrophoresis

Software Mentioned

FigTree
PAUP ( Phylogenetic Analysis Using Parsimony ,
Primer premier
DNASTAR Lasergene Core Suite
MrBayes
SeqMan
Clustal W
TreeBASE
BioEdit
MrModeltest

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