Isolation and identification of pathogen causing anthracnose disease on a rare plant species Heptacodium miconioides subsp. jasminoides

doi:10.3969/j.issn.1004-1524.2023.01.12

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (1): 112-120.DOI: 10.3969/j.issn.1004-1524.2023.01.12

• Plant Protection • Previous Articles Next Articles

Isolation and identification of pathogen causing anthracnose disease on a rare plant species Heptacodium miconioides subsp. jasminoides

ZHU Yan¹(), ZHOU Mengya¹, ZHU Yinuo¹, HE Haiye¹, BAO Honghua², ZHANG Huijuan¹, JIANG Ming¹^,^*()

1. College of Life Sciences, Taizhou University, Taizhou 318000, Zhejiang, China
2. Taizhou Municipal Ecology and Environment Bureau Luqiao Branch, Taizhou 318050, Zhejiang, China

Received:2022-04-06 Online:2023-01-25 Published:2023-02-21

Abstract

Abstract:

Anthracnose is one of the most important diseases of Heptacodium miconioides subsp. jasminoides, which seriously affects plant growth and threatens the survival of this rare species. To clarify the causative pathogen, infected leaves of H. miconioides subsp. jasminoides were collected, and the pathogen was obtained by the tissue isolation method. After determining the pathogenicity of the isolated strains, both morphological and multi-gene approaches were applied to identify the causative pathogen. Three strains were isolated, and only one isolate namely QZH could cause anthracnose disease according to Koch’s Rule. The morphological characteristics were identical with those of Colletotrichum fructicola. The multi-gene analysis results indicated that QZH and C. fructicola shared the same clade, with high support rates. Based on the morphological characteristics and multi-gene identification results, it was confirmed that the pathogen causing anthracnose disease of H. miconioidessubsp. jasminoides was C. fructicola. These data provided a start point for both disease prevention and pathogenic mechanism studies in the future.

Key words: Heptacodium miconioides subsp. jasminoides, anthraconse disease, Colletotrichum fructicola

CLC Number:

ZHU Yan, ZHOU Mengya, ZHU Yinuo, HE Haiye, BAO Honghua, ZHANG Huijuan, JIANG Ming. Isolation and identification of pathogen causing anthracnose disease on a rare plant species Heptacodium miconioides subsp. jasminoides[J]. Acta Agriculturae Zhejiangensis, 2023, 35(1): 112-120.

Figures/Tables 7

Table 1 PCR primer sequences used in this study

基因 Gene name	引物名称 Primer name	引物序列 Primer Sequence(5'→3')
ITS	ITS1	TCCGTAGGTGAACCTGCGG
	ITS4	TCCTCCGCTTATTGATATGC
CHS-1	79F	TGGGGCAAGGATGCTTGGAAGAAG
	354R	TGGAAGAACCATCTGTGAGAGTTG
ACTIN	512F	ATGTGCAAGGCCGGTTTCGC
	783R	TACGAGTCCTTCTGGCCCAT
GAPDH	GDF1	CCGTCAACGACCCCTTCATTGA
	GDR1	GGGTGGAGTCGTACTTGAGCATGT

Fig.1 Anthracnose disease of Heptacodium miconioides subsp. Jasminoides A, Infected plant; B, Infected leaf; C, Leaf lesion.

Fig.2 Result of pathogenicity detection A, Leaf inoculated with QZH; B,The front view of an inoculation site; C, The back view of an inoculation site.

Fig.3 Morphological characteristics of pathogenic fungus QZH A, Colony morphology; B, Morphology of spores under a microscope; C and D: Morphology of mycelium under scanning electron microscope.

Fig.4 PCR products of different gene fragments M, DL 2000 DNA marker; 1 and 2, ITS; 3 and 4, CHS-1 gene; 5 and 6, ACTIN gene; 7 and 8, GAPDH gene.

Fig.5 Multiple alignment of QZH ACTIN gene sequence and its homologous sequences QZH,Colletotrichum fructicola; MN273216.1, C. fructicola; KF377525.1, C. viniferum; MN685845.1, C. henanense; MT602360.1, C. karsti; MN624087.1, C. gloeosporioides; MG747694.1, C. aenigma; MT407936.1, C. tropicale; KX093967.1, C. musae; MH622614.1, C. asianum; LC186032.1, C. horii.

Fig.6 Phylogenetic trees based on gene fragments of QZH and their homologous sequences A, QZH-ACT; B, QZH-CHS-1, C: QZH-GAPDH.

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Isolation and identification of pathogen causing anthracnose disease on a rare plant species Heptacodium miconioides subsp. jasminoides

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