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Cucumber target spot disease resistance gene cca and its encoded protein and application

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: 2017-10-27
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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  • Cucumber target spot disease resistance gene cca and its encoded protein and application
  • Cucumber target spot disease resistance gene cca and its encoded protein and application
  • Cucumber target spot disease resistance gene cca and its encoded protein and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Acquisition of Cucumber Target Spot Disease Resistance Gene Cca

[0030] 1. Preliminary and fine mapping 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 an air-condi...

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-resistant 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: use WF2757 and GY14 as parents to 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 Bank.

[0071] 2. Wrap the seeds of the above-mentioned materials to be tested with gauze, soak them in warm water, sow them in 50-hole trays after accelerating germination at a constant temperature of 28°C, and raise seedlings in an air-conditioned greenhouse. Extract the genomic DNA of each material to be tested from the seedlings, and use each genomic DNA as a template to perform full-length PCR amplificati...

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Abstract

The invention discloses a cucumber target spot disease resistance gene Cca, its encoded protein and its application. The gene is the nucleotide sequence shown in SEQ ID NO: 1 in the sequence listing or has more than 90% homology with the sequence shown in SEQ ID NO: 1 in the sequence listing and has the same sequence as SEQ ID NO: 1 in the sequence listing Nucleotide sequences having the same function as the sequences shown. The disease-resistant gene Cca provided by the present invention has important application value, and it can be used as an aid in the selection of cucumber target spot disease-resistant varieties. In addition, specific molecular markers or closely linked markers can be used to identify cucumbers according to the gene sequence information. And the genotypes of offspring plants are used for molecular marker-assisted selection breeding, thereby improving breeding efficiency.

Description

technical field [0001] The invention belongs to the field of gene sequence and application, and in particular relates to cucumber target spot disease resistance gene Cca and its encoded protein and application. Background technique [0002] Cucumber (Cucumis Sativus.L) is widely cultivated in the world, and China is the country with the largest cultivation area and the highest total output of cucumber in the world. [0003] Corynesporium leaf spot of cucumber, also known as target spot, is a worldwide distribution disease. At present, it has become one of the important diseases that endanger the cultivation of cucumbers in open field and protected areas, especially in protected areas such as overwintering greenhouses, winter-spring greenhouses, and spring greenhouses. It mainly harms the leaves. The lesions are light brown at the beginning and then turn brownish green. In severe cases, the leaves die. When the disease causes the defoliation rate to be lower than 5%, the di...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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