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Novel rice gene regulating salt stress tolerance

A tolerant, salt-stressed technology, applied in a new field of genes, that can solve problems such as salt water influx, high financial or human cost, etc.

Inactive Publication Date: 2003-12-31
NAT INST OF AGROBIOLOGICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods require a lot of money or manpower
Removal of salt through irrigation results in large amounts of salt water flowing into surrounding areas, causing environmental pollution

Method used

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  • Novel rice gene regulating salt stress tolerance
  • Novel rice gene regulating salt stress tolerance
  • Novel rice gene regulating salt stress tolerance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] (Example 1: Activation of Tosl7 by culture and identification of the resulting mutants)

[0087] As described in Hirochika et al, 1996, Proc. For callus initiation culture and cell suspension culture. According to the method of Otsuki (1990) (rice protoplast culture line, Association for Agriculture, Forestry and Fisheries Technology Information), the culture conditions for activating Tosl7 for gene disruption were determined.

[0088] Briefly, mature rice seeds (Otsuki (1990), supra) were cultured in MS medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) (25°C, 1 month) , to induce callus. The resulting callus was cultured in N6 liquid medium containing 2,4-D (Otsuki (1990), supra) for 5 months, and the callus was transferred to the redifferentiation medium (Otsuki (1990), Reference supra), to obtain redifferentiated rice (first generation (R1) plants).

[0089] About 20 R1 seeds were recovered from each plant. Seeds were sterilized with 1.0% sodium hypochlor...

Embodiment 2

[0091] (Example 2: Isolation of flanking sequences of Tosl7)

[0092] To find the genes controlling the phenotypes observed in Example 1, it was necessary to isolate the Tosl7 flanking sequences that had been transferred into the genomic DNA.

[0093] DNA was prepared from the R2 rice (line ND1004) obtained in Example 1 by the CTAB method (Murray and Thompson, 1980, Nucleic Acids Res, 8, 4321-4325). Tosl7 target site sequences were amplified by inverse PCR using total DNA as described (Hirochika et al., 1996, supra; and Sugimoto et al., 1994, Plant J., 5, 863-871).

[0094] Briefly, approximately 0.5 μg of total DNA from mutant plants (ND1004- / - lines) in which Tosl7 had been inserted into the target site by transposition was first digested with XbaI. Digested DNA was extracted with phenol / chloroform, followed by ethanol precipitation for purification. Ligation was then performed overnight using T4 DNA ligase in a total volume of 300 μl at 12°C. The ligated DNA was purified...

Embodiment 3

[0095] (Example 3: Structural analysis of the priming gene in the mutant)

[0096] RNA from wild-type rice (Nipponbare) seedlings grown in soil for 11 days was prepared as follows. First, total RNA of seedlings was extracted using ISOGEN solution. Total RNA was loaded onto an oligo(dt) cellulose column included in the mRNA purification kit (Stratagene) to obtain poly(A) mRNA. cDNA is synthesized from the obtained poly(A) mRNA by a usual method. The cDNA library was constructed in the HybriZAP-2.1 vector (Stratagene). The infectious capacity of the cDNA library was 5×10 5 a plaque. The pBluescript plasmid containing the cDNA insert was lysed in vivo using the E. coli strain XL1-Blue MRF2.

[0097]The cDNA library was screened according to the method described in Molecular Cloning, A Laboratory Manual (Sambrook et al., 1989), in which the inverse PCR product of Tosl7 flanking sequence obtained in Example 2 was used as a probe.

[0098] Nine cDNA clones showing strong hybri...

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Abstract

The present invention provides genes encoding proteins capable of controlling salt stress tolerance. The present invention provides polynucleotides encoding plant genes capable of controlling salt stress tolerance. The polynucleotide includes a polynucleotide having a nucleotide sequence encoding the amino acid sequence from the 1st methionine to the 243rd asparagine of SEQ ID NO: 2 in the sequence listing, or having a coding sequence with one or more Amino acid deletions, substitutions and / or additions to the nucleotide sequence of the amino acid sequence, and polynucleotides capable of controlling salt stress tolerance.

Description

technical field [0001] The present invention relates to novel genes. More specifically, the present invention relates to novel genes encoding proteins that function to control plant tolerance to salt stress. Background technique [0002] Plants are constantly under stress, even under normal growing conditions. The stresses include the following: salt, drought, high temperature, low temperature, strong light, air pollution and the like. In agricultural production, salt stress has attracted people's attention. Salt is produced when soil and rocks break down, and there is a constant stream of salt being produced. Salt flows into rivers or seas when it rains. In desert areas where rainfall is scarce, only a small amount of salt flows out, making the salt concentration in the soil moisture quite high. Plants are absorbing salt (nutrients) at the same time as they absorb water through root osmosis. When salt concentrations are high, plants cannot absorb water. In addition, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C07K14/415C12N15/29C12N15/82
CPCC07K14/415C12N15/8273
Inventor 广近洋彦宫尾安艺雄武田真阿部清美
Owner NAT INST OF AGROBIOLOGICAL SCI
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