茶树炭疽病病原菌鉴定及其致病性分析

doi:10.16420/j.issn.0513-353x.2021-0377

摘要/Abstract

摘要：

从来自12个省份茶区的茶树炭疽病病叶分离病原菌,基于形态学和多基因聚类分析对其进行鉴定,并对不同茶区分离的纯培养物致病性进行测定。结果表明,分离到的57株菌株有明显的形态学差异,根据其在PDA上的菌落形态和分生孢子特征可划分为5种类型。选取21个代表性菌株进行多位点（ITS、TUB2、ACT、CAL、CHS-1和GAPDH）的序列和聚类分析,发现测试菌株均属于胶孢炭疽复合种（Colletotrichum gloeosporioides complex）,包括7株果生炭疽菌（C. fructicola）、8株山茶炭疽菌（C. camelliae）、3株暹罗炭疽菌（C. siamense）和3株隐秘炭疽菌（C. aenigma）,其中山茶炭疽菌和果生炭疽菌在调查茶区的炭疽病叶中分离频率较高。采用刺伤接种法将测试菌株接种于‘舒茶早’离体枝条的叶片,结果显示21个代表性菌株中17株能够侵染‘舒茶早’叶片并使其发病,但不同菌种表现出致病性差异,其中山茶炭疽菌表现出强致病力,同时发现同种病原菌不同菌株间也存在致病力差异。此外,在河南茶区首次发现由果生炭疽菌引起茶树炭疽病。

关键词: 茶树, 炭疽病, 病原菌鉴定, 聚类分析, 致病性

Abstract:

The samples were collected from tea leaves showing anthracnose symptoms from 12 different provinces,and the pathogen was isolated and identified by morphological characteristics and multi-locus phylogenetic analysis,along with the pathogenicity tests according to Koch’s postulates. The results showed that a total of 57 isolates were obtained and divided into five types based on the colony morphology and conidia characteristics. Twenty-one representative isolates were selected for further sequencing and phylogenic analysis based on six loci（ITS,TUB2,ACT,CAL,CHS-1 and GAPDH）. The results indicated that the tested isolates all belonged to Colletotrichum gloeosporioides complex,including seven isolates of C. fructicola,eight isolates of C. camelliae,three isolates of C. siamense and three isolates of C. aenigma. The separation frequency of C. camelliae and C. fructicola in tea area was higher. Pathogenicity assays were performed with 21 representative isolates from the four species of Colletotrichum. They were inoculated on the leaves of detached branches of Camellia sinensis‘Shuchazao’ by stabbing method. It was found that 17 isolates could cause necrosis at the inoculation site of tea leaves at two weeks post infection. Pathogenicity variation was observed among the four species when inoculated on‘Shuchazao’,and the isolates of C. camelliae showed strong virulence. In addition,different isolates of the same species displayed variant virulence. Moreover,this is the first report of C. fructicola causing anthracnose disease of Camellia sinensis in Henan Province,China.

Key words: Camellia sinensis, anthracnose disease, identification of pathogenic, phylogenic analysis, pathogenicity

中图分类号:

S571.1

贡长怡, 刘姣姣, 邓强, 张立新. 茶树炭疽病病原菌鉴定及其致病性分析[J]. 园艺学报, 2022, 49(5): 1092-1101.

GONG Changyi, LIU Jiaojiao, DENG Qiang, ZHANG Lixin. Identification and Pathogenicity of Colletotrichum Species Causing Anthracnose on Camellia sinensis[J]. Acta Horticulturae Sinica, 2022, 49(5): 1092-1101.

图/表 5

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菌种 Species	形态类型 Type of morphyology	采样地点 Sampling location	菌株编号 Isolate number	基因登录号 GenBank accession numbers						致病性 Pathogenicity
菌种 Species	形态类型 Type of morphyology	采样地点 Sampling location	菌株编号 Isolate number	ITS	ACT	CAL	CHS-1	GAPDH	TUB2	致病性 Pathogenicity
果生炭疽菌 C. fructicola	Ⅰ&Ⅱ	云南Yunnan	YN17	MN498091	MN525804	MN525832	MN525853	MN525874	MN525900	++
		四川Sichuan	SFD-3	MN498096	MN525800	MN525837	MN525858	MN525879	MN525902	++
		重庆Chongqing	CHQ-2	MN498088	MN525803	MN525829	MN525850	MN525870	MN525897	++
		杭州Hangzhou	ZLJ3-4	MN498090	MN525813	MN525831	MN525852	MN525873	MN525894	++
		福建Fujian	FLJ-6	MN498092	MN525814	MN525833	MN525854	MN525875	MN525898	—
		河南Henan	HXQT-2	MN498098	MN525801	MN525839	MN525860	MN525881	MN525885	+
		武汉Wuhan	HEC14-6	MN498085	MN525802	MN525826	MN525847	MN525868	MN525889	++
山茶炭疽菌 C. camelliae	Ⅲ	河南Henan	HXQT-3	MN498084	MN525809	MN525825	MN525846	MN525867	MN525890	++
		河南Henan	HXQT-5	MN498097	MN525818	MN525838	MN525859	MN525880	MN525886	++
		云南Yunnan	YN16	MN498081	MN525806	MN525822	MN525843	MN525864	MN525896	++
		福建Fujian	FWY43	MN498086	MN525810	MN525827	MN525848	MN525869	MN525895	++
		云南 Yunnan	YN5	MN498082	MN525807	MN525823	MN525844	MN525865	MN525893	++
		福建 Fujian	FWY11	MN498087	MN525811	MN525828	MN525849	MN525871	MN525899	++
		福建 Fujian	FWY41	MN498100	MN525820	MN525841	MN525862	MN525883	MN525884	++
		云南 Yunnan	YN23	MN498093	MN525815	MN525834	MN525855	MN525876	MN525888	++
暹罗炭疽菌 C. siamense	Ⅳ	云南 Yunnan	YN11	MN498094	MN525816	MN525835	MN525856	MN525877	MN525887	++
		云南Yunnan	YN22	MN498089	MN525812	MN525830	MN525851	MN525872	MN525892	+
		广东Guangdong	GHY12-3	MN498099	MN525819	MN525840	MN525861	MN525882	MN525901	—
隐秘炭疽菌 C. aenigma	Ⅴ	江苏Jiangsu	JSC-2	MN498083	MN525808	MN525824	MN525845	MN525866	MN525903	+
		江苏Jiangsu	JSC-3	MN498095	MN525817	MN525836	MN525857	MN525878	MN525904	—
		江苏Jiangsu	JYSN-2	MN498080	MN525805	MN525821	MN525842	MN525863	MN525891	—

菌种 Species	形态类型 Type of morphyology	采样地点 Sampling location	菌株编号 Isolate number	基因登录号 GenBank accession numbers						致病性 Pathogenicity
菌种 Species	形态类型 Type of morphyology	采样地点 Sampling location	菌株编号 Isolate number	ITS	ACT	CAL	CHS-1	GAPDH	TUB2	致病性 Pathogenicity
果生炭疽菌 C. fructicola	Ⅰ&Ⅱ	云南Yunnan	YN17	MN498091	MN525804	MN525832	MN525853	MN525874	MN525900	++
		四川Sichuan	SFD-3	MN498096	MN525800	MN525837	MN525858	MN525879	MN525902	++
		重庆Chongqing	CHQ-2	MN498088	MN525803	MN525829	MN525850	MN525870	MN525897	++
		杭州Hangzhou	ZLJ3-4	MN498090	MN525813	MN525831	MN525852	MN525873	MN525894	++
		福建Fujian	FLJ-6	MN498092	MN525814	MN525833	MN525854	MN525875	MN525898	—
		河南Henan	HXQT-2	MN498098	MN525801	MN525839	MN525860	MN525881	MN525885	+
		武汉Wuhan	HEC14-6	MN498085	MN525802	MN525826	MN525847	MN525868	MN525889	++
山茶炭疽菌 C. camelliae	Ⅲ	河南Henan	HXQT-3	MN498084	MN525809	MN525825	MN525846	MN525867	MN525890	++
		河南Henan	HXQT-5	MN498097	MN525818	MN525838	MN525859	MN525880	MN525886	++
		云南Yunnan	YN16	MN498081	MN525806	MN525822	MN525843	MN525864	MN525896	++
		福建Fujian	FWY43	MN498086	MN525810	MN525827	MN525848	MN525869	MN525895	++
		云南 Yunnan	YN5	MN498082	MN525807	MN525823	MN525844	MN525865	MN525893	++
		福建 Fujian	FWY11	MN498087	MN525811	MN525828	MN525849	MN525871	MN525899	++
		福建 Fujian	FWY41	MN498100	MN525820	MN525841	MN525862	MN525883	MN525884	++
		云南 Yunnan	YN23	MN498093	MN525815	MN525834	MN525855	MN525876	MN525888	++
暹罗炭疽菌 C. siamense	Ⅳ	云南 Yunnan	YN11	MN498094	MN525816	MN525835	MN525856	MN525877	MN525887	++
		云南Yunnan	YN22	MN498089	MN525812	MN525830	MN525851	MN525872	MN525892	+
		广东Guangdong	GHY12-3	MN498099	MN525819	MN525840	MN525861	MN525882	MN525901	—
隐秘炭疽菌 C. aenigma	Ⅴ	江苏Jiangsu	JSC-2	MN498083	MN525808	MN525824	MN525845	MN525866	MN525903	+
		江苏Jiangsu	JSC-3	MN498095	MN525817	MN525836	MN525857	MN525878	MN525904	—
		江苏Jiangsu	JYSN-2	MN498080	MN525805	MN525821	MN525842	MN525863	MN525891	—