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Molecular biological method for identifying three Botryosphaeria on apples

It is a technology of Vitis vinifera and molecular biology, which can be applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc. Easy and fast effects

Inactive Publication Date: 2017-05-24
ANIMAL AND PLANT & FOOD DETECTION CENTER JIANGSU ENTRY EXIT INSPECTION AND QUARANTINE BUREAU +1
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Problems solved by technology

At present, the detection of pathogenic bacteria of this genus mainly depends on the harmful symptoms of pathogenic bacteria, colony characteristics, anamorphic and sexual morphological characteristics of pathogenic bacteria, but it is difficult to distinguish them and identify the species.

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  • Molecular biological method for identifying three Botryosphaeria on apples

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[0011] The protection content of the present invention is not limited to the following examples. Without departing from the spirit and scope of the inventive concept, changes and advantages conceivable by those skilled in the art are all included in the present invention, and the appended claims are the protection scope. The process, conditions, reagents, experimental methods, etc. for implementing the present invention are general knowledge and common knowledge in the art except for the content specifically mentioned below, and the present invention has no special limitation content.

[0012] 1. Design of specific primers

[0013] In the present invention, by comparing the existing Botrytis fungi sequences ITS, SSU, LSU, EF-1 in the NCBI gene bank α， Respectively aiming at the above sequence design can simultaneously identify Botrytis ( Botryosphaeria. Sp) three important pathogenic bacteria monosporium cankerella apple shell color ( Botryosphaeria stevensii ), Botrytis ...

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Abstract

The invention provides a molecular biological method for identifying three Botryosphaeria on apples and belongs to the technical field of biology. The molecular biological method for identifying the three Botryosphaeria on the apples comprises the step of carrying out PCR (Polymerase Chain Reaction) amplification by taking DNA (Deoxyribonucleic Acid) of a to-be-detected strain belonging to Bbotryosphaeria.Sp as a template and adopting primers EF1-S and EF1-A; if an amplified product only contains 230 to 250bp of DNA fragments, the to-be-detected strain is Botryosphaeria dothidea; if the amplified product only contains 140 to 160bp of DNA fragments, the to-be-detected strain is Botryosphaeria stevensii; if the amplified product only contains 170 to 180bp of DNA fragments, the to-be-detected strain is Botryosphaeria obtusa; the sequences of the primers EF1-S and EF1-A are respectively shown as SEQ ID NO:1 and SEQ ID NO:2. The identification method provided by the invention has the advantages of simplicity, convenience, quickness, high specificity and good generality.

Description

technical field [0001] The invention relates to a molecular biology method for identifying three kinds of botrytis on apples, belonging to the field of biotechnology. Background technique [0002] Botrytis ( Botryosphaeria. Sp. Botryosphaeria stevensii ), Botrytis ( Botryosphaeria dothidea ) and Botrytis obtuseum ( Botryosphaeria obtusa ) are three important pathogenic fungi in apple diseases. These three pathogenic fungi can harm the branches and fruits of fruit trees such as apples and pears, and form symptoms such as ulcers and ring marks on the branches and fruits. They are important pathogenic bacteria that seriously affect apple production in the world (Slippers & Wingfield 2004; Wang Fan et al. 2013). Botrytis ( Botryosphaeria. Sp. B. stevensii ) is a plant quarantine pest imported into my country, with a wide range of hosts. In addition to apples and pears, it can also harm plants of 11 families and 18 genera such as grapes, oaks, elms, and loquats (Van ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12R1/645
Inventor 李彬沈宏李艳华陈战张宇杨静粟寒吴翠萍
Owner ANIMAL AND PLANT & FOOD DETECTION CENTER JIANGSU ENTRY EXIT INSPECTION AND QUARANTINE BUREAU
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