Application method of elicitor in hairy root of American ginseng
An application method, the technology of American ginseng, applied in the field of crop biology, can solve the problems of no report of synergy, few reports of adding elicitors of American ginseng hairy roots, no reports of induction of salicylic acid and methyl jasmonate, etc.
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Embodiment 1
[0048] Example 1: Production of American Ginseng Hairy Roots
[0049] (1) Transformation of American ginseng seedlings:
[0050] (1a) The influence of explant selection on the induction rate of American ginseng hairy roots: In the examples, the explants selected in this experiment are the leaves, stems and leafy young stems of the seedlings of American ginseng seedlings of 1 to 2 weeks old. To investigate the influence of R1601 infecting American ginseng and the selection of explants on the induction rate of hairy roots. Experiments show that young stems with leaves are the best explants, with an induction rate of 12.1%, which is 5.6 percentage points higher than the induction rate of stem segments, while leaves do not induce hairy roots. Other transformation conditions are pre-culture time 24h, OD 600 =0.8, the dyeing time is 20min, see Table 3.
[0051] Table 3 Effects of different explants on the induction rate of American ginseng hairy roots
[0052] Explant type
[0053] Con...
Embodiment 2
[0079] Example 2: Optimization of culture conditions for American ginseng hairy roots
[0080] (1) The effect of different media on the growth of American ginseng hairy roots and the synthesis of total saponins: 4 liquid media MS, modified White, SJ-1 and B 5 The effect on the growth of American ginseng hairy roots and the content of total saponins such as image 3 Shown. From image 3 It can be seen that the liquid SJ-1 is most conducive to the growth of hairy roots, and the monthly growth rate can reach 31.3 times, which is 1.7 times that under liquid MS culture conditions. The total saponins accumulation has the best effect on improving White (8.10%), followed by MS, and B 5 The worst (6.67%).
[0081] (2) The effect of different carbon sources on the growth of American ginseng hairy roots and the total saponins content: sugar is a necessary carbon source for plant growth in vitro, and different carbon sources have great influence on the growth of hairy roots and the synthesis o...
Embodiment 3
[0083] Example 3: The effect of salicylic acid and methyl jasmonate
[0084] (1) The addition of salicylic acid: such as Image 6 As shown, after adding different concentrations of salicylic acid, the growth of hairy roots and saponin content showed a similar trend. With the increase of salicylic acid concentration, the cell growth and saponin content increased, but the increase was not When the concentration of exogenous salicylic acid increases to 0.6mg / L, the growth of hairy roots and the saponins content both reach the maximum value of 0.63g and 2.98%. However, when the concentration of salicylic acid continues to increase, the hairy roots grow However, the amount and saponins content decreased significantly, almost reaching the control group level. This may be due to the excessively high concentration of salicylic acid, which has a dominant toxic effect on cells, causing the decline of cell proliferation, and the energy necessary for the biosynthetic pathway is difficult to o...
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