Rapid Tissue Culture And Direct Regeneration Of Ribes Nigrum L.‘Braude’

In this paper,Ribes nigrum L.’Braude’ was used as the experimental material,and the four aspects of disinfection treatment,proliferation culture,fast propagation system establishment,callus induction and direct regeneration were studied in detail.The rapid propagation system of Ribes nigrum L.’Braude’ variety was successfully established in this experiment,which laid a foundation for future breeding research.The following conclusions are drawn:1 The stem section of annual Ribes nigrum L.’Braude’ was used as explants material,and the best disinfection method was 20% sodium hypochlorite disinfection treatment for 5min,the survival rate was 83.33%,and the contamination rate was 10%.2 MS,1/2MS,1/2MS+0.5 mg/L IAA,MS+Ac and Anderson were selected for the proliferation culture of Ribes nigrum L.’Braude’.Among them,the use of Anderson medium is not only conducive to the growth of the underground part of the ’broad’ test-tube seedling ground,but also inhibits the Browning of test-tube seedlings,making the test-tube seedlings grow normally.3 In matrix I,vermiculite: perlite =1:1（V/V）,the best cultured test tube seedlings,the survival rate of transplanting reached 88.33%.The rapid propagation system of ’Braude’ was established.4 In the callus induction process,the stem segment,petiole and leaf blade were used as explants,and hormones of different concentrations and types were added to induce callus.The induction rate of NAA in the medium MS+5 mg/L KT+0.5 mg/L was the highest at 93.33%,but the callus was Browning in a small part,and the structure was too close,so it was difficult to proliferate and culture.5 Adding Ag NO3,Vc and PVP of different concentrations to control the Browning of callus,among which adding 0.5g/L PVP to the medium MS+5 mg/L KT+0.5 mg/L NAA could effectively control the Browning of callus,with the Browning rate of 6.67% and the induction rate of 96.67%.6 Orthogonal experiment was designed to explore the effect of the optimal combination ratio of hormones on callus proliferation culture,in which MS+5 mg/L KT+1 mg/L NAA+0.1mg/L 2,4-d callus had the best proliferation effect,and the highest proliferation multiple was5.3167,which could be used for regeneration research.7 In this experiment,stem segments were used as explants,MS,White,1/2 MS and Anderson were added to this medium,and hormones of different concentrations and types were added to induce regeneration,which was cultured on MS+4 mg/L KT+0.5 mg/L IAA medium,and ’broad’ regeneration of black currant could be realized.