Application of n-hexanol in induction of growth of adventitious roots of astragalus membranaceus and synthesis and accumulation of multiple active ingredients of astragalus membranaceus

An active ingredient, n-hexanol technology, applied in the field of traditional Chinese medicine Astragalus membranaceus, can solve the problems that the synthesis and accumulation of non-volatile active ingredients of medicinal plants have not been reported, and the application research has not yet been involved, so as to improve the yield of adventitious roots and various activities. The effect of ingredient content

Active Publication Date: 2021-07-30
SHANXI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Existing studies on tobacco, Arabidopsis and other plants have found that n-hexanol can mediate phytochemical defense responses, but its role in the synthesis ...

Method used

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  • Application of n-hexanol in induction of growth of adventitious roots of astragalus membranaceus and synthesis and accumulation of multiple active ingredients of astragalus membranaceus
  • Application of n-hexanol in induction of growth of adventitious roots of astragalus membranaceus and synthesis and accumulation of multiple active ingredients of astragalus membranaceus
  • Application of n-hexanol in induction of growth of adventitious roots of astragalus membranaceus and synthesis and accumulation of multiple active ingredients of astragalus membranaceus

Examples

Experimental program
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Effect test

Embodiment １

[0029] Example 1: Induction and Subculture of Adventitious Roots of Astragalus mongolica

[0030] As a starting material, Astragalus mongolica tissue culture seedlings that have been propagated for a long time (> 5 years) in the laboratory and are in a vigorous growth period are used as starting materials, such as figure 1 As shown in middle A, select well-growing terminal buds with a length of 1-2 cm, cut them aseptically, and transfer them to MS solid medium containing 0.7% agar, 30 g / L sucrose, and 1 mg / L naphthaleneacetic acid , cultured at 25°C, 16 hours (light) / 8 hours (dark) for 8 weeks, the early morphological characteristics of induced adventitious roots are shown in figure 1 In B, the morphological characteristics of adventitious roots used in liquid culture see figure 1 C in

[0031] Subsequently, the adventitious roots were aseptically cut and washed with sterile water, and transferred to 30 mL 1 / 2 MS liquid medium (100 mL Erlenmeyer flask) containing 30 g / L s...

Embodiment 2

[0032] Embodiment 2: n-hexanol, E -2-hexenal, n-hexanal treatment and sample collection

[0033] Utilize 50% ethanol solution to prepare n-hexanol, E - 2-hexenal and n-hexanal mother liquors, both at a concentration of 50 mmol / L, were sterilized by filtration and stored at 4°C for later use. The specific method of treatment is: according to the volume ratio of 1‰, add the above mother solution to 1 / 2 MS liquid culture solution containing 1 mg / L indolebutyric acid and 30 g / L sucrose; inoculate according to 10% (w:v) Inoculate the adventitious root culture of Astragalus membranaceus, weigh the quality of the culture bottle + culture solution before and after inoculation, and record it as W 01 , W 02 (g), using the deduction method that is W 02 -W 01 , the calculated inoculum size (that is, the initial biomass FW 0 ), cultured on a shaker for 14 days at 25°C and 100 rpm. At the same time, the normal culture and the culture supplemented with 1‰ of 50% ethanol were used as n...

Embodiment 3

[0035] Embodiment 3: n-hexanol, E Effects of -2-hexenal and n-hexanal on the synthesis and accumulation of astragaloside Ⅳ and calycosin glucoside in Astragalus root culture

[0036] The test solution was prepared by the previously established method, diluted 100 times with chromatographically pure methanol, filtered through a 0.45 μm filter membrane, and the contents of astragaloside Ⅳ, acteosin glucoside and formononetin were determined by UHPLC-MS.

[0037] Specific chromatography-mass spectrometry information: Thermo Fischer Scientific Ultimate 3000 HPLC chromatography-Thermo Fischer Scientific Orbitrap Q Exactive Plus mass spectrometer mass spectrometer, selective ion mode (target selected-ion-monitoring mode, SIM) quantitative detection, injection volume: 1 μL; separation column: Syncronis C18 column (2.1 mm × 100 mm, 1.7 μm; ThermoFischer).

[0038] Chromatographic conditions: flow rate is 0.2 mL / min, column temperature is 30°C, and the gradie, mobile phase is 0.1% for...

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Abstract

The invention belongs to the technical field of growth promotion of traditional Chinese medicine astragalus mongholicus, and provides application of n-hexanol in induction of growth of adventitious roots of astragalus mongholicus and synthesis and accumulation of multiple active ingredients of astragalus mongholicus. The current problems of green and sustainable development of astragalus mongholicus resources are solved. The application comprises the steps that after being filtered and sterilized by a 100 mmol/L n-hexanol aqueous solution through a sterile filter membrane, the n-hexanol aqueous solution is stored at 4 DEG C to obtain an n-hexanol inducible factor; and an n-hexanol induction factor is added into an astragalus root adventitious root liquid culture medium according to the addition amount of 0.05% by volume, an astragalus root culture is inoculated according to the inoculum size of 10% by mass volume, and culturing is carried out in a shaking table at 25 DEG C and 100 rpm for 1-2 weeks. The n-hexanol has the effect of promoting the synthesis of the calycosin-7-glucoside synthesis precursor. The dual promotion effects of adventitious root yield and active ingredient synthesis accumulation are comprehensively considered, the elicitor n-hexanol is expected to be developed into a green additive and is applied to an astragalus mongolicus adventitious root tissue culture system, and the yield of a root culture and the content of active substances are improved.

Description

technical field [0001] The invention belongs to the technical field of traditional Chinese medicine astragalus for promoting growth, and specifically relates to the use of n-hexanol in inducing the growth of adventitious roots of astragalus and the synthesis and accumulation of multiple active components of astragalus. The addition of low concentration of n-hexanol to the culture medium promoted the increase of the biomass of the adventitious root tissue culture of Astragalus membranaceus, and the synthesis and accumulation of calycocoside glucoside, formononetin and astragaloside IV in the culture. Background technique [0002] Astragalus mongolica Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) or Astragalus membranaceus Astragalus membranaceus (Fisch.) The dried root of Bge. is a common tonic medicinal material, which has the functions of invigorating qi and raising yang, solidifying the surface and antiperspirant, diuresis and swelling, promoting body...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008A01H4/005
Inventor 孙海峰盛剑张晴晴左心雨张春芬高建平曹秋芬
Owner SHANXI UNIV
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