The Function Of Scl-10 Gene In The Interaction Between Nematodes And Fungi

Caenorhabditis elegans is commonly used to study host innate immune responses to bacterial,fungal and viral pathogens.It has been reported that scl-2,scl-20 and scl-27,members of the scl family,are related to the response of C.elegans to various pathogenic infections.However,how scl-10 regulates C.elegans responses to pathogens is poorly understood.In this study,using RNA-seq technology,we found that the expression of scl-10 was significantly up-regulated in the interaction between C.elegans and predatory fungi.Therefore,scl-10,a member of the scl family,was selected as the main research object of this study,and the function of gene scl-10 in the process of resisting fungal infection was preliminarily studied.The main experimental results are as follows:1.scl-10 expression is localized to neuronsscl-10p::GFP transgenic nematodes were obtained by microinjection technology,and fluorescence observation found that scl-10 was mainly expressed in C.elegans neurons;meanwhile,scl-10p::scl-10::G overexpressing nematodes were constructed,and further observation confirmed scl-10 is mainly expressed in C.elegans head neurons.2.Predatory fungi up-regulate the expression of scl-10Comparing the fluorescence intensity under the condition of feeding on OP50,it was observed that the fluorescence intensity changed obviously after 24 hours of predatory fungi(Arthrospora oligospora)infection,suggesting that scl-10 is involved in resisting the infection of these fungi.3.Dopamine receptor signaling pathway and lysosomal pathway are involved in the nematode scl-10 response to Arthrospora oligospora infectionRNA-seq technology detected A.oligospora infected nematodes for 24 hours and found that the expression levels of dopamine receptor signaling pathway genes dop-1,dop-6 and dop-3 were increased.In addition,the response process was also up-regulated under the expression of lysosome-related genes,suggesting that the nematode increases the virulence of lysosome-regulated degradation of A.oligospora.In addition,after A.oligospora infection,the decreased expression of carbon metabolism-related genes would cause downstream changes in the methylation status of genomic DNA,thereby affecting the expression of scl-10.4.Knockout of scl-10 resulted in significantly increased mortality of C.elegans in response to fungal infection;overexpression of scl-10 compensated for the death of nematodes.Using Crispr-cas9 to obtain scl-10 mutant nematodes,the mortality of scl-10 mutant nematodes was significantly increased 24 hours after A.oligospora addition;the overexpressing nematodes(scl-10p::scl-10::G)detection showed that the mortality of C.elegans was significantly reduced by the fungus A.oligospora.RNA-seq also supported that scl-10 overexpression promoted the up-regulation of nematode lysosome and MAPK signaling pathway-related genes(mpk-2,jnk-1,etc.),so scl-10 was involved in the resistance of C.elegans to predatory fungi.In conclusion,this thesis studied the function of scl-10 in the process of C.elegans resisting fungal infection,and found a series of biological pathways related to the regulation against A.oligospora infection.The innovation of this paper: It is the first study of the function of scl-10 in the C.elegans defense against fungal infection.