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Malus xiaojinensis MxVHA-c protein and coding gene thereof, and application thereof

A technology that encodes genes and proteins, applied in applications, genetic engineering, plant genetic improvement, etc., can solve problems such as photosynthesis decline, crop yield and quality decline, premature aging, etc.

Inactive Publication Date: 2012-11-07
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Plant iron deficiency can cause leaf yellowing, premature senescence, photosynthesis decline, etc., and severe cases will cause crop yield and quality decline

Method used

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  • Malus xiaojinensis MxVHA-c protein and coding gene thereof, and application thereof
  • Malus xiaojinensis MxVHA-c protein and coding gene thereof, and application thereof
  • Malus xiaojinensis MxVHA-c protein and coding gene thereof, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1, the acquisition of the protein MxVHA-c related to plant resistance to iron deficiency and its coding gene

[0048] 1. Obtaining the full length of the MxVHA-c gene

[0049] Take Malus xiaojinensis (Malus xiaojinensis) (the public can obtain from China Agricultural University, the non-patent literature that has recorded this Malus xiaojinensis is: Cheng Minghao, Li Xiaolin, Zhang Yungui, Excellent rootstock resources for apples-Malus xiaojinensis, Journal of Southwest Agricultural University, 2000 October, 22 (5): 383-386) was used as the experimental material, the total RNA of the leaves was extracted, and it was reverse transcribed into cDNA. Using this cDNA as a template, using primer F1 (sequence 3 in the sequence listing) and primer R1 (sequence 4 in the sequence listing) as a primer pair:

[0050] Primer F1: AAA GAATTC ATGTCTTTCTTCAACCTTC, the underlined part indicates the enzyme cutting site EcoRI,

[0051] Primer R1: AAA GTC GAC CCCTCAGCTCTTGACTG...

experiment example 2

[0089] Experimental example 2, the acquisition of transgenic yeast and the analysis of its resistance to heavy metal toxicity

[0090] 1. Transform yeast with the cloned MxVHA-c gene

[0091] The plasmids of pEZS-MxVHA-c, pEZS-NL and yeast expression vector pYES2.0 (Beijing Dingguo Changsheng Biotechnology Co., Ltd.) were digested with EcoRI and XbaI respectively, and the 1218bp MxVHA-c-GFP was obtained respectively. and 720bp GFP were connected to pYES2.0 after enzyme digestion to obtain recombinant expression vectors pYES2.0-MxVHA-c-GFP and pYES2.0-GFP, the latter can be used as a positive control.

[0092] The recombinant expression vectors pYES2.0-MxVHA-c-GFP and pYES2.0-GFP were transformed into Escherichia coli competent cells, the plasmids were extracted, digested with EcoRI and XbaI, and detected by agarose gel electrophoresis to obtain MxVHA-c of about 498bp Gene fragment. The result is as Figure 5As shown, the lane M is the DNA molecular weight standard of DL1500...

Embodiment 3

[0100] Embodiment 3, the acquisition and identification of transgenic plants

[0101] 1. Construction of recombinant expression vector

[0102] 1. Digest the transient expression vector pEZS-MxVHA-c with restriction endonucleases EcoRI and XbaI, and recover the digested product (about 1218bp).

[0103] 2. Design primers F3 and R3 with EcoRI and KpnI at both ends according to the sequence of the 35S promoter. The enzyme-cut transient expression vector pEZS-MxVHA-c was used as a template, and the primer pair composed of F3 and R3 was used for PCR amplification, and about 1000bp PCR amplification product was recovered.

[0104] F3: 5'-CCGGAATTCCAATCCCCACCAAAACCTGA-3';

[0105] R3: 5'-TTCGAGCTCCGTGTCCTTCCAAATGAAA-3'.

[0106] 3. Digest the PCR amplified product of step 2 with restriction endonucleases EcoRI and KpnI, and recover the digested product.

[0107] 4. Digest pCAMBIA2300 plasmid (Beijing Dingguo Changsheng Biotechnology Co., Ltd.) with restriction enzymes EcoRI and K...

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Abstract

The invention discloses a Malus xiaojinensis MxVHA-c protein and a coding gene thereof, and application thereof. The Malus xiaojinensis MxVHA-c protein provided by the present invention is a protein (a) or (b) as follows: (a) a protein composed of an amino acid sequence shown in a sequence 1 in the sequence table; and (b) a protein derived from (a), related to iron deficiency tolerance of plant, and obtained by replacement and / or deletion and / or addition of one or more amino acid residues on an amino acid residue sequence of the sequence 1 in the sequence table. The MxVHA-c provided by the invention has important regulatory effect on plant with iron deficiency stress, and provides a good theoretical basis for further study on iron deficiency tolerance of Malus xiaojinensis.

Description

technical field [0001] The invention relates to the MxVHA-c protein of Begonia chinensis and its coding gene and application. Background technique [0002] Iron is one of the essential nutrients for plant growth. Iron deficiency in plants can cause leaf yellowing, premature senescence, and decreased photosynthesis, and in severe cases, crop yield and quality will decline. Iron deficiency in agricultural production is a worldwide problem, and there are a large number of reports about plant iron deficiency in countries all over the world. Therefore, there is a need to develop effective strategies to combat iron deficiency. Contents of the invention [0003] One object of the present invention is to provide a protein and its coding gene related to plant iron deficiency resistance. [0004] The protein related to plant iron deficiency resistance provided by the present invention is named MxVHA-c, which is derived from Malus xiaojinensis, and is the protein of (a) or (b) as ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00C12R1/865
Inventor 韩振海张倩张新忠王忆陈峰吴婷许雪峰
Owner CHINA AGRI UNIV
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