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LAMP field detection method for curvularia lunata and application of LAMP field detection method

A technology of Curvus sp. and crescent, which is applied in the field of LAMP rapid molecular detection for rapid identification of C. crescents, which can solve the problems of research and reports, lack of LAMP detection technology of C. crescents, etc.

Inactive Publication Date: 2020-10-20
HENAN INST OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These show that LAMP can be successfully applied to the detection of corn disease pathogens, but there are still no relevant studies and reports on the LAMP detection technology of Curvularia crescens

Method used

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  • LAMP field detection method for curvularia lunata and application of LAMP field detection method
  • LAMP field detection method for curvularia lunata and application of LAMP field detection method

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Embodiment Construction

[0025] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any form.

[0026] 1. Design of specific primer combination for detection of Curvularia lunata LAMP

[0027] Using the Clg2p gene sequence of Curvularia lunene, a set of Curvularia crescens-specific LAMP primer combination was designed by using the online LAMP primer design software PrimerExplorer V5, which specifically consisted of a pair of outer primers Cl-F3 / Cl-B3 and 1 For the inner primer Cl-FIP / Cl-BIP (attached figure 1 ). The primers designed in the LAMP reaction were:

[0028] Cl-F3: 5'GGGAGACGGACAACCGAT3'

[0029] Cl-B3: 5'AGGGCAAGAATTAACGGCC3'

[0030] Cl-FIP: 5'TGGCTCTGGAGATGGCACCT-CAACGGCAAGATACCGCC3'

[0031] Cl-BIP: 5'AACAAAAAACGGCTCGAGTGGC-ATGTTGACCATTTCCCTGGC3'

[0032] 2. Detection and optimization of LAMP reaction

[0033] Based on the LAMP primer combination desi...

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Abstract

The invention provides an LAMP method and a detection test kit for rapidly detecting curvularia lunata in a field and an application of the LAMP method and the detection kit, and belongs to the technical field of biology. According to the LAMP detection method, based on a Ras homologous protein Clg2p gene segment of the curvularia lunata, specific LAMP primers Cl-F3, Cl-B3, Cl-FIP and Cl-BIP are designed to establish an LAMP detection system, and a corresponding test kit is researched and developed for identification of the curvularia lunata. The technology is suitable for field detection andidentification of curvularia lunata so as to carry out early diagnosis of maize leaf curl leaf spot. The detection method disclosed by the invention has the characteristics of strong specificity, highsensitivity, strong practicability, simplicity, convenience and rapidness in operation and the like. The LAMP technology and the application of the LAMP technology are helpful for understanding the field occurrence rule of the curvularia lunata, and have important values in the layout of resistant varieties and the prevention and control of the corn leaf spot disease.

Description

technical field [0001] The present invention relates to the molecular biology detection technology of plant disease, specifically relate to the LAMP rapid molecular detection technology of rapid identification Curvularia crescens Background technique [0002] Curvularia maize leaf spot, also known as maize spot or pseudo-eye spot, is an important worldwide disease that occurs in corn-producing areas in America, Europe, Asia and Africa. In the middle and late 1980s, it was first discovered in North China. It has now expanded to most provinces in my country's corn-producing areas, especially in the northern corn-producing areas. It is another serious catastrophic disease after the large and small spot of corn in my country. The disease mainly affects leaves, but also leaf sheaths and bracts. After tasseling, the disease rapidly expands and spreads, the plants are covered with diseased spots, and the leaves dry up early, generally reducing production by 20% to 30%, and severe...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6844C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6895C12Q1/6844C12Q2531/119C12Q2563/107
Inventor 田雪亮赵松宇孙婷婷
Owner HENAN INST OF SCI & TECH