Agrobacterium-mediated Genetic Transformation Of Different Blueberry Varieties

Blueberries are perennial shrubs or shrubs of the genus Vaccinium of the family Ericaceae.Blueberry has a very high nutritional value and health care value,and the market has great potential.The cultivation of blueberry is very harsh on the cultivation environment.Therefore,it is of great significance to cultivate resistant blueberry varieties to adapt to improve the yield and quality of blueberries.Traditional breeding methods are time-consuming and labor-intensive.Using plant genetic engineering breeding can objectively improve biological traits and obtain high-quality plants.which may have potential resistance.In this experiment,the leaves of the tissue culture seedlings of 'Sierra','Brigitta','Legacy'and 'Chandler'were used as materials to explore different combinations of hormones,methods of injury,leaf position,leaf placement,seedling age Factors such as the effects on adventitious buds from leaf regeneration;The establishment of high-frequency leaf regeneration system as a receptor system,Transform VcNF-YC9 gene.explore the selection of antibiotic biotin concentration,antibacterial antibiotic concentration,Agrobacterium concentration,infestation time and co-cultivation days and other factors Bacillus-mediated Genetic Transformation of Blueberry Leaves.The main findings are as follows:1 Establishment of Adventitious Bud System for Blueberry Leaf RegenerationIn this experiment,the leaves of blueberry varieties 'Sierra','Brigitta','Legacy' and 'Chandler'were used as explants,and the best formula for the direct induction of adventitious buds by leaves was WPM+1.0 mg./LTDZ+0.1 mg/LNAA;For the larger 'Sierra' blades,the wound pattern was cut to avoid cutting the three edges along the main vein.For the smaller 'Legacy' blades,the cutting method was cut at both ends.The method of placing the leaves in contact with the culture medium was adopted;the leaves near the top were selected;the tissue culture seedlings were subcultured for about 40 days;the best medium for the shoot buds was WPM+0.5 mg/LZT+0.2 mg/LNAA.2 Agrobacterium-mediated genetic transformation of blueberry leavesIn this experiment,the established leaf genetic transformation system:the transformation of 'Sierra'leaves using 30 mg/L kanamycin,500 mg/L carbenicillin,and cultivation of Agrobacterium to OD600=0.6 min,5 min inoculation,6 d co-cultivation,4 transgenic VcNF-YC9 positive plants were obtained;the transformation of 'Brigitta' leaves using 25 mg/L kanamycin,500 mg/L carbenicillin,Agrobacterium After culturing until OD600=0.6 min,inoculation for 5 min,and co-cultivation for 6 d,5 transgenic VcNF-YC9 positive plants were obtained;the transformation of 'Legacy' leaves using 15 mg/L kanamycin,500 mg/L carbenicillin.After culturing until OD600=0.4,infection for 7 min,and co-cultivation for 4 d,4 transgenic VcNF-YC9 positive plants were obtained;the transformation of'Chandler' leaves using 25 mg/L kanamycin,500 mg/L carbenicillin.After culturing until OD600=0.4,infection for 7 min,and co-cultivation for 4 d,2 transgenic VcNF-YC9 positive plants were obtained.