Cloning and Expression Analysis of Two Key Genes of Jasmonic Acid Synthesis in Response to Endophytic Infection from Rehmannia glutinosa

doi:10.7525/j.issn.1673-5102.2021.02.017

Abstract

Abstract:

Allene oxide synthase（AOS） and 12-oxophytodienoate reductase（OPR） were the key genes of jasmonic acid biosynthesis in the plants. The genes of AOS and OPR responding to endophytic fungus infection were screened from the interaction transcriptome of Rehmannia glutinosa with endophytic fungus GG22. Specific primers were designed to the open reading frame of RgAOS and RgOPR. Bioinformatics analysis of the sequences and theirs encoded product were performed， and real-time fluorescent quantitative PCR was used to analyze the expression patterns of RgAOS and RgOPR in different tissues infection by endophytic fungus GG22. The open reading frame of RgAOS was 1 626 bp， encoding 541 amino acids with a molecular weight of 60.2 kD. The open reading frame of RgOPR was 1 197 bp， encoding 398 amino acids with a molecular weight of 44.07 kD. QPCR analysis showed that the expression of RgAOS was the highest in roots， the lowest in flowers， and the expression of RgOPR was higher in flowers and leaves， respectively. Endophytic fungus GG22 induced the expression of RgAOS and RgOPR in R.glutinosa. The RgAOS and RgOPR were successfully cloned from R.glutinosa， laying the foundation for further study on the biological activity of Jasmonic acid substances in R.glutinosa and claiming the molecular mechanism of endophytic fungi in the regulation of secondary metabolites of R.glutinosa.

Key words: allene oxide synthase, 12-oxophytodienoate reductase, bioinformatics analysis, expression analysis, Rehmannia glutinosa

CLC Number:

S567.9

Shu-Ping PENG, Cheng-Ming DONG, Yun-Hao ZHU. Cloning and Expression Analysis of Two Key Genes of Jasmonic Acid Synthesis in Response to Endophytic Infection from Rehmannia glutinosa[J]. Bulletin of Botanical Research, 2021, 41(2): 294-301.

Figures/Tables 5

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基因Gene	引物F Primer F	引物R Primer R	用途Application
AOS	TCAAAGGGCATTCACTTGCT	CACCCATGTGGATGTCTGTT	克隆Cloning
AOS	AAGCACGTGGTCTGGTCTAA	CGCCGCAACCTCAATATCAA	实时荧光定量Q-PCR
OPR	TTTCAATCATCCTCCTACCCCTCT	TGCCCAGACTCAACCAAGTTGAGCA	克隆Cloning
OPR	TGCTGATTTGGTGGCGTATG	TAAAGGCGTGATACGGGTGA	实时荧光定量Q-PCR
TIP41	ATTGGGTAGATTGCCAGGAG	CCATTGCAGCCAATTCATC	内参Reference

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