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Apple rootstock salt stress resistance related gene MdLysMe3 as well as encoding protein and application thereof

A salt stress and resistance protein technology, applied in the field of apple genes, can solve the problems of abiotic stress resistance function and mechanism of action, etc.

Active Publication Date: 2021-09-24
北京市林业果树科学研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the functional research on the identified apple LysM gene family mainly focuses on the interaction between plants and fungi and the response to heavy metal stress, etc., and there is still a gap in the research on the function and mechanism of abiotic stress resistance.

Method used

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  • Apple rootstock salt stress resistance related gene MdLysMe3 as well as encoding protein and application thereof
  • Apple rootstock salt stress resistance related gene MdLysMe3 as well as encoding protein and application thereof
  • Apple rootstock salt stress resistance related gene MdLysMe3 as well as encoding protein and application thereof

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Experimental program
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Effect test

Embodiment 1

[0028] Embodiment 1: Cloning of the gene MdLysMe3 related to salt stress resistance

[0029] Comparative analysis of the gene expression levels of apple rootstock G935 and salt-sensitive rootstock SH6 under salt stress showed that the MdLysMe3 gene positively regulated the salt stress resistance of apple rootstock ( figure 1 ). In the present invention, the root system of apple salt-resistant rootstock G935 is used as the material, the total RNA is extracted after 12 hours of salt stress treatment, and the RNA is reverse-transcribed into cDNA by a reverse transcription kit. Use cDNA as a template to amplify the target fragment. The primer sequences are:

[0030] MdLysMe3-F:5'-ATGGCATCAAGAAAACTGGCAG-3'

[0031] MdLysMe3-R:5'-TCATTCGGTAGGAGTGATCTTG-3'

[0032] The reaction system for PCR amplification is: TaKaRa LA Taq (5U / μL) 0.2μL, 10×LA PCR Buffer 2.0μL, dNTP Mixture (2.5mM each) 3.2μL, MdLysMe3-F, MdLysMe3-R primer solution (10pM / μL ) each 1.0 μL, cDNA template 1.0 μL, ...

Embodiment 2

[0036] Embodiment 2: Enhance the salt tolerance of apple sensitive salt rootstock with MdLysMe3 gene

[0037] 1. Construction of recombinant expression vectors

[0038] Using the above-mentioned recombinant vector pGEM-MdLysMe3 as a template, use specific primers containing Bgl II and BstE II linker sequences to perform PCR amplification of the MdLysMe3 gene; connect to the pGEM-T Easy vector, transform Escherichia coli, extract the plasmid for sequencing verification, and perform Bgl II and BstE II double digestion, and then the digestion product is forward inserted between the Bgl II and the BstE II restriction site after the CaMV 35S promoter of the plant expression vector pCAMBIA1304, to obtain the recombinant vector pCAMBIA1304-MdLysMe3 ( image 3 ).

[0039] The primer sequences are as follows:

[0040] MdLysMe3-OE1:5'-AGATCTATGGCATCAAGAAAACTGGCAG-3'

[0041] MdLysMe3-OE2:5'-GGTAACCTCATTCGGTAGGAGTGATCTTG-3'

[0042] 2. Obtaining genetically modified apple rootstocks

...

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Abstract

The invention belongs to the field of agricultural biotechnology engineering, and particularly relates to an apple rootstock salt stress resistance related gene MdLysMe3 as well as an encoding protein and application thereof. The sequence of the gene is as shown in SEQ ID No.1, and the amino acid sequence of the protein is as shown in SEQ ID No.2. An apple rootstock with enhanced salt stress resistance is obtained by constructing an overexpression vector of the apple rootstock salt stress related gene MdLysMe3 and transforming the overexpression vector to the apple rootstock. The apple rootstock salt stress resistance related gene MdLysMe3 can be used for improving the salt stress resistance of the apple rootstock, and has important significance in expanding the suitable growing area of apple cultivation, accelerating the stress-resistant molecular breeding process and the like.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and relates to apple genes, in particular to the application of the gene MdLysMe3 related to salt stress resistance of apple rootstocks and the encoded protein thereof. Background technique [0002] my country is a big apple producing country, and its planting area and output have ranked first in the world for many years. Apple is a fruit tree that is propagated by grafting, and the stress resistance of the rootstock directly affects its planting distribution, yield and quality. In recent years, with the acceleration of urbanization, the shortage of labor force and the increase of orchard management costs and other issues, the main apple planting areas in my country are gradually westward from emerging areas such as the Bohai Bay, the old course of the Yellow River, the Loess Plateau in Northwest China to the central and western regions of Xinjiang. shifting development process. However...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/74
CPCC07K14/415C12N15/8273
Inventor 李兴亮张军科周佳李民吉周贝贝杨雨璋孙婷婷张强魏钦平
Owner 北京市林业果树科学研究院
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