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

Inactive Publication Date: 2014-03-05
JILIN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few studies on the genetic transformation of gins

Method used

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  • Influence and application method of different strains on hairy roots of ginseng
  • Influence and application method of different strains on hairy roots of ginseng
  • Influence and application method of different strains on hairy roots of ginseng

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention relates to influence and an application method of different strains on hairy roots of ginseng, and belongs to the technical field of crop biology. The process comprises the following aspects: (1) explant treatment; (2) storage and activation of strains; (3) optimization of ginseng hairy root induction conditions; (4) acquisition and identification of ginseng hairy roots; (5) determination of the ginseng hairy root liquid, the solid culture growth amount and the content of total saponins; (6) establishment and condition optimization of a ginseng hairy root regeneration system; (7) study of physiological-biochemical characteristics in the hairy root regenerating plant pathway; and (8) histological observation and karyotype analysis on ginseng hairy roots and culture thereof. According to the influence and application method, ginseng is infected by agrobacterium rhizogene to induce hairy roots, a plurality of influence factors of hairy root induction and the hairy root regenerating plant process can be provided, a novel path is provided to germplasm resource storage and creative storage of ginseng, and a basis is established for ginseng breeding.

Description

technical field [0001] The invention relates to the influence of different bacterial strains on hairy roots of ginseng and an application method thereof, belonging to the field of crop biotechnology. Background technique [0002] Ginseng (Panax ginseng C.A.Mey.) is a plant of the genus Panax of the family Araliaceaae. It has been shining with infinite charm and sacred brilliance since ancient times. my country is the birthplace of ginseng. As early as two thousand years ago, our ancestors discovered and used ginseng to prevent and treat diseases. my country's earliest pharmacy classic "Shen Nong's Materia Medica" stated that ginseng "mainly nourishes the five internal organs, calms the spirit, calms the soul, relieves panic, eliminates evil spirits, improves eyesight, improves intelligence, and long-term use lightens the body and prolongs life." There are detailed descriptions in medical books such as "Bie Lu", "Treatise on Febrile Diseases", "Tang Materia Medica" and "Compe...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 王义张美萍李维蒋世翠孙春玉王康宇杨广顺
Owner JILIN AGRICULTURAL UNIV
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