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Plant salt resistance related protein, and coding gene and application thereof

A coding and genetic technology, applied in the direction of plant gene improvement, application, plant peptides, etc., can solve problems such as complex mechanisms

Inactive Publication Date: 2014-03-12
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the complexity of its mechanism, many important issues in plant salt resistance remain to be explored

Method used

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  • Plant salt resistance related protein, and coding gene and application thereof
  • Plant salt resistance related protein, and coding gene and application thereof
  • Plant salt resistance related protein, and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1. Obtaining the full-length cDNA sequence and genome sequence of the gene Lc9230 related to salt resistance of Leymus chinensis

[0059] 1. Cloning of the 5' end sequence of Lc9230, a salt-resistance-related gene of Leymus chinensis

[0060] 1. Plant material processing and total RNA extraction

[0061] The seedlings of Leymus chinensis (Trin.) variety Zhongke No. 2 (Institute of Botany, Chinese Academy of Sciences) were used as materials, and total RNA was extracted after 6 hours of salt stress treatment, and detected by 1% agarose gel electrophoresis. The results are as follows: figure 1 shown. The extracted RNA has two obvious electrophoresis bands, which are 28S RNA and 18S RNA from top to bottom, indicating that the total RNA with higher purity and integrity has been obtained.

[0062] 2. Cloning of the 5' end sequence of Lc9230, a salt-resistance-related gene of Leymus chinensis

[0063] Using the total RNA of Leymus chinensis seedlings subjected to sa...

Embodiment 2

[0083] Example 2. Expression pattern and tissue specificity analysis of Lc9230 gene under salt stress conditions

[0084] Leymus chinensis (Zhongke No. 2) seedlings grown normally for 4 weeks were subjected to salt stress (400mM NaCl). The stress treatment time was 0, 1, 3, 5, 8, 12, 24 hours. The total RNA of Leymus chinensis seedlings treated above was extracted, and the expression patterns of Lc9230 gene under different stress conditions were analyzed by RT-PCR method. The sequence of primers for amplifying the Lc9230 gene is 5'-GGACGACGACCATCAATC-3' (position 5-22 of sequence 2) and 5'-CTTCACGCCGAAAGGACA-3' (reverse complementary sequence of position 851-868 of sequence 2); reaction The conditions are 94°C for 4 minutes; 94°C for 30s, 56°C for 30s, 72°C for 30s, 30 cycles; 72°C for 10min. Using Actin as the internal reference gene, the primer sequences for amplifying internal reference Actin were 5′-GTGCTTTCCCCTCTATGCAAGTGGT-3 and 5′-CTGTTCTTGGCAGTCTCCAGCTC-3′; the react...

Embodiment 3

[0088] Example 3, Subcellular Localization of Lc9230 Gene

[0089] The Lc9230 gene cDNA (sequence 2) amplified in the above-mentioned Example 1 was used as a template, and the upstream and downstream primers 5'-G with restriction sites Bgl II and Spe I at both ends were respectively used AAGATCT GATGGGGGGACGTGTCG-3' (the underlined part is the Bgl II recognition site, and the following sequence is the 145th-160th position of sequence 2) and 5'GG ACTAGT CCACCGCTTGTCGCC-3' (the underlined part is the Spe I recognition site, and the subsequent sequence is the reverse complementary sequence of the 731-745th position of sequence 2) was amplified by PCR to obtain the target band with a size of 617bp. The target band was double-digested with restriction endonucleases Bgl II and Spe I. With the expression vector pCAMBIA 1302 (Robert et al. (2006) A hypermorphic mutation in the protein phosphatase2C HAB1 strongly affects ABA signaling in Arabidopsis FEBS Lett.580, 4691–4696, which c...

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Abstract

The invention discloses a plant salt resistance related protein, and a coding gene and an application thereof. The protein is a protein (a) composed of an amino acid sequence represented by sequence 1 in a sequence table, or is a protein (b) obtained by substituting and / or deleting and / or adding one or more amino acid residues to the amino acid sequence represented by the sequence 1, relating to the plant salt resistance and derived from the sequence 1. The plant salt resistance related protein and its coding gene Lc9230 are induced by high salt, and transgenic plants of the protein and its coding gene have a strong salt resistance. Experiment results show that after 150mM NaCl tolerant culture is carried out for 3 weeks, the survival rate of wild Arabidopis thaliana plants is 25%, and the survival rate of the transgenic plants with the Lc9230 gene is about 90%, so the Lc9230 can be used as a candidate gene of the plant salt resistance gene engineering, and can be used for improving the plant salt resistance ability.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a protein related to plant salt resistance and its encoding gene and its application, in particular to a protein and its encoding gene derived from Leymus chinensis and related to plant salt resistance. Use in transgenic plants with increased salinity. Background technique [0002] The area of ​​saline soil in the world exceeds 800 million hectares 2 , the total area of ​​various saline soils in my country is about 99 million hm 2 . The continuous expansion and deepening of salinized land has seriously restricted the development of agriculture and animal husbandry. Salt damage is the main factor affecting plant growth and crop yield, which can reduce the average yield per mu by more than 50%. Therefore, improving the adaptability of crops and various livestock grasses to salt is a research hotspot in the field of modern agriculture and animal husbandry technology, and it is also a ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/21C12N15/82C12N15/84A01H5/00
CPCC07K14/415C12N15/8273
Inventor 刘公社李晓霞程丽琴高琼马甜
Owner INST OF BOTANY CHINESE ACAD OF SCI
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