Relationship Between BcPDR1 Gene and MAPK Pathway Genes BcBMP1 and BcBMP3 in Botrytis cinerea

doi:10.7668/hbnxb.20192641

Abstract

Abstract:

To clarify the relationship between BcPDR1 gene and MAPK pathway genes BcBMP1 and BcBMP3 of Botrytis cinerea.We analyzed the expression levels of BcBMP1 and BcBMP3 in mutants of BcPDR1 by Real-time PCR and the expression levels of BcPDR1 in RNAi mutants of BcBMP1 and BcBMP3.In the background of mutant ΔBcpdr1,strains ΔBcpdr1/BcBMP1-OE and ΔBcpdr1/BcBMP3-OE over-expressing BcBMP1 and BcBMP3 were constructed,and the phenotype and pathogenicity of the over-expressing strains were analyzed.It was found that the expression levels of BcBMP1 and BcBMP3 in BCt89 and ΔBcpdr1,gene mutants of BcPDR1,were significantly better than those of the wild-type strain and the revertant strain,and the expression levels of BcPDR1 in the RNAi mutants of BcBMP1 and BcBMP3 gene were obviously lower than those of the wild-type strain.The colony morphology,hyphal morphology,growth rate,sporulation,and pathogenicity of the over-expression isolates ΔBcpdr1/BcBMP1-OE and ΔBcpdr1/BcBMP3-OE were obviously different from ΔBcpdr1,and closer to the wild-type BC22.These results indicated that the BcPDR1 was closely related to the expression levels of BcBMP1 and BcBMP3 in B.cinerea.The BcPDR1 gene negatively regulates the expression of BcBMP1 and BcBMP3,while BcBMP1 and BcBMP3 genes positively regulate the expression of the BcPDR1.

Key words: Botrytis cinerea, BcPDR1, MAPK pathway genes, BcBMP1, BcBMP3

LIU Xiaoying, WEI Yadi, LI Bai, CAO Hongzhe, ZANG Jinping, ZHANG Kang, XING Jihong, DONG Jingao. Relationship Between BcPDR1 Gene and MAPK Pathway Genes BcBMP1 and BcBMP3 in Botrytis cinerea[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(S1): 318-323. doi: 10.7668/hbnxb.20192641.

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http://www.hbnxb.net/EN/Y2022/V37/IS1/318

Figures/Tables 6

Tab.1 Real-time PCR primers design

基因名称 Gene name	引物名称 Primer name	序列 (5'-3') Sequence(5'-3')
BcBMP1	rtBcBMP1-F	ATCTCTTTCAATGTCAGCGAGCAAT
	rtBcBMP1-R	ATGGAGTAATCTTCTTGATAGCGAC
BcBMP3	rtBcBMP3-F	ACAACATGTCCGTGTTCAACCTG
	rtBcBMP3-R	CACGCCAGCTTGCTTGTTCT
BcPDR1	rtBcPDR1-F	CATTCCGGGGTGCCAAGACTCCTGA
	rtBcPDR1-R	ACATGATTTTCATCATAAGGCTTCG
Tubulin	Tubulin-F	TCTGGCGAGCACGGTCTTGACGGTT
	Tubulin-R	TGGCTCCAAATCGACGAGGACGGCA
BcBMP1	BcBMP1-F	ATGACAGCTCGTGCGCCTAATCC
	BcBMP1-R	TCTCATGATCTCATCATAAATC
BcBMP3	BcBMP3-F	ATGGCAGACCTGCAAGGAAGAAAGG
	BcBMP3-R	CGATCGCATAGCATCCAAACCACCT

Fig.1 Expression levels of BcBMP1 and BcBMP3 in BcPDR1 gene mutants A.Expression of BcBMP1 in BcPDR1 gene mutant;B.Expression of BcBMP3 in BcPDR1 gene mutant.The different lowercase letters in the figure show significant differences(P<0.05).The same as Fig.2-5.

Fig.2 Expression levels of the BcPDR1 in RNAi mutants of BcBMP1 (A)and BcBMP3 (B)genes A.Expression of BcPDR1 in RNAi mutants of the BcBMP1 gene;B.Expression of BcPDR1 in RNAi mutants of BcBMP3.

Fig.3 Construction of over-expression isolates of BcBMP1 and BcBMP3 in ΔBcpdr1 background A.Expression analysis of BcBMP1 in strains;B.Expression analysis of BcBMP3 in strains;C.GFP fluorescence detection of strains.

Fig.4 Phenotypic analysis BcBMP1 and BcBMP3 over-expressing strains in the background of mutant ΔBcpdr1 A.Colony morphology observation; B.Observation of hyphal morphology; C.Growth rate assay; D.Conidial yield analysis.

Fig.5 Pathogenicity analysis of BcBMP1 and BcBMP3 over-expressing strains in the background of mutant ΔBcpdr1 A.Analysis of the pathogenicity of the overexpression strains;B.Statistics on lesion area.

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