Establishment Of Rapid Propagation For Sterile Seedlings Of Saxifraga Stolonifera And The Response To Elicitors

Saxifraga stolonifera Meerb.is a perennial evergreen herb of Saxifragaceade which has the effects of dispelling wind,clearing heat,cooling blood,removing toxic material and so on.Modern pharmacological study shows that the extract of S.stolonifera has pharmacological activities of anti-inflammatory,anti-bacteria,bactericide,anti-cancer,and anti-oxidant which indicates it has good medicinal value.Moreover,it is also used as a type of ornamental plants and has good effect in purifying sewage as well.There have been many studies on SA,NO（nitric oxide donor）and MeJA elicitors regulating medicinal components in plants,which can increase the content of secondary metabolites in many plants.Hence in this study,the leaves and stolons of S.stolonifera were used as explants for sterile culture based on the media with different ratios of hormones and the tissue culture conditions was explored,which laid the foundation for the cell culture system of S.stolonifera.Meanwhile,SA,SNP and MeJA in different concentrations were suppled separately in callus and tissue culture seedlings of S.stolonifera to improve the contents of medicinal components,such as bergenin,gallic acid and protocatechuic acid,which provided a clue of improving the contents of secondary metabolites in the cultivation of S.stolonifera.The main results are described as follows.1.Tissue culture conditions for S.stolonifera.The results of the combinations of different explants and ratios of hormones show that.（1）the better combination for leaves sterilization was 70%alcohol for 40 s plus 0.1%Hg Cl₂ for 10 min.（2）The better treatment for leaf callus induction was MS+mg/L 6-BA+0.15 mg/L NAA.（3）The better treatment for leaf callus proliferation was MS+mg/L 6-BA+0.3 mg/L NAA.（4）The better treatment combination for proliferation of stolon clump buds was MS+0.2 mg/L 6-BA+0.1 mg/L NAA.（5）The better rooting condition of stolon clump buds was 1/2 MS+30 g/L sucrose+0.04 mg/L 2,4-D.2.Influence of different elicitors on callus of S.stolonifera.The results of SA,SNP and MeJA on six components in BB leaf callus were compared.Among those treatments,the contents of gallic acid and quercitrin in callus treated with 60μmol/L SA were the highest,which were 0.1514%and0.0132%,separately.The contents of protocatechuic acid and bergenin in callus treated with 40μmol/L SA and SNP were the highest,which were 0.0149%and 0.1554%,respectively.The highest content of chlorogenic acid was 0.0533%in callus treated with 80μmol/L SNP.The highest content of quercetin was 0.0106%in callus of S.stolonifera treated with 20μmol/L SNP.Within range of the appropriate concentration,SA promoted the accumulation of gallic acid,protocatechuic acid and quercetin in callus S.stolonifera,which is better than SNP and MeJA.However,SNP promoting the accumulation of bergenin,chlorogenic acid and quercetin in callus S.stolonifera is better than SA and MeJA.3.Effects of different elicitors on S.stolonifera tissue culture seedlings.The tissue culture seedlings of S.stolonifera were sprayed with SA,SNP and MeJA in different concentrations for 14 d or 28 d,respectively.The results showed that among all the treatments of SA,SNP and MeJA on S.stolonifera tissue culture seedlings,the chlorogenic acid content of tissue culture seedlings treated with 200μmol/L SNP for 28 d was the highest,which was 1.80 times of that of the control.The gallic acid content and quercitrin content of tissue culture seedlings treated with 50μmol/L SNP for28 d were the highest,which were 7.50 and 1.22 times of which of control,respectively.The contents of protocatechuic acid and extract of tissue culture seedlings treated with 100μmol/L SA for28 d were the highest,which were 1.25 and 1.16 times of that of the control,individually.The content of bergenin in the tissue culture seedlings of Trifolium pratense treated with 300μmol/L MeJA for 14 d was the highest,which was 2.94 times that of the control group.Spraying appropriate concentrations of elicitors during the cultivation of S.stolonifera tissue culture seedlings was conducive to improving the contents of gallic acid,protocatechuic acid,bergenin,chlorogenic acid and quercitrin in S.stolonifera tissue culture seedlings.SNP has the better effect on promoting the accumulation of gallic acid,chlorogenic acid and quercitrin in S.stolonifera tissue culture seedlings.SA plays an important role in promoting the accumulation of protocatechuic acid in S.stolonifera tissue culture seedlings.MeJA has the better influence on the accumulation of bergenin in S.stolonifera tissue culture seedlings.