The Establishment Of In Vitro Regeneration System Of Bitter Gourd And Preliminary Study On The Mechanism Of Adventitious Bud Regeneration

In this study,bitter gourd was used as the research object,and the main factors affecting the in vitro regeneration of bitter gourd were disinfection method,exogenous honnone types and ratio,genotype and explant type,etc.,in order to establish an efficient and stable bitter gourd.The body regeneration system;a series of physiological and biochemical indicators during the regeneration of adventitious buds were measured,and various explants that are prone to produce adventitious bud genotypes and three genotypes with high,medium and low adventitious bud regeneration rates were analyzed.The physiological and biochemical changes of hypocotyls at various stages of adventitious buds are aimed at exploring the mechanism of action of various factors in the process of adventitious buds,in order to provide a theoretical basis for the regeneration of adventitious buds in hypocotyls.The results are as follows:1.The seeds are dehulled,soaked in tap water for 15 min,soaked in 75%alcohol for 30 s in a clean bench,then immersed in 4%NaCIO for 6 min,and rinsed with sterile water for 4 to 5 times.Disinfection method;the best explant is hypocotyl of 12-14 d seedling age of.Haiyan 2,;the medium for optimal callus and adventitious bud differentiation is MS+2.5 mg·L-1 6-BA+0.01 mg·L-1 IBA;the optimal elongation medium is MS+1.0 mg·L-1 6-BA+0.005 mg·L-1 IBA;the best rooting medium is MS+0.1 mg·L-1 IBA;the transplanting method of the test tube seedlings is:when the root length of the test tube seedlings reaches about 2.5?3cm,the seedling transplanting is carried out.Water was poured once a day,and the statistical survival rate was 74%after two weeks.2.The occurrence of adventitious buds during the regeneration of adventitious buds of hypocotyls of "Haiyan 2" from 12 to 14 days was closely related to the changes of endogenous hormone content and balance.The occurrence of adventitious buds requires higher levels of CTK.IAA,ETH,and GA,and requires a certain amount of SA,JA,BR,SLS,and ABA.ABA/GA is low,about 0.72 is conducive to callus expansion and expansion and elongation of adventitious bud;ABA/IAA is between 0.82 and 1.49,low value is conducive to callus enlargement,adventitious bud formation and expansion The higher value is conducive to the elongation of adventitious buds;CTK/IAA is beneficial to the expansion of callus and the expansion and elongation of adventitious buds at around 1.00,and the lower value is beneficial to the initiation of bud primordium.3.The occurrence of adventitious buds in the process of adventitious bud regeneration of hypocotyls of "Haiyan 2" from 12 to 14 days was closely related to the changes of physiological indexes.The occurrence of adventitious buds is accompanied by the production of a large amount of protein.In the germination phase and the adventitious bud expansion period,the soluble protein content will increase;the bud primordium starts,the expansion and elongation of adventitious buds require a large amount of energy to maintain,a certain amount Soluble sugar is beneficial to the occurrence of adventitious buds.In the early stage of adventitious bud expansion,the higher SOD activity is beneficial to the formation of bud primordium and the differentiation of adventitious buds.In the early stage of adventitious bud expansion and late stage of adventitious bud differentiation,higher specific activity of POD is beneficial to the development of adventitious buds,while higher H2O2 plays a positive role in the formation and growth of adventitious buds.