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• W4205640168 abstract "HomePlant DiseaseVol. 106, No. 6First Report of Epicoccum layuense Causing Leaf Brown Spot on Camellia oleifera in Hefei, China PreviousNext DISEASE NOTE OPENOpen Access licenseFirst Report of Epicoccum layuense Causing Leaf Brown Spot on Camellia oleifera in Hefei, ChinaXiang Xie, Shiqiang Zhang, Qingjie Yu, Xinye Li, Yongsheng Liu, and Lihuan WangXiang XieSchool of Horticulture, State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, ChinaSearch for more papers by this author, Shiqiang ZhangSchool of Horticulture, State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, ChinaSearch for more papers by this author, Qingjie YuSchool of Horticulture, State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, ChinaSearch for more papers by this author, Xinye LiSchool of Horticulture, State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, ChinaSearch for more papers by this author, Yongsheng Liu†Corresponding authors: Y. Liu; E-mail Address: liuyongsheng1122@ahau.edu.cn, and L. Wang; E-mail Address: Wanglihuan0928@ahau.edu.cnSchool of Horticulture, State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, ChinaMinistry of Education Key Laboratory for Bio-resource and Eco-environment, College of Life Science, State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu 610064, ChinaSearch for more papers by this author, and Lihuan Wang†Corresponding authors: Y. Liu; E-mail Address: liuyongsheng1122@ahau.edu.cn, and L. Wang; E-mail Address: Wanglihuan0928@ahau.edu.cnhttps://orcid.org/0000-0002-7522-4629School of Horticulture, State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, ChinaSearch for more papers by this author AffiliationsAuthors and Affiliations Xiang Xie1 Shiqiang Zhang1 Qingjie Yu1 Xinye Li1 Yongsheng Liu1 2 † Lihuan Wang1 † 1School of Horticulture, State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, China 2Ministry of Education Key Laboratory for Bio-resource and Eco-environment, College of Life Science, State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu 610064, China Published Online:21 Apr 2022https://doi.org/10.1094/PDIS-08-21-1726-PDNAboutSectionsView articlePDFSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat View articleCamellia oleifera is a major tree species for producing edible oil from China. Its oil, also called “eastern olive oil,” has high economic value due to its richness in a variety of healthy fatty acids (Lin et al. 2018). However, leaves are susceptible to leaf spot disease (Zhu et al. 2014). In May 2021, we found circular to irregular reddish-brown lesions, 4 to 11 mm in diameter, near the leaf veins or leaf edges on 30 to 50% of leaves of one-third of C. oleifera in a garden of Hefei City, Anhui Province, China (31.86°N, 117.27°E). To isolate the causal agents, symptomatic leaves were cut from the junction of diseased and healthy tissues (5 × 5 mm), treated with 70% alcohol for 30 s and 1% NaClO for 5 min, and inoculated onto PDA medium for culture. After 3 days, hyphal tips were transferred to PDA. Five isolates were obtained and cultured on PDA at 25°C for 7 days. The mycelia appeared yellow with a white edge and secreted a large amount of orange-red material to the PDA. Twenty days later, the mycelium appeared reddish-brown, and subcircular (3 to 10 mm) raised white or yellow mycelium was common on the Petri dish with occasional black particles. The colonies on the PDA produced abundant conidia. Microscopy revealed that conidia were globular to pyriform, dark, verrucose, multicellular, and 14.2 to 25.3 μm (x¯ = 19.34 μm, n = 30) in diameter. The morphological characteristics of mycelia and conidia from these isolates were similar to those of Epicoccum layuense (Chen et al. 2020). To further determine the species, DNA was extracted from 7-day-old mycelia cultures and the PCR-amplified fragments were sequenced for internal transcribed spacer (ITS), beta-tubulin, and 28S large subunit ribosomal RNA (LSU) gene regions ITS1/ITS4, Bt2a/Bt2b, and LR0R/LR5, respectively, followed by sequencing and molecular phylogenetic analysis of the sequence analysis (Glass and Donaldson 1995; Vilgalys and Hester 1990; White et al. 1990). Sequence analysis revealed that ITS, beta-tubulin, and LSU divided these isolates into two groups. The isolates AAU-NCY1, representing the first group (AAU-NCY1 and AAU-NCY5), and AAU-NCY2, representing the second group (AAU-NCY2, AAU-NCY3, and AAU-NCY4), were used for further studies. Based on BLASTn analysis, the ITS sequences of AAU-NCY1 (MZ477250) and AAU-NCY2 (MZ477251) showed 100 and 99.6% identity with E. layuense accessions MN396393 and KY742108, respectively. The beta-tubulin sequences (MZ552310; MZ552311) showed 99.03 and 99.35% identity with E. layuense accessions MN397247 and MN397248, respectively. The LSU sequences (MZ477254; MZ477255) showed 99.88 and 99.77% identity with E. layuense accessions MN328724 and MN396395, respectively. Phylogenetic trees were built by the maximum likelihood method (1,000 replicates) using MEGA v.6.0 based on the concatenated sequences of ITS, beta-tubulin, and LSU. Phylogenetic tree analysis confirmed that AAU-NCY1 and AAU-NCY2 are closely clustered with E. layuense strains. To test pathogenicity, a conidial suspension of AAU-NCY2 (106 spores/ml) was prepared and sterile water was used as the control. Twelve healthy leaves (six per treatment) on a C. oleifera tree were punched with a sterile needle (0.8 to 1 mm) and the sterile water or spore suspension was added dropwise at the pinhole. The experiment was repeated three times. By 10 days post inoculation, leaves infected by the conidia developed reddish-brown necrotic spots similar to those observed in the garden, while control leaves remained asymptomatic. DNA sequences from the strain reisolated from the infected leaves were identical to those of the original strain. E. layuense has been reported to cause leaf spot on C. sinensis (Chen et al. 2020), and similar pathogenic phenotypes were reported on Weigela florida (Tian et al. 2021) and Prunus × yedoensis Matsumura in Korea (Han et al. 2021). To our knowledge, this is the first report of E. layuense causing leaf spot on C. oleifera in Hefei, China.The author(s) declare no conflict of interest.References:Chen, Y. J., et al. 2020. Plant Dis. 104:2029. https://doi.org/10.1094/PDIS-09-19-1906-PDN Link, ISI, Google ScholarGlass, N. L., and Donaldson, G. C. 1995. Appl. Environ. Microbiol. 61:1323. https://doi.org/10.1128/aem.61.4.1323-1330.1995 Crossref, ISI, Google ScholarHan, V. C., et al. 2021. Plant Dis. 105:2734. https://doi.org/10.1094/PDIS-12-20-2711-PDN Link, ISI, Google ScholarLin, P., et al. 2018. Int. J. Mol. Sci. 19:118. https://doi.org/10.3390/ijms19010118 Crossref, Google ScholarTian, Y., et al. 2021. Plant Dis. 105:2243. https://doi.org/10.1094/PDIS-07-20-1498-PDN Link, ISI, Google ScholarVilgalys, R., and Hester, M. 1990. J. Bacteriol. 172:4238. https://doi.org/10.1128/jb.172.8.4238-4246.1990 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. Google ScholarZhu, H., et al. 2014. Plant Dis. 98:1427. https://doi.org/10.1094/PDIS-11-13-1166-PDN Link, ISI, Google ScholarFunding: This study was supported by grants from the National Natural Science Foundation of China (31900257).The author(s) declare no conflict of interest.DetailsFiguresLiterature CitedRelated Vol. 106, No. 6 June 2022SubscribeISSN:0191-2917e-ISSN:1943-7692 Download Metrics Downloaded 409 times Article History Issue Date: 2 Jun 2022Published: 21 Apr 2022First Look: 12 Jan 2022Accepted: 2 Jan 2022 Page: 1755 Information© 2022 The American Phytopathological SocietyFundingNational Natural Science Foundation of ChinaGrant/Award Number: 31900257KeywordsCamellia oleiferaEpicoccum layuensefungipathogen detectionThe author(s) declare no conflict of interest.PDF download" @default.
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