Screening DNA Barcode of Three Pathogenic Fungi within Genus Botryosphaeria in Apple

doi:10.14088/j.cnki.issn0439-8114.2018.02.031

Abstract

Abstract: Species of Botryosphaeria are the important pathogenic fungi on many kinds of fruits. They are very always difficult to distinguish from each other based on morphological characters. In this study, to identificate of three important apple-pathogens Botryosphaeria stevensii, Botryosphaeria dothidea, Botryosphaeria obtuse. 4 gene fragments were selected, which encoded ITS、LSU、SSU、EF-1α,to be the candidate DNA barcodes. By using of PCR amplification, Sanger sequencing and data analysis,we aimed to find the molecular markers in the identification of these species. The results showed that, based on the PCR and sequencing success,it was ranked as follow：ITS>SSU>LSU=EF-1α,and ITS primer worked best（100%）. According to the acquired sequences,ITS and EF-1α were quite conserved among different strains of the same species,and have the significant differences between the species. Meantime,a region of EF-1α （approx. 100 bp） was exclusively existed in the investigated strains of B. dothidea,which therefore could be used for the distinction of this species. However,the sequences of LSU and SSU have the same variation within and between the species. Combining the available ITS sequences in GeneBank,we constructed maximum likelihood tree to identify the phylogenetic relationships of Botryosphaeria. The results showed that three investigated species belonged to three monophyletic clades. These results were congruent with the taxonomic units based on the morphological characters. Accordingly, we recommended the usage of ITS and EF-1α to be the efficient molecular makers in the identification of B. stevensii、B. dothidea、B. obtuse.

Key words: Botryosphaeria, apple pathogen, barcode, EF-1α;, ITS

CLC Number:

S432.4

CHEN Zhan, SHEN Hong, XU Ping-ping, WANG Zhen-hua, WU Cui-ping, LI Bin. Screening DNA Barcode of Three Pathogenic Fungi within Genus Botryosphaeria in Apple[J]. HUBEI AGRICULTURAL SCIENCES, 2018, 57(2): 126-131.

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