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Calcium-dependent protein kinase gene in rice, and its application

A protein and rice technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve problems affecting rice quality and reducing rice yield, etc.

Active Publication Date: 2014-01-22
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Rice is an important food crop in the world, and the occurrence of rice diseases seriously affects the quality of rice and reduces the yield of rice

Method used

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  • Calcium-dependent protein kinase gene in rice, and its application
  • Calcium-dependent protein kinase gene in rice, and its application
  • Calcium-dependent protein kinase gene in rice, and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1. Screening and Cloning of Rice CDPK Gene Related to Disease Resistance Defense Response

[0053] 1. Preparation of Magnaporthe grisea inducer

[0054] The preparation method of blast fungus induced crude extract is as follows:

[0055] (1) The strains of Magnaporthe grisea are preserved in the form of filter paper sheets covered with fungal hyphae. When inoculating, the filter paper sheets are placed on the tomato-oat solid medium, and 3-4 filter paper sheets are placed on each plate.

[0056] (2) Place the plate in an incubator at 30°C for 4-5 days. At this time, a large number of hyphae will grow centered on the paper sheet.

[0057] (3) Add 4-5mL of sterile water to each plate, and use a coating stick to spread back and forth on the plate to scrape the hyphae.

[0058] (4) Wash the mycelium with sterile water, add it to the tomato-oat liquid medium, and culture it with shaking at 28°C and 200rpm in the dark for 4-5 days.

[0059] (5) Collect the culture,...

Embodiment 2

[0096] Embodiment 2, preparation and phenotypic analysis of transgenic rice

[0097] 1. Cloning of OsCPK10 and OsCPK20 genes

[0098] (1) The primers for the OsCPK10 gene were designed as follows:

[0099] OsCPK10-sense pair: 5'-ATGGGGAACACGTGCGTC-3'

[0100]OsCPK10-antisense pair: 5'-TCAAGACATCCTCAAGGCCTC-3'

[0101] The primers for the OsCPK20 gene were designed as follows:

[0102] OsCPK20-sense pair: 5'-ATGGGCAACTGCTGCGTGAC-3'

[0103] OsCPK20-antisense pair: 5'-CTATTGGGTAGTTGTTAACTGCAATG-3'

[0104] (2) Extract the RNA of the Nipponbare rice callus treated with the blast fungus inducer for 1 h in Example 1, obtain cDNA after reverse transcription, use the cDNA as a template, and perform PCR amplification with the primer pairs of OsCPK10 and OsCPK20 genes respectively, The OsCPK10 and OsCPK20 genes were obtained.

[0105] The PCR reaction system is as follows:

[0106]

[0107] (3) Agarose gel electrophoresis to detect the size of the PCR amplified fragment, the ...

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PUM

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Abstract

The invention discloses a calcium-dependent protein kinase (CDPK) gene in rice, and its application. A protein disclosed in the invention is a protein represented by first to 390th amino acids from the N terminal in SEQ ID No.2 or a protein obtained by substituting and / or deleting and / or adding one or more amino acid residues to the protein represented by first to 390th amino acids from the N terminal in SEQ ID No.2 and having same functions. The CDPK gene is of great significance to using the CDPK responsive to pathogens in rice in molecular breeding for enhancing the disease resistances of plants.

Description

technical field [0001] The invention relates to rice calcium-dependent protein kinase gene and application thereof. Background technique [0002] Rice is an important food crop in the world, and the occurrence of rice diseases seriously affects the quality of rice and reduces the yield of rice. Preventing rice diseases by improving the plant's own disease resistance is the most economical, durable and environmentally friendly method. An effective way to improve rice disease resistance is to use genetic engineering to improve the expression of key genes in the process of plant disease resistance defense response. [0003] Calcium-dependent protein kinase (CDPK) is a new type of calcium ion sensor that combines protein kinase structure and calmodulin-like structure (1). CDPK is a plant-specific (and also present in two protozoans, ciliates and Eimeria tenella), a serine / threonine kinase that is absent in animals. CDPKs exist in plants as large gene families: 34 CDPKs in Ara...

Claims

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Application Information

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IPC IPC(8): C12N9/12C12N15/54C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/82A01H5/00
CPCC12N9/12C12N15/8282C12Y207/11001
Inventor 安成才付力文于祥春
Owner PEKING UNIV