Screening Of Clubroot Effect Factor And Endogenous Interaction Proteins In Brassica Napus L.

Plasmodiophora brassicae Wor.(Pb)is a soil-borne disease with a wide range of obligate parasitic cruciferous species.With the continuous improvement of mechanization level,it has become a major problem that plagues rapeseed production in China.The molecular mechanism of the work has great practical significance.Pathogenic bacteria secreting effector factors can disrupt or interfere with the immune response of plant PTI or ETI immune defense pathways,and play a vital role in the pathogenicity of pathogens.It is important to study the molecular mechanism of the interaction between resistance genes and effector factors of clubroot to analyze the molecular mechanism of plant disease resistance.In this study,the effect factor secreted by clubroot was used as the entry point,Study on endogenous proteins interacting with plants,and the following results were obtained:1.Greenhouse inoculation system was established.The stable greenhouse inoculation system was optimized and established.The pathogenicity difference of the same resistant materials in different regions was compared and analyzed.The results showed that the 409 R material was immunologically resistant to Sichuan,and further to the clubroots of different regions.At the molecular level,it is found that it is feasible to classify with effect factors as markers.The sequence analysis of the clubroot effectors PBRA6449,PBRA9214,PBRA6069 and PBRA0619 in different regions of the isolates revealed a single base mutation site(SNP site)in the same effector.2.Screening of 10 candidate effective factors of clubroot by resistance gene.In this study,23 clubroot effector factors were successfully isolated,and 3 tobacco clubroot effector factors were identified by tobacco leaf transient expression system to cause necrosis of tobacco leaves.The co-expression of clubroot effector and disease resistance gene CRa was screened 10 to cause PCD in tobacco leaves.The expression level of 10 candidate clubroot effect factors was analyzed and found to be expressed in root tissue.3.Screening of 23 endogenous proteins interacting with Brassica napus by using clubroot effect factor as bait.The transcriptional activity of the candidate 10 effector factors in yeast cells was found PBRA9214 to be transcriptionally active.PBRA6449 effector has weak transcription active,and the other 8 have no transcription active.Further,the clubroot effect factor PBRA6449 was used as the bait protein to screen the molecular targets of the interaction.As a result,a total of 192 monoclonal clones were grown in the QDO medium,and BLAST was found to have 23 proteins encoding complete amino acid fragments.Proteins with a higher number of occurrences were tested point-to-point and found to interact.