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Identification and characterization of the Enterobacter complex causing mulberry (Morus alba) wilt disease in China

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Abstract

Mulberry wilt disease (MWD) was recently identified in Hangzhou, Zhejiang province, China. Typical symptoms of the disease are browning of vascular tissues, leaf wilt, defoliation, and tree decline. Unlike the symptoms of bacterial wilt disease caused by Ralstonia solanacearum, symptoms of MWD generally started from the bottom of the plants and moved upward. In inoculation experiments, four selected MWD strains caused mulberry shoot leaf wilt, discoloration, and defoliation. They also induced whole plant leaf wilt, defoliation and dark brown discoloration of vascular tissue. Based on Biolog metabolic profiles, fatty acid methyl ester analysis (FAME) and sequence analysis of the partial 16S rDNA and rpoB genes four MWD strains were identified as members of the genus Enterobacter. The 16S rDNA and rpoB gene sequences revealed a close relationship among two isolates, R2-2 and R6-2, and the E. asburiae type strain JCM6051. The isolates showed >98% similarity to E. asburiae JCM6051 in their rpoB gene. These results indicated that isolates R2-2 and R6-2 belonged to E. asburiae. No similarity in 16S rDNA sequences above 97% was found between either of the remaining isolates, R11-2 or R18-2, and any recognized Enterobacter species, suggesting that the two isolates may represent novel Enterobacter species. rpoB gene similarity values between the isolates and Enterobacter spp. type strains were <98%, providing further evidence that the two isolates may represent a novel species within the Enterobacter. The causal agent for MWD was previously reported to be E. cloacae, however, this study found that other Enterobacter spp. (E. asburiae and Enterobacter sp.) also cause MWD.

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Acknowledgements

We gratefully acknowledge the support of the National Natural Science Foundation of China (30871655, 30671397), Agricultural Ministry of China (nyhyzx07-056) and Zhejiang Provincial Agricultural Departments of China (2008C22072, SN200811). We also thank Fu-Shou Xu and Qin Zhou for providing some samples and mulberry seedlings. We also thank Dr. Bin Li, Dr. Ivan Kurtovic and Mrs. Miao-Lin Li for manuscript revisions and Dr. Steve ShearDown for access to DNAstar package 5.00.

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Authors and Affiliations

  1. State Key Laboratory of Rice Biology, Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Ministry of Agriculture, Institute of Biotechnology, Zhejiang University, P. O. Box 3-398, Hangzhou 310029, Zhejiang province, People’s Republic of China

    Guo-Fen Wang, Guan-Lin Xie & Bo Zhu

  2. Institute of Environmental and Plant Protection, Chinese Academy of Tropical Agricultural Sciences, Danzhou 571737, Hainan province, People’s Republic of China

    Guo-Fen Wang & Jun-Sheng Huang

  3. Agricultural Bioresource Research Institute, Fujian Academy of Agricultural Science, Fuzhou 350003, Fujian province, People’s Republic of China

    Bo Liu

  4. Faculty of Natural Resources and Agro-Industry Kasetsart University, Sakonnakhon Province, 47000, Thailand

    Praphat Kawicha

  5. USDA-ARS-USHRL, Fort Pierce, FL, 34945, USA

    Lesley Benyon & Yong-Ping Duan

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  1. Guo-Fen Wang
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Correspondence to Guan-Lin Xie.

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Wang, GF., Xie, GL., Zhu, B. et al. Identification and characterization of the Enterobacter complex causing mulberry (Morus alba) wilt disease in China. Eur J Plant Pathol 126, 465–478 (2010). https://doi.org/10.1007/s10658-009-9552-x

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  • DOI: https://doi.org/10.1007/s10658-009-9552-x

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