JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2023, Vol. 47 ›› Issue (4): 123-130.doi: 10.12302/j.issn.1000-2006.202108040

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Cloning and expression of GPS gene induced by methyl jasmonate in Sanghuangporus baumii

LIU Zengcai(), WANG Shuting, TONG Xinyu, ZOU Li()   

  1. College of Forestry, Northeast Forestry University, Harbin 150040, China
  • Received:2021-08-23 Revised:2022-04-29 Online:2023-07-30 Published:2023-07-20

Abstract:

【Objective】 To further explore the molecular mechanisms of triterpenoid biosynthesis of Sanghuangporus baumii, in this study, the induced expression of the key enzyme geranyl pyrophosphate synthase (GPS) gene, which is involved in triterpenoid biosynthesis, was cloned and analyzed. 【Method】The full-length cDNA and promoter of the GPS gene were cloned by PCR amplification, and the sequence was analyzed by bioinformatics software. Then, the qRT-PCR technique was used to analyze the effects of different concentrations of MeJA on the GPS gene transcript level of S. baumii, and the change in triterpenoid content of S. baumii induced by different concentrations of MeJA was determined by spectrophotometer. 【Result】The cloned GPS gene and promoter from S. baumii were named SbGPS and SbGPS promoter, respectively. The cDNA sequence of the SbGPS gene was 1 617 bp, encoding 538 amino acids. Signal peptides and transmembrane structures were not detected. The results of SbGPS promoter analysis showed that, in addition to the typical promoter action elements, such as CAAT-box and TATA-box, a methyl jasmonate response element was included. The results of qRT-PCR analysis and triterpenoid content determination showed that the expression of the SbGPS gene and triterpenoid content initially increased and then decreased with an increase in MeJA concentration. The change trend was basically the same, showing a significant positive correlation. 【Conclusions】 The SbGPS gene plays an important role in the biosynthesis of triterpenoids by cloning and expression analysis, which lays a foundation for further study on the regulation mechanisms of the SbGPS gene in the biosynthesis of triterpenoids of S. baumii.

Key words: Sanghuangporus baumii, geranyl pyrophosphate synthase, gene cloning, promoter cloning, methyl jasmonate(MeJA)

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