Study On The Transformation And Accumulation Of Salidroside And Verbascoside In Two Medicinal Plants In Vitro Culture Syste

With the anti-radiation,anti-fatigue,immune regulation and othe pharmacological activities,salidroside has been widely used in cosmetics,food,military,aerospace,and health care,and demand for it rose year by year.Verbascoside has anti-aging,enhance immunity and other biological activities.It mainly exists in plants such as Cistanche deserticola,Rehmannia glutinosa,and plantain.Among them,Rehmannia glutinosa leaves is the highest,and the content of the target ingredient of Rehmannia glutinosa in the Pharmacopoeia is not less than 0.02%.Based on the previous research,the in vitro culture system of medicinal plants was used to study the transformation and accumulation of salidroside and verbascoside in this research.The experimental contents and results are as follows:1.Establish the suspension cell line of Motherwort and add the precursor tyrosol to investigate the transformation and accumulation of salidroside.The experimental results showed that two transformation products（salidroside and icariin D₂）were obtained by transforming tyrosol from motherwort suspension cells.The addition of 3～5 mmol/L tyrosol to the suspension culture system of Motherwort was carried out for 48-84 h.The conversion rate of salidroside was 69.53%and the highest yield of salidroside was 189.76 mg/L.2.To study the effect of elicitor on the conversion rate of salidroside in the transformation system of Leonurus in suspension（adding 3 mmol/Ltyrosol）.MJ（methyl jasmonate）and SA（salicylic acid）were added into the above system respectively.The results showed that the addition of elicitors had no significant effect on the transformation of Leonurus suspension cells into salidroside.3.This study also used the young leaves of Rehmannia glutinosa as the material to establish the liquid culture system of Rehmannia glutinosa.The technical route was determined as follows:the young leaves of the rehmannia-the loose callus-the adventitious root-the adventitious roots were connected to the liquid medium.Studies have shown that the back of the young leaves is the best way to access;the callus of the leaves of Rehmannia glutinosa is:MS+0.5 mg/L2,4-D+1.0 mg/L6-BA;the regeneration and purification formula of Rehmannia callus is:0.4 mg/L2,4-D+1.0 mg/L6-BA;callus-induced adventitious root and adventitious root solidification formula:MS+1.0 mg/LIBA+0.3 mg/L6-BA;optimal conditions for liquid culture of adventitious roots of Rehmannia glutinosa:the formula was 1.5 g of Rehmannia adventitious roots inoculated into 60 m L of MS+1.0 mg/LNAA+0.3 mg/L6-BA liquid medium.4.The transformation accumulation of verbascoside and salidroside was studied using the adventitious root culture system which the elicitor and the precursor were added into.The results showed that the content of verbascoside in the adventitious root of Rehmannia glutinosa was0.65 mg/g,and the content of verbascoside in the root of fresh rehmannia root was 0.04mg/g.After 25μmol/L MJ was added and cultured for 10 days,the content of verbascoside was2.29 mg/g.While after 25μmol/LMJ+3 mmol/L tyrosol was added and cultured 6 days,the content of verbascoside was determined up to 2.86 mg/g.This study aimed to investigate the transformation and accumulation of salidroside and verbascoside in medicinal plant in vitro culture system,and provide experimental basis for solving the problem of salidroside and verbascoside resources.As well as,provide basis for further study on specific pathways for the synthesis and key genes of salidroside and mullein.