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Method for fast detection of lily bulb rot pathogen fusarium oxysporum

A technology for Fusarium oxysporum and bulb rot, which is applied in the field of specific primers and can solve problems such as direct detection of Fusarium oxysporum.

Inactive Publication Date: 2017-01-11
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Abstract
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AI Technical Summary

Problems solved by technology

The present invention aims to solve the problem that the existing universal primer pair cannot directly detect Fusarium oxysporum from mixed fungal DNA samples, and provides a species-specific primer pair for rapid detection of Fusarium oxysporum in order to quickly and accurately Molecular identification of Fusarium oxysporum in lily

Method used

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  • Method for fast detection of lily bulb rot pathogen fusarium oxysporum
  • Method for fast detection of lily bulb rot pathogen fusarium oxysporum
  • Method for fast detection of lily bulb rot pathogen fusarium oxysporum

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Embodiment Construction

[0044] 1. Extraction of fungal DNA by CTAB method

[0045] (1) Liquid culture of pathogenic bacteria

[0046] On the ultra-clean workbench, use a sterilized inoculation loop to pick the mycelium at the edge of the colony and culture it in the potato liquid medium for 5-7 days with shaking (temperature 26°C, rotation speed 150r / min). Filter the bacterial solution with sterile gauze, and collect the mycelia for DNA extraction.

[0047] (2) DNA extraction by CTAB method

[0048] ①Add 650 μL of 2% CTAB buffer solution to a 1.5 mL centrifuge tube and preheat at 65°C. Thoroughly grind the mycelium in liquid nitrogen, quickly take 0.2 g of the ground mycelium powder and put it into the preheated CTAB buffer solution, mix well, put it in a water bath at 65°C for 1 hour, and shake it gently once every 15 minutes.

[0049] ②After the 1h water bath, place at 4°C or room temperature to cool down to room temperature.

[0050] ③ Add 650 μL of chloroform: isoamyl alcohol (24:1), turn it ...

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Abstract

The invention relates to a method for fast detection of lily bulb rot pathogen fusarium oxysporum based on a DNA molecule detection technology. The invention relates to specific primers for lily fusarium oxysporum amplification. The method solves the problem that the existing method needs separation and purification of pathogens from bulbs suffering from rot, realizes identification of pathogen types through primer amplification, sequencing and comparison after DNA extraction and cannot realize identification of lily fusarium oxysporum through the existing general primers directly based on an amplification result. The specific primer pair for lily fusarium oxysporum amplification is named as FO-lilii F / FO-lilii R. The primers realize fast and accurate amplification of a desired fragment of the lily bulb rot pathogen fusarium oxysporum.

Description

technical field [0001] The invention relates to a pair of specific primers for rapid detection of Fusarium oxysporum lily, belonging to the field of detection and identification of pathogens. Background technique [0002] Lily is a perennial bulbous flower of the genus Lilium in the family Liliacea. It has beautiful flowers and bright colors, and is deeply loved by people. It is one of the top five cut flowers in the world, and it is also a high-grade potted flower and an excellent garden flower. However, lily is also a kind of flower that is more susceptible to diseases. With the expansion of cultivation area and the prolongation of cultivation time, the occurrence of lily diseases is becoming more and more serious. Among lily diseases, lily bulb rot (also known as stem rot and wilt) caused by Fusarium oxysporum is one of the most serious soil-borne fungal diseases in lily production (Wu Zhuhua et al., 2013). Fusarium oxysporum can harm all organs of lily plants, mainly un...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/77
CPCC12Q1/6895
Inventor 明军袁素霞杨盼盼曹雨薇徐雷锋
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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