Research On Key Techniques Of Tissue Culture Of Qinghai Rhubarb

Rhubarb is a perennial herb of Rheum L.in the Polygonaceae family.It is a traditional Chinese herbal medicine in my country and has extremely high medicinal value.In recent years,the demand for rhubarb has been increasing day by day.Due to the limitations of natural planting conditions,traditional methods of planting and raising seedlings can no longer meet the needs of the market.Qinghai rhubarb is the top grade among many high-quality varieties of rhubarb,and palm leaf rhubarb is a common variety of Qinghai rhubarb.The Qinghai rhubarb used in this study is palm leaf rhubarb(Rheum palmatum L.).At present,research data on the tissue culture of Qinghai rhubarb is scarce.Therefore,in order to meet the development needs of the rhubarb market,this article will treat Qinghai rhubarb seed characteristics,sterilization treatment plan screening,Qinghai rhubarb hypocotyls,cotyledons,leaves and other plant parts.Injury tissue induction,proliferation and differentiation,and systematic research on overcoming callus browning.Research indicates:(1)Use SPSS19.0 to perform K-means cluster analysis on 6 types of indicators,and sort according to the F value,germination potential(241.015)>thousand grain quality(30.182)>germination rate(17.998)>Clarity(17.785)> Viability(16.966)> water content(4.93),germination potential,1000-grain quality and germination rate are the main indicators of Qinghai rhubarb seed quality classification.Qinghai rhubarb seeds used in the experiment can be divided into 3 categories,QHDH2 is the first category,and QHDH1 is the first Category 2 and QHDH3 are Category 3.(2)The best disinfection method for seeds is: 75% ethanol for 30 s + 2% sodium hypochlorite for 8 min;the best disinfection method for Qinghai rhubarb explants is: 75%ethanol for 20 s + 2% sodium hypochlorite for 8 min.(3)The best medium for sterile buds is(4.43 g/L)MS+(1.0 mg/L)6-BA+(0.1 mg/L)NAA+30g/L sucrose+3g/L agar,the germination rate is(96.667±5.774)%.(4)The optimal p H of Qinghai rhubarb tissue culture is 5.8.(5)The optimal medium for callus induction is(4.43 g/L)MS+(1.5 mg/L)6-BA+(1.5mg/L)NAA+30 g/L sucrose+3 g/L agar.(6)The best medium for callus proliferation is(4.43 g/L)MS +(1 mg/L)KT +(0.5 mg/L)6-BA +(1.5 g/L)NAA + 3 g/L agar +30 g/L sucrose.(7)The best bud differentiation medium is(4.43 g/L)MS+(1.5 mg/L)6-BA+(0.5mg/L)NAA+3 g/L agar+30g/L sucrose.(8)The root differentiation medium is(2.215 g/L)MS+15 g/L sucrose+5 g/L agar.The rooting situation is relatively good,but the rooting rate is low,which is(6.667±2.887)%.(9)Adding 100 mg/L Vc and 500 mg/L activated carbon can effectively reduce the degree of browning of Qinghai rhubarb hypocotyls,and the browning rate is(48.333±2.887)%.This research is the first to use tissue culture technology to carry out a comprehensive and systematic study on the establishment of Qinghai rhubarb aseptic system,and to explore the solution to the browning problem,so as to speed up the process of artificial rapid breeding of Qinghai rhubarb,break the restriction of natural environment,and hope that Qinghai rhubarb will propagate faster The in-depth study of education provides theoretical basis.