First Report of Neopestalotiopsis piceana Causing Gray Blight in Camellia sinensis in China

HomePlant DiseaseVol. 107, No. 7First Report of Neopestalotiopsis piceana Causing Gray Blight in Camellia sinensis in China PreviousNext DISEASE NOTE OPENOpen Access licenseFirst Report of Neopestalotiopsis piceana Causing Gray Blight in Camellia sinensis in ChinaQiaomei Wang, Ruijuan Yang, Yanmei Yang, Jie Lv, Wenshu Peng, Liang Yan, and Xianqi HuQiaomei Wanghttps://orcid.org/0000-0002-9352-1467College of Plant Protection, Yunnan Agricultural University, Kunming 650201, ChinaPuer Institute of Pu-Erh Tea, Pu’er 665000, ChinaCollege of Pu’er Tea, West Yunnan University of Applied Sciences, Pu’er 665000, ChinaSearch for more papers by this author, Ruijuan YangCollege of Food Science and Technology, Yunnan Agricultural University, Kunming 650201, ChinaSearch for more papers by this author, Yanmei YangCollege of Plant Protection, Yunnan Agricultural University, Kunming 650201, ChinaSearch for more papers by this author, Jie LvCollege of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, ChinaSearch for more papers by this author, Wenshu PengPuer Institute of Pu-Erh Tea, Pu’er 665000, ChinaCollege of Pu’er Tea, West Yunnan University of Applied Sciences, Pu’er 665000, ChinaSearch for more papers by this author, Liang Yan†Corresponding authors: L. Yan; E-mail Address: [email protected], and X. Hu; E-mail Address: [email protected]Puer Institute of Pu-Erh Tea, Pu’er 665000, ChinaCollege of Pu’er Tea, West Yunnan University of Applied Sciences, Pu’er 665000, ChinaSearch for more papers by this author, and Xianqi Hu†Corresponding authors: L. Yan; E-mail Address: [email protected], and X. Hu; E-mail Address: [email protected]College of Plant Protection, Yunnan Agricultural University, Kunming 650201, ChinaSearch for more papers by this authorAffiliationsAuthors and Affiliations Qiaomei Wang1 2 3 Ruijuan Yang4 Yanmei Yang1 Jie Lv5 Wenshu Peng2 3 Liang Yan2 3 † Xianqi Hu1 † 1College of Plant Protection, Yunnan Agricultural University, Kunming 650201, China 2Puer Institute of Pu-Erh Tea, Pu’er 665000, China 3College of Pu’er Tea, West Yunnan University of Applied Sciences, Pu’er 665000, China 4College of Food Science and Technology, Yunnan Agricultural University, Kunming 650201, China 5College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China Published Online:8 Jul 2023https://doi.org/10.1094/PDIS-07-22-1721-PDNAboutSectionsView articlePDFSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat View articleTea (Camellia sinensis L.) is a major cash crop grown worldwide (Palanisamy et al. 2014). Tea gray blight is an important tea plant disease as it can reduce quality and reduce yields by 20 to 30% (Sanjay et al. 2008). In August 2018, a disease survey was conducted on 400 ha of organic tea plantations in the Puʼer area of Yunnan Province (22.48°N, 100.58°E). Disease was causing damage to 40% of the tea plantations. The most affected variety was Yunkang No. 10, with an average disease incidence of 30 to 35%. Affected leaves had small yellow-green spots on their tips or margins in the early stage that expanded into round or irregular brown spots with distinct concentric whorls and black conidial disks arranged in whorls when the humidity was high, which is consistent with tea gray blight disease (Zheng et al. 2021). Twenty-four diseased leaf samples were collected from four different tea plantations and taken to the Pu-Erh Tea Research Laboratory. Leaves with disease spots were cut into 4- × 4-mm square pieces, surface sterilized with 75% alcohol for 1 min, disinfected with 1% NaOCl for 3 min, and washed thrice with sterile water. The tissue pieces were placed on PDA plates containing 100 μg/ml of chloramphenicol (Wang et al. 2021). After 3 days in the dark at 28°C, 20 pure cultures with similar morphology were obtained; two representative isolates, C-5 and B-3, were transferred to new PDA. After 7 days, circular fungal colonies with dense aerial mycelium produced black, wet spore masses that grew on the PDA. Conidia were spindle shaped with four septa and 25.0 (21.0 to 26.0) × 6.0 (4.5 to 7.0) μm (n = 15). Conidia had three median cells, two of which were dark brown with unclear separations, with one basal hyaline appendage 3.8 (3.5 to 4.5) μm (n = 30) long and two to three apical hyaline appendages 31 (27 to 35) μm long (n = 30), similar to the conidia of Neopestalotiopsis piceana (Maharachchikumbura et al. 2014). Two isolates were selected for DNA extraction. The internal transcribed spacer (ITS) region, partial translation elongation factor 1-alpha (tef1-α) gene, and partial β-tubulin (tub2) gene were amplified using the primer sets ITS1F-ITS4 (White et al. 1990), EF-1α-F and EF-1α-R (Li et al. 2018), and tub1 and tub2, respectively (Chauhan et al. 2007). The ITS (OP535632 to OP535632), tef1-α (OP589285, OP589287), and tub2 (OP589286, OP589288) sequences were submitted to GenBank. BLAST analysis found that the sequences were 100% similar to those of N. piceana isolates in GenBank. The sequences were compared using the Mafft software package, and sequences with the same ID were concatenated using scripts. A maximum likelihood phylogenetic tree was constructed using the MEGA (ver. 5.1) software package based on the concatenated sequences (ITS, tef1-α, and tub2). Phylogenetic analysis revealed that C-5 and B-3 showed 95% bootstrap support with N. piceana isolates in references. According to the morphology and molecular characterization, C-5 and B-3 were identified as N. piceana. Pathogenicity of the two isolates was tested using 36 healthy tea plants. Leaves were scratched slightly with sterile toothpick tips, and pathogen cakes (6-mm diameter) were placed on the wounds with the mycelial side facing down and covered with sterile absorbent cotton to maintain moisture. Control leaves were wounded and covered with sterile PDA plugs (three replicates per treatment, three plants per replicate). Seven days later, inoculated leaves had similar symptoms to those observed under natural conditions, and control leaves had no symptoms. The same isolates as the introduced strains were isolated from the diseased tea leaves, completing Koch’s postulates. To our knowledge, this is the first report of N. piceana causing gray blight on tea leaves in China. These results are valuable for the prevention and management of gray blight on tea leaves.The author(s) declare no conflict of interest.References:Chauhan, J. B., et al. 2007. Indian J. Biotechnol. 6:404. Google ScholarLi, D. X., et al. 2018. J. Trop. Crops. 39:1827. Google ScholarMaharachchikumbura, S. N., et al. 2014. Stud. Mycol. 79:121. https://doi.org/10.1016/j.simyco.2014.09.005 Crossref, ISI, Google ScholarPalanisamy, S., and Mandal, A. K. A. 2014. Appl. Biochem. Biotechnol. 172:216. https://doi.org/10.1007/s12010-013-0529-z Crossref, ISI, Google ScholarSanjay, R., et al. 2008. Crop Protect 27:689. https://doi.org/10.1016/j.cropro.2007.09.014 Crossref, ISI, Google ScholarWang, Q. M., et al. 2021. Front. Microbiol. 12:654058. https://doi.org/10.3389/fmicb.2021.654058 Crossref, ISI, Google ScholarWhite, T. J., et al. 1990. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA. Crossref, Google ScholarZheng, S., et al. 2021. Plant Dis. 105:3723. https://doi.org/10.1094/PDIS-02-21-0318-A Link, ISI, Google ScholarFunding: Funding was provided by the Major Science and Technology Projects in Yunnan Province (2018ZG011) and by the Project of Yunnan Provincial Science and Technology Department (202104BI090008).The author(s) declare no conflict of interest.DetailsFiguresLiterature CitedRelated Vol. 107, No. 7 July 2023SubscribeISSN:0191-2917e-ISSN:1943-7692 Download Metrics Article History Issue Date: 26 Jul 2023Published: 8 Jul 2023First Look: 26 Dec 2022Accepted: 22 Dec 2022 Page: 2229 Information© 2023 The American Phytopathological SocietyFundingMajor Science and Technology Projects in Yunnan ProvinceGrant/Award Number: 2018ZG011Project of Yunnan Provincial Science and Technology DepartmentGrant/Award Number: 202104BI090008KeywordsCamellia sinensisNeopestalotiopsis piceanapathogentea gray blightThe author(s) declare no conflict of interest.PDF downloadCited byEffects of Pu-erh and Dian Hong tea polyphenols on the gut-liver axis in mice2 June 2023 | AMB Express, Vol. 13, No. 1