油茶果炭疽病病原菌分离鉴定及防治药剂筛选

Isolation and identification of anthracnose pathogens in Camellia oleifera fruit and fungicides screening

摘要

为明确引起油茶果炭疽病的病原菌种类并筛选高效防治药剂，本研究从湖南省长沙市、浏阳市、永州市、常德市、益阳市、衡阳市和贵州省贵阳市 7 个样地采集病果，经组织分离与纯化，获得 132 株炭疽菌。通过形态学观察、致病性测定及基于 ApMat 基因序列的系统发育分析，共鉴定出 4 种炭疽菌:山茶炭疽菌 Colletotrichum camelliae Massee 124 株，占 93.9%；果生炭疽菌 C. fructicola Prihastuti., L. Cai and K.D. Hyde 5 株，占 3.8%；暹罗炭疽菌 C. siamense Prihastuti., L. Cai and K.D. Hyde 2 株，占 1.5%；胶孢炭疽菌 C. gloeosporioides (Penz.) Penz. & Sacc 1 株，占 0.8%。致病力测定结果显示:4 种菌均可侵染油茶果实和叶片，其中山茶炭疽菌对果实致病力最强，果生炭疽菌对叶片致病力最强。采用菌丝生长速率法，测定多菌灵、苯醚甲环唑和戊唑醇对优势种山茶炭疽菌的毒力，多菌灵和苯醚甲环唑的 EC₅₀ 均值分别为 0.072 5、0.183 8 μg·mL⁻¹，抑菌活性强；戊唑醇毒力稍弱，EC₅₀ 为 0.365 6 μg·mL⁻¹。研究结果表明，山茶炭疽菌是湖南省和贵州省油茶果炭疽病的优势致病菌，多菌灵和苯醚甲环唑可作为田间防治的候选药剂。

Abstract

This study aimed to clarify the species of Colletotrichum causing anthracnose on Camellia oleifera fruits and to evaluate the in vitro toxicity of selected fungicides against the dominant pathogenic species，thereby providing a basis for targeted disease management. Diseased fruits were collected from seven sites in Hunan(Changsha，Liuyang，Yongzhou，Changde，Yiyang，Hengyang)and Guiyang，Guizhou Province. Fungal isolates were obtained through tissue isolation and purification. Species were identified by combining morphological characteristics，pathogenicity tests using Koch’s postulates，and phylogenetic analysis based on ApMat gene sequences with the neighbor-joining method. The in vitro toxicity of three fungicides against the dominant species was determined using the mycelial growth inhibition method. A total of 132 Colletotrichum isolates were obtained，and four species were identified: C. camelliae Massee(124 isolates，93.9%)， C. fructicola Prihastuti.，L. Cai and K.D. Hyde(5 isolates，3.8%)， C. siamense Prihastuti.，L. Cai and K.D. Hyde(2 isolates，1.5%)，and C. gloeosporioides(Penz.)Penz. & Sacc(1 isolate，0.8%). All four species infected both fruits and leaves. C. camelliae exhibited the highest virulence on fruits，while C. fructicola was the most virulent on leaves. Toxicity assays against C. camelliae showed that carbendazim and difenoconazole had strong inhibitory effects，with average EC ₅₀ values of 0.072 5 and 0.183 8 μg·mL ⁻¹，respectively，whereas tebuconazole was less effective，with an average EC ₅₀ of 0.365 6 μg·mL ⁻¹. The results show that C. camelliae is the dominant pathogenic species causing fruit anthracnose on C. oleifera in Hunan and Guizhou and exhibits the highest virulence. Carbendazim and difenoconazole are recommended as candidate fungicides for field efficacy trials against this disease.

关键词

炭疽病 / 油茶 / 病原菌 / 种类鉴定 / 致病性 / 毒力测定

Key words

anthracnose / Camellia oleifera / pathogen / species identification / pathogenicity / toxicity test

参考文献

[1] 陈守常, 田泽君, 郭隆錫. 1965. 油茶炭疽病的发生与蔓延[J]. 植物保护学报(3):207-218.

[2] Chen S C, Tian Z J, Guo L X, et al. 1965. Development and spread of the anthracnose of Camellia oleosa(lour.)rehd [J]. Journal of Plant Protection(3):207-218.(in Chinese)

[3] 刁春玲, 刘芳, 宋宝安. 2006. 农用杀菌剂作用机理的研究进展[J]. 农药(6):374-377.

[4] Diao C L, Liu F, Song B A . 2006. Research progress on the mechanism of action of agricultural fungicides[J]. Agrochemicals(6):374-377.(in Chinese)

[5] 范昆, 张雪丹, 余贤美, 等. 2013. 无花果炭疽病菌的生物学特性及 8 种杀菌剂对其抑制作用[J]. 植物病理学报, 43(1):75-81.

[6] Fan K, Zhang X D, Yu X M, et al. 2013. Biological characteristics of Colletotrichum gloeosporioides and inhibitory effects of eight fungicides [J]. Acta Phytopathologica Sinica, 43(1):75-81.(in Chinese)

[7] 靳爱仙, 周国英, 李河. 2009. 油茶炭疽病的研究现状、问题与方向[J]. 中国森林病虫, 28(2):27-31.

[8] Jin A X, Zhou G Y, Li H . 2009. Progress，problem and prospect of oil camelliae anthracnose( Colletotrichum gloeosporioides) research [J]. Forest Pest and Disease, 28(2):27-31.(in Chinese)

[9] 李河, 李司政, 王悦辰, 等. 2019. 油茶苗圃炭疽病原菌鉴定及抗药性[J]. 林业科学, 55(5):85-94.

[10] Li H, Li S Z, Wang Y C, et al. 2019. Identification of the pathogens causing anthracnose of Camellia oleifera in nursery and their resistence to fungicides [J]. Scientia Silvae Sinicae, 55(5):85-94.(in Chinese)

[11] 李河, 李杨, 蒋仕强, 等. 2017. 湖南省油茶炭疽病病原鉴定[J]. 林业科学, 53(8):43-53.

[12] Li H, Li Y, Jiang S Q, et al. 2017. Pathogen of oil—tea trees anthracnose caused by Colletotrichum spp. in Hunan Province [J]. Scientia Silvae Sinicae, 53(8):43-53.(in Chinese)

[13] 李河, 周国英, 徐建平. 2015. 一种新油茶炭疽病原多基因序列鉴定[J]. 植物保护, 41(2):92-96.

[14] Li H, Zhou G Y, Xu J P . 2015. Pathogen identification of a new anthracnose of Camellia oleifera in China based on multiple—gene sequences [J]. Plant Protection, 41(2):92-96.(in Chinese)

[15] 李河, 周国英, 徐建平, 等. 2014. 一种油茶新炭疽病原的多基因系统发育分析鉴定[J]. 植物保护学报, 41(5):602-607.

[16] Li H, Zhou G Y, Xu J P, et al. 2014. Pathogen identification of a new anthracnose of Camellia oleifera in China based on multiple—gene phylogeny [J]. Acta Phytophylacica Sinica, 41(5):602-607.(in Chinese)

[17] 李河. 2018. 油茶炭疽病菌群体遗传及 MAPK 基因 CfPMK1 功能研究 [D]. 长沙: 中南林业科技大学.

[18] Li H. 2018. Population genetic analyses of the fungal pathogen colletotrichum on tea—oil trees in China and characterization of a MAPK Gene CfPMK1 in the pathogen [D]. Changsha: Central South University of Forestry and Technology.(in Chinese)

[19] 李科澄, 李河. 2025. 湖南省杉木炭疽病病原菌鉴定及抗药性研究[J]. 树木医学, 2(3):45-55.

[20] Li K C, Li H . 2025. Identification and fungicide resistance of anthracnose pathogen in Cunninghamia lanceolata in Hunan Province [J]. Tree Health, 2(3):45-55.(in Chinese)

[21] 梁子超, 岑炳沾, 戴品初. 1964. 油茶炭疽病菌的有性世代、越冬方式和侵入途径[J]. 植物保护学报(3):316-317.

[22] Liang Z C, Cen B Z, Dai P C . 1964. Studies on anthracnose of Camellia oleosa(Lour.)Rehd. I. the perfect stage, mrode of overwintering and ingress of the pathogen [J]. Acta Phytophylacica Sinica(3):316-317.(in Chinese)

[23] 刘宏玉. 2024. 油茶叶斑病原菌的分离鉴定及重要种属室内药剂筛选[D]. 长沙: 中南林业科技大学.

[24] Liu H Y . 2024. Isolation and identification of pathogenic fungi causing leaf spot on Camellia oleifera and fungicides screening of important species in laboratory [D]. Changsha: Central South University of Forestry and Technology.(in Chinese)

[25] 帅小春, 阮成江, 熊朝伟, 等. 2019. 油茶炭疽病新致病菌 Colletotrichum kahawae 的多基因序列鉴定 [J]. 分子植物育种, 17(18):5939-5945.

[26] Shuai X C, Ruan C J, Xiong C W, et al. 2019. Identification of pathogen sequence of new pathogenic strain Colletotrichum kahawae of Camellia oleifera anthracnose [J]. Molecular Plant Breeding, 17(18):5939-5945.(in Chinese)

[27] 宋丹丹, 张伊莹, 张琳婧, 等. 2016. 杨树炭疽病菌对多菌灵及 3 种 DMIs 杀菌剂的敏感性[J]. 农药学学报, 18(5):567-574.

[28] Song D D, Zhang Y Y, Zhang L J, et al. 2016. Sensitivities of poplar anthracnose fungi isolates to carbendazim and three C—14α—demethylation inhibitors[J]. Chinese Journal of Pesticide Science, 18(5):567-574.(in Chinese)

[29] 孙伟, 袁会珠, 陈淑宁. 2022. 油茶炭疽病病原鉴定及绿色防治药剂筛选[J]. 中国生物防治学报, 38(4):880-889.

[30] Sun W, Yuan H Z, Chen S N . 2022. Identification of Colletotrichum species associated with anthracnose of Camellia oleifera and screening of fungicides [J]. Chinese Journal of Biological Control, 38(4):880-889.(in Chinese)

[31] 谭晓风, 马履一, 李芳东, 等. 2012. 我国木本粮油产业发展战略研究[J]. 经济林研究, 30( 1):1-5.

[32] Tan X F, Ma L, Li F D, et al. 2012. Industrialization development strategy of woody grain and oil in China[J]. Non—wood Forest Research, 30( 1):1-5.(in Chinese)

[33] 王静, 叶敏, 范黎明, 等. 2013. 菌丝生长速率法筛选纤维素降解菌的研究[J]. 中国农学通报, 29(33):323-326.

[34] Wang J, Ye M, Fan L M, et al. 2013. Screening of cellulose degrading fungi by Mycelium growth rate method [J]. Chinese Agricultural Science Bulletin, 29(33):323-326.(in Chinese)

[35] 杨光道. 2009. 油茶品种对炭疽病的抗性机制研究[D]. 合肥: 安徽农业大学.

[36] Yang G D . 2009. Resistance mechanism of Camellia oleifera cultivars to Colletotrichum gloeosporioides [D]. Hefei: Anhui Agricultural University.(in Chinese)

[37] 叶滔, 马志强, 毕秋艳, 等. 2012. 植物病原真菌对甾醇生物合成抑制剂类(SBIs)杀菌剂的抗药性研究进展[J]. 农药学学报, 14( 1):1-16.

[38] Ye T, Ma Z Q, Bi Q Y, et al. 2012. Research advances on the resistance of plant pathogenic fungi to SBIs fungicides[J]. Chinese Journal of Pesticide Science, 14( 1):1-16.(in Chinese)

[39] 袁嗣令, 张能唐, 翁月霞, 等. 1963. 油茶炭疽病的研究[J]. 植物保护学报(3):253-262.

[40] Yuan S L, Zhang N T, Weng Y X, et al. 1963. The anthracnose disease of Camellia oleosa(lour)behd [J]. Journal of Plant Protection(3):253-262.(in Chinese)

[41] Cannon P F, Buddie A G, Bridge P D. 2008. The typification of Colletotrichum gloeosporioides [J]. Mycotaxon, 104: 189-204.

[42] Dickens J S W, Cook R T A. 1989. Glomerella cingulata on Camellia [J]. Plant Pathology, 38(1):75-85.

[43] Kumar S, Stecher G, Tamura K. 2016. MEGA7:Molecular evolutionary genetics analysis version 7.0 for bigger datasets[J]. Molecular Biology and Evolution, 33(7):1870-1874.

[44] Li H, Zhou G Y, Qi X Y ,et al. 2018. First report of Colletotrichum henanense causing anthracnose on tea—oil trees in China [J]. Plant Disease, 102(5):1040.

[45] Li L L, Li H. 2022. First report of Colletotrichum aeschynomenes causing anthracnose on Camellia oleifera in China [J]. Forest Pathology, 52(5):e12770.

[46] Li S Z, Li H. 2020. First report of Colletotrichum nymphaeae causing anthracnose on Camellia oleifera in China [J]. Plant Disease, 104(6):1860.

[47] Peng X J, Wang Q C, Zhang S K ,et al. 2023. Colletotrichum species associated with Camellia anthracnose in China [J]. Mycosphere, 14(2):130-157.

[48] Prihastuti H, Cai L, Chen H ,et al. 2009. Characterization of Colletotrichum species associated with coffee berries in northern Thailand [J]. Fungal Diversity, 39: 89-109.

[49] Shiori Y, Naoki E, Ken—ichi K ,et al. 2017. Phylogenetic relationship and fungicide sensitivity of members of the Colletotrichum gloeosporioides species complex from apple [J]. Journal of General Plant Pathology, 83(5):291-298.

[50] Silva D N, Talhinhas P, Várzea V ,et al. 2012. Application of the Apn2/MAT locus to improve the systematics of the Colletotrichum gloeosporioides complex:An example from coffee (Coffea spp.)hosts [J]. Mycologia, 104(2):396-409.

[51] Wang Y X, Chen J Y, Xu X W ,et al. 2020. Identification and characterization of Colletotrichum species associated with anthracnose disease of Camellia oleifera in China [J]. Plant Disease, 104(2):474-482.

[52] Weir B S, Johnston P R, Damm U. 2012. The Colletotrichum gloeosporioides species complex [J]. Studies in Mycology, 73: 115-180.

基金资助

国家重点研发计划项目(2023YFD1401301)