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successful momilactone biosynthetic pathway reconstitution in Nicotiana benthamiana expressing the biosynthetic gene cluster (BGC), transient coexpression of CpMAS with CpDTC1/HpDTC1, CpCYP970A14, and CpCYP964A1 genes encoding diterpene cyclase 1, pimaradiene oxidase 1, and pimaradiene oxidase 2, respectively. The potential enzymatic activity of momilactone A synthesis from 3OH-syn-pimaradienolide exists endogenously in Nicotiana benthamiana leaves, based on in vitro assays with crude protein and in planta feeding assays. Eventually, coexpression of CpMAS with other clustered genes increases the production of momilactone A, and in planta conversion of 3OH-syn-pimaradienolide to momilactone A is apparently enhanced by CpMAS expression in Nicotiana benthamiana as compared to that in the vector control plant |
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