Study On Effect Of Genotype And2,4-D Concentration On Explant In Vitro Culture Traits

The establishment of in viro tissue culture line is the basis of plant hereditarytransformation cell engineering breeding. As experimental materials, culture characteristicsand influencing factors of four common type wheat varieties’(lines’) immature inflorescence,anther, immature embryo and mature embryo tissue were researched on Xiaoyan NO.22,Xinong NO.1376, Xinong NO.1012and Xinong NO.1013, aimed at probing the effect ofgenotype, explants and culture condition on induction, differentiation and regeneration ofwheat in vitro tissues and callus and selecting best in vitro tissue, genotype and culturecondition to provide theory basis for the establishment of efficient genetic transformation ofwheat cytogenetic decorated change of wheat. The main conclusions are as follows:1. When wheat immature inflorescence was explant in vitro material, Xinong NO.1376had the highest callus induction rate, as high as98.97%. Average callus induction rate ofXiaoyan NO.22, Xinong NO.1012and Xinong NO.1013was98.17%,87.27%and86.00%,respectively. Xinong NO.1376had the highest frequency of regenerated differentiation plants,as high as12.83%. Average frequency of regenerated differentiation plants of Xiaoyan NO.22, Xinong NO.1012and Xinong NO.1013was8.40%,6.30%and5.10%, respectively.2. When wheat anther was explant in vitro material, Xinong NO.1376had the highestcallus induction rate, as high as3.85%. Average callus induction rate of Xinong NO.1013,Xinong NO.1012and Xiaoyan NO.22was2.31%,1.85%and1.68%, respectively. XinongNO.1376had the highest frequency of anther culture，as high as2.40%. Average frequency ofanther culture of Xinong NO.1013, Xiaoyan NO.22and Xinong NO.1012was1.11%,0.93%and0.79%, respectively.3. When wheat immature embryo was explant in vitro material, Xinong NO.1376hadthe highest callus induction rate, as high as99.27%.Average callus induction rate of XiaoyanNO.22, Xinong NO.1013and Xinong NO.1012was97.63%,97.07%and97.03%,respectively. Xinong NO.1376had the highest frequency of regenerated plants, as high as51.03%. Average frequency of regenerated differention plants of Xinong NO.1013, XiaoyanNO.1376and Xiaoyan NO.22was49.01%,39.93%and33.77%, respectively.4. When wheat mature inflorescence was explant in vitro material, Xiaoyan NO.22hadthe highest callus induction rate, as high as93.22%. Average callus induction rate of Xinong NO.1376, Xinong NO.1012and Xinong NO.1013was89.78%,86.98%and83.64%.Xinong NO.1376had the highest frequency of regenerated plants, as high as19.78%.Average frequency of regenerated differention plants of Xiaoyan NO.22, Xinong NO.1012and Xinong NO.1013was14.34%,10.06%and8.90%, respectively.5. In explants induction frequence of four wheat varieties were closely related to2,4-Dconcentration, especially1.0-3.0mg/L2,4-D had better effect on inducing the generation anddifferentiation of four tissues of wheat varieties. Differences of best2,4-D inductionconcentration were observed among different tissues of different or same genotype. Whenwheat immature embryo was explant in vitro, best induction and differentiation concentrationfor Xiaoyan NO.22, Xinong NO.1012and Xinong NO.1013was2.0mg/L. Xinong NO.1376had the highest frequency of regenerated plants，as high as51.03%. Others descend in theorder of Xinong NO.1013(49.01%), Xiaoyan NO.1376(39.93%) and Xiaoyan NO.22(33.77%). Best2,4-D induction and differentiation, regeneration concentration for XinongNO.1376was1.0mg/L and2.0mg/L respectively.When wheat anther was explants in vitro, best induction and differentiationconcentration for Xiaoyan NO.22, Xinong NO.1012and Xinong NO.1013was2.0mg/L.Xinong NO.1376had the highest frequency of regenerated plants，as high as51.03%. Othersdescend in the order of Xinong NO.1013(49.01%), Xiaoyan NO.1376(39.93%) and XiaoyanNO.22(33.77%). Best2,4-D induction and differentiation, regeneration concentration forXinong NO.1376was1.0mg/L and2.0mg/L respectively. When wheat immature embryowas explants in vitro, best induction and differentiation concentration for Xiaoyan NO.22,Xinong NO.1376and Xinong NO.1012was2.0mg/L. Xinong NO.1013had the best2,4-Dinduction and differentiation, regeneration concentration of2.0mg/L and1.0mg/L,respectively.When wheat mature embryo was explants in vitro, best induction and differentiation,regeneration concentration for Xiaoyan NO.22were2.0mg/L and1.0mg/L, respectively. bestinduction and differentiation, regeneration concentration for Xinong NO.1376, XinongNO.1012and Xinong NO.1013were all1.0mg/L.6.Vitro tissue culture regeneration characteristics of the four tested wheat showed thatregeneration rate of vitro immature embryo is the highest, followed by mature embryo, whileyoung panicle and anther had the lowest regeneration rate.