Method for culturing and producing cffeetive ingredients in polyose of saussurca hairy roots of saussurea medusa maxim

A hairy root and polysaccharide technology, applied in the field of plant biotechnology engineering, can solve the problems of natural resource destruction, lack of natural resources of snow lotus, etc., and achieve the effects of stable yield and content, reduction of cost and price, and low pollution rate.

Inactive Publication Date: 2005-06-15
INST OF BOTANY CHINESE ACAD OF SCI
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  • Description
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AI Technical Summary

Problems solved by technology

[0006] In order to solve and overcome the lack of natural resources of

Method used

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  • Method for culturing and producing cffeetive ingredients in polyose of saussurca hairy roots of saussurea medusa maxim
  • Method for culturing and producing cffeetive ingredients in polyose of saussurca hairy roots of saussurea medusa maxim
  • Method for culturing and producing cffeetive ingredients in polyose of saussurca hairy roots of saussurea medusa maxim

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: Establish high-frequency regeneration system of jellyfish snow lotus

[0042] 1.1 Seed disinfection treatment

[0043] Source of seeds: Dried jellyfish snow lotus (S.medusa Maxim) picked in Qilian Mountains in 2001, peeled off the seeds from the inflorescence

[0044] 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.

[0045] Disinfection: Soak in 70% ethanol for 30s, act on 0.01% mercuric chloride for 10-14min, wash with sterile water 4-6 times.

[0046] Sterile seed culture: use hormone-free MS medium, 0.6% agar. 25°C, light intensity 2000lux, continuous light, the seeds germinated successively after 3 days.

[0047] 1.2 Inducing cotyledons to produce clustered seedlings

[0048] Explant treatment: select the green seedlings with cotyledons unfolded after 3 days of germination, cut at the cotyledon petiole, and leave a cot...

Embodiment 2

[0057] Embodiment 2: establish jellyfish snow lotus transformation system

[0058] 2.1 Transformation preparation experiment 1-determining the strain of Agrobacterium rhizogenes:

[0059] Determination of Agrobacterium strains: The infection effect of Agrobacterium rhizogenes LBA9402, R1601, and R1000 was tested with tobacco, and it was found that R1601 was the most toxic, and the time it took for the infected tobacco leaves to root was the shortest, and the induced hair roots had typical Morphological characteristics of hairy roots (Kanamycin 50mg / l, slender, many hairs, weak gravitropism, growth along the bottle wall, oblique secondary roots), and the frequency of 90% of the same family plant Annua annua R1601 Hairy roots were induced, so we determined to use Agrobacterium rhizogenes R1601 to infect Saussurea jellyfish.

[0060] 2.2 Transformation preparation experiment 2--determination of Km screening pressure

[0061] The clustered seedlings were cultured in the maintena...

Embodiment 3

[0074] Example 3: Screening of Hairy Roots

[0075] Delayed screening: insert the leaves with roots into 1 / 2MS+500mg / LCef+0.5-1.0mg / L IAA+0.5mg / L NAA+0.5mg / L GA containing 20mg / L sucrose 3 Delayed selection medium, kept in low light for 5-8 days to promote the induction and growth of hairy roots.

[0076] Km screening: Cut off the 1.0-2.5cm long root tips of all roots and insert them into the screening medium of 1 / 2MS+300mg / L Cef+50mg / L Km containing 20mg / L sucrose, culture in low light, and cut after 10 days The 1.0-2.5 cm root tips of the Km-resistant root system were cultured in liquid.

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Abstract

A process for culturing the hairy root of medusa saussura to obtain saussura polyose includes providing the aseptic test-tube plantlet of medusa saussura, infecting it with bacterial strain R1000, R1601, or LBA 9402, and suspending culture in the 1/2 MS culture medium. Its advantages are high content of polyose (5-10%), and high output rate of polyose (1-2 g/L).

Description

technical field [0001] The invention belongs to the field of plant biotechnology engineering, and specifically relates to a method for cultivating hairy roots of Saussurea jellyfish with high polysaccharide yield, and also includes polysaccharide separation, extraction and preparation process methods. technical background [0002] Snow lotus, also known as snow lotus, is one of the three rare plants 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, the name of the native plant varies with the place of origin. For example, S.tridactyla Sch.Bip. is used in Tibet and other places; S.lanicps Hand.-Mazz. is used in Sichuan, Yunnan and other places; Qinghai and other places use jellyfish snow lotus S.medusa Maxim; Qinghai also uses snow rabbit S.gossypiphora D.Don.; It belongs to the subgenus Subgen Eriocorye of Saussurea, and is gener...

Claims

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Application Information

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IPC IPC(8): A01H4/00B01D29/00
Inventor 赵德修杨睿付春祥
Owner INST OF BOTANY CHINESE ACAD OF SCI
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