Knockout And Phenotype Analysis Of Phenotype Related Genes In Aspergillus Oryzae

Aspergillus oryzae,one of the most important industrial strains in filamentous fungi,has been widely applied to food and industrial production due to its strong ability of extracellular protein secretion.It is acknowledged that Aspergillus oryzae shows colony rapid growth on its phenotype,the color of which is white in the first phrase and then yellow and green.Conidia spores were spherical,loose radial.Mycelial cells on the formation of conidiosis,the top of the formation of the top of the expansion of the capsule,the top of the hemisphere for the secretion of protein.The gene that affects its phenotype,however,has not ever been reported.In this paper,Rho GTPase family gene knockout strain of RacA,CftA,RhoA,RhoB,RhoC and RhoD were used to construct the Rho GTPase family gene of Aspergillus oryzae4177 strain.The approach to compare those by phenotype turned out that the growth rate of RacA and CftA knockout strains was slower than that of wild type;RhoA knockout strain was slow in colony growth,and also demonstrates a clear difference from wild type On the forced medium.The changes on the phenotype of the rest are rather slimmer.The effects of CotA,CrzA,GasA,Mpg1 and OrlA on the cell wall-related genes of Aspergillus oryzae 4177 were compared by homologous recombination.The results delivered the fact that(the knockouted)Gasl and Mpgl are different from those of wild type in colony formation and growth rate.In the meanwhile,At the same time,homologous recombination was used to construct the multi-knockout strain of Aspergillus oryzae 4177,which was a direct knockout of the amylase gene AmyB,AmyC and the neutral protease gene AplA.The phenotypic comparison showed that the knockout was slightly spherical in the liquid culture.There was a significant difference in the growth status of wild type paste.a relatively big difference in growing status.The paper showed that RacA,CftA,Gasl,Mpgl have a great effect on the phenotype of Aspergillus oryzae in the 4177 family genes.Among which,the effect of RhoA is the greatest.And the other genes had little effect on Aspergillus oryzae phenotype.Furthermore,whether the genes will change the phenotype of Aspergillus oryzae by affecting cell walls or thepolarity of cells,is still unknown and in need of further exploration.