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A method for improving saponin production of 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: 2017-06-20
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • A method for improving saponin production of Panax notoginseng tissue culture seedlings
  • A method for improving saponin production of Panax notoginseng tissue culture seedlings
  • A method for improving saponin production of Panax notoginseng tissue culture seedlings

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Step 1: Establishment of the rapid propagation system of Panax notoginseng tissue culture seedlings.

[0028] The two-year-old Panax notoginseng plants were washed with alkaline soapy water to remove the surface sludge, then rinsed with clean water, rinsed with running water for 30 minutes, and rinsed with distilled water for 4-5 times. Take the leaves and petioles and soak them in 75% ethanol solution for 1 min, rinse with sterile distilled water 3-4 times, blot dry with sterile filter paper, and place them in 0.1% mercuric chloride (HgCl). 2 ) solution for 40min, rinsed with sterile water for 5-6 times, and dried with sterile filter paper. Cut the leaves into 0.5cm in a sterile ultra-clean bench 2 The block, the petioles were cut into 0.5cm strips. Inoculated in B5+KT 0.5mg / L+NAA0.6mg / L+2,4D2mg / L solid medium, at 25±2℃, under dark conditions, callus induction and culture were carried out for 20 days. The induction rate was 98.3%. The well-grown callus was transfer...

Embodiment 2

[0039] Methyl jasmonate was dissolved in 20% absolute ethanol to prepare a 50 mmol / L ethanol stock solution, which was filtered and sterilized with a 0.22 μm microporous membrane. The sterilized methyl jasmonate mother solution was added to the test medium that was sterilized and cooled to 35±5°C, and allowed to stand and cool at room temperature. Solid medium with methyl jasmonate concentration of 0, 150, 200, 250 and 300 μmol / L was prepared. Accurately weigh the obtained Panax notoginseng tissue culture seedlings in the step (1) of Example 1 under aseptic conditions, and transfer to the above-mentioned culture medium (fresh weight 5g / bottle of seedlings, each concentration gradient of methyl jasmonate is set to 10 repetitions, And take the Panax notoginseng group culture seedlings as the control group in the solid medium with the methyl jasmonate concentration of 0), placed in a light intensity of 30 μmol m -2 s -1 , light / dark 8 / 16, subculture for 5 days under the conditi...

Embodiment 3

[0044] The notoginseng tissue culture seedlings of robust growth and consistent weight per plant obtained in the step (1) of Example 1 wash the culture medium on the surface of the root with running water, and are transplanted to the perlite after sterilization: vermiculite (2:1 ) in the seedbed paved with the substrate, the planting density is 10cmX 10cm, with 0.5m 2 As an experimental plot, 50 tissue culture seedlings were planted in each plot. Under the conditions of substrate humidity of 25-28%, air humidity of 60%, temperature of 23±2°C, and shade transmittance of 17%, and supplemented with MS macroelements as top dressing, it was cultured to 8 months. Me-JA was formulated into 150, 200, 250 and 300 μmol / L aqueous solutions, and sprayed evenly on the leaves of Panax notoginseng after 8 months of transplanting until the leaves dripped (spray once in the morning, in the middle and in the evening) , Harvest Panax notoginseng on the 5th day after spraying. And dried at 45°C...

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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

[0001] Technical field: [0002] The invention relates to the field of plant biotechnology, in particular to a new method for improving saponin yield of Panax notoginseng tissue culture seedlings by applying methyl jasmonate. [0003] technical background: [0004] Panax notoginseng (Burk.) F.H.Chen is a plant of the Araliaceae ginseng genus. It is a traditional and precious traditional Chinese medicine in my country. It is mainly produced in Yunnan and is also an important characteristic botanical medicine in Yunnan Province. With the continuous clarification of the active ingredients and pharmacological effects of Panax notoginseng, the products of Panax notoginseng are continuously developed, and the market demand is increasing day by day. As of the end of 2013, more than 400 Chinese patent medicine preparations containing Panax notoginseng have been approved by the SFDA, more than 3,300 related drug production approval numbers and more than 1,300 pharmaceutical companies. T...

Claims

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Application Information

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Patent Type & Authority Patents(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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