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Application of brassica napus BnMAPK1 gene in improving plant shade tolerance and method

A technology of Brassica napus and shade tolerance, which is applied in the field of plant genetic engineering and can solve problems such as effects that have not been reported yet

Active Publication Date: 2021-06-25
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the role of Brassica napus BnMAPK1 gene in the process of plant light response has not been reported

Method used

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  • Application of brassica napus BnMAPK1 gene in improving plant shade tolerance and method
  • Application of brassica napus BnMAPK1 gene in improving plant shade tolerance and method
  • Application of brassica napus BnMAPK1 gene in improving plant shade tolerance and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1, Cloning of BnMAPK1 promoter and analysis of cis-acting elements

[0036] The 3-4 week-old Brassica napus Zhongyou 821 seedlings were selected, and the genomic DNA (gDNA) of the roots, stems, and leaves were extracted respectively, and mixed at equal masses to prepare a mixed gDNA template for future use. Referring to the Brassica napus sequence in the GENOSCOPE database, specific primers ProBnMAPK1-F and ProBnMAPK1-R were designed, and the promoter of the BnMAPK1 (BnaC05g07560D) gene was cloned using the mixed gDNA of Zhongyou 821 plant as a template. A promoter with a length of 1389bp was obtained and named as ProBnMAPK1. The cloning primer sequences are as follows:

[0037] ProBnMAPK1-F SEQ: 5'-ATACTATTGTTTAAATGCATGTACTG-3' (SEQ ID NO.2);

[0038] ProBnMAPK1-R SEQ: 5'-TAGTTTCCTCTCCTTCTTCATCACA-3' (SEQ ID NO.3);

[0039] The cloned promoter sequence is shown in SEQ ID NO.4.

[0040] Using the PlantCARE online tool to analyze the cis-acting elements of Pr...

Embodiment 2

[0044] Embodiment 2, the photoresponse curve determination of Brassica napus Zhongyou 821

[0045] In order to explore how BnMAPK1 regulates the light response process, the photosynthetic curves of Brassica napus Zhongyou 821DH line plants were firstly measured. When the seedlings grew to 3-4 weeks, three healthy plants with basically the same growth were taken, and the net photosynthetic rate Pn (Netphotosynthetic rate). will CO 2 Concentration set to ambient CO 2 Concentration, photosynthetically active radiation PAR (Photosynthetically active radiation) gradient from high to low is set to 1400, 1200, 1000, 800, 500, 400, 300, 200, 100, 50, 20, 0μmol m –2 ·s –1 , respectively measure the corresponding Pn under each PAR intensity of the leaves. The photoresponse curve (http: / / photosynthetic.sinaapp.com / index.html) was fitted online using the rectangular hyperbolic correction model of Ye et al., and the light saturation point LSP (Light saturation point) and light compens...

Embodiment 3

[0049] Example 3. Effects of BnMAPK1 Overexpression on Photosynthetic Characteristics of Brassica napus in Weak Light Environment

[0050] Brassica napus BnMAPK1 overexpression (OE) and Zhongyou 821DH wild-type (WT) plants were used as test materials, and 300 μmol·m –2 ·s –1 Low light treatment. The relative content of chlorophyll, photosynthetic parameters, chlorophyll fluorescence parameters and RUBP carboxylase (Ribulose-1,5-bisphosphate carboxylase / oxygenase) activity were measured in different periods of low light treatment.

[0051] 3.1 The effect of overexpression of BnMAPK1 on the relative content of chlorophyll in low-light environment

[0052] Select 3 stable expression T 3 The BnMAPK1 overexpression lines (OE-1, OE-2, OE-3) and Zhongyou 821DH line WT of the first generation were selected. Three healthy plants with the same growth were selected for each line, and they were treated with low light at 0, 7, 14, After 21 and 28 d, the relative chlorophyll content (SP...

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Abstract

The invention discloses an application of a brassica napus BnMAPK1 gene in improving plant shade tolerance and a method. Under the stress of weak light, BnMAPK1 improves the shade tolerance of brassica napus by positively regulating photosynthetic parameters such as chlorophyll content, net photosynthetic rate, stomatal conductance, intercellular CO2 concentration and transpiration rate in plant photosynthesis and activating a PSII photochemical reaction center; Molecular mechanism research shows that the BnMAPK1 plays a role in regulating and controlling a light system II complex LHCB gene through a photosynthesis-antenna protein approach, and the BnMAPK1 positively regulates and controls the shade tolerance of the brassica napus by negatively regulating and controlling expression of LHCB1.3, LHCB3 and LHCB4.2 genes in a light system II, so that reference data is provided for functional research of MAPKs cascade, and meanwhile, a theoretical foundation is laid for development of shade-tolerant molecular markers and breeding of excellent varieties of the shade-tolerant molecular markers.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to the application of BnMAPK1 gene of Brassica napus in improving the shade tolerance of plants, and also relates to the method for improving the shade tolerance of plants of Brassica napus BnMAPK1 gene. Background technique [0002] Rapeseed is one of the most important crops in the world. It ranks first in my country's oil crops in terms of planting area, yield, vegetable oil supply, and protein source, and occupies a pivotal position in agricultural production. With the growth of population, the improvement of living standards and the development of aquaculture, the demand for edible vegetable oil and protein feed continues to increase. However, the area of ​​cultivated land in our country continues to decrease, the space for expanding crop planting area is relatively limited, the contradiction between grain and oil land use is prominent, and water and soil reso...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N15/84A01H5/00A01H6/20
CPCC12N9/12C12N15/8269C12N15/8261C12N15/8205C12Y207/11024
Inventor 王珍梁颖王晓改王淑文李加纳
Owner SOUTHWEST UNIVERSITY
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