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Corynespora cassiicola anti-disease gene Cca as well as encoding protein and application thereof

A cucumber target spot disease and disease-resistant gene technology, applied in the field of gene sequence and application, to achieve the effect of improving breeding efficiency

Active Publication Date: 2015-09-23
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, a large number of plant genes have been isolated by using map-based cloning technology, especially genes related to disease resistance (Yan Qitao, Map-based cloning technology for plant gene isolation, Molecular Plant Breeding, 2005, 585-590), but no A report about the successful cloning of the resistance gene of cucumber target spot disease

Method used

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  • Corynespora cassiicola anti-disease gene Cca as well as encoding protein and application thereof
  • Corynespora cassiicola anti-disease gene Cca as well as encoding protein and application thereof
  • Corynespora cassiicola anti-disease gene Cca as well as encoding protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Obtainment of the Cucumber Target Spot Resistance Gene Cca

[0030] 1. Preliminary localization and fine localization of cucumber target spot disease resistance gene Cca

[0031] The specific positioning method includes the following steps:

[0032] A. Mapping research on resistance genes of cucumber target spot disease Parents and genetic groups:

[0033] Select WF2757 (female parent) and Xintaimici (male parent) as resistant parents, and construct 150 strains of F 2:3 populations and more than 2000 strains of F 2 large groups. Among them, the male parent Xintaimici and the female parent WF2757 were purchased from the Beijing Crop Germplasm Bank.

[0034] B. Investigation of Disease Index of Cucumber Target Spot Disease Resistance Gene Mapping Research:

[0035] The seeds of parents and each group were wrapped with gauze, soaked in warm soup, and then sowed in 50-hole hole trays after germination at a constant temperature of 28°C. Seedlings were raised in...

Embodiment 2

[0059] Example 2: Identification and Analysis of Cucumber Target Spot Disease Resistance Gene Cca cDNA and Protein Sequence

[0060] 1. The cDNA (mRNA) full-length amplification of the target spot disease gene Cca of cucumber wilt:

[0061] Using the resistant and susceptible parental materials WF2757 and Xintaimici, young leaf tissues were selected to extract total RNA, and reverse transcription was performed to synthesize the first strand of mRNA (cDNA). Using the first strand of mRNA as a template, the full-length Cca gene was used Amplify the upstream and downstream primers SEQ ID NO: 7 and SEQ ID NO: 8 for PCR amplification to obtain the full-length cDNA (mRNA) sequences SEQ ID NO: 3 and SEQ ID NO: 4 in the anti- / sensitivity material, respectively. Using DNAMAN software to compare and analyze the full length of the Cca gene in the resistant / susceptible materials, it was found that the gene has only one exon and no intron structure in the resistant / susceptible materials, b...

Embodiment 3

[0068] Example 3: Functional verification of disease resistance gene Cca and its application in breeding cucumber target spot disease resistant varieties

[0069] 1. Functional verification of cucumber target spot disease resistance gene Cca

[0070] 1. Materials to be tested: Using WF2757 and GY14 as parents, construct F 2 Genetic population, from which 120 individual plants were randomly selected for target spot disease resistance identification and sequencing. Among them, the male parent GY14 and the female parent WF2757 were purchased from the Beijing Crop Germplasm Resource Bank.

[0071] 2. The seeds of the above-mentioned materials to be tested were wrapped with gauze, soaked in warm water, sown in a 50-hole plug tray after germination at a constant temperature of 28°C, and the seedlings were raised in an air-conditioned greenhouse. The seedling substrate was sterilized perlite or nutrient soil. The genomic DNA of each material to be tested is extracted from the seedl...

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Abstract

The invention discloses a corynespora cassiicola anti-disease gene Cca as well as an encoding protein and an application thereof. The gene is a nucleotide sequence shown as SEQ ID NO.1 in a sequence list or a nucleotide sequence with 90% or more homology with the sequence shown in SEQ ID NO.1 and with a function identical to that of the sequence shown in SEQ ID NO.1 in the sequence list. The anti-disease gene Cca has an important application value and can be used for assisting the breeding of corynespora cassiicaloa anti-disease species; in addition, a specific molecular marker generated according to the gene sequence information or a tightly linked marker can be used for identifying the gene type of a cucumber and progeny plants, which can be used for the molecular marker to assist the selection and seed breeding, so that the seed breeding efficiency can be improved.

Description

technical field [0001] The invention belongs to the field of gene sequences and applications, in particular to a cucumber target spot disease resistance gene Cca and its encoded protein and applications. Background technique [0002] Cucumber (Cucumis Sativus.L) is widely cultivated in the world. China is the country with the largest cucumber cultivation area and the highest total output in the world. [0003] Cucumber corynebacterium leaf spot, also known as target spot disease, is a worldwide disease. At present, it has become one of the important diseases that endanger the cultivation of cucumbers in open fields and protected areas, especially in winter greenhouses, winter and spring greenhouses, spring greenhouses and other protected areas. It mainly damages the leaves, the lesions are light brown at first and then turn brownish green, and the leaves die in severe cases. When the defoliation rate caused by the disease is lower than 5%, the disease spreads slowly, about...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/11C12Q1/68C07K14/415
Inventor 温常龙毛爱军赵泓王永勤董从娟许勇
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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