| Chinese traditional medicine——Glycyrrhiza uralensis and Astragalus membranaceus were transformed by 6 different types of Agrobacterium rhizogenes(1000,ATCC15834,R1601,1025,A4,9402)for hairy root.There were 7 factors putted in this experiment to analysis how to increase the rate of induction.They were different types of explants,different Agrobacterium rhiaogenes,different pre-culture time,different culture mediums,different induced method,different time for inducing,different content of acetosyringone.Ultrasonic wave and microwave were first used in the hairy root induction of Glycyrrhiza uralensis and Astragalus membranaceus,and the effect of using ultrasonic wave and microwave to induce was remarkable.Otherwise,the author studied the callus of the Glycyrrhiza uralensis and Astragalus membranaceus, reached the effect of different hormone playing in the induce of that two plants.And tried to ues callus to induce the hairy roots,but failed.1.Seeds were cultivated in the medium,which was sterilizing 15min.After 6 days,took that seedlings as explants.Seedlings were cut into 4 different parts:seed leaf,hypocotyl,stem,leaf as explants to analysis the effect of different plants on hairy root induction.Glycyrrihiza uralensis:basic culture medium B5;pre-culture time 3 days;Agrobacterium rhiaogenes 15834; AS 100μm/L;25±1℃;co-culture time 3days;highest inducing rate 60%2.Took the Astragalus membranaceus seeds with the same.The best condition of inducing Astragalus membranaceus:basic culture medium MS/N;pre-culture time 3 days;Agrobacterium rhiaogenes R1061; AS 100μm/L;25±1℃;co-culture time 3days;highest inducing rate 40%3.The callus of Glycyrrhiza uralensis and Astragalus membranaceus were regulated by phytohormone. 6-BA,2,4-D,NAA were used to induct callus.Glycyrrhiza uralensis:6-BA 1.0mg/L,2,4-D 1.0mg/L,NAA0.2mg/L,Astragalus membranaceus:MS+2,4-D(1mg/L)+6-BA(0.5 mg/L)+NAA(0.5mg/L).The experiment indicated that hypocotyl was the best explants on callus induction. |
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