The Preliminary Exploration Of Catalitaxol Biosynthetic Mechanism By Transcriptomics

Catalitaxol is secondary metabolite of endophytic fungus FSN002,which can inhibit the growth of liver cancer cells and has a strong inhibitory effect on liver cancer.Its ability to suppress cancer is equivalent to 10 times the taxol's,but the molecular mechanism of biosynthesis is still blank.In this study,some differentially expressed genes were found through transcriptome sequencing of FSN002 samples in different growth periods.Some mutant strains of FSN002 were got by UV mutagenesis and Agrobacterium-mediated transformation.They are used test the related genes of Catalitaxol synthesis,which laid the theoretical basis for its biosynthetic pathway and efficient regulation of production.Some genes related to the Catalitaxol synthesis were found by transcriptome sequencing.A UV mutagenesis method was established to get FSN002 mutant strains.When spores are cultured for 13 h,4-6 cells are germinated and the UV irradiation time is 6 s.The UV-light intensity is90 000?J/cm~2,the lethal rate of the fungi between 90%-95%is the optimum range for getting mutant strains.In this study,a mutant strain b4which did not secrete Catalitaxol was got.The Agrobacterium tumefaciens AGL-1 was successfully transformed into FSN002,and the p450 gene was successfully knocked out to obtain p450-deficient mutant strain.The successes of ultraviolet mutagenesis and Agrobacterium tumefaciens-mediated transformation laid a basis for getting FSN002mutant strains and testing gene function related to Catalitaxol synthesis.These methods are helpful in finding Catalitaxol synthesis.