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Detection primer for banana wilt germina number-four biological strain and method for detecting same

A technique for detecting primers for Fusarium wilt of bananas, which is applied in the field of detection primers for plant pathogenic fungi, and can solve the problems of large-area outbreak loss and long survival period

Inactive Publication Date: 2008-01-30
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the long survival period and rapid spread of the bacteria, there is no specific drug to prevent the disease. Therefore, once it occurs in banana production areas, it is likely that a large-scale outbreak will cause heavy losses.

Method used

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  • Detection primer for banana wilt germina number-four biological strain and method for detecting same
  • Detection primer for banana wilt germina number-four biological strain and method for detecting same
  • Detection primer for banana wilt germina number-four biological strain and method for detecting same

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Embodiment 1

[0032] 1. Establishment of PCR system

[0033] Design the primer concentration, template, dNTPs, TaqDNA polymerase and PCR annealing temperature gradient to detect the effect of each reaction condition on the amplification effect, as follows:

[0034] Template DNA 1ul

[0035] 10×PCR Buffer 2.5ul

[0036] Mg 2+ (25mmol / l) 2.5ul

[0037] dNTPs (25mM) 2ul

[0038] Primer RF4-1 1ul

[0039] Primer RF4-2 1ul

[0040] Taq enzyme (5U / μL) 0.2ul

[0041] Sterilized ddH 2 O 14.8ul

[0042] Total volume 25ul

[0043] Prepare the reaction system on ice, mix well, and perform amplification according to the following PCR program: 94°C for 4min; 94°C for 30s, 52°C for 30s, 72°C for 45s, repeating 35 cycles; 72°C for 10min. Store at 4°C.

[0044] After the PCR reaction, 5 μl of the PCR product was taken for 1% agarose gel electrophoresis detection, and then the specific fragment was recovered, cloned, and sequenced to obtain the nucleic acid sequence of the specific fragment of th...

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Abstract

The invention relates to the biological technical field and discloses a detection primer of plant pathogenic fungi and a detection method thereof. The invention adopts a special primer RF4 and optimizes action conditions to invent a PCR detection method for banana fusarium wilt physiological races (Race No. 4). The method can especially, sensitively and effectively detect banana fusarium wilt physiological races (Race No. 4) and mycelium even weighting 0.1 Mug can be detected and pathogen can even be detected from the root and the stem of bananas by adopting the method.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a detection primer for plant pathogenic fungi and a detection method thereof. Background technique [0002] Banana fusarium wilt, also known as banana Panama disease and yellow leaf disease, is a soil-borne disease that causes vascular necrosis caused by Fusarium infection. The pathogen is Fusarium oxysporum f.sp.cubense( E.F. Smith) Snyder et Hansen] (FOC). The pathogen invades from the injured rhizome, develops to the stem through the host vascular bundle, and further spreads to the false leaves. When the plant dies, the pathogenic bacteria remains in the soil with the remains, and then spreads naturally in the banana orchard along with the water flow or the infected agricultural tools, and its chlamydospores can survive in the soil for 8-10 years. Because the bacteria has a long survival period and spreads rapidly, there is no specific drug to prevent the disease. Therefore, onc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N27/26
Inventor 王振中廖林凤
Owner SOUTH CHINA AGRI UNIV
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