| Rhododendron hybrids is a perennial ever-green bush in the rhododendron family, andis a precious woody flower; Cutting propagation is the conventional breeding ways, but itis affected by the material of female parent and the breeding season, which can not meetthe needs of the martet; it is of great significance and good for the introduction andpromotion of fine varieties that using tissue culture techniques in propagation ofornamental plants,which own high propagation speed from seasonal impact.In this research, the micpropagation procedure of Rhododendron hybridscn.’Dr.Tjebbe’had been researched, including explant implantation, initial culture,subculture breeding, callus induction of stems and leavies, differentiation,stronger nurslingbreeding, transplantation breeding, preventing from browning and vitrificating, tosummarize and establish the technich system of tissue culture and rapid propagation.The main findings are as follows:（1） Temperature for tissue culture of Rhododendron lapponicum cn.‘Dr.Tjebbe’:Temperature of the culture room:（23±1）℃,light intensity:2000-3000lx,12hoursillumination every day. the unit of plant growth regulator in medium:mg·L-1;concentrations of sucrose:30g·L-1; concentrations of barits condensate:10g·L-1;PH:6.0～6.3.（2） The season for selecting explants had a larger impact on their survival, using thebuds as the explant,the best season for selecting material was December; using the stems asthe explant,the best season for selecting material was late March to early April.（3） After being treated by alcohol for30seconds, the concentration of disinfectantmercuric chloride for buds was0.1%, The optimizing time of disinfeetion was60seconds.The live explant rate was74.4%. The polluted rate was11.1%. After being treated byalcohol for30seconds, the concentration of disinfectant mercuric chloride for stems was0.1%. The optimizing time of disinfeetion was9minutes. The live explant rate was64.4%.The polluted rate was17.7%.（4） The ability that different hormones on the germinating rate and amount of growingof buds was different. The sequence was: KT>2-ip>6-BA>TDZ. The germinating rate andamount of growing of buds reaches its maximum in the medium included KT, which were 74.5%and1.6cm. The optimizing hormones was KT.（5） Using the buds as the explant, the optimizing medium of initial culture wasAnderson+KT12.0mg·L-1+IAA1.0mg·L-1. After60days culture, the germinating ratewas75.0%. Using the stems as the explant, the optimizing medium of initial culture wasWPM+2-ip8.0mg·L-1+IAA1.0mg·L-1. After60days culture, the germinating rate was64.4%.（6） The optimizing medium was Anderson+KT3.0mg·L-1+IAA1.0mg·L-1in themultiplication of shoot clump, the increasing coefficient was6.11.（7） The stems with buds of tube seedings culture and callus culture: The optimingmedium for the stems with buds of tube seedings culture and callus induction were WPM+2-ip8.0mg·L-1+IAA1.0mg·L-1, After40days culture, the germinating rate was80.00%,Average number for buds formation was5.33, The optiming medium for callus inductionwas WPM+2-ip3.0mg·L-1+IAA1.0mg·L-1, The regeneration rate was72.22%, Averagenumber for buds formation was4.11.（8） Leaf callus culture: Callus regeneration ability was different in different parts ofthe same leaf. The sequence of regeneration ability was: leaf apex>leaf base>leaf midst.The sites of recation was face up. The optiming medium for callus induction wasAnderson+6-BA2.0mg·L-1+2,4-D0.5mg·L-1.（9） The process through direct differentiation from leaves culture: The optimingmedium was WPM+2-ip8.0mg·L-1+IAA1.0mg·L-1, The regeneration rate was70.00%.（10） Stronger nurslings culture, The optiming medium of Stronger nurslings culturewas the Anderson medium with nothing, the leaf was dark green, the leaf area wasincreased, the stems was thicker, some treatment produce roots.（11） Rooting culture: The optiming medium of rooting was1/2Anderson+IAA4.0mg·L-1+AC3.0mg·L-1, The rooting rate was96.7%.（12） The optimizing soil as culture substrat was Peat: Pearlite=3:1, the ratio oftransplant viability was above86.7%. |
PDF Full Text Request