Method for improving yield of saponin in panax notoginseng tissue culture seedlings

A technology of tissue culture seedlings and Panax notoginseng, applied in the field of plant biology, to achieve the effect of increasing yield, reducing production cost, and not being restricted by seasons

Active Publication Date: 2015-09-23
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report on the application time and concentration of suitable methyl jasmonate to improve the yield of notoginseng saponin in the prior art

Method used

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  • Method for improving yield of saponin in panax notoginseng tissue culture seedlings
  • Method for improving yield of saponin in panax notoginseng tissue culture seedlings
  • Method for improving yield of saponin in panax notoginseng tissue culture seedlings

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Step 1: Establishment of the rapid propagation system of Panax notoginseng tissue culture seedlings.

[0024] The two-year-old Panax notoginseng plants are washed with alkaline soap water to remove surface sludge, rinsed with clean water, rinsed with running water for 30 minutes, and rinsed with distilled water for 4-5 times. Soak its leaves and petioles in 75% ethanol solution for 1 min, rinse them with sterile distilled water for 3-4 times, dry them with sterile filter paper, and place them in 0.1% mercuric chloride (HgCl 2 ) solution for 40 minutes, washed with sterile water for 5-6 times, and blotted dry with sterile filter paper. Cut the leaves into 0.5cm in a sterile ultra-clean workbench 2 block, petiole cut into 0.5cm strips. Inoculate in B5+KT 0.5mg / L+NAA0.6mg / L+2,4D2mg / L solid medium, induce and culture callus for 20 days at 25±2°C in the dark, the callus The induction rate was 98.3%. Transfer the well-grown callus to MS+6-BA1mg / L+IBA0.1mg / L solid medium, ...

Embodiment 2

[0035] Methyl jasmonate was dissolved in 20% absolute ethanol to prepare a 50 mmol / L ethanol mother liquor, which was sterilized by filtration with a 0.22 μm microporous membrane. Add the sterilized methyl jasmonate mother liquor to the test medium that has been sterilized and cooled to 35±5°C, let stand at room temperature, and cool down. Prepare solid medium with methyl jasmonate concentrations of 0, 150, 200, 250 and 300 μmol / L. Accurately weigh the Panax notoginseng tissue culture seedlings obtained in step (1) of Example 1 under sterile conditions, and transfer to the above-mentioned medium (the fresh weight of the seedlings is 5g / bottle, and each concentration gradient of methyl jasmonate is set for 10 repetitions, And take the Panax notoginseng group cultured seedlings in the solid medium with the concentration of methyl jasmonate as 0 as the control group), placed in a light intensity of 30 μmol m -2 the s -1 , light / dark 8 / 16, temperature 23 ± 2 ° C under the condit...

Embodiment 3

[0040] The Panax notoginseng tissue culture seedlings with consistent growth and consistent weight per plant obtained in embodiment 1 step (1) wash the culture medium on the root surface with running water, and transplant to perlite after sterilization: vermiculite (2:1 ) in the seedbed paved with substrate, the planting density is 10cm X 10cm, with 0.5m 2 It is an experimental plot, and 50 tissue culture seedlings are planted in each plot. Cultivate under the conditions of 25-28% substrate humidity, 60% air humidity, 23±2°C temperature, and 17% light transmittance of the awning, and use MS macronutrients as top dressing, and cultivate for 8 months. Make Me-JA into 150, 200, 250 and 300 μmol / L aqueous solutions, and spray evenly on the leaves of Panax notoginseng 8 months after transplanting until the leaves drip (spray once in the morning, middle and evening) , Harvest notoginseng on the 5th day after spraying. And dried at 45 DEG C, weighed, crushed and passed through a 60...

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Abstract

The invention provides a method for improving the yield of saponin in panax notoginseng tissue culture seedlings by applying jasmonic acid methyl ester. The method comprises the following steps: establishing a fast breeding system of panax notoginseng tissue culture seedlings, applying jasmonic acid methyl ester to pseudo-ginseng tissue culture seedlings, cultivating pseudo-ginseng tissue culture seedlings and applying and spraying jasmonic acid methyl ester to the leaf surfaces of pseudo-ginseng tissue culture seedlings. The method is simple, convenient and easy to implement, can enable a great number of good pseudo-ginseng fast breeding seedlings to be formed in a short time, and has the advantages of high breeding speed and short period and the like; the externally applied jasmonic acid methyl ester can effectively improve the yield of total panax notoginseng saponins by 15 times within 5-10 days, and can improve the yield of main monomer ginsenoside by 2-5 times. According to the method for improving yield of saponin in panax notoginseng tissue culture seedlings, the yield of notoginsenoside can be effectively improved, the panax notoginseng tissue culture seedlings are bred by adopting a solid cultivation method, expensive fermentation equipment is not needed, the investment is reduced, the culture method is simple to operate, the breeding coefficient is high, the breeding speed is high, the contamination rate is low, seasonal limitation is avoided, the production cost of notoginsenoside is effectively reduced, and a practical guiding significance is provided for mass production of notoginsenoside.

Description

Technical field: [0001] The invention relates to the field of plant biotechnology, in particular to a new method for increasing saponin production of Panax notoginseng tissue culture seedlings by applying methyl jasmonate. technical background: [0002] Panax notoginseng (Burk.) F.H.Chen is a plant of the genus Ginseng in Araliaceae. It is a traditional precious Chinese medicine in my country. It is mainly produced in Yunnan and is also an important characteristic herbal medicine in Yunnan Province. With the continuous elucidation of the active ingredients and pharmacological effects of Panax notoginseng, the products of Panax notoginseng have been continuously developed, and the market demand is increasing day by day. As of the end of 2013, there were more than 400 Chinese patent medicine preparations containing Panax notoginseng approved by SFDA, more than 3,300 related drug production approval numbers and more than 1,300 pharmaceutical companies. The annual output value o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 朱宏涛张颖君王东许敏杨崇仁
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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