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Protein VaPBP2-L for enhancing drought resistance of plants and application of protein VaPBP2-L

A drought resistance, vapbp2-l technology, applied in the direction of application, plant peptides, plant products, etc., to achieve the effect of promoting the cultivation process, improving plant drought tolerance, and improving drought resistance

Active Publication Date: 2021-12-21
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, while screening the drought-resistant germplasm resources of adzuki bean, looking for new drought-resistant genes, and mining and analyzing the drought-resistant genes, they can be used as effective drought-resistant gene resources for the research on drought-resistant breeding of other crops. less

Method used

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  • Protein VaPBP2-L for enhancing drought resistance of plants and application of protein VaPBP2-L
  • Protein VaPBP2-L for enhancing drought resistance of plants and application of protein VaPBP2-L
  • Protein VaPBP2-L for enhancing drought resistance of plants and application of protein VaPBP2-L

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 - Acquisition of protein VaPBP2-L and its coding gene and recombinant expression vector

[0023] (1) Using 9.0% mannitol stress treatment for 36h as material, extract total RNA, and obtain cDNA by reverse transcription, using the cDNA as a template, under the guidance of primer VaPBP2-L-F1 and primer VaPBP2-L-R1 , amplified by the conventional PCR method, after the reaction, the PCR amplified product was detected by 1% agarose gel electrophoresis, and a DNA fragment of about 1511bp was recovered and purified, such as figure 1 shown;

[0024] (2) Using LIC (non-ligation reaction cloning) reaction, the gene fragment is connected to the carrier PVX-LIC (the T-DNA fragment of the carrier PVX-LIC contains the lethal gene ccdB, and its two sides contain the recognition sequence of the LIC reaction, The vector is owned by the laboratory, literature: Zhao J, Liu Q, Hu P, et al (2016) An efficient Potato virus X-basedmicroRNA silencing in Nicotiana benthamiana. Sci Re...

Embodiment 2

[0056] Example 2 - Acquisition of recombinant Agrobacterium tumefaciens

[0057] The recombinant vector PVX-LIC-VaPBP2-L was transformed into Agrobacterium tumefaciens GV3101 by freeze-thaw method to obtain Agrobacterium tumefaciens GV3101 containing the recombinant vector PVX-LIC-VaPBP2-L, and the recombinant Agrobacterium was named GV3101 / PV X- LIC-VaPBP2-L; (freeze-thaw method refers to Amanda M Davis, Anthony Hall, Andrew J Millar, ChiarinaDarrah and Seth J Davis, Protocol: Streamlined sub-protocols for floral-dip transformation and selection of transformants in Arabidopsis thaliana, 2009, public available obtained from Yangtze University).

[0058] The empty vector PVX-LIC was transformed into Agrobacterium tumefaciens GV3101 by freeze-thaw method to obtain Agrobacterium tumefaciens GV3101 containing the empty vector PVX-LIC, and the recombinant Agrobacterium was named GV3101 / PVX-LIC.

Embodiment 3

[0059] Example 3 - Acquisition and Identification of Transient Expression Transgenic Tobacco

[0060] (1) Obtaining genetically modified tobacco

[0061] Two kinds of recombinant Agrobacterium GV3101 / PVX-LIC-VaPBP2-L and GV3101 / PVX-LIC obtained in Example 2 were used to prepare an Agrobacterium suspension, and the volume ratio of the culture medium to the cell in the suspension was 1:1. The seeds of Nicotiana benthamiana were sown in the culture medium (peat: vermiculite: perlite mixed at a volume ratio of 1:3:0.5) and cultivated in an artificial greenhouse. When the tobacco grows to 4-5 leaves, start injecting the topmost fully expanded new leaves. Use a disposable syringe to absorb 1mL of the bacterial solution, remove the needle of the syringe, hold the lower part of the leaf with your fingers, and gently force the bacteria in the syringe to hydraulically send and penetrate into the leaf tissue. Inject 2 leaves per tobacco, GV3101 / PVX- Five plants were injected with LIC-V...

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Abstract

The invention provides a protein VaPBP2-L for enhancing drought resistance of plants as well as a coding gene and application of the protein VaPBP2-L. A protein VaPBP2-L gene is separated by taking red-bean germinated seeds as materials, an amino acid sequence of the protein VaPBP2-L gene is shown as SEQ ID NO.1, and after a virus expression vector is constructed by using the gene and is excessively expressed and transformed into tobacco, the drought resistance of the plants can be remarkably improved; and various plant expression vectors are constructed by utilizing the protein VaPBP2-L gene, the protein VaPBP2-L gene can be widely applied to cultivation of new drought-resistant varieties of transgenic plants and crops, is applied to drought-resistant breeding of plants as a drought-resistant genetic resource, and a cultivation process of the new drought-resistant varieties (lines) of the crops and the plants is promoted.

Description

technical field [0001] The invention relates to the technical field of biogenetic engineering, in particular to a protein VaPBP2-L for enhancing plant drought resistance, its coding gene and its application. Background technique [0002] As the global warming continues to intensify in recent years, drought stress has become one of the main abiotic stresses that cause crop yield reduction. The grain loss caused by drought accounts for more than 50% of all natural disasters in my country. Therefore, solving the drought problem is an important challenge to achieve sustainable agricultural development. [0003] Traditional crop drought-resistant breeding has a long cycle, large investment, and limited drought-resistant germplasm resources, which lead to slow progress in current drought-resistant breeding. Biotechnology breeding can break the limitation between species and provide a new way for efficient drought-resistant breeding. Therefore, identifying and screening drought-re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/00A01H6/82
CPCC07K14/415C12N15/8273C12N15/8205
Inventor 沙爱华陈银华王燕娟蒋浩中黄林涛向艳涛魏正欣
Owner HAINAN UNIVERSITY
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