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Rice disease resistance related gene, encoding protein thereof and preparation method for strain for improving rice broad spectrum disease resistance

A disease-resistance-related gene and rice technology, applied in botany equipment and methods, biochemical equipment and methods, genetic engineering, etc., to achieve the effect of reducing rice pests and diseases and improving broad-spectrum disease resistance

Inactive Publication Date: 2012-08-08
ANHUI AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, OsMOS in rice has not been reported so far, and the present invention found that for OsMOS (modifier of scn1 in rice) in rice, the same as OsMOS in Arabidopsis Compared with i>MOS3, they have similar sequences, and both contain a Nup96 nucleoporin, which controls the nuclear export and nuclear transport of materials and information, and shows broad-spectrum resistance to various pathogenic bacteria

Method used

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  • Rice disease resistance related gene, encoding protein thereof and preparation method for strain for improving rice broad spectrum disease resistance
  • Rice disease resistance related gene, encoding protein thereof and preparation method for strain for improving rice broad spectrum disease resistance
  • Rice disease resistance related gene, encoding protein thereof and preparation method for strain for improving rice broad spectrum disease resistance

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Embodiment 1

[0019] 1. Trizol method to extract Nipponbare total RNA

[0020] (1) Weigh 0.1g of fresh and tender Nipponbare leaves ( Oryza. Sativa L. spp. japonica, var nipponbare, AA genome, The appraiser, Professor Cheng Beijiu), was quickly ground with liquid nitrogen, and transferred to a 1.5mL centrifuge tube (pre-cooled with liquid nitrogen). And add 1mL Trizol to the centrifuge tube, place on ice for 10min;

[0021] (2) 4°C, 13000rpm, centrifuge for 20min;

[0022] (3) Transfer the supernatant to a new tube, add 250uL of 5M NaCl, 250uL of chloroform / isoamyl alcohol, shake vigorously for 15S, and place on ice for 3 min;

[0023] (4) 4°C, 13000rpm, 15min;

[0024] (5) Transfer the clear to another new centrifuge tube, add 500uL isopropanol to mix, and place at room temperature for 10 minutes;

[0025] (6) 4°C, 13000rpm, 15min;

[0026] (7) Discard the supernatant, add 500uL 75% ethanol, and wash;

[0027] (8) 4°C, 7500rpm, 5min, pour off the ethanol, put it on paper, and l...

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Abstract

The invention discloses a method for improving the rice disease resistance by using a broad spectrum disease resistance related gene modifier of scnl n rice (OsMOS) and belongs to the technical field of rice genetic engineering. The method comprises the following steps of: cloning a full-length complementary deoxyribonucleic acid (cDNA) of the OsMOS gene from rice nipponbare, connecting the full-length cDNA to a plant carrier, and building overexpression of the OsMOS gene by initiating the expression of the gene by using a constitutive promoter CaMV35S; obtaining a transgenic plant by agrobacterium tumefaciens-mediated transformation; and selecting a marked gene hygromycin and a target gene OsMOS through multiplex polymerase chain reaction (PCR) detection, detecting free salicylic acid (FreeSA) in the transgenic rice, and performing inoculation identification on rice bacterial leaf blight resistance. Results show that the content of the FreeSA in the OsMOS overexpression transgenic plant is obviously higher than that in a contrast plant, and the OsMOS overexpression transgenic plant is higher in disease resistance to rice bacterial leaf blight. Therefore, the method for obtaining the strain for improving the rice broad spectrum disease resistance is provided.

Description

technical field [0001] The invention relates to the technical field of rice genetic engineering, specifically cloned from rice Nipponbare OSMOS gene full-length cDNA 。 Through genetic transformation of rice, the OSMOS Transgenic plants with gene overexpression were analyzed for bacterial blight resistance and salicylic acid content of transgenic plants and non-transgenic plants respectively. The results show OSMOS Genes are significantly correlated with broad-spectrum disease resistance in rice. Background technique [0002] Rice is not only an important food crop, but also an important model organism in the field of plant molecular biology research. In the process of rice production, due to the impact of diseases such as rice blast, sheath blight, and bacterial blight, it has always been an important problem faced by grain production. Under the current control conditions, the national average annual rice production reduction due to various rice diseases reaches 20 bill...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/63C12N5/10C12N15/82C12N15/84A01H5/00
Inventor 江海洋吴青青程备久朱苏文周玉琼
Owner ANHUI AGRICULTURAL UNIVERSITY
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