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- Title
Isopentenyl Diphosphate Isomerase (IPI) Gene Silencing Negatively Affects Patchouli Alcohol Biosynthesis in Pogostemon cablin.
- Authors
Yan, Wuping; Yang, Yuzhang; Wu, Yougen; Yu, Jing; Zhang, Junfeng; Yang, Dongmei; Muhammad, Zeeshan Ul Haq
- Abstract
Pogostemon cablin is a medicinally and economically important plant because it contains a high content of patchouli oil. Patchouli oil is composed of more than 24 unique sesquiterpene compounds, with the most abundant being patchouli alcohol (PA). PA is a sesquiterpene and is mainly synthesized through the mevalonate (MVA) pathway in plants in which isopentenyl diphosphate isomerase (IPI) is a key enzyme. Here, a full-length cDNA encoding IPI (designated as PcIPI) was cloned from P. cablin through the rapid amplification of cDNA ends (RACE). Bioinformatics analysis revealed that the full-length PcIPI includes an 879 bp open reading frame (ORF) that encodes a 292 amino acid protein with a theoretical isoelectric point of 5.55 and a molecular mass of 32.8 kDa. Phylogenetic analysis demonstrated that PcIPI shared the most recent genetic relationship with the IPI of Isodon rubescens (86.43%), Salvia miltiorrhiza (85.38%), Plectranthus barbatus (86.54%), and Prunella vulgaris (86.31%). To verify the function of the PcIPI gene in PA biosynthesis, the P. cablin plants were infected with Agrobacterium tumefaciens bearing a pTRV2-PcIPI construct to induce virus-mediated PcIPI silencing. Quantitative real-time polymerase chain reaction (qRT-PCR) and gas chromatography mass spectrometry (GC-MS) analysis were applied to detect changes in the PcIPI catalytic products and transcription levels. The trend of decreasing transcript level was found to be similar to that of decreasing sesquiterpene content. These results revealed that PcIPI silencing had a negative effect on PA biosynthesis.
- Subjects
GENE silencing; BIOSYNTHESIS; MOLECULAR weights; COMPLEMENTARY DNA; SALVIA miltiorrhiza; ISOMERASES
- Publication
Plant Molecular Biology Reporter, 2021, Vol 39, Issue 3, p557
- ISSN
0735-9640
- Publication type
Article
- DOI
10.1007/s11105-020-01269-0