Method for increasing tropane alkaloid content in belladonna hairy roots by using calcium signal inhibitor

An inhibitor and calcium signal technology, applied in the field of plant biology, can solve the difficult problems of TAs synthesis regulation mechanism to provide access, reduce, increase the total content of TAs, etc.

Active Publication Date: 2022-06-24
GUIZHOU UNIV
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Problems solved by technology

Although digital expression profiling based on transcriptome sequencing can provide valuable clues for the study of TAs metabolic regulation, it is difficult to provide an entry point for in-depth study of TAs synthesis regulation mechanism if there is a lack of data on gene-induced expression or content-induced synthesis o

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  • Method for increasing tropane alkaloid content in belladonna hairy roots by using calcium signal inhibitor
  • Method for increasing tropane alkaloid content in belladonna hairy roots by using calcium signal inhibitor
  • Method for increasing tropane alkaloid content in belladonna hairy roots by using calcium signal inhibitor

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Embodiment 1

[0057] 1. Calcium treatment of belladonna hair roots

[0058] Belladonna hair roots were expanded and cultured in liquid medium to prepare for treatment, and were subcultured twice on MS solid medium. After the first subculture 7-10 days, the newly grown tender white and The multi-branched hair roots were subcultured for the second time. After about 5-7 days, the physiological state and growth of the root tips of the hair roots were basically the same. Cut three root tips with a length of 3-4 cm and inoculate them in 50 mL of MS liquid medium at 110 rpm. , 25 ℃ shaker dark culture for subsequent processing.

[0059] Calcium treatment: hair roots were inoculated with different CaCl 2 Concentration of calcium treatment medium, harvested after culturing for 25 days, the medium was extracted with alkaloids, and the roots were dried at 40°C and ground into powder to prepare alkaloids;

[0060] Treatment with various calcium signal inhibitory reagents: Remove the old medium from h...

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Abstract

The invention discloses a method for increasing tropane alkaloid content in belladonna hairy roots by using a calcium signal inhibitor, which is characterized in that extracellular Ca < 2 + > concentration is reduced by using a Ca < 2 + > specific chelating agent, or a Ca < 2 + >/CaM signal pathway in the belladonna hairy roots is inhibited by using a cell membrane Ca < 2 + > channel inhibitor or an intracellular Ca < 2 + > receptor calmodulin inhibitor, so that the content of tropane alkaloid in the belladonna hairy roots is increased. Therefore, the expression of related genes PMT, TRI, CYP80F1, H6H and ArAT4 in the synthetic route of the tropane alkaloids is induced, and the synthesis and accumulation of the tropane alkaloids hyoscyamine, anisodamine or scopolamine are promoted.

Description

technical field [0001] The invention relates to the technical field of plant biology, in particular to a method for increasing the content of tropane alkaloids in the hairy roots of belladonna by utilizing a calcium signal inhibitor. Background technique [0002] As an anticholinergic drug, tropane alkaloids (TAs) are widely used in clinical practice, including analgesia, anesthesia, anti-motion sickness, cough, asthma, treatment of Parkinson's disease, and can also be used to improve microcirculation, detoxification and detoxification. addiction, treatment of agricultural poisoning, etc. Since the supply of tropane alkaloids mainly based on atropine and scopolamine currently on the market mainly depends on the extraction from medicinal source plants, researching the molecular biology of TAs biosynthesis in plants and carrying out metabolic engineering research to improve the content of TAs are currently cultivated One of the most effective methods for high-content and high...

Claims

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Application Information

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IPC IPC(8): A01H1/00A01H6/82
CPCA01H1/101A01H6/82
Inventor 强玮张明生谭艾娟敖雯雯穆德会刘艳红
Owner GUIZHOU UNIV
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