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Method for culturing xinjiang saussurea involucrata fuzzy root and regenerated seeding and method for producing lilac saponin

A cultivation method, the technology of syringin, applied in the field of plant biotechnology engineering, to achieve the effect of good quality, high yield and rapid growth

Inactive Publication Date: 2006-03-08
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And the research about the Xinjiang snow lotus hairy root and its regenerated seedling cultivation has not been seen at home and abroad.

Method used

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  • Method for culturing xinjiang saussurea involucrata fuzzy root and regenerated seeding and method for producing lilac saponin
  • Method for culturing xinjiang saussurea involucrata fuzzy root and regenerated seeding and method for producing lilac saponin
  • Method for culturing xinjiang saussurea involucrata fuzzy root and regenerated seeding and method for producing lilac saponin

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] 1. Establish a high-frequency regeneration system for Xinjiang snow lotus

[0067] 1.1. Seed disinfection treatment

[0068] Source of seeds: the dried Xinjiang snow lotus (S. Involucrata) picked in Bogda Peak, Tianshan Mountain, Xinjiang in 2002, and the seeds were stripped from the inflorescence.

[0069] Cleaning: Choose healthy, full-bodied, mildew-free seeds, rinse them with tap water 4 times, soak them overnight at room temperature, and rinse them with running water for 1 hour.

[0070] Disinfection: Soak in 70% ethanol for 30s, act on 0.1% mercuric chloride for 20-25min, wash with sterile water 4-6 times.

[0071] Seed germination: hormone-free MS medium, 3% sucrose, 0.8% agar, 25°C, light intensity 2000lux, continuous light, and seeds germinated after 3 days. (Figure 1 germination and seedling of Xinjiang snow lotus)

[0072] 1.2. Inducing cotyledons to produce clustered seedlings

[0073] Explant treatment: select the green seedlings with cotyledon unfolded...

Embodiment 2

[0081] Embodiment 2: establish the transformation system of Snow Lotus in Xinjiang

[0082] 2.1. Transformation preparation experiment 1-determining the strain of Agrobacterium rhizogenes:

[0083] Determination of Agrobacterium strains: The infection effect of Agrobacterium rhizogenes LBA9402, R1601, R1000, and A4 was tested with tobacco, and it was found that R1601 was the most toxic, and the tobacco leaves infected by it took the shortest time to root, and the induced The hairy root has typical hairy root morphological characteristics (Kanamycin 100mg / L, slender, many hairs, weak gravitropism, growth along the bottle wall, oblique secondary root), and there are plants of the same family Artemisia annua, Saussurea jellyfish is induced by R1601 to obtain hairy roots at a rate of 90%. Therefore, the present invention prefers Agrobacterium rhizogenes R1601 to infect Xinjiang Saussurea jellyfish.

[0084] 2.2. Transformation preparation experiment 2--determining Km screening pr...

Embodiment 3

[0104] Example 3: Screening of Hairy Roots

[0105] The 1.0-2.5 cm long root tip of the completely sterilized hairy root was cut and placed on the selection medium 1 / 2MS (3% sucrose+300mg / l Cef+100mg / l Km) and cultured in low light. After 10-12 days, the 1.0-2.5 cm root tip of the Km-resistant root system was cut for liquid culture.

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Abstract

The method for culturing hairy root and regenerative seedling of saussurea involucrata and producing syringin includes the following steps: placing the green seed leaf of saussurea involucrata germinated seed on the induced comatus seedling culture medium to make culture to obtain comatus bud, placing the comatus bud on the seedling growth culture medium to make single plant culture, placing the single plant on the root-inducing culture medium to make culture to obtain the regenerative seedling whose lower end has root from seedling, cutting explant of seedling with root from seedling, placing the obtained leaf segment, leaf stalk segment and root segment on the preculture culture medium to make dark culture, using root-growing agrobacterium mediation to make conversion, culturing and inducing explant to promote root growth, placing the obtained hairy root on the screening culture medium to make subculture, and making liquid amplification culture of root tip to obtain regenerative seedling, drying hairy root and its regenerative seedling culture, pulverizing, separating by using organic solvent, extracting to obtain its syringin.

Description

technical field [0001] The invention belongs to the field of plant biotechnology engineering, in particular to a method for cultivating hairy roots and regenerated seedlings of Xinjiang Saussurea involucrata Kar.et Kir. [0002] The present invention also relates to a method for producing syringin by using the above-mentioned hairy roots and regenerated shoots. technical background [0003] Snow lotus, also known as snow lotus, is a second-class rare protected plant in my country. As early as in "Northwest Territories" and "Xiangyuan Xiaozhi", there are records about snow lotus, and the two are combined into one, collectively called snow lotus. As snow lotus, there are many kinds of native plants, such as the three-fingered snow rabbit (Saussurea tridactyla) used in Tibet and other places; the cotton-headed snow rabbit (Saussurea lanicps) used in Sichuan, Yunnan and other places; medusa); Qinghai also uses Saussurea agossypiphora; Xinjiang uses Saussurea involucrata, etc., ...

Claims

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Application Information

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IPC IPC(8): A01H3/02
Inventor 赵德修付春祥
Owner INST OF BOTANY CHINESE ACAD OF SCI
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