Screening Maize Endogenous Insect - Resistant Genes By Means Of Maize Borer Feeding And MeJA - Induced Double Stress

In the long-term evolution, agricultural pests and crop formed a relationship of mutual restraint, and adaptation, clearly knowing pest resistant crop characteristics of its own defense protein and utilized them, and has important significance and application of scientific value to promote the sustainable development of agriculture. Clearing corn insect resistant defense proteins and methyl jasmonate(MeJA) induced by insect feeding and relevance of stress, which has important guiding significance to the rational use of thead vantages of agricultural pests of plant defense. Early experimental results show that: the corn under the induction of MeJA to the growth and development of the corn borer have a significant impact. Using proteomics method to clear that with MeJA induced, we know that the law of difference protein of corn, screening the defence protein gene, and constructed expression vector to Ostriniafurnacalis have good anti-feeding activity. Considering the kind of defense protein gene in plant transcription level should have significant difference, the experimental mainly use RNA-Seq technology method as the main line, researching on the system of MeJA and insect herbivory under double stress change law of corn defense protein gene transcription, the maize genome database and GeneBank database information data as the bridge to constructed corn, insect resistant defensive protein gene mapping database, sequence analysis of clone screening valuable defensive proteins, to elucidate the relationship between induction and corn borer feeding the protein and MeJA at transcription level in corn by fluorescence quantitative PCR method. Comprehensive analysis of plant induced defense mechanism, ultimately for the use of plant insect resistance itself to provide the scientific foundation for environmental friendly agriculture pest control strategy. The experimental results are as follows:1, complete the sequencing RNA-Seq of Asian corn borer feeding(CO group), MeJA induced(CJ group) andthe Asian corn borer feeding induced+ MeJA(JO group) and the control group transcriptome, identified the gene of significant different expression.2, Venn diagram of differential gene profile revealed that MeJA induction is associated with insect feeding in synergies.3. the GO annotation of differentially expressed genes show that methyl jasmonate induced the effect of defense enhancement of maize self-defense system similar to the Asian corn borer feeding, there are certain things in common.4. the KEGG pathway of differentially expressed genes was used to screen 15 genes of corn defense. Datas show that the defense protein gene is mainly affected by jasmonate signal pathway.5, the cloned sequence size of 9 corn defense protein gene(LOX5、 BBTI2、 BBTI11、 BBTI12、 BBTI13、 Cl-1B、 TPS10、 DOX、 A20/AN1) were consistent to sequencing results, and it has been clear about the informationg of the cloning of candidate gene.6, through the qRT-PCR analysis of the candidate genes of insect resistant gene as a reference gene, GAPDH. The result shows that: the amount of increased differential expression of all candidate gene analysis, consistent with the average fold change and transcriptome sequencing results showed similar gene expression patterns; there’s a synergistic effect on corn gene LOX5, BBTI2, BBTI13, CI-1B, DOX by Methyl jasmonate induction and Asian corn borer insect feeding.