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Protein ErNAC7 related to drought and salt resistance of plants and coding gene and application thereof

A technology related to proteins and plants, applied in the field of genetic engineering, can solve problems that have not been reported yet

Inactive Publication Date: 2010-12-08
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, through genetic engineering, there are no relevant reports on cloning drought resistance and salt tolerance genes and constructing plant expression vectors to transform wheat and other food crops from Thinopyrum xinjiang

Method used

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  • Protein ErNAC7 related to drought and salt resistance of plants and coding gene and application thereof
  • Protein ErNAC7 related to drought and salt resistance of plants and coding gene and application thereof
  • Protein ErNAC7 related to drought and salt resistance of plants and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1: cDNA Cloning of ErNAC7 Gene Related to Drought Resistance and Salt Tolerance of Echinopsis Xinjiang.

[0067] The 45-day-old Thinopyrum seedlings were subjected to drought treatment for 5 hours, and the total RNA was extracted with Trizol. 5'RACE kit (5'RACE System for Rapid Amplification of cDNA Ends Kit) (GIBCOBRL, CAT.NO.18374-058) and 3'RACE kit (3'RACE System for Rapid Amplification of cDNA Ends Kit) (GIBCOBRL, CAT.NO.18373-019) obtained the full-length sequence of 894bp of ErNAC7 gene.

[0068] Trizol was used to extract the total RNA of Tritium xinjiang seedlings, and cDNA was obtained by reverse transcription with superscript II (invitrogen) reverse transcriptase. Primers P1 and P2 were designed according to the sequence of ErNAC7 gene coding region. Using the cDNA obtained by reverse transcription as a template, PCR amplification was performed with primers P1 and P2. The sequences of primers P1 and P2 are as follows:

[0069] P1: 5'-ATGAGCGGCGGAC...

Embodiment 2

[0073] Example 2: Expression characteristics of ErNAC7 gene in Xinjiang Thinopyrum under adversity stress treatment

[0074] The seeds of Thinopyrum xinjiang are planted in pots, and after 3 weeks of growth, the seedlings are taken and subjected to the following stress treatments: 1) drought treatment: the seedlings of Henpyrum xinjiang are taken out from the soil to absorb the moisture on the roots, and placed on dry filter paper; 2) Salt treatment: Put the seedlings of Echinopsis xinjiang in 200mM NaCl solution; samples were taken at 0, 1, 2, 5, 10, and 24 hours after the above treatments, quick-frozen with liquid nitrogen, and stored at -80°C for later use.

[0075] Total RNA was extracted from samples taken 0, 1, 2, 5, 10, and 24 hours after drought treatment and salt treatment, respectively, and the ErNAC7 DNA was used as a probe for quantitative fluorescence analysis.

[0076] Fluorescence quantitative analysis results see figure 2 . A is the sample treated with drought...

Embodiment 3

[0078] Embodiment 3: use ErNAC7 gene to enhance the drought resistance, salt tolerance of plants

[0079] 1. Construction of recombinant expression vector

[0080] 1) Construction of 35S-ErNAC7 recombinant expression vector

[0081] Using the cDNA obtained by reverse transcription of the total RNA of Echinopsis xinjiang as a template, PCR amplification was performed with specific primers containing SmaI and XbaI linker sequences; then SmaI and XbaI double-enzyme-digested the PCR product, recovered, and inserted the enzyme-cleaved product in the forward direction Between the SmaI and XbaI restriction sites behind the CaMV 35S promoter of the vector pBI121, a recombinant vector 35S-ErNAC7 was obtained.

[0082] The primer sequences are as follows:

[0083] ErNAC7[SmaI]5'-GCGCCCGGGATGAGCGGCGGACAGGAGCT-3'

[0084] ErNAC7[XbaI]5'-TGCTCTAGAGAGATTGAACGGCTTGCCCCAGT-3'

[0085] 2. Acquisition and identification of genetically modified tobacco

[0086] 1) Obtaining genetically modi...

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Abstract

The invention relates to the field of genetic engineering, in particular to protein ErNAC7 related to drought and salt resistance of plants and a coding gene and application thereof. The protein has an amino acid sequence shown as SEQ ID NO.1 and a gene sequence shown as SEQ ID NO.2. The protein related to the drought and salt resistance and the coding gene thereof have important theoretical and actual meanings for improving and enhancing the stress resistance of tobacco, improving yield, accelerating the breeding process of stress-resisting molecules and effectively saving water resources.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular, the invention relates to a plant drought-resistant and salt-tolerant related protein ErNAC7 and its coding gene and application. Background technique [0002] As one of the important food crops in our country, wheat occupies a very important position in the national economy. However, the arable land area in my country's arid areas is about 570 million mu, and the saline-alkali land is about 554 million mu. Every year, due to adversity such as drought and saline-alkali, the production of wheat in my country is reduced by 70-80 billion kilograms, which seriously affects the production and production of wheat. Quality restricts the rapid development of my country's wheat grain production. [0003] With the development of modern molecular biology, genetic engineering technology is used to study the relationship between plants and abiotic stress at the molecular level, reveal the mole...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63A01H5/00
CPCC12N15/8213C12N15/8261C12N15/8266Y02A40/146
Inventor 高世庆赵昌平唐益苗杨颖
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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