Molecular identification of ‘Candidatus phytoplasma asteris’ related strain (16SrΙ-B) associated with Broussonetia papyrifera in Nanjing, China
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A new disease associated with paper mulberry (Broussonetia papyrifera) leaf yellowing and curling symptoms was observed in Nanjing, Jiangsu Province, China, in 2014 and 2015. The disease aetiology was investigated by direct and nested polymerase chain reaction (PCR) with phytoplasma-specific primers, sequencing and phylogenetic analysis. Total DNA extracted from symptomatic paper mulberries and healthy plants was used for PCR amplification. Fragments of 1.2 kb for 16S rDNA, 1.2 kb for rp (ribosomal protein) and 1.1 kb for tuf (translation elongation factor EF-Tu) were obtained in symptomatic paper mulberries, whereas these fragments were absent in healthy plants. The resulted sequences were identified using BLAST search tool as belonging to phytoplasmas. Comparison results showed that 16S rDNA fragment of this phytoplasma was homologous with the members of 16SrΙ (AY) group and shared 100 % identity with the sequences of ‘Candidatus Phytoplasma asteris’-related strain AY-27 (HM467127.1)(16SrΙ-B group). Virtual restriction fragment length polymorphism (RFLP) also showed the same results. In addition, rp and tuf fragments of this phytoplasma shared 99 % identity with the members of 16SrΙ subgroup B. This is the first report of a phytoplasma member from AY group (16SrΙ group) infecting paper mulberry.
KeywordsBroussonetia papyrifera; Phytoplasma; Leaf yellowing and curling; 16SrΙ group
This work was supported in part by grants from the National Natural Science Foundation of China (J1210056).
Compliance with ethical standards
Conflict of interest
The authors declare no conflict of interest.
- Hoshi, A., Oshima, K., Kakizawa, S., Ishii, Y., Ozeki, J., Hashimoto, M., Komatsu, K., Kagiwada, S., Yamaji, Y., & Namba, S. (2009). A unique virulence factor for proliferation and dwarfism in plants identified from a phytopathogenic bacterium. Proceedings of the National Academy of Sciences of the United States of America, 106, 6416–6421.CrossRefPubMedPubMedCentralGoogle Scholar
- Lee, I. M., Gundersen-Rindal, D. E., Davis, R. E., Bottner, K. D., Marcone, C., & Seemuller, E. (2004). ‘Candidatus phytoplasma asteris’, a novel phytoplasma taxon associated with aster yellows and related diseases. International Journal of Systematic and Evolutionary Microbiology, 54(Pt 4), 1037–1048.CrossRefPubMedGoogle Scholar
- Lee, I., Zhao, Y., Davis, R. E., Weintraub, P., & Jones, P. (2009). Prospects of multiple gene-based systems for differentiation and classification of phytoplasmas. Phytoplasmas: Genomes, plant Hosts and Vectors, 51–63.Google Scholar
- Marcone, C., Lee, I. M., Davis, R. E., Ragozzino, A., & Seemuller, E. (2000). Classification of aster yellows-group phytoplasmas based on combined analyses of rRNA and tuf gene sequences. International Journal of Systematic and Evolutionary Microbiology, 50(Pt 5), 1703–1713.CrossRefPubMedGoogle Scholar
- Peng, X., Teng, L., Wang, X., Wang, Y., & Shen, S. (2014). De novo assembly of expressed transcripts and global transcriptomic analysis from seedlings of the paper mulberry (broussonetia kazinoki × broussonetia papyifera). PloS One, 9, e97487. doi: 10.1371/journal.pone.0097487.CrossRefGoogle Scholar
- Richards, E., Reichardt, M., & Rogers, S. (2001). Preparation of genomic DNA from plant tissue. Current Protocols in Molecular Biology, Chapter 2, Unit2.3, doi:10.1002/0471142727.mb0203s27.Google Scholar
- Sugio, A., Kingdom, H. N., MacLean, A. M., Grieve, V. M., & Hogenhout, S. A. (2011). Phytoplasma protein effector SAP11 enhances insect vector reproduction by manipulating plant development and defense hormone biosynthesis. Proceedings of the National Academy of Sciences of the United States of America, 108(48), E1254–E1263.CrossRefPubMedPubMedCentralGoogle Scholar
- Suzuki, S., Oshima, K., Kakizawa, S., Arashida, R., Jung, H. Y., Yamaji, Y., et al. (2006). Interaction between the membrane protein of a pathogen and insect microfilament complex determines insect-vector specificity. Proceedings of the National Academy of Sciences of the United States of America, 103(11), 4252–4257.CrossRefPubMedPubMedCentralGoogle Scholar
- Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., & Kumar, S. (2011). MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28(10), 2731–2739.CrossRefPubMedPubMedCentralGoogle Scholar