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Research method for American ginseng hairy root induction and plant regeneration

A technology of American ginseng and hairy roots, applied in the field of hairy root induction and plant regeneration research of American ginseng, can solve the problems of difficult root differentiation, few roots, weak growth ability and the like

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

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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for the induction of American ginseng hairy roots and plant regeneration resear

Method used

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  • Research method for American ginseng hairy root induction and plant regeneration
  • Research method for American ginseng hairy root induction and plant regeneration
  • Research method for American ginseng hairy root induction and plant regeneration

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0098] Embodiment 1: the optimization of the induction condition of American ginseng hairy root

[0099] (1) Effects of different strains on the induction rate of American ginseng hairy roots: In the examples, four strains of R1601, 8196, A4, and rolA were selected to infect American ginseng seedling tissue sections of 2 to 3 weeks seedling age, and the effects of different strains on American ginseng were investigated. The effect of hairy root induction rate is shown in Table 1.

[0100] Table 1 Effects of different bacterial species on the induction rate of American ginseng hairy roots

[0101]

[0102] There are great differences in the pathogenicity of different Agrobacterium rhizogenes strains. The 2-3 week-old American ginseng seedlings infected by Agrobacterium rhizogenes R1601 have white particles at the incision on the 20th day of infection Tissue appeared, some granular tissue appeared in the browned part of the wound, and some granular tissue appeared on the enl...

Embodiment 2

[0130] Example 2: Acquisition and identification of American ginseng hairy roots

[0131] (1) Obtaining American ginseng hairy roots: the co-cultivated materials were rinsed with 500 mg / L cephalosporin (Cef) aqueous solution for 3 to 5 times, the surface of the materials was blotted with sterile filter paper, and then the materials were connected to Bacteria-free culture was carried out on MS solid medium of L cephalosporin (Cef). The medium was replaced once a week, and after 5 to 6 weeks, the material was transferred to MS solid medium without antibiotics to continue culturing. In this example, the earliest occurrence of American ginseng hairy roots is about 30 days after infection (Condition: OD600=0.6, seedslings with stem explants, preconditioning time=48h, AS100 μmol / L). Hairy roots appear at the wound, and several or even dozens of hairy roots can be produced on a piece of explant material, while some explant materials first dedifferentiate to produce loose callus, and...

Embodiment 3

[0133] (3) Dynamic curves of American ginseng hairy root growth and saponin content: under liquid culture conditions, American ginseng hairy root growth and total saponin content curves basically conform to the "S" type (see Figure 10 ). After a short lag period, the growth of American ginseng hairy roots began to enter the logarithmic growth phase on the 9th day, and reached the highest growth on the 27th day, and almost stopped growing after the 27th day, and the dry weight basically did not increase. The content of total saponins increased with the growth of hairy roots, and the total saponins content also reached the maximum on the 27th day, and then the yield began to decline gradually. Example 3: Induction of American Ginseng Hairy Root Callus (1) Effects of Different Growth Regulating Substances on American Ginseng Hairy Root Callus: The analysis of the induction rate of American ginseng hairy root callus is shown in Table 9 and Table 10. According to the order of ext...

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Abstract

The invention relates to a research method for American ginseng hairy root induction and plant regeneration, belonging to the field of crop biotechnologies. The method comprises the steps of (1) treating an explant; (2) preserving, activating and culturing a strain; (3) optimizing conditions for American ginseng hairy root induction; (4) obtaining and identifying American ginseng hairy roots; (5) determining the liquid culture growth and total saponin content of the American ginseng hairy roots; (6) establishing a regeneration system of the American ginseng hairy roots and performing condition optimization. According to the method, the American ginseng hairy roots are used as a material to induce a regenerated plant in an embryogenesis way, so that a new direction for breeding of American ginseng is opened up. By adopting Ri plasmids, an exogenous gene can be guided in more simply and conveniently; besides, the regenerated plant obtained from hairy roods is a transgenic plant to ensure that the transgenic plant has a characteristic of rooting very easily, so that the difficult problems of difficult root differentiation, or weak growing ability and small amount of formed roots in plant regeneration can be well solved.

Description

technical field [0001] The invention relates to a research method for inducing hairy roots of American ginseng and plant regeneration, and belongs to the field of crop biotechnology. Background technique [0002] American ginseng (Panax quinque folium L.) American Ginseng is a traditional and precious medicinal material in my country. It belongs to the perennial herb of the Araliaceae ginseng genus. The virgin forests in North America are now mainly produced in France, Canada and the United States. The best environment for growth is in hilly areas at an altitude of about 2000 meters, and it belongs to the oceanic climate. °~125° in the big forest. Since the 1980s, my country has successfully introduced American ginseng in a large area. After decades of hard work, my country has developed into the world's third Atlantic ginseng producer and the world's largest consumer country after the United States and Canada. [0003] American ginseng contains a variety of physiologically ...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H4/00
Inventor 王义张美萍宫秀杰谢海云孙春玉蒋世翠王康宇杨广顺
Owner JILIN AGRICULTURAL UNIV
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