Screening And Genetic Analysis Of Key Gene Mutants In ?-Ocimene Signaling Pathway

As a new plant communication signaling molecule,?-basilene can induce the overall defense response of plants,and simultaneously activate the expression of indicator genes(PDF1.2 and PR1)of JA and SA pathways.The key components are not clear.In this study,using forward genetics methods,PDF1.2pro::Luciferase and PR1pro::Luciferase transgenic Arabidopsis seeds(M0)were used as background materials,and a mutant library was constructed by EMS mutagenesis,using luciferase-luciferin The in vivo fluorescence imaging detection system screened and identified mutants related to ?-Ocimene signaling from the mutant library,and further conducted genetic analysis on the mutants to obtain the following research results:1.Successfully screened mutants induced by ?-basilene from EMS-mutated M1materials(non-fluorescence means that the ?-basilene signaling pathway is blocked,indicating that the mutation site is located in basil Ene signaling pathway),12 mutant plants(Ef-1?12)were screened from the mutagenic material with PDF1.2pro::Luciferase expression box from 7681 strains,and the PR1pro::Luciferase expression box was induced from 7537 strains.Ten mutant plants(E1-1?10)were selected from the variable materials.Twenty-two mutants were selfed,and in the M3 generation,stable mutant plants that were basically consistent with the appearance phenotype of Col-0 wild type were selected.2.The M3 generation of Ef and E1 series mutant materials were tested for the expression of the downstream indicator genes(PDF1.2 and PR1)of the ?-Ocimene signaling pathway,and screened the downstream indicator genes induced by ?-Ocimene without screening.Individuals: 9 strains of Ef materials and 3 strains of E1 materials.The integrity of the PDF1.2pro::Luciferase and PR1pro::Luciferase expression cassettes in the above 12 mutant plants was sequenced,and the promoter sequence of the PDF1.2 gene of the Ef-8strain was found(467 base position T base deletion)Mutation occurred,and the expression frame sequences of other materials were unchanged.Ef-8,which mutated the fluorescent expression cassette itself,was eliminated,and it was confirmed that 11 mutants related to the?-basilene signaling pathway were obtained,including 8 strains of Ef material and 3 strains of E1 material.3.Further,a total of 11 mutant materials of Ef and E1 series were crossed with transgenic Arabidopsis materials with PDF1.2pro::Luciferase and PR1pro::Luciferase expression boxes.The results showed that: under the induction of ?-basilene,the F1 generation plants of the mutant all showed ?-basilene-induced “fluorescence” phenotype,indicating that the mutant genes of these 11 materials were recessive;Separation of fluorescent and non-fluorescent phenotypes,statistical data shows that in the F2 generation from Ef-7 and E1-2,the separation ratio of fluorescent and non-fluorescent phenotypes is about 9:7(85:65,91:69),This indicates that the fluorescence traits of these two mutant strains are controlled by two pairs of alleles.In the F2 generations of the other 9 mutant strains,the separation ratio of fluorescence traits was close to 3:1,indicating that the fluorescence traits of these 9 mutants were controlled by each pair of alleles.The above results indicate that we have successfully obtained mutants of key components in the ?-basilene signaling pathway;this is to advance the analysis of the?-basilene signaling pathway and to locate and clone key genes in the ?-basilene signaling pathway.Laid a good foundation.