油茶果生刺盘孢液泡分选蛋白CfVps26的功能

doi:10.11707/j.1001-7488.20210809

Abstract

Abstract:

Objective: Colletotrichum fructicola is a major epidemic and causes anthracnose of Camellia oleifera. This paper aims to study the biological functions of the vacuolar protein sorting Vps26 in C. fructicola to provide a theoretical basis for the prevention and control of oil-tea tree anthracnose, and also for further analysis of Vps26 action mechanism. Method: The over-lap method was used to construct the CfVPS26 gene knockout vector fragment which was transformed into the protoplast of C. fructicola by using PEG-mediated transformation method to obtain the mutant strain of ΔCfvps26. The PCR-amplified CfVPS26 gene-containing complement of the promoter was taken to construct a complementary vector. Then the complementary vector was transformed into the mutant protoplasts. The complemented strain was screened by fluorescence. The biological phenotypes, such as vegetative growth, appressorium formation, external environmental pressure, glycogen metabolism and pathogenicity, of the wild-type strain, mutant strain, and the complemented strain were measured. Result: The results showed that the mycelial growth rate of ΔCfvps26 was significantly lower than that of the wild type strain and the complemented strain. And the aerial hyphae were significantly decreased, and more sensitive to 200 μg·mL^-1 calcofluor white and 0.01% sodium dodecyl sulfate. Compared with the wild type and the complemented strain, the tolerance of the mutant strain ΔCfvps26 to dithiothreitol was enhanced. The conidia production and appressorium formation rate of CfVPS26 gene deletion mutant were significantly reduced, and glycogen metabolism was blocked. The mutant strain almost lost pathogenicity on the leaves of C. oleifera. Conclusion: This study reveals that the vacuolar sorting protein CfVps26 plays critical roles in growth and development, condiation, appressorium formation, tolerance to external stresses, glycogen metabolism and pathogenicity in C. fructicola on C. oleifera. These results provide potential target sites for the development of new fungicides against C. fructicola.

Key words: Camellia oleifera, Colletotrichum fructicola, CfVps26, pathogenicity

CLC Number:

S718.46

Xiya Li,Shengpei Zhang,He Li. Function of Vacuolar Protein Sorting CfVps26 in Colletotrichum fructicola on Camellia oleifera[J]. Scientia Silvae Sinicae, 2021, 57(8): 94-101.

Figures/Tables 7

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引物名称 Primer	序列 Sequence(5’—3’)	目的 Purpose
CfVPS26-1F	GATAGAATAAGCGGATGCGG	扩增CfVPS26 5′侧翼序列 Amplify CfVPS26 5′ flank sequence
CfVPS26-2R	TTGACCTCCACTAGCTCCAGCCAAGCCCTTGGCTTCGGTGGTTACCC	扩增CfVPS26 5′侧翼序列 Amplify CfVPS26 5′ flank sequence
CfVPS26-3F	CAAAGGAATAGAGTAGATGCCGACCGAGCTGTGTTGCAATGCTGCA	扩增CfVPS26 3′侧翼序列 Amplify CfVPS26 3′flank sequence
CfVPS26-4R	GAGATGCTTAGTCGCCAGTT	扩增CfVPS26 3′侧翼序列 Amplify CfVPS26 3′flank sequence
CfVPS26-5F	GCATGCATTCAATTGCCTGC	CfVPS26基因缺失验证 Validation of CfVPS26 gene deletion
H855R	GCTGATCTGACCAGTTGC	CfVPS26基因缺失验证 Validation of CfVPS26 gene deletion
CfVPS26-7F	CGGCAAGCGACTAGAGCATA	扩增CfVPS26基因序列 Amplify CfVPS26 gene sequence
CfVPS26-8R	ATCCCGGAATGTAGGGGTCA	扩增CfVPS26基因序列 Amplify CfVPS26 gene sequence
CfVPS26-9F	ACTCACTATAGGGCGAATTGGGTACTCAAATTGGTTACACCGACAACCAGGTTGTT	扩增回补序列 Amplify complemented sequence
CfVPS26-10R	CACCACCCCGGTGAACAGCTCCTCGCCCTTGCTCACCGCTTGCGAGGGCTGTGGCA	扩增回补序列 Amplify complemented sequence
Hyg F	GGCTTGGCTGGAGCTAGTGGAGGTCAA	扩增HPH序列 Amplify HPH sequence
Hyg R	CGGTCGGCATCTACTCTATTCCTTTG	扩增HPH序列 Amplify HPH sequence
GFP-R	GACACGCTGAACTTGTGGCCGTT	验证回补序列 Validation of complemented sequence

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Function of Vacuolar Protein Sorting CfVps26 in Colletotrichum fructicola on Camellia oleifera

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