Influence and application method of different strains on hairy roots of ginseng
An application method, hairy root technology, applied in the field of crop biology, can solve the problem of less genetic transformation of ginseng
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Embodiment 1
[0155] Example 1: Optimization of Ginseng Hairy Root Inducing Conditions
[0156] (1) Effects of different strains on the induction rate of ginseng hairy roots: In this example, six strains of R1601, 8196, A4, rolA, rolB, and rolC were selected to infect the leaves of ginseng seedlings with a seedling age of 2 to 3 weeks, and different strains were investigated. See Table 3 for the effects of strains on the induction rate of ginseng hairy roots.
[0157] Table 3 Effects of different bacterial species on induction rate of ginseng hairy roots
[0158]
[0159] There are great differences in the pathogenicity of different strains. On the 16th day after the infection of the 2-3 week-old ginseng seedling leaves infected by Agrobacterium rhizogenes A4, white granular tissue appeared at the incision of the leaves. Some appeared on the browned part of the leaf wound, and others appeared on the enlarged callus of the leaf and were not obvious. As time goes by, the white granular t...
Embodiment 2
[0184] Example 2: Identification of Ginseng Hairy Roots
[0185] After co-cultivation, the materials were rinsed with 500mg / L Cef aqueous solution for 3 times, the surface of the materials was blotted dry with sterile filter paper, and inoculated on MS solid medium containing 500mg / L Cef for sterilized culture. Replace the sterilized medium every 7 days, and transfer to MS solid medium without any antibiotics for culture after 4 to 5 weeks.
[0186] The hairy roots of ginseng in this example were produced at the earliest 27d after infection (Condition: OD 600 =0.6, seedlings with stem explants, preconditioning time=48h, AS100μmol / L). Hairy roots appear at the wound, and one or several hairy roots can be produced on a piece of explant material, while some materials first produce loose callus, and then hairy roots gradually grow out of the callus . All the hairy roots produced showed rapid growth, white root hairs, many branches, no need to add outer edge hormones, attached t...
Embodiment 3
[0190] Example 3: Induction of ginseng hairy root callus
[0191] (1) Effects of different growth-regulating substances on callus induction: using L 9 (3 4 ) Orthogonal experiment to study the influence of BA, KT and 2,4-D on the induction of ginseng hairy root callus. The factor levels are shown in Table 11. The number of samples in each experiment is more than 30, and the experiment is repeated 3 times. The test results are recorded by scoring method. The average results of the 3 experiments are analyzed with SPSSV13.0, see Table 12 and Table 13.
[0192] Table 11 Factor level design of callus induction experiment
[0193]
[0194] Table 12 Orthogonal test scheme and test results
[0195]
[0196]
[0197] Table 13 Analysis of variance
[0198] Dependent Variable: callus induction rate of ginseng hairy roots
[0199]
[0200] a R Squared = .980 (Adjusted R Squared = .919)
[0201] Note: (P<0.01) represents extremely significant level; (P<0.05) represents si...
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