Method for improving plant adverse resistance and application thereof

A plant stress resistance, plant technology, applied in the directions of botanical equipment and methods, biochemical equipment and methods, applications, etc., can solve problems such as improving plant stress resistance

Inactive Publication Date: 2015-09-02
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is still little research on the information related to AKT1 genes in soybean, and there is no method to improve plant stress resistance using AKT1 genes

Method used

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  • Method for improving plant adverse resistance and application thereof
  • Method for improving plant adverse resistance and application thereof
  • Method for improving plant adverse resistance and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The acquisition of embodiment 1GmAKT1

[0039] 1. Cloning and bioinformatics analysis of GmAKT1

[0040] The homologous gene of Arabidopsis thaliana AKT1 was obtained from the Phytozome database, and the two gene copies with the highest homology were obtained from Dongnong 50 by homologous cloning method, which were located on chromosome 17 and chromosome 5 respectively, and the amino acid homology rate 69.41% and 68.00%, respectively, named GmAKT1, GmAKT2. The cloned GmAKT1 of the present invention contains an open reading frame with a length of 2628 bp, encoding 875 amino acids.

[0041] Soybean (Glycine max) extracted with Trizol reagent (Dongnong 50, Wu Tianlong, Yang Qingkai, Ma Zhanfeng, Wu Zongpu, Zhao Shuwen, Gao Fenglan, Li Wenbin, Zhang Guodong, Yang Qi, Meng Qingxi, Wang Jinling. Dongnong Xiaodoudou No. 1 new soybean variety selection Report, [J]. Northeast Agricultural University Journal, 1994, (25) 1:104; the public can obtain from Northeast Agricultural ...

Embodiment 2

[0050] Embodiment 2 Construction of GmAKT1 gene plant expression vector and cultivation of GmAKT1 gene Arabidopsis

[0051] 1. Construction of plant expression vector pCXSN-GmAKT1

[0052] The pCXSN vector is a plant expression vector for TA cloning. The pCXSN vector fragment with a T-terminal was obtained by single-digestion with XcmⅠ, and the GmAKT1 fragment with an A-terminal was inserted into the pCXSN plant expression vector. After Hind III identified the positive connection, it was transformed into a TOP10 sensor After the positive colonies were identified, they were transformed into Agrobacterium to obtain the pCXSN-GmAKT1 plant expression vector.

[0053] 2. Cultivation of Transgenic GmAKT1 Arabidopsis

[0054] 1. Obtaining of transgenic GmAKT1 Arabidopsis

[0055] 1) The plant expression vector pCXSN-GmAKT1 obtained above was transformed into Agrobacterium tumefaciens EHA105 by freeze-thaw method and sent for sequencing. As a result, the Agrobacterium with plasmid p...

Embodiment 3

[0066] Example 3 Transgenic T4 generation function analysis of GmAKT1 Arabidopsis

[0067] 1. Phenotype analysis of transgenic Arabidopsis thaliana

[0068] Seeds of wild type, GmAKT1-transformed wild-type Arabidopsis, AKT1 mutant, and GmAKT1-transferred AKT1 mutant Arabidopsis were planted on MS medium, and moved to soil after 1 week to observe their growth and development process and count the plant height , rosette leaf number, flowering days, pod number, calculate the average value, the results are shown in Table 1 and Figure 4 shown.

[0069] Table 1 The growth conditions of four kinds of Arabidopsis

[0070]

[0071] The plant height of GmAKT1-transformed wild-type Arabidopsis was significantly higher than that of wild-type, mutant and GmAKT1-transformed mutants. It had fewer rosette leaves, earlier flowering days, and more pods in the growth cycle.

[0072] 2. ABA treatment of transgenic Arabidopsis

[0073] Seeds of wild-type Arabidopsis thaliana, AKT1 mutant, ...

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Abstract

The invention discloses a method for improving plant adverse resistance and an application thereof, and belongs to the technical field of plant transgenic breeding. According to the method, after a stress resistance gene is obtained through PCR amplification, a plant expression vector is established and then transformed to an agro-bacterium; the agro-bacterium containing the plant expression vector infects a target plant, and infected target plant is cultivated until a seed containing the stress resistance gene is obtained. The GmAKT1 gene is transformed to Arabidopsis, so that the Arabidopsis containing the GmAKT1 gene shows the characteristics that the number of rosette leaves is small, blooming is early, the number of pads is large and the plant height is increased, and the Arabidopsis is superior to wild type Arabidopsis to a certain extent in physiology. Meanwhile, the Arabidopsis containing the GmAKT1 gene is improved in ABA resistance, drought resistance and slat resistance. In addition, the over-expression of the GmAKT1 can remove reactive oxygen in the plant in an adverse situation to some extent.

Description

technical field [0001] The invention relates to a method for improving stress resistance of plants and an application thereof, belonging to the technical field of plant transgenic breeding. Background technique [0002] Soybean is very sensitive to adversity stresses such as drought and salinity, and adversity leads to a decline in soybean quality and yield. Therefore, it is very important to study stress-related genes, clarify the molecular mechanism of stress resistance, and improve soybean stress resistance. With the development of molecular biotechnology, it has become the focus of current research to transfer stress-resistant genes into related crops so that they can be efficiently expressed in target crops, thereby improving the adaptability of crops to adverse environments and improving crop yield and quality. However, there is still little research on the information about AKT1 genes in soybean, and there is no method to use AKT1 genes to improve plant stress resist...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H5/00
Inventor 李文滨李永光王英琪
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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