Plant stress resistance related protein PvNAC52 as well as coding gene and application thereof

A plant and genetic technology, applied in the direction of plant genetic improvement, botany equipment and methods, applications, etc., can solve problems affecting plant growth, affecting plant growth and development, productivity, oxidative damage, etc.

Active Publication Date: 2022-07-12
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, saline-alkali stress can lead to abnormal plant growth, dwarfing, yellowing, wilting, etc.; drought stress affects the growth and productivity of plants, resulting in cellular energy consumption, redox imbalance and oxidative damage, etc., which will affect the emergence rate and growth of kidney beans , leading to a decline in the yield of kidney beans, and in some places there may even be zero yield due to drought; low temperature is also a situation that often affects plant growth, and low temperature stress will affect the growth and yield of kidney beans, etc.

Method used

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  • Plant stress resistance related protein PvNAC52 as well as coding gene and application thereof
  • Plant stress resistance related protein PvNAC52 as well as coding gene and application thereof
  • Plant stress resistance related protein PvNAC52 as well as coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Cloning of PvNAC52 gene

[0059] The total RNA of kidney bean leaves was extracted with TRIzol reagent (Invitrogen), and the product cDNA was obtained by reverse transcription with ReverTr aAce qPCR RTMaster Mix with gDNA Remover (TOYOBO).

[0060] Using cDNA as a template, PCR amplification was performed using the following primers:

[0061] PvNAC52-F: 5′-ATTTCATTTGGAGAGAACACGGGGGACTCTAGAGGATCCCCGGGATGGGAGTTCCAGAG-3′;

[0062] PvNAC52-R: 5'-TTGCTCACCATAAGGGACTGACCACCCGGGGAATGCCTGAACCCGAATCCCACCGGCTG-3'.

[0063] The amplification system is shown in Table 1, and the procedure is shown in Table 2.

[0064] Table 1PCR reaction system

[0065]

[0066] Table 2 PCR reaction conditions of bacterial liquid

[0067]

[0068] cloned products such as figure 1 , the obtained PCR amplification product was subjected to agarose gel electrophoresis, and then recovered by gel recovery kit (OMEGO).

Embodiment 2

[0070] Obtaining transgenic plants

[0071] 1. Construction of recombinant plasmids

[0072] PBI121-EGFP was digested and recovered by restriction endonuclease SmaI, such as figure 2 . The obtained linearized PBI121-EGFP and PvNAC52 cloned PCR products were ligated with T4 ligase, the reaction system was as shown in Table 3, and the reaction was performed at 22 °C for 10 min to obtain the recombinant plasmid PBI121-EGFP-PvNAC52. At the same time, sequencing verification was performed.

[0073] Table 3T4 ligase ligation system

[0074]

[0075] The acquisition of transgenic plants

[0076] 1. The recombinant plasmid PBI121-EGFP-PvNAC52 was introduced into Agrobacterium GV3101 to obtain recombinant Agrobacterium.

[0077] 2. Suspend Agrobacterium to prepare Agrobacterium infection solution, adjust OD 600 is 0.6-1. The Colombian wild-type Arabidopsis thaliana plants were taken, and the inflorescences were completely immersed in the transfection solution for 15 s. After...

Embodiment 3

[0084] Example 3 Resistance identification of transgenic plants

[0085] In this example, by adding NaHCO 3 , NaCl, ABA and Mannitol stress to study the resistance of the gene to abiotic stress.

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PUM

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Abstract

The invention discloses a plant stress resistance related protein PvNAC52 as well as a coding gene and application thereof. The amino acid sequence of the protein PvNAC52 provided by the invention is as shown in SEQ ID NO: 1. Experiments prove that the stress resistance of the arabidopsis thaliana can be improved by over-expressing the PvNAC52 gene in the arabidopsis thaliana, and the improvement of the stress resistance is shown as follows: under salt / alkali / drought / osmotic stress, the stress resistance is improved; the survival rate of the transgenic plant is increased, the root length is increased, the fresh weight is increased, the water loss rate is reduced, the cell membrane damage is reduced, the malondialdehyde content is reduced, the relative conductivity is reduced, the H2O2 content is reduced, superoxide anions are reduced, the ROS content is reduced, the catalase activity is increased, the SOD enzyme activity is increased, the peroxidase activity is increased, and the proline content is increased. Therefore, the protein PvNAC52 and the coding gene thereof can regulate and control the stress resistance of plants, and have important theoretical significance and practical value in cultivation of plants with enhanced stress resistance.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a plant stress resistance-related protein PvNAC52 and its encoding gene and application. Background technique [0002] Plant abiotic stress, including drought stress, low temperature stress, salinity stress, high temperature stress and heavy metal stress, will have a great impact on plant growth and development. For example, saline-alkali stress can cause plants to fail to grow normally, dwarf, yellow, wilt, etc.; drought stress affects plant growth, development and productivity, resulting in cellular energy consumption, redox imbalance and oxidative damage, which will affect the emergence rate and growth of kidney beans. , resulting in a decline in the yield of kidney beans, and even zero yield in some places due to drought; low temperature is also a common situation that affects plant growth, and low temperature stress will affect the growth and yield of kidney beans. ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/84C12N1/21A01H5/00A01H6/20C12R1/01
CPCC07K14/415C12N15/8273C12N15/8271
Inventor 于崧刘梦红翟玲侠秦猛张翼飞于立河
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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