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Rice RDB1 drought resistant gene

A rice and genetic technology, applied in genetic engineering, plant genetic improvement, plant peptides, etc., can solve problems such as uninvolved and insufficient

Inactive Publication Date: 2010-09-29
SHANGHAI AGROBIOLOGICAL GENE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its related research is still not enough, and its role in the stress resistance mechanism has not been involved, especially in rice, the most important food crop in my country, no one has carried out related research

Method used

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  • Rice RDB1 drought resistant gene

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Cloning of Rice RDB1 Gene from Drought-resistant Rice

[0065] 1. Tissue separation (isolation)

[0066] The drought-resistant rice Huhan 3 was obtained from the Shanghai Agricultural Biological Gene Center. The drought-resistant rice Huhan 3 was germinated at 25°C for 24 hours and planted in a liquid medium in a greenhouse. The leaves of the drought-resistant rice Huhan 3 were 3- At 5 slices, it is ready to extract DNA or RNA.

[0067] 2. RNA isolation (RNA isolation)

[0068] Take the seedlings out of the water first, expose the roots to the air for 1 hour to simulate drought conditions, then take the leaves and put them into liquid nitrogen, grind them with a mortar, add a 1.5mL EP tube filled with lysate, shake them fully, and then transfer them into in a glass homogenizer. After homogenization, transfer to 1.5mL EP tube, and extract total RNA (TRIzol Reagents, GIBCO BRL, USA). The quality of total RNA was identified by formaldehyde denaturing gel electrophoresi...

Embodiment 2

[0085] Copy number analysis of rice RDB1 derived from drought-resistant rice in the drought-resistant rice Huhan 3

[0086] Adopt conventional method to extract DNA from the leaf of drought-resistant rice Huhan No. 3 (with reference to "molecular cloning", Sambrook et al., 1989), digest DNA [20 μ g (microgram) / sample] with EcoR I and PstI respectively, transfer DNA to After hybridization membrane (nylon membrane). Using GeneImages from Amersham Pharmacia TM Contents CDP-Star TM labeling module (PRN3540), we labeled the coding region of RDB1 gene as a probe, and then performed hybridization (hybridization at 60°C for 16 hours). The membrane was taken out, placed in washing solution I (1*SSC, 1% SDS), and rinsed three times at 60° C. for 15 minutes each time. Transfer to washing solution II (0.1*SSC, 1% SDS) and rinse at 60°C for 3 times, each time for the same 15 minutes. Press the X-ray film for 60-90 minutes, then develop and fix (refer to the Roche DIG labeled kit manu...

Embodiment 3

[0088] Bioinformatics analysis of rice RDB1 protein derived from drought-resistant rice

[0089] In order to understand the conserved functional region of the rice RDB1 protein, its protein sequence was searched on the professional protein family classification website Pfam (http: / / www.sanger.ac.uk / Software / Pfam / search.shtml) , and found that the most reliable region of amino acids 361-584 of the protein sequence is the BURP region, which is named after the first letter of the four protein names of rapeseed BnBNM2, broad bean VfUSP, Arabidopsis AtRD22 and tomato LsPG1β, including The proteins in this region are plant-specific and play an important role in plant development and stress resistance. Its Pfam-B region number is: Pfam-B22373. Tables 3 and 4 below are search results and descriptions provided on the Pfam website.

[0090] table 3

[0091] Domain

start

end

Bits

value

Alignment

mode

BURP

361

584

348.60

9.1e-102

...

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PUM

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Abstract

The invention discloses an OsRDB1 drought-resistant gene sequence of Huhan No.3 of drought-resistant rice in the technical field of genetic engineering. The code of the gene sequence has the active polypeptide nucleotide sequences of the rice RDB1 protein of Huhan No.3 of the drought-resistant rice, and the nucleotide sequences can be hybridized with the nucleotide sequences from the orders of 60to 1817 of nucleotides in SEQ ID NO.3 and have the homology by at least 70 percent with the nucleotide sequences from the orders of 60 to 1817 of the nucleotides in the SEQ ID NO.3. By applying the rice RDB1 protein sequence and the nucleotide sequence thereof in a research on the improvement of plant stress resistance by a transgenic technique, the plants can have obvious drought resistance performance and the loss on the biological yield of different crops planted in arid lands can be reduced.

Description

technical field [0001] The invention relates to a gene protein coding sequence used in the technical field of genetic engineering. More specifically, it relates to a rice drought resistance gene. Background technique [0002] Plants will encounter a variety of natural environments during their growth, and some natural disasters often cause a large reduction in crop production, such as drought, stagnant waterlogging, and pests and diseases. Breeding stress-resistant crop varieties is one of the important goals of agricultural science and technology research. An important breeding method at present is to genetically engineer crops with various stress-resistant genes to obtain new varieties of stress-resistant crops. In order to resist the stress of the adverse external environment, plant cells sense the changes in the external environment and transmit signals to the cells, which will induce the cells to express various response genes to resist the damage of the adverse envir...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415
Inventor 余舜武吴金红罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT