Cloning And Functional Analysis Of Ab-Gpero Effector From Aphelenchoides Bessseyi

Plant Parasitic Nematodes (PPNS), a kind of nematodes which can attack plant tissue, make plant dysplasia and cause some diseases of plant. It secretes effector proteins to modify host cells by the stylet and ingests nutrients to support their growth and development. Effectors control the expressions of the Effector Proteins, widespread in the Plant Parasitic Nematodes. It plays an important role in the development of Plant Parasitic Nematodes. The depth functional studies are important for the control of nematode and minimizing economic damage. Currently, the molecular function of nematode effectors is the subject of intense investigation. The study cloned Ab-Gpero from Aphelenchoides bessseyi Christie second instar larvae using RACE. Use of In Situ Hybridization technique to understand the function of the gene location. Use of RNAi technology to analyze its function, in order to understand the effect factors in Aphelenchoides bessseyi Christie infection. The study may provide a new idea for biological control of nematodes in the future.Cloing objective gene. Selecting target genes by bioinformatics methods. Extraction of RNA, building cDNA for the study of gene partial sequence by some technique, such as PCR, Sequencing, NCBI. According to the obtained sequence, design new primers, using RACE cDNA cloning for purpose gene sequence and genetic analysis.Analysis of Ab-Gpero gene expression by In Situ Hybridization. According to the design of the purpose gene probe, In Situ Hybridization is used to determine the position of gene expression. Micrographs showed that the sites of Ab-Gpero gene expressions were almost in one same position. Ab-Gpero expression site is the nematode esophagus.Analysis of Ab-Gpero gene function by RNAi. Using of T7RNA Polymerase to produce dsRNA. Will dsRNA into nematodes by soak method. Putting the dsRNA treated nematodes and untreated nematodes into carrot medium to observe the infection of callus. Found that the growth of the treated is much butter than the untreated. Results indicated that Ab-Gpero can play an important role in the infection and development of Aphelenchoides bessseyi Christie.