Studies On The Establishment Of Rapid Propagation System In Vitro Of Lonicera Confusa

Lonicera confusa DC.,is named as shanyinhua,dayinhua,tuyinhua in chinese,a plant in woodbine,Caprifoliaceae,is described in Chinese Pharmacopoeia(2005).It's the alabastrum or the new-blown flower which are used for medicinal purposes in china,has the efficacy for heat-clearing and detoxicating,dispelling wind and heat.It is used to treat above 40 diseases clinically,such as respiratory infection,bacterial infection,hyperthermia,chronic pharyngitis,children pneumonia and parasynanche.It is also broad-spectrum in edible,beverage,health products and daily chemical series,productsBecause of the expanding development and increasing market demand,the wild resources of Lonicera eonfusa were damaged destructivly and became shortage.And when the seeds of Lonicera confusa were hard to find,so the supply of seeds was difficult to meet production needs.The artificial cultivated resources of Lonieera eonfusa were mainly from cutting propagation of wildlife resources,and the survival rate of artificial cultivated seedlings was low and the growth cycle was long.In addition,because of lack of selection and optimization,the artificial cultivated resources were prone to growth inconsistencly, and the standardized production management and cultivation became difficult.So,it is important to establish the standardized of rapid propagation of Lonicera confusa.1.Research summary in domestic and foreignThere are massively literature in domestic and foreign about this research,and they had summaried a lot of Lonicera confuse.The contents refer to its history,current situation of his resources,chemical compostition and pharmacologic action.They also had summaried the culture in vitro of its sibling plant and the quality evaluation about tissue culture plants.2.The establishment of rapid propagation system of Lonicera confusa in vitro.Intitial culture:the different distinfection way,times and lignifications level explants have different effect on the induction of rosette bud.Lignifying inadequacy explants is optimum. The optimum sampling period is spring and summer.In the wet weather days,the pollution frequency is 100%.The optimum time to sterilize the explants is 15seconds in 75%alcohol and 9min in 0.2%Hg₂Cl₂ with TW-80.Shoot-induction:in the course of shoot-induction,different combination of plant growth regulators and explants were compared,results showed that:lignifying inadequacy explants is optimum.Different combination of plant growth regulators have different effect on shoot-induction of Lonicera confuse.,the optimal combination of hormones for shoot formation were 1.0 mg·L^-1 6-BA and IBA 0.2 mg·L^-1.Shoot-proliferation:in this course,different combination of plant growth regulators was compared.And the factors of 6-BA,IBA,and NAA on the shoot proliferation were investigated by orthogonal experiment,In result,the optimal conditions for shoot proliferation were 6-BA1.0 mg·L^-1+ IBA0.02mg·L^-1.The test result also indicated that,the optimum medium was one which the macro-element of MS had changed,the concentration of ammonium nitrate should be 0 mg·L^-1,and halve the concentration of the other four.Rooting-induction:The test result indicated that,the optimum plant growth regulator was NAA,which concentration was 0.02-0.03 mg·L^-1.And active carbon which concentration was 0.2-0.3.g·L^-1 can greatly promote the growth of roots.The optimal conditions for rooting-induction was 1/2MS+NAA0.03 mg·L^-1 and 0.3 g·L^-1 active carbon and sugar 15g·L^-1.The transplantation of regenerated plantlets:By study the number of days of transplanting and matrix,which affect the survival rate and growth trend,the results shown that it was the best management mode that we open the cap 5-6 days to exercise,and the most suitable management of transplantion was transplanting the seedlings into the matrix containingl∶1∶2 of sand,mud and perlite.Two methods were compared,the new one is better..3.The change of chlorogenie acid content of Lonicera confusa in vitro. in this course,the chlorogenic acid content of different plant tissue culture in vitro were compared,what were from different culture medium and period.The test result indicated that,there is no inherent law between the chlorogenic acid content of plant and the culture time.