| Double flower trait is a very important ornamental characters, this research to improve the formation mechanism of ornamental plants have great significance. Petunia hybrida is a model plant of floral development in recent years, because of its nature petals varieties, the study of plena trait is an ideal experimental material. Discussion of the formation mechanism of double flowers that are mostly in respect of Class C gene mutation caused stamens petals and floral meristem termination procedures postponed. The relevant regulation mechanism of double flower Petunia morphogenesis network that is conducive to the cultivation of commercial petals varieties, can also provide guidance on the construction of male sterile line, and can be further enriched the theory of flower development.CRISPR-Cas9 system as a new gene fixed-point editing techniques has been successful applied in many plants. Targeted genes can be targeting knock-out and knock-in by the technology of CRISPRs. The application of this technique has a great significance in the study of the formation mechanism of double flower.To study the related action to the formation mechanism of flower petals, we choose Petunia hybrida as the research material. This experiment based on the theoretical basis of ABCDE model of floral development, select class PhAP2 A, Ph WUS gene, C gene PMADS3, FBP6, PhARF3 gene doing differential expression analysis. We isolated two genes: PMADS3, FBP6. Then use CRISPR-Cas9 plant gene knockout construct editing system constructs vector of PMADS3 and vector of FBP6 to edite genome of petunia. The main findings are as follows:Obtained the results of differences expression in genes of PhAP2 A, PhWUS, PMADS3, and Ph ARF3, by RT-PCR; Cloned and isolated partial genomic sequence of PMADS3, FBP6; Successfully constructed knockout vector of gene of PMADS3 and FBP6 in univalve petunia, and transformated into callus of petunia by Agrobacterium-mediated. |
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