Application of SHR-SCR in fate determination of cortex cells of leguminous plants and modification of division potential of cortical cells of non-leguminous plants

A cortical cell and cytokinin technology, applied in the field of cell attribute transformation, can solve the problem of poor understanding of the regulation mechanism of cell division

Active Publication Date: 2020-09-08
CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, little is known about the regulatory mechanisms of cell division

Method used

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  • Application of SHR-SCR in fate determination of cortex cells of leguminous plants and modification of division potential of cortical cells of non-leguminous plants
  • Application of SHR-SCR in fate determination of cortex cells of leguminous plants and modification of division potential of cortical cells of non-leguminous plants
  • Application of SHR-SCR in fate determination of cortex cells of leguminous plants and modification of division potential of cortical cells of non-leguminous plants

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0118] Preparation of Agrobacterium rhizogenes competent cells

[0119] 1) Medium and solution

[0120] Ultrapure water, LB medium, 10% glycerol (v / v).

[0121] 2) Competent preparation

[0122] Step 1: Take the preserved strain, streak it on the LB plate containing the corresponding antibiotic, and incubate at 28°C for 24-48hrs.

[0123] Step 2: Pick a single clone in 3mL LB liquid medium, shake it overnight at 28°C, inoculate it into anti-LB medium at 1:100, and incubate at 28°C at 200rpm until OD600=0.5~1.0.

[0124] Step 3: Ice bath for 10 minutes, then centrifuge at 2,500 g for 10 minutes at 4°C.

[0125] Step 4: Remove the supernatant, first gently suspend the cells in 5 mL of ice-cold ultrapure water, then add 200 mL of ice-cold ultrapure water, and centrifuge at 2,500 g for 10 min at 4°C.

[0126] Step 5: Repeat step 4 once.

[0127] Step 6: Remove the supernatant, first lightly suspend the cells in 5 mL of ice-cold 10% glycerol, then add 200 mL of ice-cold 10% glycerol, and centrif...

Embodiment 1

[0175] Example 1. SCR is conservatively expressed in cortical cells of legumes

[0176] 1. MtSCR extended expression in root cortex and epidermal cells

[0177] The inventors found that the Medicago truncatula SCARECROW (MtSCR) gene is not only expressed in the quiescent center and endodermis of Medicago truncatula roots, but also in the cortex and epidermal cells of the roots, which is specific to Arabidopsis thaliana AtSCR. Sexual expression is very different in the resting center and endothelial layer of Medicago truncatula root ( figure 1 A~ figure 1 B).

[0178] 2. AT1 and Enhancer control the expression of MtSCR promoter in root cortex cells

[0179] The inventors performed a series of truncation experiments on the MtSCR promoter (2899 upstream of ATG of MtSCR; called pMtSCR(2899bp)), and combined the prediction analysis of cis-elements (http: / / plantpan2.itps.ncku.edu.tw / ) ), it was found that when AT1 Box (AT1 for short) and Enhancer (En for short) are missing in the promoter,...

Embodiment 2

[0188] Example 2: MtSCR participates in nodule symbiosis

[0189] 1. MtSCR participates in nodule symbiosis

[0190] The present inventors obtained plant materials of the Mtscr-1 (NF11026) and Mtscr-2 (NF20550) insertion mutants of Medicago truncatula Tnt1, which were grown at 24°C, 16h light / 22°C, 8h dark environment, and wild-type Medicago truncatula (WT) served as a control.

[0191] Sm1021 Rhizobium was inoculated when the plants grew to 3 days. After that, the nodule growth of the plants was measured on the 7th day, the 14th day, the 21st day, and the 28th day after the inoculation.

[0192] The result is image 3 As shown in A, the wild-type Medicago truncatula can produce nodules normally, while the Tnt1 insertion mutant Mtscr-1 (NF11026) and Mtscr-2 (NF20550) of Medicago truncatula can produce very low nodules on the 7th day after inoculation with rhizobia. The number of nodules on the 14th, 21st, and 28th day afterwards was also much lower than that of the wild type.

[0193...

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Abstract

The invention belongs to the field of biotechnology and botany, relates to a method for modifying cell attributes and regulating root nodule symbiosis, and in particular to an application of a plant in-vivo new mechanism formed on the basis of SHR-SCR in modification of division potential of cortex cells. The invention further provides a novel way for identifying the traits of plants, so that a feasible method is provided for effective identification of plants, and a powerful tool is provided for breeding and screening of plants.

Description

Technical field [0001] The present invention belongs to the fields of biotechnology and botany; more specifically, the present invention relates to a method for modifying cell attributes and regulating nodule symbiosis. [0002] technical background [0003] Nitrogen-fixing symbiosis is a mutually beneficial symbiosis between plants and nitrogen-fixing microorganisms. According to the different symbiotic bacteria, it can be divided into cyanobacteria symbiosis, actinomycete symbiosis and rhizobia symbiosis. The symbiosis of rhizobia is the symbiosis between legumes and rhizobia. The new organ formed by rhizobia in plants-root nodules convert free nitrogen in the air into nitrogen-containing compounds for plant growth, enhance the ability of legumes to adapt to low-nitrogen fertilizer soil, and at the same time, legumes provide suitable rhizobia Carbohydrates necessary for the environment and growth. In addition, when the legumes die, the fixed nitrogen will be released into the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895A01H1/04C12N15/82C12N15/29A01H5/06A01H6/54
CPCC12Q1/6895A01H1/04C12N15/8227C12N15/8261C07K14/415C12Q2600/156C12Q2600/13
Inventor 王二涛董文涛朱亚云常会中
Owner CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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