| Asparagus cochinchinensis the important active substances of amino polysaccharide,days total saponins,winter has clear heat decrease internal heat,the effect such as antioxidation,antitumor,as one of the authentic Chinese herbal medicine in our country.With the development of the market,aspidistra is widely used in medicinal,edible,ornamental,feeding and other fields,forming a market shortage situation,artificial cultivation of aspidistra is an effective means to solve the shortage of aspidistra resources.It is difficult for aspartame seeds to germinate under natural conditions,which leads to the limitation of large-scale cultivation of aspartame by conventional propagation methods.The rapid propagation system of aspartate tissue culture has not been applied to the actual production of aspartate due to its long cycle and low proliferation rate.It is not clear whether there are differences between the two kinds of breeding methods and the quality of aspartic seedlings and medicinal materials.Therefore,this study focuses on the above issues,optimizes the two breeding methods of Aspidate,preliminarily identifies the differences between the two breeding methods,and provides a reference basis for the selection of breeding methods of Aspidate artificial planting.The main research results are as follows:1.The stem segments with buds in different parts of the newly cored aspartes were used as explants to induce cluster buds.It was found that the induction rate of the stem segments with buds in the middle reaches 73.33%,which is suitable for explants.However,the contamination rate of the stem segments with buds in the top,middle and lower parts was found to be high by routine disinfection.After disinfection with different concentrations of sodium hypochlorite,it was found that the contamination rate of 2%sodium hypochlorite for10min could be reduced to less than 10%,and the bud point was not damaged.2.Taking the tuft buds induced by the stem segment with buds as experimental materials,the effects of different plant growth regulators on the proliferation and rooting of the tuft buds of aspitic winter were screened.The results showed that 6-BA had the most obvious effect on the proliferation of tuft buds,and the proliferation ratio was 4.13 times when the concentration of 6-BA was 0.5 mg·L-1.IAA and IBA were more conducive to rooting induction of Asparagus cochinchinensis.The rooting rate was 66.63%when IBA 2.0 mg·L-1was contained in the medium for rooting induction.The taproot roots were strong and fibrous roots were vigorous.The orthogonal experiment combined with the practical results,the growth medium of cluster buds was MS+0.5 mg·L-1 6-BA+0.5 mg·L-1 IAA+0.5 mg·L-1 KT+sucrose 30 g·L-1+AGAR 7 g·L-1;Root induction medium:1/2MS+0.5 mg·L-1 IBA+0.5mg·L-1 IAA+sucrose 25 g·L-1+AGAR 7 g·L-1.3.The collected aspartate seeds were used as experimental materials,and the seed structure of Aspartate seeds was observed by using hard tissue sections and saffron solid green staining.TTC was used to detect Aspartate seed vitality.The results showed that a large amount of red and green staining between seed coat and embryo indicated that there was a large amount of cellulose in the middle or the cell wall formed keratinization.The TTC vitality test showed that the seed embryos were all dyed red,which proved that the seed embryos had vitality.Combined with the section observation,the dense seed coat was an important factor for the formation of seeds that were difficult to germinate under natural conditions.The effects of water absorption time,hormone,acid,alkali and salt on seed germination were investigated.The results showed that water absorption time of 6h could improve the germination of aspartate seeds,and the germination rate was 40%.6-BA treatment can increase to 80%;After treating seeds with acid,alkali and salt,it was found that concentrated sulfuric acid could not improve seed germination but reduce germination rate.0.5%potassium nitrate could promote seed germination and shorten the germination period,and the germination rate was 86.66%.Combined with orthogonal experiment,the pretreatment methods were as follows:the seed skin was softened by water at 35℃for 2h,and the 0.5%potassium nitrate treatment for 4h was followed by germination at 25℃.The germination rate was high and the period was short.4.Tissue culture seedlings and seedling seedlings were used as experimental materials.After transplanting,growth and physiological indexes such as plant height,stem diameter and antioxidant enzyme activity were compared and analyzed.The results of growth index showed that the plant height,stem diameter and aboveground（underground）biomass of tissue cultured aspartate seedlings were higher than those of seedling after transplanting.Root analysis showed that the root length,root area and root diameter of tissue cultured seedlings were significantly higher than those of seedling.The results of physiological indexes showed that the chlorophyll content was the same,which indicated that the tissue culture seedlings had stronger photosynthesis and grew faster than the seedling.The activity of antioxidant enzymes in the surface part of tissue culture seedling was significantly higher than that in the seedling（P<0.05）,but there was no significant difference in the underground part.The osmotic substances（soluble sugar,soluble protein）in different tissues（root,stem,leaf）were higher than those of the seedling,and the tolerance and resistance of the two seedlings were better than that of the seedling under the same environment.5.The growth,yield and pharmacodynamic components of different medicinal materials were compared and analyzed with the tissue culture of 3 years old and raw aspartate medicinal materials as experimental materials.The results showed that the average number,weight and biomass of root tubers in tissue culture propagation mode were higher than those in growth-growth mode.Combined with previous experiments,the stronger photosynthesis of tissue culture seedlings was more conducive to the accumulation of biomass,resulting in higher yield than growth-growth mode.The comparative analysis of the pharmacodynamic components in the two propagation methods showed that the content of polysaccharide in the tissue culture method was significantly higher than that in the growth method,and the monosaccharide composition was the same,but the monosaccharide content in the growth method was higher than that in the tissue culture method,which might be due to its different stability.The contents of total saponins and secondary metabolites（flavonoids and phenols）were similar.The types of free amino acids were the same in the two propagation modes,but the content of different amino acids in tissue culture mode was higher than that in growth mode.It shows that the two different methods have their own advantages.Different propagation methods can be selected according to the cost and required materials,and the comprehensive analysis of tissue culture propagation methods is more economical and cost-effective. |
PDF Full Text Request