New anti-drought and cold-resistant gene CaDHN1 in caragana korshinskii

A caragana and gene technology, applied in genetic engineering, plant gene improvement, recombinant DNA technology, etc., can solve the problems of destroying free radical metabolism of plant cells, increasing active oxygen free radicals, and cell membrane system damage

Inactive Publication Date: 2010-09-01
INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
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  • Summary
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AI Technical Summary

Problems solved by technology

Low temperature, drought and other adversities often destroy the balance of free radical metabolism in plant cells, increase the production of

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] Embodiment 1, the construction of twisted bar leaf full-length cDNA library

[0010] The caragana plant was cultivated in a cultivation pot, cultivated in a greenhouse for 60 days, and then subjected to drought treatment until the whole plant wilted. The young leaves were quick-frozen in liquid nitrogen, ground, and total RNA was extracted with TRIzole and dissolved in water. The mRNA was processed by the mRNA purification kit. The mRNA is taken for reverse transcription, and then the second strand is synthesized. Ligate with pDNR-LIB vector after homogenization (see Creator for details TM SMART TM cDNA Library Construction Kit (Clontech Cat. No. 634903) and transform Escherichia coli. Single clones were obtained by smearing the plates and stored separately.

Embodiment 2

[0011] Example 2. Random EST sequencing of cDNA library: 3000 single clones were randomly selected from the library for 5'-end EST sequencing, and the successfully sequenced sequences were subjected to BLAST to select ESTs similar to verified drought-resistance-related genes.

Embodiment 3

[0012] Example 3 Select ESTs similar to dehydrin by BLAST, and pass through their corresponding clones to obtain the full-length DNA sequence of the candidate stress resistance gene. If the full-length sequence cannot be obtained, use the EST to design primers, and obtain the full-length sequence by 5' or 3' RACE.

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PUM

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Abstract

The invention discloses caragana korshinskii dehydrin and gene thereof. The dehydrin consists of 243 amino acid residues, and comprises two Y stricture regions and a K structure region; and compared with the other dehydrin, the dehydrin is abundant in histidine (H) and glycocoll (G). The whole length of the gene of the dehydrin is 1165bp, the dehydrin is a 731bp opened reading frame from 5'end to 106-837 position, and the dehydrin has intense express under the condition that the plant is stressed by the drought and the cold, so that the dehydrin reduces the damage to plant cells, improves the anti-drought and cold-resistant capability, and is the key element of the anti-drought mechanism of the plant. The similarity between the amino acid sequence of the gene of the caragana korshinskii dehydrin and that of the gene of the pea dehydrin is 34%, and the gene of the dehydrin is a new gene. If the plant expression vector of the gene is built, and the anti-drought gene is leaded into various plants, the anti-drought and cold-resistant capability of the plant can be improved.

Description

technical field [0001] A new drought-resistant and cold-resistant gene CaDHN1 in caragana, relates to a plant dehydrin and its coding gene, and its function is to improve the drought-resistant and cold-resistant performance of plants, which belongs to the field of plant genetic engineering. Background technique [0002] Various abiotic stresses, including drought, salinity, low temperature and other abiotic stresses, are important factors restricting the production and development of crops, forest grasses and vegetables. my country's arid and semi-arid arable land totals 570 million mu, accounting for 38% of the country's arable land area. Our country's annual grain production is reduced by 70-80 billion kg due to water shortage. Cultivating and promoting stress-resistant plant varieties is the most effective way to increase their yield and quality, ensure national food security and improve the ecological environment. Although the use of plant's own disease resistance genes...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N15/82C12N5/10A01H5/00
Inventor 郭九峰孙国琴白晨李晓东乔慧蕾
Owner INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
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