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SiMYB107 protein related to plant adverse resistance, as well as related biomaterial and application of SiMYB107 protein

A biological material and stress-resistant technology, applied in the biological field, can solve problems affecting farmers' income and waste of nitrogen fertilizer

Inactive Publication Date: 2015-11-18
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the absorption and utilization of nitrogen fertilizer by plants is far less than half of its applied amount, and most of the nitrogen fertilizer is due to N 2 and NH 3 The volatilization of other gases, nitrification and denitrification denitrification, leaching, etc. are wasted
Moreover, nitrogen fertilizer has now become the most important cost required for crop production, and it will also affect farmers' income to a greater extent

Method used

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  • SiMYB107 protein related to plant adverse resistance, as well as related biomaterial and application of SiMYB107 protein
  • SiMYB107 protein related to plant adverse resistance, as well as related biomaterial and application of SiMYB107 protein
  • SiMYB107 protein related to plant adverse resistance, as well as related biomaterial and application of SiMYB107 protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Embodiment 1, the acquisition of SiMYB107 gene

[0097] 1. Processing of plant material

[0098] The seedlings of millet variety H214 at the seedling stage were treated with 0.2mM low nitrogen medium for 48h.

[0099] 2. RNA extraction

[0100] The total RNA of the millet variety H214 treated with the low-nitrogen culture solution was extracted, and the obtained RNA was used as a template for reverse transcription to obtain cDNA.

[0101] 3. PCR amplification

[0102] Using the cDNA obtained in step 2 as a template, PCR amplification was performed using primers F and R to obtain a PCR amplification product.

[0103] F: 5'-ATGGGGAGGTCGCCGTGCT-3' (SEQ ID NO: 3);

[0104] R: 5'-TCATAGTGGCAAGCTGCTGCTCTCT-3' (SEQ ID NO: 4).

[0105] The PCR amplification product was detected by 1% agarose gel electrophoresis, recovered and purified to obtain a DNA fragment with a size of 948bp. It was connected with the pEasyBlunt vector purchased from Tiangen Biochemical Technology Co...

Embodiment 2

[0107] Example 2, analysis of subcellular localization of SiMYB107 protein in plant cells

[0108] In order to locate the position of SiMYB107 protein in plant cells, the subcellular localization analysis of SiMYB107 protein was carried out. Specific steps are as follows:

[0109] 1. Using the reverse transcription product of the total RNA of millet H214 as a template, the full-length CDS region of the SiMYB107 gene was amplified by PCR using gene-specific primers to obtain PCR amplification products. The primer sequences are as follows:

[0110] F: 5'-ATGGGGAGGTCGCCGTGCT-3';

[0111] R: 5′-TCATAGTGGCAAGCTGCTGCTCTCT-3′

[0112] 2. Ligate the PCR amplification product with the pJIT16318-GFP vector to obtain the pJIT16318-SiMYB107-GFP fusion expression vector.

[0113] 3. The pJIT16318-SiMYB107-GFP fusion expression vector was transformed into protoplasts of millet leaves by PEG method, and the expression of green fluorescent protein was observed by laser confocal microscope...

Embodiment 3

[0115] Embodiment 3, the expression pattern analysis of SiMYB107 gene under the condition of low nitrogen stress

[0116] In order to study the expression of SiMYB107 gene under low-nitrogen stress conditions, the expression of SiMYB107 gene in millet H214 seedlings under low-nitrogen stress conditions was analyzed by using fluorescence quantitative technology. The specific steps are as follows:

[0117] 1. Select millet H214 seedlings and treat them with low nitrogen stress. The method of low nitrogen treatment: plant the millet seeds in nutrient soil, and after two weeks, move the seedlings to 0.2mM low nitrogen medium for low nitrogen treatment.

[0118] 2. Take the leaves of the seedlings at 0h, 1h, 6h, 12h, 24h and 48h of the low-nitrogen treatment, extract the total RNA, and reverse transcribe to obtain cDNA.

[0119] 3. Using the cDNA obtained in step 2 as a template, use SiMYB107-F and SiMYB107-R, Si001873m.g-F and Si001873m.g-R primers respectively, and detect the exp...

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Abstract

The invention discloses SiMYB107 protein related to plant low-nitrogen tolerance, as well as a related biomaterial and application of the SiMYB107 protein. The SiMYB107 protein is a), b) or c); a) protein of which the amino acid sequence is shown in the sequence 2 in the sequence table; b) fusion protein obtained by connecting labels to the N end and / or the C end of the protein of which the amino acid sequence is shown in the sequence 2 in the sequence table; c) the protein related to plant low nitrogen tolerance and obtained through replacement and / or deletion and / or addition of one or more amino acid residues based on the amino acid sequence shown in the sequence 2 in the sequence table. Experimental results show that the provided protein can be used for improving the low-nitrogen tolerance, low-potassium tolerance and low-phosphorous tolerance of a plant, and has an important application value in the aspect of plant breeding.

Description

technical field [0001] The invention relates to a SiMYB107 protein related to plant stress resistance and related biological materials and applications, belonging to the field of biotechnology. Background technique [0002] Nitrogen is a mineral nutrient element that needs the most in plant growth and development. The nitrogen content in plant dry matter and plant total nitrogen is 1.5%-2% and 16%, respectively. Elemental nitrogen plays a very important role in crop growth. It is a component of amino acids in plants, a component of protein, and a component of chlorophyll that plays a decisive role in photosynthesis in plants. [0003] In the second half of the 20th century, the demand for nitrogen fertilizer and other nutrients in major high-yielding crops increased dramatically in order to increase crop productivity, leading to a tenfold increase in global nitrogen fertilizer use. However, the absorption and utilization of nitrogen fertilizer by plants is far less than hal...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N5/10C12N15/11A01H5/00
CPCC07K14/415C12N15/8271
Inventor 陈明马有志徐兆师李连城周永斌胡利芹王二辉薛飞洋
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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