Two promoters of asymmetric cell division regulation pathway of root tip stem cell of plant and application of two promoters

A promoter and stem cell technology, which is applied in the field of preparation of promoters for the asymmetric division regulation pathway of plant root tip stem cells, can solve the problems that there are no reports of promoters for the ACD regulation pathway of stem cells with baicalein and wild scutellarein

Inactive Publication Date: 2017-12-12
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

[0005] At present, there is no report in the prior art that baicalein and scutellarein are used as promoters of stem cell ACD regulatory pathways

Method used

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  • Two promoters of asymmetric cell division regulation pathway of root tip stem cell of plant and application of two promoters
  • Two promoters of asymmetric cell division regulation pathway of root tip stem cell of plant and application of two promoters
  • Two promoters of asymmetric cell division regulation pathway of root tip stem cell of plant and application of two promoters

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Preparation of Baicalein:

[0029] Name: Baicalein / Scutellarin / 4-Hydroxybaicalein-7-O-glucuronide

[0030] (scutellarin / Scutellarein 4'-methyl ether 7-glucuronide)

[0031] Molecular formula: C 21 H 18 O 12

[0032] Structural formula:

[0033]

[0034] Preparation: Crushing of Erigeron breviscapus (vant) Hand Mass → 70% ethanol percolation → concentrated extract → concentrated solution with macroporous resin → 50% ethanol elution → concentrated eluent → concentrated solution adjusted with hydrochloric acid PH value to 1-2 → stand for 24 hours for crystallization → filter crystal → methanol reflux recrystallization (multiple times) → obtain pure scutellarin. (3g of pure scutellarin B prepared from 1kg of short pavilion Feipeng medicinal materials)

[0035] Preparation of scutellarin:

[0036] Name: scutellarin / 4',5,6,7-tetrahydroxyflavone

[0037] (scutellarein / 4',5,6,7-Tetrahydroxyflavone)

[0038] Molecular formula: C 15 H 10 O 6

[0039] Structural f...

Embodiment 2

[0043] Construction of three plasmids, SHR-YN, RBR-YN, and SCR-YC:

[0044] (1) BIFC experiments are a classic method to identify the localization and interaction of target proteins in living cells. The principle is to fuse and express the two complementary fragments of the fluorescent protein with the target protein respectively. If the target protein interacts, the two complementary fragments of the fluorescent protein can be brought closer to restore the function of the fluorescent protein to generate fluorescence under excitation light. Construct a plasmid fused with two complementary fragments of target protein and fluorescent protein. The root RNA of Colombia ecotype Arabidopsis thaliana (Col) was extracted and reverse transcribed to obtain cDNA. SHR upstream primer (5'-CGCGGATCCATGGATACTCTCTTTAGACTAGTC-3') and downstream primer (5'-CCGCTCGAGCGTTGGCCGCCACGC-3'), RBR upstream primer (5'-CGCGGATCCATGGATACTCTCTTTAGACTAGTC-3') were used respectively. '-CGCGGATCCATGGAAGAAGTT...

Embodiment 3

[0046] Three plasmids SHR-YN, RBR-YN and SCR-YC were transfected into tobacco:

[0047] The three successfully constructed plasmids SHR-YN, RBR-YN and SCR-YC were transformed into Agrobacterium GV3101 by electric shock method, and positive clones were selected to extract plasmids. After sequence determination and comparison, it was proved that the three target genes were successfully transformed into Agrobacterium tumefaciens. Bacillus. Pick three single colonies of Agrobacterium-positive clones of the target gene, inoculate them into 100 mL LB medium containing 50 μg / mL kanamycin, gentamycin and rifampicin, respectively, and culture in a shaker at 28°C and 200 rpm. Overnight, 200 μM acetosyringone was added in the next morning to continue the culture, and the OD value of the bacterial solution was measured every two hours until the OD value reached 0.8-1.0. The bacterial solution was centrifuged at 5000g for 10 minutes, the bacteria were collected, the supernatant was discar...

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Abstract

The invention provides two flavonoid compounds, namely baicalein and scutellarein, of promoting a asymmetric cell division (ACD) regulation pathway of a root tip stem cell of a plant. The two compounds have effects of promoting SCR-SHR (scarecrow-short root) combination of an ACD active regulation complex of a root tip stem cell, does not influence RBR-SHR (retinoblastoma-related-short root) combination of a non-active complex and can activate NUC (nutcraker) expression of an ACD downstream key gene. At the same time, the invention provides an application of baicalein and scutellarein in preparing an SHR-SCR protein binding promoter and an NUC gene expression promoter. A secondary metabolism product of promoting ACD regulation of the root tip stem cell of the plant is found for the first time; a new biological effect of baicalein and scutellarein is found; an important tool is provided for an in-depth study of the stem cell regulation of the plant; and a new clue is provided for the development and application of a plant stem cell technology in industries of modern agriculture, biological medicine, hairdressing, health and the like.

Description

Technical field: [0001] The invention belongs to the field of plant stem cell regulation, and in particular relates to two flavonoids baicalein and scutellarin that promote the regulation pathway of asymmetric division of plant root tip stem cells, and their use in the preparation of a promoter for the regulation pathway of plant root tip stem cell asymmetric division Applications. Background technique: [0002] Every multicellular organism in nature originates from a single cell. During the development process, the offspring produced by the division of a single cell differentiate into various cell types required for life functions, and then can form different tissues, organs, and systems, and finally develop into a complete shape. A fully functioning organism. Stem cells are the cell source and signal regulation center of biological development, and are a kind of primitive undifferentiated cells with multi-directional differentiation potential and self-renewal and replicat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04
CPCC12N5/04C12N2500/30
Inventor 李唯奇张旭东黄星
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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