Cloning, molecular properties and differential expression analysis of the isopentenyl diphosphate isomerase gene in Sanghuangporus baumii

Abstract

Sanghuangporus baumii (Pil at) L.W. Zhou & Y.C. Dai, a popular medicinal fungus, has been used to treat several diseases by its triterpenoids bioactive ingredient for many years. However, the triterpenoids content of S. baumii is low and their biosynthesis mechanisms are not clearly understood. In order to reveal the regulation mechanism of triterpenoids biosynthesis in S. baumii, the cDNA encoding isopentenyl diphosphate isomerase (IDI) involved in triterpenoids biosynthesis was cloned and named as SbIDI (GenBank number MK955885). The open reading frame of the SbIDI gene cDNA comprised 792 bp and encoded a polypeptide of 263 amino acids with a predicted protein molecular weight of 29.80 kDa. The transcript level of the SbIDI gene and triterpenoids content of S. baumii were detected at different developmental stages. The results showed that the transcript level of the SbIDI gene and triterpenoids content had almost the same trend of change, increased first and then decreased dynamically. The highest transcript level of the SbIDI gene occurred earlier than the highest triterpenoids content, indicating that the SbIDI gene may play an important role in triterpenoids biosynthesis in S. baumii. Moreover, the SbIDI gene cDNA was successfully expressed in Escherichia coli BL21 (DE3), and the protein bands were consistent with the prediction.The results will lay a foundation for further study of the function of the SbIDI gene.

Cloning and expression analyses of a Pyrabactin Resistance 1 (PYR1) gene from Magnolia sieboldii K. Koch

“…The monoclones were selected and inoculated in fresh LB liquid medium (containing 50 mg/L kanamycin) for approximately 4 h at 37°C, and the OD 600 was approximately 0.6. IPTG with a final concentration of 0.5 mM was added to induce the expression of PYR1 [ 19 ].…”

Molecular cloning and characterization of a gene encoding HMG-CoA reductase involved in triterpenoids biosynthetic pathway from Sanghuangporus baumii

“…The vector peT-32a and pMD18-T were obtained from Takara Biotechnology (Takara, Dalian, China). The mycelia of S. baumii were inoculated into 500 mL erlenmeyer flasks containing potato dextrose (PD) medium (200 g potato, 20 g glucose, 1 L H 2 o) and grown in a shaking incubator at 170 rpm and 25 °C for 6, 8, 10, 12 and 14 days [23]. The primordia and fruiting body of S. baumii were maintained on bags filled with substrate (77% sawdust, 20% wheat bran, 2% corn flour, 0.5% gypsum and lime, based on the dry weight of ingredients).…”

“…As the key genes in triterpenoids biosynthetic pathway, the transcript level of SbMVD, SbIDI [23], SbLSD were also measured. Transcript level of the three genes were almost analogical, increased, then decreased and peaked on 12th or 14th day in the mycelium stage.…”

Metabolome and transcriptome profiling reveals that terpenoid biosynthesis dominates the growth and development of Sanghuangporus baumii

“…Dai is a precious medicinal fungus with medicinal effects including improving immunity, lowering blood sugar, and anti-cancer and anti-tumour activities (Huang et al, 2020;Zheng et al, 2021). The market demand for S. baumii is increasing, and this is gradually exhausting wild resources (Liu, Sun, Wang, & Zou, 2020). Wild-type (WT) S. baumii is host-specific and grows only on Syringa reticulata .…”

“…Terpenoids are important molecules medicinal research, as exemplified by artemisinin (Ro et al, 2006), taxol (Ajikumar et al, 2010) and ganoderic acid (Zhang et al, 2017), and they are also involved in growth and development, including Cis-Zeatin (CZ), Trans-Zeatin (TZ; Kaori et al, 2006), Abscisic Acid (ABA; Tan, Schwartz, Zeevaart, & McCarty, 1997) and Gibberellin A4 (GA4; Schomburg, Bizzell, Lee, Zeevaart, & Amasino, 2003). Terpenoids can be categorised into monoterpenoids, sesquiterpenoids, diterpenoids, triterpenoids and steroids (Huang et al, 2020;Liu, Sun, Wang, & Zou, 2020;Zheng et al, 2021). They are synthesised by the mevalonate (MVA) pathway, and geranyl pyrophosphate synthase (GPS), famesyl pyrophosphate synthase (FPPS), geranylgeranyl pyrophosphate synthase (GGPS), squalene synthase (SQS) and lanosterol synthase (LS) are direct precursor genes for the biosynthesis of monoterpenoids, sesquiterpenoids, diterpenoids, triterpenoids and steroids (Ali et al, 2017).…”