Screening of taxol-producing endophytic fungi from Taxus chinensis var. mairei.
A total of 38 endophytic fungus strains were isolated from Taxus chinensis var. mairei by aseptic technique. Genomic DNA was extracted from isolated endophytic fungi and subjected to polymerase chain reaction (PCR) analysis for the presence of Taxus taxadiene synthase (TS) gene, a rate-limiting enzyme gene in the taxol biosynthetic pathway. Twelve out of 38 isolated endophytic fungus strains showed PCR positive for the ts gene. Subsequently, taxol and its related compounds were extracted from culture filtrates and mycelia of the PCR positive strains, separated by column chromatography and analyzed by High Performance Liquid Chromatography and Mass Spectrum. The analysis result showed that 3 strains could produce taxol and its related compounds at the detectible level. This study indicates that molecular detection of the ts gene is an efficient method for primary screening of taxol or its related compounds-producing endophytic fungi which can improve prominently screening efficiency.
Zhou
X
X
Plant Biotechnology Research Center, School of Agriculture and Biology, Fudan-SJTU- Nottingham Plant Biotechnology R&D Center, Shanghai Jiao Tong University, Shanghai 200030, China.
Wang
Z
Z
Jiang
K
K
Wei
Y
Y
Lin
J
J
Sun
X
X
Tang
K
K
D002851:Chromatography, High Pressure Liquid; D004271:DNA, Fungal / Q000378:metabolism; D005658:Fungi / Q000145:classification / Q000302:isolation & purification* / Q000378:metabolism; D016681:Genome, Fungal; D007535:Isomerases / Q000378:metabolism; D013058:Mass Spectrometry; D017239:Paclitaxel / Q000096:biosynthesis*; D020946:Taxus / Q000382:microbiology*
D004271:DNA, Fungal; D007535:Isomerases; C095926:taxa-4(5),11(12)-diene synthase; D017239:Paclitaxel