Functional Analysis Of Cmbzip-like And Cmapp In Development And Conidiation Of Coniothyrium Minitans

Coniothyrium minitans is a sclerotial mycoparasite of the plant-pathogenic fungus Sclerotinia sclerotiorum, and an effective biocontrol agent against Sclerotinia disease. Conidia are the main way of survival, parasitism, and dissemination, also is the main component in biocontrol agents. Conidia plays an important role in growth and development of C. minitans. Understanding signaling pathways that modulate conidiation of mitosporic fungi is of both practical and theoretical importance. In this research, two conidiation deficient mutants ZS-1T11329and ZS-1T21656of C. minitans were studied. The results are as follows:Mutant ZS-1T11329cannot produce conidia on PDA, with faster growth rate, increased mycelial branch, declined capacity of parasitic rot to Sclerotinia hyphae sclerotia.Some of mutants in the mutant library such as ZS-1T21656could restore conidiation.Mutant ZS-1T21656could not produce conidia with similar growth rate, parasitism and rot sclerotia capacity as strain ZS-1. The parasitic Sclerotinia mycelium ability is declined and the production of antifungal substances ability increased.Southern blot analysis confirmed that T-DNA inserted in mutant ZS-1T11329and ZS-1T21656by single site. The promoter region of CmbZIP-like gene was inserted by T-DNA in mutant ZS-1T11329, and promoter of CmAPP gene was inserted by T-DNA in mutant ZS-1T21656. The CmbZIP-like is similar to the basic leucine zipper (bZIP) transcription factor, and CmAPP highly homologous to aminoacyl prolyl aminopeptidase. Both CmbZIP-like and CmAPP are single copy in ZS-1genome.Used of knockout, complementary approaches, Agrobacterium mediated transformation (ATMT) technology analysized the function of genes. Through knockout CmAPP gene and research biological characteristics of knockout transformants, CmAPP gene was proved to be irrelevant to the characteristics of mutant ZS-1T21656. Since it is impossible to get knockout transformants of CmbZIP-like gene, the complementary vector of the CmbZIP-like gene has been build. Through ATMT technology complementary research of CmbZIP-like gene in the mutant ZS-1T11329is been doing.