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Cucumber Fusarium wilt resistance gene foc‑4 and its encoded protein and application

A cucumber fusarium wilt and disease-resistant gene technology, applied in the field of gene sequence and application, can solve the problem of successful isolation and cloning of the disease-resistant gene of physiological race 4 of cucumber fusarium wilt, and achieve the effect of improving breeding efficiency

Active Publication Date: 2017-03-29
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 the genes related to disease resistance (Zeng ZB, 1994; Yimai, 1998), but there is no disease resistance related to cucumber wilt physiological race 4. Reports of genes being successfully isolated and cloned

Method used

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  • Cucumber Fusarium wilt resistance gene foc‑4 and its encoded protein and application
  • Cucumber Fusarium wilt resistance gene foc‑4 and its encoded protein and application
  • Cucumber Fusarium wilt resistance gene foc‑4 and its encoded protein and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Obtainment of Cucumber Fusarium Wilt Resistance Gene Foc-4

[0033] 1. Preliminary localization and fine localization of the resistance gene Foc-4 of cucumber Fusarium wilt race 4

[0034] The specific positioning method includes the following steps:

[0035] A. Cucumber Fusarium Wilt Resistance Gene Mapping Research Parents and Genetic Populations:

[0036] WI2757 (male parent) and Jinyan No. 2 (female parent) were selected as anti-infective parents to construct 90 RIL-F 8 strains, F of 130 strains 2:3 populations and more than 2000 strains of F 2 large groups. Among them, the male parent WI2757 and the female parent Jinyan 2 were purchased from the Beijing Crop Germplasm Resource Bank.

[0037] B. Cucumber Fusarium Wilt Disease Resistance Gene Location Research Disease Index Survey:

[0038] The seeds of the parent and each group were wrapped with gauze, soaked in warm soup, sown in 50-hole plug trays after germination at a constant temperature of 28°...

Embodiment 2

[0062] Example 2: Identification and Analysis of Cucumber Fusarium Wilt Resistance Gene Foc-4 cDNA and Protein Sequences

[0063] 1. Amplification of the full-length cDNA (mRNA) of the resistance gene Foc-4 of Fusarium wilt in cucumber:

[0064] Using the anti-infective parent materials WI2757 and Jinyan No. 2, the young leaf tissue was selected to extract total RNA, and reverse transcription was performed to synthesize the first strand of mRNA (cDNA). Amplify the upstream and downstream primers SEQ ID NO:7 and SEQ ID NO:8 for PCR amplification, and obtain the full-length cDNA (mRNA) sequence SEQ ID NO:3 and SEQ ID NO:4 in the anti / sense material, respectively. The DNAMAN software was used to compare and analyze the full length of the Foc-4 gene of the resistant / susceptible materials. For size differences, see Figure 1. Compared with the susceptible material, the cDNA sequence of the resistant material showed a deletion between 1915-2184 bp. It is speculated that the existenc...

Embodiment 3

[0071] Example 3: Functional verification of the disease-resistant gene Foc-4 and its application in the breeding of cucumber Fusarium wilt disease-resistant varieties

[0072] 1. Functional verification of cucumber Fusarium wilt resistance gene Foc-4

[0073] 1. Materials to be tested: Using WI2757 and GY 14 as parents, construct F 2 Genetic population, from which 100 individual plants were randomly selected for disease resistance identification and sequencing of Fusarium wilt. Among them, the male parent WI2757 and the female parent GY 14 were purchased from the Beijing Crop Germplasm Bank.

[0074] 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 was extracted from ...

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Abstract

The invention discloses a cucumber blight disease resistance gene Foc-4 as well as an encoding protein and application thereof. The gene is a nucleotide sequence shown in SEQ ID NO:1 in a sequence table. The resistance gene Foc-4 provided by the invention has important application value, and can generate a specific molecular marker or a tightly linked marker according to the gene sequence information; genotype of cucumber and descendant plants thereof can be identified by using these markers; the cucumber blight disease resistance gene Foc-4 is applied to the molecular marker to assist selective breeding, so as to improve the breeding efficiency. In addition, the cucumber blight disease resistance gene Foc-4 disclosed by the invention also can be used for selectively breeding scab disease-resistant variety of the cucumber.

Description

technical field [0001] The invention belongs to the field of gene sequences and applications, and particularly relates to a cucumber fusarium wilt disease resistance gene Foc-4 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 Fusarium Wilt (Fusarium Wilt), also known as wilt disease, undercutting disease, and dead seedling disease, is a systemic disease (ductal fusarium wilt) that is transmitted by soil, invades from roots or root necks, and parasites in vascular bundles. ), is one of the three major diseases that are difficult to control in cucumber production, often causing large losses, and is currently the main soil-borne disease of cucumbers around the world, especially in greenhouses and other protected areas. It overwintered in the body and immature farmyard ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/11C07K14/415C12Q1/68
Inventor 温常龙许勇于拴仓毛爱军王永健张丽蓉赵泓董从娟
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES