Study On Tissue Culture And Artificial Seeds Of Curculigo Orchioides

Curculigo orchioides is blindly using a thousand years of traditional Chinese medicine in China,the main treatment of impotence,bones and muscles Withers softly,waist and knee cold paralysis,deficiency cold diarrhea and other symptoms,it is very valuable Chinese medicine.Wild currant is due to man-made and livestock damage,resulting in a sharp decline in the number;but as a raw material of Chinese herbal medicines and health care products,the market demand is growing,the amount of wild currants has been unable to meet people's needs;Has shown great economic value.Traditional cultivation of Curculigo orchioides,Mango way of work,the survival rate is low,and wasted a lot of high quality currants.This paper studied the more efficient and convenient breeding methods of Curculigo orchioides,and explored the problem of insufficient yield of Curculigo orchioides,to a certain extent by using plant tissue culture and artificial seed technology,which had great practical significance.This paper comes from the national forestry public welfare industry research project "Curculigo,Millettia resources cultivation and undergrowth cultivation technology research",project number 201304807.Currently on the market a lot of varieties of Curculigo.This study is based on the cultivation in Guangzhou Conghua wild curculigo yibin in sichuan province.By means of single factor test and orthogonal test,three ways to obtain Cordyceps sinensis quickly were explored.The whole plant was obtained from callus of Curculigo orchioides,and the whole plant was obtained from the adventitious buds.The artificial seeds were induced by cell suspension culture of Curculigo orchioides to induce the whole plant.The experimental results show that the three methods are feasible:Orthogonal test and single factor test showed that 0.1 mg/L 2,4-D,1.5 mg/L KT,3%carbendazim and 8mg/L citric acid were added to MS medium to induce The callus of Curculigo chinensis was the best,up to 96%.The results showed that the effects of 2,4-D,KT and carbendazim on the curative rate of Curculigo were significantly different.The difference of 6-BA concentration and NAA concentration was significant.Subculture medium for the addition of 0.3mg/L NAA,2mg/L of 6-BA,0.3mg/L 2,4-D,the best subculture effect,the highest proliferation rate,the best callus quality,Easier to induce adventitious buds.In the culture medium to add 0.3mg/L NAA is conducive to rooting,transplanted to nutrient soil survival rate can reach more than 90%.In the artificial endosperm,0.05 mg/L 6-BA and 0.8 mg/L NAA were used to produce artificial seed germination rate of 93%.When the addition of 3 mg/L carbendazim,the seed germination rate was the lowest.Comparative experiments show that the artificial seed of the two layers of film is better than the artificial seed,water retention and mechanical properties of the film.When the concentration of sodium alginate 3%,the production of artificial seed quality is the best,the same size,soft and hard appropriate.With 2%CaCl2as curing agent,the reaction twice,the first curing 5 minutes,the second curing 10 minutes,made of artificial seed morphology is the best,uniform particles,soft and hard moderate,spherical,transparent and flexible.When the callus was subcultured,the sucrose concentration was 15g/L,the concentration of 6-BA was 2mg/L,and the NAA concentration was 0.5mg/L,the callus could be obtained as loose and fragile.The effects of non-hormonal levels on cell suspension culture were studied by orthogonal test.The results showed that sucrose had a significant effect on the PH value of suspended cells,and the results showed that different concentrations of sucrose,hydrolyzed casein,MES and proline had significant differences in the suspension culture of Curculigo.To investigate the effect of hormones on cell suspension culture,it was found that 2,4-D affected the structure of suspended cells.When 2,4-D concentration was 0.5mg/L,the number of embryogenic cells reached the maximum.The growth of cells showed "S" curve with the change of culture time.The inoculation amount had the greatest effect on the suspension culture,and the inoculation amount was 15%,which was the optimum inoculation amount.In this paper,the tissue culture and breeding system of Curculigo was established successfully,which could be applied to the large-scale production of Curculigo,and the current situation of shortage was solved.And on this basis,the use of Curculigo buds,successlfully developed a two-layer cinnamon artificial seed,to a certain extent,to solve the traditional artificial seed compressive strength and poor water retention problems.Successful establishment of plant cell suspension system of Curculigo,for the future at the cellular level to study the theoretical basis of Curculigo.