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Rice root development and stress resistance related OsSAUR11 genes and encoding protein and application

A technology of stress resistance and root development, applied in the field of genetic engineering, can solve problems such as mRNAs instability

Active Publication Date: 2018-01-05
SHANGHAI AGROBIOLOGICAL GENE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The mRNAs encoded by SAUR are extremely unstable and will be degraded within minutes

Method used

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  • Rice root development and stress resistance related OsSAUR11 genes and encoding protein and application
  • Rice root development and stress resistance related OsSAUR11 genes and encoding protein and application
  • Rice root development and stress resistance related OsSAUR11 genes and encoding protein and application

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Experimental program
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Effect test

Embodiment 1

[0032] The cloning of embodiment 1 rice OsSAUR11 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]RNA extraction: freeze the sample in a mortar and grind it into a powder, add 1mL TRNzol-A+ reagent to a 2mL EP tube, shake fully, place at room temperature for 5min, then add 0.2mL chloroform, shake vigorously After 15 seconds, place at room temperature for 3 minutes; centrifuge at 12,000 rpm for 10 minutes at 4°C, transfer the supernatant to a new 2 mL EP tube, add an equal volume of isopropanol to precipitate RNA, and 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.

[0037] 3. Synthesis of first-strand cDNA by reverse transcription

[0038] (1) Before reverse transcript...

Embodiment 2

[0047] Example 2 Sequence information and homology analysis of rice OsSAUR11 protein

[0048] According to the novel rice OsSAUR11 ORF of the present invention, the amino acid sequence of rice OsSAUR11 is deduced, with a total of 190 amino acids and a molecular weight of 21043 Daltons. See SEQ ID NO: 2 for the detailed sequence. According to the comparison of the BLASTP program on the NCBI website (https: / / blast.ncbi.nlm.nih.gov / Blast.cgi), the protein encoded by OsSAUR11 has an auxin-induced SAUR domain and belongs to the SAUR subfamily.

[0049] Through the multiple sequence alignment of some SAUR-encoded proteins in rice ( figure 1 ), we found that all of the proteins contain a conserved SAUR domain.

Embodiment 3

[0050] Example 3 Expression analysis of rice gene OsSAUR11 under adversity stress

[0051] 1. Adversity stress treatment

[0052] Select plump Nipponbare seeds, wash them with distilled water, sterilize with 3% NaClO for 10 minutes and then clean them, accelerate germination at 30°C, transfer the seeds to the germination box for hydroponic growth after the seeds are white, and apply nutrient solution (standard nutrient solution of the International Rice Institute) after the three-leaf stage . Cultivate in 28°C, 16h / 8h light culture room, and carry out various stress and hormone treatments at the four-leaf stage: 20% PEG6000, 60mM hydrogen peroxide, 100μM abscisic acid (ABA) and 10μM auxin (IAA). The stress and hormone-treated materials were sampled before stress and 0.5, 1, 2h, 4h, 8h, 12h, 24h after stress. All processing and sampling were performed under continuous light conditions.

[0053] 2. RNA extraction and first-strand cDNA synthesis

[0054] With embodiment 1.

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Abstract

The invention discloses rice root development and stress resistance related OsSAUR11 genes which are isolated and cloned from rice DNA fragments. Protein encoded by the genes contains SAUR family conserved structural domains. The rice root development and stress resistance related OsSAUR11 genes belong to the field of rice cloning genes. The sequence of the OsSAUR11 genes is as shown in followingsequences: a DNA sequence as shown in SEQ ID NO: 1, a DNA sequence at least 90% homologous to SEQ ID NO: 1 and a subfragment functionally equivalent to the sequence as shown in SEQ ID NO: 1; the expression of the OsSAUR11 genes is the root development and stress resistance. The rice root development and stress resistance related OsSAUR11 genes disclosed by the invention have the beneficial effectsthat the OsSAUR11 genes are in inducible expression of osmotic stress, abscisic acid and auxins, and overexpression of OsSAUR11 can enhance the deep root ratio of rice and enhance drought resistanceof the rice, is related to rice stress resistance, and has obvious response to adverse environments, thereby being applied to plant drought-resistance breeding, improving the root configuration and enhancing plant stress resistance.

Description

technical field [0001] The invention relates to a gene related to rice root growth and stress resistance, in particular to a new rice deep root growth and stress resistance related gene OsSAUR11 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 important food crops. Rice needs 65% of the water used for agricultural irrigation. Modern biological technology is used to study the physiological and biochemical reactions of plant drought resistance, clone important drought resistance genes, and then transfer them to plants to cultivate stress-resistant New crop varieties, especially cultivating new water-saving and drought-resistant rice varieties are an effective way to alleviate the pressure on food production brou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00
Inventor 徐凯楼巧君陈亮罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT
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