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Application of the thick rattan gene iperd15

A gene and amino acid technology, which is applied in the field of application of the salt-tolerant and drought-resistant gene IpERD15, can solve the problems that breeding methods cannot meet people's needs, and achieve the effect of improving salt tolerance

Active Publication Date: 2019-03-05
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Today, with the rapid development of science and technology, the improvement of crop varieties is undergoing new changes, and the traditional breeding methods have become increasingly unable to meet people's needs

Method used

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  • Application of the thick rattan gene iperd15
  • Application of the thick rattan gene iperd15
  • Application of the thick rattan gene iperd15

Examples

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experiment example 1

[0046] Experimental Example 1: Cloning of the IpERD15 gene of Austenitum vine and construction of the IpERD15-pYES2 vector.

[0047] Specifically, the method comprises the following steps: extraction of total RNA, construction of cDNA library and gene cloning. Each step is mainly summarized as follows: (1) total RNA extraction

[0048] Take about 1g of the young roots, leaves, vines and leaf buds of the thick rattan, add liquid nitrogen and grind them thoroughly, use the MAGEN RNA extraction kit, and extract according to the instructions. The quality and concentration of RNA were determined with a Nanodrop micro-spectrophotometer.

[0049] (2) Construction of cDNA primary library

[0050] Use the Promega MMLV reverse transcription kit to synthesize the first strand of cDNA, take about 2 μg of the total RNA of the thick rattan, add 0.5 μg Oligo(dT)Primer and RNase-free H 2 O to a total volume of 15μL; 70°C for 5min, add 5μL M-MLV 5X Reaction Buffer and 5μL 10mM dNTP, 25units...

experiment example 2

[0055] Experimental example 2: Overexpression of IpERD15 gene in yeast improves the tolerance of yeast to salt and oxidative stress

[0056] (1) Yeast strain transformed with IpERD15-pYES2

[0057] Adjust the concentration of the constructed and sequence-verified IpERD15-pYES2 recombinant plasmid to 0.1 μg / μL, and use the lithium acetate transformation method to transform the yeast salt-sensitive mutant AXT3 and the corresponding wild-type yeast strain (W303), and transform the yeast to H 2 o 2 Sensitive mutants yap1Δ and skn7Δ and corresponding wild-type yeast strains (WT). At the same time, the yeast expression vector empty vector pYES2 was used as a control, and the above-mentioned yeast strains were respectively transformed to construct the Saccharomyces cerevisiae recombinant expression vector IpERD15-pYES2 (see figure 2 ).

[0058] (2) Overexpression of IpERD15 in yeast salt-sensitive mutant strain AXT3 and wild-type strain W303 improves salt tolerance of transgenic...

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Abstract

The invention belongs to the field of bioengineering of plant genes and discloses an application of an ipomoea pes-caprae gene IpERD15. An ipomoea pes-caprae protein IpERD15 with an amino acid sequence as shown in SEQ ID NO. 4 and / or an ipomoea pes-caprae gene IpERD15 are applied to improvement of salt resistance and drought resistance of plants, a cDNA reading frame of the ipomoea pes-caprae geneIpERD15 has a nucleotide sequence as shown in SEQ ID NO. 3 or a nucleotide sequence completely complementary and paired with the SEQ ID NO. 3, or the ipomoea pes-caprae gene IpERD15 has a nucleotidesequence for encoding the amino acid sequence as shown in SEQ ID NO. 4. The gene IpERD15 can be used for improving the salt resistance and drought resistance of the plants and can also be used for cultivating salt-resistant and drought-resistant plants in seed breeding.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and relates to the application of a salt-tolerant and drought-resistant gene IpERD15 of the thick rattan. Background technique [0002] With the increase of population and the continuous decrease of arable land, the environment continues to deteriorate, and various agricultural natural disasters such as drought and the formation of saline-alkali land have seriously threatened crop production. Increasing crop yield has attracted more and more attention. If there is no solution to the food problem, billions of people will face food shortages in the future. Therefore, it is of great strategic significance and necessity to understand the formation mechanism of plant stress resistance, discover stress-resistant gene resources, and then cultivate new stress-resistant crop varieties to increase food production and ensure food security. Today, with the rapid development of science and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/81C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8273
Inventor 罗鸣张美张会
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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