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Rapid trace detection reagent for Botrytis cinerea based on LAMP method, method and application thereof

A technology of botrytis cinerea and detection reagents, which is applied in the field of fast trace detection reagents for botrytis cinerea, and can solve the problems of missed control period, unsatisfactory disease prevention effect, and delayed application of medicines.

Inactive Publication Date: 2019-01-18
YANGZHOU UNIV
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Problems solved by technology

The existing disease forecasting methods mainly rely on experts to conduct systematic large-scale investigations in the field according to disease symptoms after the disease occurs, and then publish disease information based on experience. Medication often misses the proper period for prevention and treatment, which leads to late application of the medicine, and the effect of disease prevention is not ideal

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  • Rapid trace detection reagent for Botrytis cinerea based on LAMP method, method and application thereof
  • Rapid trace detection reagent for Botrytis cinerea based on LAMP method, method and application thereof
  • Rapid trace detection reagent for Botrytis cinerea based on LAMP method, method and application thereof

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Embodiment Construction

[0024] 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.

[0025] 1. Design and screening of specific primers

[0026] (1) According to the Lac1 gene sequence of Botrytis cinerea (accession number: U20192.1), the online software PrimerExplorer V was used to design a LAMP detection primer set, and 30 pairs of BcB3 / BcF3 primers were selected for specificity determination (Table 1).

[0027] Table 1 Part of the LAMP primer set designed according to the sequence of Botrytis cinerea Lac1 gene

[0028]

[0029]

[0030]

[0031] From left to right, from top to bottom are SEQ ID No.1-120. For example, the sequence of primer combination 01 is SEQ ID No.1-4 from left to right.

[0032] (2) Use 30 pairs of BcF3 / BcB3 primers to perform PCR amplification on Botrytis cinerea genomic DNA. A total of 17 pairs of primers can amplify a singl...

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Abstract

The invention relates to a rapid trace detection reagent for Botrytis cinerea based on LAMP method, a method and application thereof. A specific primer combination is used. The primer combination wasdesigned according to the sequence of Botrytis cinerea Lac1 gene and could amplify the genomic DNA of Botrytis cinerea without amplification of other common pathogenic fungi. The optimum reaction conditions were 63 DEG C for 40 min, and the amount of DNA of Botrytis cinerea could reach pg. The method could be used for the rapid detection of Botrytis cinerea.

Description

technical field [0001] The invention relates to a rapid detection technology of plant pathogenic bacteria, belongs to the field of disease prediction research and application in the discipline of plant protection, and specifically relates to a rapid trace detection reagent, method and application of Botrytis cinerea based on a LAMP method. Background technique [0002] Botrytis cinerea can cause gray mold of vegetables, fruit trees, flowers and other plants, especially in the production of fruits and vegetables in protected areas. It often causes damage to plant stems, leaves, flowers, and fruits, and has a serious impact on yield and quality. In the field, botrytis cinerea is explosive. If chemical pesticides are not applied timely and effectively, not only will it be difficult to control the occurrence of the disease, but the pathogenic bacteria will develop resistance due to the frequent use of chemical pesticides in large quantities, which will lead to pesticide residues ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6895
Inventor 陈夕军李丽丽陈银凤
Owner YANGZHOU UNIV
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