Functional Study On Gene Perg-22in Trichoderma Harzianum T88

As a biological pesticide to prevent and treat a variety of pathogenic fungi,Trichoderma has caused a growing interest of worldwide researchers. This study SelectTrichoderma harzianum T88as the object, coding sequence of perg-22has beenobtained by RT-PCR and PCR, this gene encodes616amino acids, it’s theoreticalmolecular weight is155.5kD.By using the EST sequence of perg-22gene and the original plasmid pSilent-1andpCambia1301, the gene silencing vector is constructed, and Agrobacterium tumefaciensEHA105is successfully transformed. Transformants of T. harzianum T88were obtainedby Agrobacterium-mediated transformation system.The liquid co-cultured method isused in the experiment for the first time, and it’s conversion efficiency is about5-10times of transfer film culture method and complex medium method.Real Time PCR test results showed that the expression of perg-22gene in thetransformants decreased by99.3%, and the expression of some gene related to energymetabolism have a different decline (53%-90%), this showed that the gene is related tothe energy metabolism of T. harzianum T88. The phenomenon that the amount ofergosterol in transformants decreased by93.52%was detected, this means that the geneassociated with the synthesis of ergosterol in T. harzianum T88. This result has greatsignificance for industrialization of the fungal ergosterol production and Trichodermainocula.