| Rice blast is one of the most important fungal diseases and occurs repeatedly in rice growing areas all over the country,threatening the food security of rice in the world.Every year,rice blast causes 3 billion kilograms of food losses in China,the annual global rice production losses can feed up to 60 million people.The developed countries through the cultivation of resistant varieties,the use of fungicides,to achieve control of the rice blast fungus to a certain extent,but can't achieve long-term effective control;while in developing countries,because the popular rice blast occurred,often lead to crop failure,which means that farmers huge economic losses,and even the occurrence of famine.Therefore,it is helpful to establish a new mechanism to prevent and control rice blast.There are many membrane enclosed organelles in eukaryotic cells,which communicate mainly through the process of vesicle transport.The transport of vesicles usually involves the formation of vesicles on the former membrane,vesicle trafficking,and the fusion process with the target.This series of processes is very complex,first of all,the size and shape of the organelles are different,and the formation and fusion of vesicles is regulated by many protein complexes.The inner membrane fusion reaction is controlled by the macromolecular complex,which is assembled at the time of need,and is separated after the completion of the task.Many proteins are involved in this process,and SNARE has been the center of membrane fusion since its discovery.It is very important for the growth and development of rice blast fungus that SNAREs regulates the fusion of cell membrane and the process of vesicle transport.For infection,the rice blast fungus can secrete a large number of effectors into the host cells through the transfer function of SNARE protein,which interferes with the host immune response.We show that syntaxin 8(MoSyn8),a Qc-SNARE protein homolog,plays important roles in secretion.Our data shows that MoSyn8 regulats the secretion of avirulence effectors Avr-Pia and Avr-Piz-t,interfered the interaction between rice blast and rice.In addition,we through comparative secretome dentified 12 secreted proteins that regulated by MoSyn8.The knockout and functional analysis of these genes was performed to identify a pathogenic factor regulated by MoSyn8.We further analyzed the function of MoPrs5,and found that MoPrs5 can be secreted into the host cells,suppressing host immune response.The rice cDNA library screening found that MoPrs5 targets chlorophyll a/b binding protein Lhcb5 in rice.We also found Lhcb5 protein responses to the infection of Magnaporthe oryzae.In the interaction of rice blast and rice,Lhcb5 is induced to be phosphorylated,which regulates cell death and inhibits the infection of Magnaporthe oryzae.While MoPrs5 can hijack the phosphate group of Lhcb5,inhibiting the immune response regulated by Lhcb5 and promoting the infection of Magnaporthe oryzae.Comparative vesicle proteomics revealed that MoVam7 interacts with MoSsal to regulate clathrin-mediated endocytosis and mediates the cytoplasm to vacuole targeting pathway(Cvt).We also found the auxilin like protein MoSwa2,a disassemble factor of clathrin coated vesicle,has dominant role in pathogenicity.We proved that both MoVam7 and MoSwa2 regulate clathrin dynamics in vivo,and are crucial for the secretion of effectors,so the uncoating process of vesicles not only play a role in endocytosis,but also in effector secretion.That sheds us a new sight on effectors are packaged and transported in vesicles.In conclusion,this paper reveals the important role of SNARE protein MoSyn8 and MoVam7 in the process of secretion and vesicle transport through the analysis of comparative secretome and vesicle proteomics.And find out the novel virulence factors for pathogenicity of M.oryzae. |
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