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Cloning and application of R gene TN2 for regulating and controlling powdery mildew resistance of plants

A powdery mildew, plant technology, applied in the direction of plant genetic improvement, application, plant peptides, etc., can solve problems such as infection and economic losses in agricultural production

Inactive Publication Date: 2015-04-08
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Worldwide, there are more than 500 species of powdery mildew infecting more than 10,000 plants, causing huge economic losses to agricultural production

Method used

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  • Cloning and application of R gene TN2 for regulating and controlling powdery mildew resistance of plants
  • Cloning and application of R gene TN2 for regulating and controlling powdery mildew resistance of plants
  • Cloning and application of R gene TN2 for regulating and controlling powdery mildew resistance of plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1, TN2 mutation inhibits exo70B1 mutant powdery mildew resistance

[0052] 1. Obtaining of mutant exo70B1tn2

[0053] A mutant library of exo70B1 was obtained by EMS mutagenesis. In the M2 generation, a mutant exo70B1tn2 that can inhibit exo70B1-mediated powdery mildew resistance was screened.

[0054] The mutant exo70B1tn2 can also be prepared according to the following method: the mutant exo70B1 (N717829) is crossed with the mutant tn2 to obtain a hybrid F1 generation, and the homozygous double mutant exo70B1tn2 can be obtained by screening the F2 generation obtained by selfing of the F1 generation.

[0055] Wild-type Arabidopsis Col-0 (hereinafter referred to as wild-type Arabidopsis), recorded in the following documents: Catherine A.Frye, Dingzhong Tang, and Roger W.Innes (2001).Negative regulation of defense responses in plants by a conserved MAPKK kinase. PNAS 98(1):373-378; publicly available from Institute of Genetics and Developmental Biology, Chin...

Embodiment 2

[0069] Example 2, TN2 mutation inhibits the expression of PR1 gene (AY064023, submitted on November 19, 2001) in exo70B1 mutant

[0070] In order to verify whether the TN2 gene is involved in plant disease resistance, real-time quantitative PCR was used to detect the expression patterns of PR1 gene in Col-0, mutant exo70B1 and double mutant exo70B1tn2 before and after inoculation with powdery mildew. The instrument used is Germany Eppendorf Mastercyclereprealplex 4S, details are as follows:

[0071] Arabidopsis wild-type plants Col-0(WT), mutant exo70B1 and double mutant exo70B1tn2 grown for 4 weeks were inoculated with powdery mildew for 3 days, and the leaves of the uninoculated and 3-day-inoculated plants were taken respectively and put into the liquid Quick-frozen in nitrogen, then stored at -70°C, and extracted RNA for later use.

[0072] The total RNA of leaves was extracted with Trizol (Invitrogen) reagent, the contaminated DNA was digested with DNase I (Promega) for h...

Embodiment 3

[0075] Example 3, TN2 mutation inhibits the accumulation of free salicylic acid in the exo70B1 mutant

[0076] Salicylic acid is a central molecule regulating plant disease resistance. Salicylate-dependent signaling is critical for the establishment of local and systemic resistance in plants. The content of free salicylic acid in Col-0, mutant exo70B1 and double mutant exo70B1tn2 before and after inoculation with powdery mildew was detected.

[0077] Seedlings of Arabidopsis wild-type plant Col-0 (WT), mutant exo70B1 and double mutant exo70B1tn2 were inoculated with powdery mildew in large quantities. The leaves of each material were quick-frozen in liquid nitrogen, and then stored at -70°C to extract salicylic acid for later use.

[0078]The content of salicylic acid was determined by high performance liquid chromatography, and the method described in the literature Xin Li, Yuelin Zhang, Joseph D Clarke, Yan Li, Xinnian Dong (1999) Identification and cloning of a negative r...

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Abstract

The invention discloses cloning and application of an R gene TN2 for regulating and controlling powdery mildew resistance of plants. The invention provides a method for regulating and controlling the resistance of plants on powdery mildew. An amino acid sequence of TN2 protein is a sequence 2 in a sequence table; premature translation termination is caused by single amino acid change of seven sites and mutation of two sites; and the resistance of an exo70B1 mutant on the powdery mildew and powdery mildew-induced cell death can be restrained, but no obvious effect is generated on growth and development of plants, thus, the TN2 can be applied to the technical field of genetic engineering, so as to regulate and control resistance reaction of the plants on the powdery mildew.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the cloning and application of an R gene TN2 regulating plant powdery mildew resistance. Background technique [0002] In the natural environment, plants are constantly infected by various pathogens, including bacteria, fungi, and viruses. In order to resist the invasion of pathogenic bacteria, plants have evolved a complex defense system, which is divided into two levels: the immune response PTI induced by pathogen-associated molecular patterns (PAMPs) and the immune response ETI induced by effectors. First, pathogenic bacteria enter the plant through stomata, epidermal gaps or even injured parts, and pathogen-related molecular patterns, such as flagellin, elongation factor, and chitin, are recognized by pattern recognition receptors (PRR) located on the cell membrane , to trigger PTI immune response; however, during the long-term evolution of pathogenic bacteria, they have evolved...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/82C12N15/8282
Inventor 芮璐赵婷唐定中
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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