| Dracocephalum rupestre Hance is a perennial herb.It is the raw material of Shanxi medicinal tea D.rupestre tea.Its flowers are bright blue-purple.It is recorded in the Dictionary of Traditional Chinese Medicine that the whole herb can be used as a medicinal material,which has medicinal effects such as removing internal heat and eliminating inflammation.D.rupestre has a fragrance,hot water brewing has a unique flavor,rich in tea functional ingredients such as flavonoids,caffeine,polyphenols,etc,the folks have always made it into tea for drinking.However,at present,the wild resources of D.rupestre are scarce,and the traditional propagation method is vulnerable to environmental influence,with low yield and long cycle,which is difficult to meet the production needs of D.rupestre Tea enterprises.This experiment conducted a preliminary study on the establishment of D.rupestre tissue culture system,and at the same time measured and compared some functional components of D.rupestre tea and other teas,tissue cultured D.rupestre and wild D.rupestre.First,a system of tissue culture and rapid propagation of D.rupestre seeds was established.The optimal disinfection plan of D.rupestre seeds is0.1% mercury dichloride disinfection for 4 minutes,the germination rate was45.99%,and the contamination rate was as low as 5.02%.WPM medium is the most suitable medium for the germination of D.rupestre,with a germination rate of 61.11%.Different concentrations of plant hormones have certain effects on the germination of D.rupestre seeds.Auxin IBA,cytokinin6-BA and KT all promote seed germination,among which IBA 0.6 mg/L,6-BA 3.0 mg/L and KT 3.0 mg/L group had the highest germination rate,and NAA concentration of 0.9 mg/L could promote seed germination.The optimal subculture medium for sterile seedlings of D.rupestre is: MS+0.5mg/L 6-BA+0.05 mg/L NAA,the proliferation factor is 2.60,and the optimal rooting medium is: 1/2 MS+NAA 0.1 mg/ L+IBA 0.6 mg/L,the rooting rate was 66.67%.Secondly,the induction and differentiation of callus of D.rupestre were studied.The influential factors of callus induction were screened,including explants,medium and plant hormone combinations.The results showed that the stems of the sterile seedlings of D.rupestre were the most suitable explant for callus induction.The optimal medium for callus induction was MS+6-BA 0.5 mg/L+NAA 0.5 mg/L.On this basis,the adventitious bud differentiation of the callus of D.rupestre was found to be difficult to differentiate into adventitious buds in the four hormone combinations.The differentiation rate was low,the highest differentiation rate was 6.66%,and the rooting rate was high..Finally,the five functional components of flavonoids,caffeine,polyphenols,polysaccharides and free amino acids from 7 materials of D.rupestre seeds,wild D.rupestre,tissue cultured D.rupestre,D.rupestre tea,Pu'er,Wuyi rock tea and green tea were carried out Determination.The results showed that among the four kinds of tea,the content of polysaccharide in D.rupestre tea was higher,reaching 2.95%,while the content of the other four components was lower than that of the other three kinds of tea.Green tea has the highest content of flavonoids,polyphenols,and free amino acids,which are 237.44 mg/g,25.34% and 2.79%,respectively;Pu'er has the highest caffeine content,which is 6.68%.The content of caffeine and polysaccharides in tissue cultured D.rupestre was not significantly different from that of wild D.rupestre,and both were higher than that of D.rupestre seeds;free amino acids and flavonoids were higher than wild D.rupestre and D.rupestre seeds,which were 3.06%,128.82 mg/g;there were significant differences among the three;the content of polyphenols was not significantly different from of the D.rupestre seeds,which was 3.56%,lower than that of the wild D.rupestre. |
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