基于ITS和β-tubulin基因分析的居间疫霉菌系统发育

doi:10.11707/j.1001-7488.20180308

Abstract

Abstract: [Objective] A survey of diseases caused by the genus of Phytophthora were carried out in the subtropical forests of China. This study aimed at exploring the genetic diversity of Phytophthora and providing a theory basis for controlling the disease caused by Phytophthora.[Method] The healthy leaves were used to bait Phytophthora isolates from stream water.The symptomatic leaf tissues were isolated and Phytophthora was preliminarily determined based on its hyphal branching structure under a microscope,then ITS and β-tubulin gene of strains were amplified with PCR and sequenced.The spliced sequence was used for phylogenetic analysis with MAFFT 7,PAUP 4 beta10,MrBayes 3.2.6 and PhyML 3 software.[Result]Forty-six isolates of Phytophthora were obtained, and their morphological characteristics were consistent with P. intercalaris, a new record species in China. All isolates produced ITS amplicons of 847 to 849 bp in size, which differed from each other in one to three positions. These sequences had identity from 99.29% to 99.53% with the reference sequences of P. intercalaris type culture (KT163268). All the β-tubulin amplicons were 882 bp, which shared 99.43% to 99.66% identity with reference sequences of P. intercalaris (KT163336). Phylogenetic analysis showed that the tested strains were clustered with P. intercalaris with a high bootstrap support value (100%). This study not only increased the number of P. intercalaris isolates, but also expanded the distribution range of P. intercalaris, and enriched the number of Phytophthora species in China as well.[Conclusion]The result of gene sequence analysis showed that there was high sequence consistency among isolates of P. intercalaris from different regions, but there were variations in the base site, which formed different gene types. The present study provides a theoretical basis for the further development of the epidemiology and control strategy for the tree blight diseases.

Key words: Forest Phytophthora, Phytophthora intercalaris, ITS, β-tubulin, phylogenetic analysis

CLC Number:

S718.81

Li Wenwen, Zhao Wenxia, Lin Ruozhu, Yao Yanxia, Li Juan, Huai Wenxia. Phylogenetic Relationships of Phytophthora intercalaris Based on ITS and β-tubulin Gene Sequence[J]. Scientia Silvae Sinicae, 2018, 54(3): 73-82.

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Phylogenetic Relationships of Phytophthora intercalaris Based on ITS and β-tubulin Gene Sequence

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