The Study Of Tissue Culture Response Of Different Wheat Types And Genotypes And Effect Of Embryo Dipping With Agrobacterium To Seedling Growth

1. Using immature embryo and mature embryo of21different types of wheat as explantsfor tissue culture, elected suitable wheat genotypes for tissue culture. At the same time, Inorder to compare the callus of different wheat genotypes and different explants, and primarilyidentify the physiological and biochemical traits related to callus differentiation and seedling,using split-plot design, genotypes as the subplot, explant types as main plot, determined thethe physiological and biochemical traits of callus, and correlation analysis was made. Theresults showed that:(1) Color, status and embryogenic callus rate of immature embryo callus were better thancallus from mature embryos. The average differentiation rate of immature embryo callus was51.37%, and seedling rate was20.82%on average. The best genotype for immature embryocallus culture were Ningchun16and Ei15, Xiaoyan22, followed by Durum332and Tr256.The average differentiation rate of mature embryos callus was19.32%, and seedling rate was4.37%on average. The best genotype for mature embryo callus culture was Ying4286,Yunyin01and Xiaoyan22.(2) Fresh weight, total phenol content and the differentiation rate of immature embryocallus were significantly higher than that of mature embryo callus, and dry weight, PODactivity and seedling rate were very significantly higher than mature embryo callus, whileSOD activity of immature embryo callus was significantly lower than mature embryo callus.Other traits were not significantly different. Physiological and biochemical traits of the callusfrom different genotypes demonstrated significant or very significant difference. Theinteraction effect between explant type and genotype of the physiological and biochemicaltraits were all very significant.(3) The results of correlation analysis showed that the total phenol content and solublesugar content were significantly correlated to callus differentiation rate and seedling rate.2. Preliminarily compared the immature embryo callus culture response of95commonwheat genotypes in Huanghuai wheat area, correlation analysis and cluster analysis weremade, and elected genotypes suitable for immature embryo callus culture. The results showedthat: (1) The best genotype for immature embryo callus culture were Xinong1376, followedby Shanqu26, and Lunhui201, Xuzhou26, CYL8, Zheng9023, Lankao90(6), CYL2,Ningchun16and durum wheat.(2) The results of correlation analysis showed that the color and status, embryogenic rateand callus greening rate of the common wheat immature embryo callus were verysignificantly positively correlated to seedling rate. And the differentiation rate and seedlingrate were significantly positively related.(3) Clustering analysis showed that95genotypes can be divided into7groups, and thefirst group including53genotypes was better for immature embryo callus culture.3. Wheat genotypes 'Xiaoyan22','Xinong1376' and 'Xinong889' as main plot,Agrobacterium tumefaciens bacterial concentration0,0.5,1.0and1.5OD₆₀₀as subplot, usingsplit plot design, studied the influences of Agrobacterium tumefaciens infection on theregeneration of large immature embryos, the seedling growth and development related traits.The results showed that sensitivity of different genotypes to the infection of Agrobacteriumtumefaciens was different. The influence of Agrobacterium tumefaciens infection toregeneration stage of wheat seedlings was not significant, seedling regeneration rate, freshweight, plant height and root length did not change with the changes of bacterialconcentration. However, in the seedling growth and development process, with the increasingof Agrobacterium tumefaciens bacterial concentration, chlorophyll content and SOD activitythe seedlings showed a downward trend, POD activity, CAT activity and MDA contentshowed an increasing trend, and demonstrated significant difference with the control(0OD₆₀₀)when bacterial concentration reached1.0OD₆₀₀. In conclusion, when bacterial concentrationreached1.0OD₆₀₀, Agrobacterium tumefaciens will significantly influence the growth anddevelopment of wheat regeneration seedlings.