Rice stress resistance-related OsHMGB3 gene, and encoding protein and application thereof

A technology of genetic coding and rice stress resistance, which is applied in the fields of application, genetic engineering, plant gene improvement, etc., to achieve the effect of improving stress resistance

Inactive Publication Date: 2012-07-18
SHANGHAI AGROBIOLOGICAL GENE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the relationship between rice high mobility group box (HMGB) protein and stress resistance

Method used

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  • Rice stress resistance-related OsHMGB3 gene, and encoding protein and application thereof
  • Rice stress resistance-related OsHMGB3 gene, and encoding protein and application thereof
  • Rice stress resistance-related OsHMGB3 gene, and encoding protein and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Cloning of Rice OsHMGB3 Gene

[0032] 1. Seedling cultivation

[0033] The rice is germinated at 30°C for 48 hours, and then sown in the greenhouse. When the rice leaves are 3-5 pieces, the DNA or RNA is ready to be extracted.

[0034] 2. RNA extraction and first-strand cDNA synthesis

[0035] Extraction of RNA: The sample was frozen in a mortar with liquid nitrogen and ground into powder, and added to 1ml TRNzol-A + For the 2mL EP tube of the reagent, after fully shaking, place at room temperature for 5min, then add 0.2ml chloroform, shake vigorously for 15s, then place at room temperature for 3min; centrifuge at 12000rpm at 4°C for 10min, and transfer the supernatant to a new 2mL EP tube , add an equal volume of isopropanol to precipitate RNA, add 100 μl RNase-free ddH 2 O dissolved. The quality of total RNA was identified by electrophoresis, and then the RNA content was determined on a spectrophotometer.

[0036] Before reverse transcription, the extracted sampl...

Embodiment 2

[0049] Sequence information and homology analysis of rice OsHMGB3 protein

[0050] The ORF length of the novel rice OsHMGB3 full-length cDNA of the present invention is 399 bp, and the detailed sequence is shown in SEQ ID NO:3.

[0051] The amino acid sequence of rice OsHMGB3 was deduced according to the full-length ORF, with a total of 132 amino acid residues, a molecular weight of 14002.37 Daltons, and an isoelectric point (pI) of 7.80. See SEQ ID NO:3 for the detailed sequence. By comparison with the BLASTP program, it was concluded that the encoded protein of OsHGMB3 had a HMGB-UBF_HMG-box domain and belonged to the HMGB family.

[0052] Through the multiple sequence alignment of the HMGB encoded proteins of rice, Arabidopsis, corn, wheat, barley, soybean, upland cotton and Physcomitrella patens ( image 3 ), we found that, except for the core region of the protein, HMG-box, which is highly conserved among different species, its acidic C-terminus is also relatively conse...

Embodiment 3

[0054] Expression analysis of rice gene OsHMGB3 in various tissues and organs

[0055] 1. Select material

[0056] Seedling stage: select plump IRAT109 seeds, wash them with distilled water, sterilize with 3% NaClO for 10 minutes, wash them clean, accelerate germination at 30°C, transfer the seeds to the germination box for hydroponic growth after the seeds turn white, and apply nutrient solution after the three-leaf stage. Cultivate in a 16h / 8h light culture room at 28°C, and take the materials of new leaves and roots at the four-leaf stage for analysis;

[0057] Adult plant stage: IRAT109 was planted in paddy fields, and the flag leaves, leaf sheaths and ears of the plants were collected for analysis at the booting stage.

[0058] 2. RNA extraction and first-strand cDNA synthesis

[0059] With embodiment 1.

[0060] 3. Quantitative PCR Analysis

[0061] Quantitative analysis of gene expression was performed using Takara's Premix Ex Taq TM (Perfect Real Time) kit, and ...

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Abstract

The invention provides a rice stress resistance-related OsHMGB3 gene which is separated and cloned from a rice DNA (deoxyribonucleic acid) fragment. The gene contains the conserved domain of the HMG (high-mobility group) gene family, and is an HMGB (high-mobility group box) type member of the gene family. The OsHMGB3 gene is expressed after being induced by polyethylene glycol (PEG), high salinity, low temperature, high temperature and abscisic acid (ABA), and the over-expressed OsHMGB3 can enhance the osmotic stress resistance of rice at the seedling stage, and is related to rice stress resistance. The rice gene can notably respond to stress, can be applied to plant breeding for stress resistance, and can enhance the stress resistance of plants.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to a novel rice stress-resistance-related gene OsHMGB3 and its coded protein and application thereof, which can be used to cultivate transgenic plants with enhanced drought resistance. Background technique [0002] Drought and water shortage are one of the main factors affecting crop yield, and rice, as one of the main food crops, consumes a lot of water in its production, which has formed a sharp contradiction with the increasingly serious shortage of water resources in the world in recent years. It is one of the effective ways to deal with water shortage and ensure food security by using genetic engineering technology to transfer stress-resistant genes into currently widely used excellent crop varieties, and to breed new crop varieties that are both stress-resistant and high-yield. In-depth exploration and study of plant water-saving and drought-resistant genes is the basi...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/63C07K14/415A01H4/00A01H5/00
Inventor 刘鸿艳陈守俊徐凯罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT
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