Rapid LAMP detection method for watermelon fusarium oxysporum

A watermelon fusarium wilt and detection method technology, applied in the field of watermelon fusarium wilt molecular detection, can solve the problem that the watermelon fusarium wilt LAMP detection technology has not yet been reported and the like

Inactive Publication Date: 2021-07-30
HUNAN AGRI BIOTECH RES CENT
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Problems solved by technology

At present, the LAMP detection technology of Fusarium wilt of watermelon has not been reported

Method used

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  • Rapid LAMP detection method for watermelon fusarium oxysporum
  • Rapid LAMP detection method for watermelon fusarium oxysporum
  • Rapid LAMP detection method for watermelon fusarium oxysporum

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Embodiment

[0040] 1 Materials and methods

[0041] 1.1 Materials

[0042] Tested strains: including Fusarium oxysporum watermelon specialization type 0, 1, 2 physiological races (FON-0, FON-1, FON-2), Fusarium oxysporum specialization type bitter melon (F.f.sp.momordicae) , Fusarium oxysporum melon specialization (F.f.sp.melonis), Fusarium oxysporum cucumber specialization (F.f.sp.cucumerinum), Fusarium moniliforme (F.moniliforme), F.f.sp. F.f.sp.conglutinans) and Trichoderma harzianum T2-16 (Trichoderma harzianum, T2-16) are both preserved by the Institute of Agricultural Biotechnology, Hunan Academy of Agricultural Sciences. All tested strains were identified through morphological observation and ITS sequence comparison. Zaojia '8424' watermelon seeds (Xinjiang Nongren Seed Technology Co., Ltd.).

[0043] Reagents: 50mmol / L sodium hydroxide (Sinopharm Group Chemical Reagent Co., Ltd.), pH=7.5 Tris-HCl (Beijing Suolaibao Technology Co., Ltd.), 2×LAMP reaction buffer, Bst DNA polymera...

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Abstract

The invention discloses a rapid LAMP detection method for watermelon fusarium oxysporum. According to the method, a set of sensitive and specific LAMP primers are designed by taking a specific DNA fragment of fusarium oxysporum watermelon specialized type as a target, and a rapid LAMP detection system of the watermelon fusarium oxysporum is established by combining a rapid DNA extraction method. The lower detection limit of the LAMP system on FON-1 genome DNA is 10 pg/[mu]L, and the lower detection limit of the LAMP system on the plant bacterium carrying amount is 5 * 10<2> spores/g. According to the system, only about 90 minutes are needed from DNA extraction to detection result acquisition, and pathogenic bacteria hyphae and plant tissues can be used as detection materials. The reaction result can be distinguished by naked eyes. The method has the characteristics of simplicity in operation, high specificity, high sensitivity, high stability and wide applicability, and can provide technical support for rapid diagnosis of field watermelon fusarium oxysporum.

Description

technical field [0001] The invention belongs to the technical field of molecular detection of watermelon Fusarium wilt, in particular to a rapid LAMP detection method for watermelon Fusarium wilt. Background technique [0002] Watermelon wilt is an important disease of watermelon (Citrullus lanatus), which occurs worldwide. The pathogenic bacterium of watermelon wilt is the soil-borne fungus Fusarium oxysporum watermelon specialization. The pathogen invades the plant from the root wound or between the cells at the top of the root hair, and gradually occupies and blocks the plant vascular vessels, affecting water transport and causing the plant to wilt due to water shortage. eventually dies. Different watermelon-specific strains of Fusarium oxysporum have different pathogenicity to different watermelon varieties, and there is physiological race differentiation. There are mainly 4 physiological races of Fusarium oxysporum watermelon that have been reported at present, respec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6844C12Q1/6806C12Q1/04C12R1/77
CPCC12Q1/6895C12Q1/6844C12Q1/6806C12Q2531/119C12Q2563/107C12Q2523/32C12Q2527/125
Inventor 肖姬玲梁志怀张屹杨科向吉方
Owner HUNAN AGRI BIOTECH RES CENT
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