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Rice stress resistance related gene OsPP2C44 and coded protein and application thereof

A rice stress resistance and gene coding technology, applied in the field of genetic engineering, can solve the problems of ABA sensitivity reduction and increase

Inactive Publication Date: 2014-11-26
SHANGHAI AGROBIOLOGICAL GENE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The protein phosphatase activity they exhibit is increased by the influence of ABA, and when they are highly expressed or their activity is increased, the sensitivity of the plant to ABA will be significantly reduced

Method used

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  • Rice stress resistance related gene OsPP2C44 and coded protein and application thereof
  • Rice stress resistance related gene OsPP2C44 and coded protein and application thereof
  • Rice stress resistance related gene OsPP2C44 and coded protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Rice OsPP2C44 gene cloning

[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. RNA isolation:

[0036] Extraction of RNA: The sample was frozen with liquid nitrogen in a mortar, ground into powder, added to a 2mL EP tube containing 1mL TRNzol-A+ reagent (Tiangen Biochemical Technology Co., Ltd.), shaken fully, 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 2mL EP tube, add an equal volume of isopropanol to precipitate RNA, add 100μL 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.

[0037] 3. First-strand cDNA synthesis by reverse tr...

Embodiment 2

[0045] Example 2 Rice OsPP2C44 Protein sequence information and homology analysis

[0046] According to the new rice of the present invention OsPP2C44 ORFs derived from rice OsPP2C44 The amino acid sequence has a total of 321 amino acids and a molecular weight of 34700 Daltons. See SEQ ID NO: 2 for the detailed sequence. Comparing with the BLASTP program on the NCBI website (http: / / www.ncbi.nlm.nih.gov / Structure / cdd / wrpsb.cgi?RID=6ACDWHRB016

[0047] &mode=all), OsPP2C44 The encoded protein has a protein phosphatase PP2C catalytic domain and belongs to the PP2C subfamily.

[0048] By multiple sequence alignment of some PP2C encoded proteins in rice ( figure 1 ), we found that the protein has a highly conserved PP2C catalytic domain.

Embodiment 3

[0049] Example 3 Rice Gene OsPP2C44 Expression analysis in various tissues and organs

[0050] 1. Select material

[0051] IRAT109 was planted in paddy fields, and the roots, stems, flag leaves, leaf sheaths and ears of the plants were collected at the booting stage for analysis.

[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 carried out using SYBR® Premix Ex TaqTM (Perfect Real Time) kit from Takara Company and ABI PRISM® 7000 quantitative PCR instrument from the United States. according to OsPP2C44 Quantitative primers were designed for the full-length cDNA sequence. rice housekeeping genes actin (GenBank accession No. AY212324) was used as a reference gene, and primers were designed according to its cDNA sequence. Preparati...

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Abstract

The invention discloses an OcPP2C44 gene separated and cloned from rice deoxyribonucleic acid (DNA) fragments and related to rice stress resistance. The protein of the genetic code contains protein phosphatase family conservative catalytic structural domain and is a member of a PP2C type of the gene family. The OsPP2C44 gene is performed with inducible expression of high salinity, low temperature, high temperature and abscisic acid (ABA), and overexpression OsPP2C44 can improve osmotic stress resistance capability of rice during seedling stage and is related to rice stress resistance. The rice gene of the OcPP2C44 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 related to stress resistance of rice OsPP2C44 The invention and its application belong 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 agricultural water use. 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 glob...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/55C12N9/16C12N15/63A01H5/00
Inventor 刘鸿艳徐凯陈守俊罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT
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