1.Somatic embryogenesis from petiole in Eleutherococcus senticosus
Zhaobin XING ; Fengyun LAO ; Chunying TIAN ; Jianshi WANG
Chinese Traditional and Herbal Drugs 1994;0(08):-
Objective The petiole of Eleutherococcus senticosus was used as somatic embryo,which is widely original plants with the pharmacological active components whose contents could be determined,the somatic embryo in E.senticosus was studied,the aim of this study is to provide the proof of E.senticosus species which have the higher yield of pharmacological active components.Methods Using the petiole of E.senticosus of three years old plants germinated somatic embryos within 15 d to observe the somatic embryogenesis of E.senticosus with the 2,4-D+BA medium.Results After cultured for 28 d with 2,4-D 1.5 mg/L+BA 1.0 mg/L,71.4% of the petiole somatic embryos were directly produced or 8.5 embryos in total were produced via callus.Both of the two methods could be used in the elicitation medium,but the percentage of indirect production was smaller.After transforming into the same or the lower concentration of 2,4-D medium,the somatic embryos gradually matured.At the same time,those of the new somatic embryos were also produced,the percentage of the somatic embryos which were produced by indirect way was increased with it.Conclusion Using the petiole of E.senticosus germination within 15 d could make somatic embryogenesis.It confirms that the somatic embryogenesis and the bodybmeryos inductivity depend on the 2,4-D and BA concentration.
2.Effect of endophytic fungi on expression amount of key enzyme genes in saponins biosynthesis and Eleutherococcus senticosus saponins content.
Zhaobin XING ; Yuehong LONG ; Fengyun LAO ; Shan HE ; Nengsong LIANG ; Baocai LI
China Journal of Chinese Materia Medica 2012;37(14):2041-2045
OBJECTIVETo analyze the effect of endophytic fungi on expression amount of key enzyme genes SS (squalene synthase gene), SE (squalene epoxidase gene) and bAS (beta-amyrin synthase gene) in saponin biosynthesis and saponins content in Eleutherococcus senticosus.
METHODWound method was used for back meeting the endophytic fungi to E. senticosus. With GAPDH as internal control gene, the expression of key enzyme genes was detected by real time PCR method. E. senticosus saponins content was measured by spectrophotometry method.
RESULTWhen wound method back meeting P116-1a and P116-1b after 30 d, the expression content of SS improved significantly (P < 0.05), however the back meeting of P109-4 and P312-1 didnt change the expression of SS. After that SS expression showed reduction-equality-reduction varying trend. Thirty days after back meeting P312-1, the expression content of SE improved significantly (P < 0.05). Ninty days after back meeting P116-1b and P312-1, the expression content of SE improved significantly to 130%,161%, respectively (P < 0.05). After 120 d, back meeting four endophytic fungi, the expression of SE were significantly higher than the control (P < 0.05). Back meeting four endophytic fungi form 60 d to 120 d, the expression of bAS was significantly higher than the control (P < 0.05). The back meeting four endophytic fungi improved E. senticosus saponins content significantly (P < 0.05).
CONCLUSIONEndophytic fungi P116-1a, P116-1b, P1094 and P312-1 significantly effected the expression of key enzyme genes SS, SE and bAS and then affected E. senticosus saponins content. Among the genes, bAS was key target gene.
Eleutherococcus ; chemistry ; metabolism ; microbiology ; Endophytes ; physiology ; Farnesyl-Diphosphate Farnesyltransferase ; genetics ; Fungi ; physiology ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Plant ; Intramolecular Transferases ; genetics ; Saponins ; analysis ; biosynthesis ; Squalene Monooxygenase ; genetics