Medicago ruthenica late-embryogensis abundant protein gene and application thereof

An embryogenesis, lentil bean technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problems of gene cloning and stress resistance function identification without the late embryonic rich protein of lentil bean, and achieve the improvement of salt resistance. Effect

Inactive Publication Date: 2014-07-09
CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on the cloning of the gene enriched in late embryogenesis and the identification of the stress resistance function of L. alfalfa L.

Method used

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  • Medicago ruthenica late-embryogensis abundant protein gene and application thereof
  • Medicago ruthenica late-embryogensis abundant protein gene and application thereof
  • Medicago ruthenica late-embryogensis abundant protein gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 The late embryogenesis rich protein gene of lentil, the gene encoding MrLEA2, is derived from lentil, and it has the nucleotide sequence shown in SEQ ID NO.1 from position 1 to position 1181 in the sequence table.

[0034] 1. Cloning of MrLEA2 gene, a protein rich in late embryogenesis of lentil.

[0035] 1. Low temperature stress treatment and transcriptome sequencing of lentil seedlings:

[0036] ⑴Lentulina faecalis low temperature stress treatment:

[0037] After the lentil seeds were treated with concentrated sulfuric acid for 10 minutes, they were washed with tap water to remove residual sulfuric acid. The treated seeds are placed in a petri dish with double filter paper and germinated under the conditions of 21℃ and 16h / 8h photoperiod; 14 days later, the seedlings are transplanted into vermiculite to continue cultivation under the above conditions. Water once with 1 / 2MS nutrient solution; after 14 days, transfer the seedlings to a -4℃ light culture incubator f...

Embodiment 2

[0114] Example 2 Enriched protein gene in late embryogenesis of lentilus. This gene is the MrLEA2 coding gene. It has the formation of one or more base deletions, insertions or substitutions in SEQ ID NO.1 from position 1 to 1181 in the sequence table. Functionally active alleles.

[0115] The cloning of the late embryogenesis-rich protein MrLEA2 gene of lentil is the same as the prokaryotic expression of the late embryogenesis-rich protein MrLEA2 gene of lentil Example 1 .

Embodiment 3

[0116] Example 3 Enriched protein gene in late embryogenesis of lentil, this gene is MrLEA2 coding gene, it has figure 2 The protein with the amino acid residue sequence shown in SEQ ID NO. 2 from position 1 to position 321 shown in the sequence table.

[0117] The cloning of the late embryogenesis-rich protein MrLEA2 gene of lentil is the same as the prokaryotic expression of the late embryogenesis-rich protein MrLEA2 gene of lentil Example 1 .

[0118] The protein product encoded in the reading frame of MrLEA2 cDNA consists of 322 amino acid residues, of which aspartic acid, isoleucine and lysine account for 11.8% of the total number of amino acid residues respectively, and the molecular weight is 36.1kD, and the isoelectric point It is 4.55 and the average hydrophobicity (Grand average hydropathy, GRAVY) index is -0.4276, which has strong hydrophilicity.

[0119] The Pfam database (http: / / pfam.janelia.org) searched for the domain of the amino acid sequence encoded by MrLEA2, wh...

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Abstract

The invention relates to a medicago ruthenica late-embryogensis abundant protein gene and application thereof. The gene is an MrLEA2 coding gene, derived from medicago ruthenica, and has a nucleotide sequence as shown in a sequence table SEQ ID NO.1 or an allelomorphic gene with functional activity formed by deleting, inserting or substituting one or more basic groups in SEQ ID NO.1 in the sequence table, or the protein with an amino acid residue sequence as shown in SEQ ID NO.2 in the sequence table, or the protein derived from SEQ ID NO.2, which is formed by substituting, deleting or adding one or more amino acid residues for the amino acid residue sequence as shown in SEQ ID NO.2 in the sequence table, and provided with activity the same as that of the amino acid residue sequence in the SEQ ID NO.2. The gene obtained by the invention can be used for improving salt resistance, freezing resistance and heat resistance of plants and microorganisms.

Description

technical field [0001] The invention relates to the technical field of biogenetic engineering, in particular to a late embryogenesis-rich protein gene of alfalfa bean and its application. Background technique [0002] Late embryogenesis abundant protein (LEA) was first discovered in cotton seeds and was named for its large accumulation in late embryonic development (Dure et al., 1981). As well as decarboxylation (ABA) also induces the accumulation of late embryogenesis-abundant proteins in plant vegetative organs. Subsequent studies have found that late embryogenesis-rich proteins are not only present in plants, but also in Deinococcus radiodrurana, Bacillus subtilis, Artemia, nematodes, and rotifers. Evidence for an abundant protein in embryogenesis. According to the differences in amino acid sequence characteristics and amino acid composition, late embryogenesis enrichment can be divided into different groups. [0003] Intracellular functions of late embryogenesis-abund...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/70C12N1/21
Inventor 王海庆马超沈迎芳窦全文陈志国
Owner CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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