Tissue Culture Of Alcea Rosea And It’s Natural Active Substance Content Research

Hollyhock,it is perennial herb which belongs to hollyhock malvaceae.It has great ornamental value.Meanwhile,hollyhock has a strong resistance to sulfur dioxide and hydrogen chloride,.It is an excellent urban landscape plant.The whole hollyhock used in medicine,in which root,stem,leaf and flower,there are rich variety of plant polysaccharides and flavonoids.It has great medicinal value.This paper attempts to explore the impact the culture conditions of anther,the preparation of haploid,the suspension culture of cell,the culture of adventitious root and the elicitor conditions on its growth and mainly secondary metabolites content through tissue culture methods to accumulate the basic parameters for industrialized exploitation of hollyhock.In the process of the preparation of callus,we found that the optimum conditions for sterilization of hollyhock anther is soaking 20 min with 4%sodium hypochlorite;The best inducing hormone combination and concentration is 1.5 mg/L 6-BA,2 mg/L 2,4-D and 1 mg/L NAA;Optimal sucrose concentration is 30 g/L;The best medium was MS medium.Moreover,We detect the ploidy of callus cells by flow cytometry and find that most of the callus cells are haploid and diploid cell hybrid.We do suspension culture to callus and establish suspension culture system.then measure growth curve and dynamic changes of pH and conductivity.We found that the growth curve was S-shaped,the pH and conductivity show the overall downward trend.We use young stems of hollyhock induce adventitious roots and find its best inducing hormone concentration is 2 mg/L IBA.We carry on suspension culture to the adventitious root of hollyhock and found that the growth curve was S-shaped,the pH and conductivity show the overall downward trend.The results of elicitor experiment show that methyl jasmonate and salicylic acid both inhibit their growth,low concentration of yeast extract promote the growth of adventitious.Methyl jasmonate has no effect on its uronic acid content.Salicylic acid and yeast extract promote the uronic acid content in suspension cells and adventitious roots.When salicylic acid is 5mg/L,yeast extract islg/L respectively,the uronic acid content reaches the maximum;Yeast extract is better,reaches 1.47 times and 1.38times of the control respectively,reached 47.53mg/g and 75.34mg/g;followed by salicylic acid,is 1.34 times and 1.27times of the control respectively,reached 43.57mg/g and 67.94mg/g;Three kinds of elicitors all promote total flavonoids.Respectively,suspension cells and the total flavonoid content of adventitious reach the maximum,when methyl jasmonate concentration is 0.2 mg/L,salicylic acid is 1 mg/L,yeast extract is 0.1 g/L.Of which the best is methyl jasmonate,increased by 1.64 times and 1.70 times,reached 8.91mg/g and 10.95mg/g;followed by salicylic acid,increased by 1.52 times and 1.58 times,reached 8.26mg/g and 10.19mg/g;The last one is yeast extract,increased by 1.40 times and 1.38 times,reached 7.61mg/g and 8.88mg/g.Add elicitors can effectively improve the content of natural active substances.