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Leymus chinensis fructan hydrolases, and encoding genes and use thereof

A technology of fructan hydrolase and encoding gene, which is applied to Leymus chinensis fructan hydrolase and its encoding gene and application field, and achieves the effect of high practical application value, broad application prospect and improvement of stress resistance.

Inactive Publication Date: 2009-04-22
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, only a few plant FEHs have been cloned, namely chicory (C.intybus) 1-FEH I (Claessens et al, 1990) and 1-FEH II (De Roover et al, 1999), Jerusalem artichoke (H. 1-FEH (Marx et al, 1997a), ryegrass (Lolium perenne) 6-FEH (Marx et al, 1997b), oat (Avena sativa) FEH with β(2→6) bond specificity (Henson and Livingstone, 1996), β(2→1)-specific barley (H.Vulgare) FEH (Henson and Livingstone, 1998), although FEH was obtained in ryegrass, little is known about fructan metabolism in Leymus chinensis, The research on the gene of fructan exohydrolase FEH is still blank

Method used

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  • Leymus chinensis fructan hydrolases, and encoding genes and use thereof
  • Leymus chinensis fructan hydrolases, and encoding genes and use thereof
  • Leymus chinensis fructan hydrolases, and encoding genes and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1. Obtaining the Complete Sequence of Leymus fructan Exohydrolase Gene Lc1-FEH cDNA

[0040] 1. Obtaining the full-length cDNA sequence of Leymus chinensis fructan exohydrolase gene Lc1-FEH

[0041] 1. Extraction of total RNA from plant materials

[0042] Leymus chinensis (Trin.) Tzvel (Jisheng No. 1, Jilin Province Jisheng Leymus chinensis Fine Variety Station)) seedlings were used as materials, and total RNA was extracted for 1% agarose gel electrophoresis detection. The detection results were as follows: figure 1 As shown, the extracted RNA has two obvious electrophoresis bands (swimming lane 1 and swimming lane 2), which are 28S RNA and 18S RNA respectively from top to bottom, indicating that the total RNA with high purity has been obtained.

[0043] 2. Cloning of Leymus chinensis fructan exohydrolase gene Lc1-FEH sequence fragment

[0044] According to the amino acid residue sequence of plant FEH, find the conserved region, and design primers according t...

Embodiment 2

[0065] Example 2, Bioinformatics analysis of Lc1-FEH and its encoded protein Lc1-FEH

[0066] 1. Bioinformatics analysis of Lc1-FEH gene and its encoded protein

[0067] Utilize DNAMAN to carry out bioinformatics analysis to the full-length cDNA of Lc1-FEH that embodiment 1 obtains, and this sequence full-length 1803bp (has the 1st-1803rd nucleotide sequence of the 5 ' end of sequence 2 in the sequence listing), coded by A protein composed of 600 amino acid residues, the amino acid sequence is the amino acid residue sequence of Sequence 1 in the sequence table, its estimated molecular weight is 66.8kDa, and the isoelectric point pI value is 5.49. The protein contains three typical NDPNG, FRDP and [WEC(V / P)D] domains, and the 72nd-76th amino acid residues at the amino terminal (N) of Sequence 1 in the sequence listing are conserved NDPSG domains , the 196th-199th is the FRDP domain, and the 251st-255th is the WECPD domain. Use the Signal3P (http: / / www.cbs.dtu.dK / services / Sign...

Embodiment 3

[0070] Example 3, Tissue-specific expression analysis of Lc1-FEH gene

[0071] To analyze the tissue-specific expression pattern of Lc1-FEH, the specific experimental method is: extract Leymus chinensis (Trin.) Tzvel (Jisheng No. 1, Jilin Province Jisheng Leymus chinensis Fine Breed Station)) seedlings (the seedlings grown for 8 weeks after germination) ), the total RNA of the four tissues of roots, rhizomes, leaf sheaths, and leaves, and the roots, rhizomes, leaves, pedicels, stem second nodes, flowering inflorescences, filling inflorescences, The highlights of milk ripening period use primer 3: TGA ATG GCT ACA GTG CTG C ​​and primer 4: CCA GTC CAC CGT TAT TGC C for RT-PCR analysis, and Actin gene (amplification primers are: primer 5: GCC AAC AGA GAG AAG ATG ACC and primer 6: ATA GAG GGA AAG CAC CGC CT) were used as internal standards to perform quantitative PCR on the cDNA template. After the reaction, the expression level was detected, and the results were as follows: Fi...

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Abstract

The present invention discloses a Chinese wildrye levan hydrolase, and an encoded gene and an application thereof. The albumen is Chinese wildrye levan hydrolase, which is the protein having one of the following amino acid residue sequence: 1) the amino acid residue sequence of the sequence 1 in the sequence table; and protein that is obtained by substituting and / or deficiency and / or addition of one or more amino acid residue to the amino acid residue sequence of the sequence 1 in the sequence table and has [beta]-2,1 glycosidic bond hydrolase function. The encode gene of the low temperature proof albumen can be transferred to a plant to improve the stress-resistance of the plant. The gene has a significant theoretical and practical significance for the research of the stress-resistance molecular mechanism of plant and the breeding of stress-resistance varieties of plant. The gene provides an economical, fast and effective approach for improving the stress-resistance of crop. The gene has wide applications and market prospect in the agriculture field.

Description

technical field [0001] The present invention relates to Leymus chinensis fructan hydrolase and its encoding gene and its application, in particular to the use of Leymus chinensis fructan hydrolase and its encoding gene in cultivating plants with improved stress resistance and hydrolyzing fructan β-2,1 glucoside application in the key. Background technique [0002] Fructan is a linear or / and branched polymer composed of fructose, and is an important storage carbohydrate of about 15% of angiosperms. With the discovery of fructan-metabolizing enzymes, fructan not only serves as short-term or long-term storage of plant heterotrophic organ roots, stems, grains and autotrophic organ leaves, but also has a variety of important physiological functions, such as regulating plant adaptation. Low-temperature photosynthesis, regulation of plant sucrose allocation, and adaptation of plants to water deficits, high concentrations of CO 2 , Hypoxic conditions, etc. In addition, fructan ha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/24C12N15/56C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H1/00A01H5/00C12P19/14
Inventor 刘公社王丽娟齐冬梅李晓峰
Owner INST OF BOTANY CHINESE ACAD OF SCI
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