TaLEA Overexpressed Poplus Simonii×P. Nigra Dwarf Mutation Identified And Flanking Sequences Of Key Gene Expression Analysis

Plant genetic mutants is the good material to carry out studies and to determine gene function. Therefore, it is the use of physical and chemical methods of plant mutations induced obtained mutants. However, these mutants mutant loci strong randomness, it is difficult to determine the specific location of the cloning of genes. With the development of new technologies for genome-wide insertional mutagenesis using the known plant traits caused by mutation, then clone, so in the past difficult to accomplish things very easy. It can be amplified by TAIL-PCR sequences flanking the inserted fragment, found similar genes and gene function analysis to determine the mutant gene.Eleven TaLEA overexpressed Poplus simonii×P. nigra transgenic lines and wild-type control were studied for growth traits and leaf anatomy. Among the transgenic lines, one line performed dwarf trait, which was given a name of XL11.The results showed that XL11lines were significantly lower than the other tested lines on leaf area, stomatal density and leaf thickness. On the contrary, XL11lines were significantly higher than in the other lines on epidermis, palisade and spongy tissue ratios and pore size. Focusing on XL11lines, TAIL-PCR method was used for cloning flanking sequences of insertion position, and found AP1/PthRAV （XM₀02311402.1）. Fluorescent quantitative analysis of the relative expression of the AP2/PthRAV lines in dwfl was7.78times significantly higher than other transgenic lines. A preliminary view of dwfl lines in the growth and anatomical structure of leaf traits change may be related to the upregulation of AP2/PthRAV genes. Furthermore, expression profiling of the AP2transcription factor superfamily to the dwarf strain with the wild type. With structure and function analysis of the family genes, the results were closely related to plant growth and development. Then we build a overexpressed vector35S::AP2and RNA interference plant expression vector35S: Amir-AP2. We carried out Poplus simonii×P. nigra genetic transformation, and obtained resistant plants with inhibiting the expression.According to above results, we found that transgenic plants performed low survival rate, lack of green, slow root growth, more lateral root, short and stout, difficulty on rooting.