Rice stress resistance related gene OsDTH1 related to and its encoded protein and application thereof

A rice stress resistance and gene technology, applied in the field of genetic engineering, can solve the problems of complex expression regulation mechanism, large number and variety of transcription factors, etc.

Inactive Publication Date: 2019-02-01
SHANGHAI AGROBIOLOGICAL GENE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The large number and variety of transcription factors in plants

Method used

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  • Rice stress resistance related gene OsDTH1 related to and its encoded protein and application thereof
  • Rice stress resistance related gene OsDTH1 related to and its encoded protein and application thereof
  • Rice stress resistance related gene OsDTH1 related to and its encoded protein and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Cloning of embodiment 1 rice OsDTH1 gene

[0033] 1. Seedling cultivation

[0034] 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.

[0035] 2. Isolation of RNA:

[0036] Extraction of RNA: The sample was ground into powder after being frozen with liquid nitrogen in a mortar, added to a 2mL EP tube filled with 1mL TRNzol-A+ reagent (Tiangen Biochemical Technology Co., Ltd.), fully oscillated, and placed at room temperature for 5min. Then add 0.2mL chloroform, shake vigorously for 15s, and place at room temperature for 3min; centrifuge at 12000rpm for 10min at 4°C, transfer the supernatant to a new 2mLEP tube, add an equal volume of isopropanol to precipitate RNA, add 100μL RNase-free wxya 2 O dissolved. The quality of total RNA was identified by electrophoresis, and then the RNA content was determined on a spectrophotometer.

[0037] 3. Synthesis of first-stra...

Embodiment 2

[0047] Example 2 Sequence information and homology analysis of rice OsDTH1 protein

[0048] According to the novel rice OsDTH1 ORF of the present invention, the amino acid sequence of rice OsDTH1 is deduced, with a total of 274 amino acids and a molecular weight of 249394 Daltons. See SEQ ID NO: 2 for the detailed sequence. Through the comparison of the BLASTP program on the NCBI website ( http: / / www.ncbi.nlm.nih.gov / Structure / cdd / wrpsb.cgi? RID=6ACDWHRB016 &mode=all), the OsDTH1 encoded protein has a bZIP domain and belongs to the bZIP family. Through the multiple sequence alignment of some bZIP encoded proteins in rice ( figure 1 ), we found that the protein has a highly conserved bZIP DNA-binding domain.

Embodiment 3

[0049] Example 3 Expression Analysis of Rice Gene OsDTH1 in Various Tissues and Organs

[0050] 1. Select material

[0051] Nipponbare rice is planted in paddy fields, and the roots, stems, flag leaves, leaf sheaths and ears of the plants are analyzed at the booting stage.

[0052] 2. RNA extraction and first-strand cDNA synthesis

[0053] According to the method in Example 1, RNA was extracted and reverse transcription was performed to synthesize the first-strand cDNA.

[0054] 3. Quantitative PCR Analysis

[0055] Quantitative analysis of gene expression was performed using Takara's Premix Ex TaqTM (Perfect RealTime) kit, and the United States Bio-Rad CFX96 quantitative PCR instrument. Quantitative primers were designed according to the full-length cDNA sequence of OsDTH1 (F: GCACGGAAGCAGGCATA, R: TCGACGCAGCTAGGGAA). The rice housekeeping gene actin (GenBankaccession No. AY212324) was used as a reference gene, and primers were designed according to its cDNA sequence. ...

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Abstract

The invention discloses an OsDTH1 gene separated and cloned from paddy rice (DNA) fragments and related to rice stress resistance. The gene encodes a transcription factor, which belongs to the basic region-leucine zipper (bZIP) protein family. The OsDTH1 gene is performed with inducible expression of high salinity, low temperature, high temperature and abscisic acid (ABA), and overexpression of OsDTH1 can improve osmotic stress resistance capability of paddy rice during seedling stage and is related to rice stress resistance. The rice gene has obvious response to adverse situation, can be applied to plant stress resistance breeding, and improves stress resistance of plants.

Description

technical field [0001] The invention relates to a gene related to stress resistance of rice, in particular to a new gene OsDTH1 related to stress resistance of rice and its application, belonging to the field of genetic engineering. Background technique [0002] Drought and water shortage are one of the important factors causing the reduction of global food production. Drought stress seriously affects plant growth and development, and even causes plant death. Rice is one of the most important food crops. Rice needs 65% of the water used in agriculture. Modern biological technology is used to study the physiological and biochemical reactions of plants against drought, clone important drought resistance genes, and then transfer them to plants to cultivate crops with enhanced stress resistance. New varieties, especially cultivating new water-saving and drought-resistant rice varieties are an effective way to alleviate the pressure on food production brought about by global dro...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8271
Inventor 徐凯杨诗勤夏辉罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT
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