Screening And Functional Identification Of CFEM Effector Of Camellia Anthracis

Camellia oleifera is an important woody edible oil species endemic to China.It is widely cultivated in south China.Camellia oleifera anthracnose is a common disease of camellia oleifera,resulting in falling flowers and fruit,or even the death of the whole plant,resulting in huge economic losses.In two systematic and large-scale studies,it was pointed out that Colletotrichum fructicola had the highest isolation rate of Colletotrichum fructicola in the leaves of oil camellia anthracnose,up to 70%.Colletotrichum camelliae was the dominant pathogen on fruits,accounting for more than 90%.CFEM(Common in several fungal extracelluar membrane)domain is a fungal specific domain,and the CFEM domain in pathogenic fungi is often related to the pathogenicity of fungi,some of which are effectors directly affecting the plant immune response.This study is based on the existed C.fructicola’s transcriptome and posted C.camelliae’s genome on the NCBI,extract contains CFEM field effect of the structure,and through YTK12 yeast secretory system transient expression validation and tobacco bioinformatics analysis results and to study the effect of the child for the influence of the plant immune response,The results will lay a foundation for future research.The main results are as follows:1.Screening and structural analysis of candidate CFEM effectors of C.fructicola and C.CamelliaeBioinformatics analysis showed that there were 7 CFEM effectors in Colletotrichum fruticola and 6 CFEM effectors in Colletotrichum camellia.Through the construction of evolutionary tree,it was found that 5 pairs of CFEM effectors of the two anthrax had 100%homology,and sequence alignment showed that there were only 6 amino acids difference between the sequences of two pairs of effectors.2.Functional identification and subcellular localization of candidate CFEM effector signal peptidesIn the experiment of defective YTK12 yeast,it was found that all effectors were secreted proteins,and transient expression of effectors in tobacco showed that most of the full length effectors were located on the cell membrane of tobacco cells,while several effectors showed subcellular localization different from the full length of effectors.3.CFEM effect sub-function analysisTransient expression of effectors in benben tobacco showed that the full length of all CFEM effectors and the sequence of de-signalizing peptide could not cause the necrosis of Nicotiana benthamiana.The results showed that CfCFEM6,19 and 21,CcCFEM8,10,13 and 31 simultaneously inhibited PCD induced by BAX or INF1 in tobacco leaves.However,only CfCFEM6_NS could inhibit THE PCD induced by BAX or INF1 in N.benthamiana leaves.At the same time,after the full length of effectors were expressed in bunyi tobacco leaves and inoculated with Phytophthora capsici,CfCFEM6、12、19、21 and 24 and CcCFEM10、13 and 15 could weaken the disease resistance of Bunyi tobacco and promote the infection of P.capsici.In conclusion,this study found that some CFEM effectors can affect the immune response of plants,and may play an important role in the interaction between pathogens and plants.