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Application and method of Brassica napus bnmapk1 gene in improving plant shade tolerance

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: 2022-02-18
SOUTHWEST UNIV
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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 and method of Brassica napus bnmapk1 gene in improving plant shade tolerance
  • Application and method of Brassica napus bnmapk1 gene in improving plant shade tolerance
  • Application and method of Brassica napus bnmapk1 gene in improving plant shade tolerance

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 the application and method of Brassica napus BnMAPK1 gene in improving the shade tolerance of plants. Under weak light stress, BnMAPK1 positively regulates the chlorophyll content, net photosynthetic rate, stomatal conductance, intercellular CO 2 Concentration, transpiration rate photosynthetic parameters, activate PSⅡ photochemical reaction center at the same time, improve the shade tolerance of Brassica napus; molecular mechanism research shows that BnMAPK1 regulates the LHCB gene of the photosystem Ⅱ complex through the photosynthesis-antenna protein pathway, and BnMAPK1 regulates the LHCB gene of the photosystem Ⅱ complex through the negative Regulate the expression of LHCB1.3, LHCB3 and LHCB4.2 genes in photosystem Ⅱ, and then positively regulate the shade tolerance of Brassica napus, which provides a reference for the functional study of the MAPKs cascade, and at the same time provides a reference for the development of shade tolerance molecular markers. It has laid a theoretical foundation for the selection and breeding of its fine varieties.

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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Patent Type & Authority Patents(China)
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 UNIV
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