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Method for detecting overwintering amount of plasmopara viticola in soil by real-time fluorescence quantitative PCR

A real-time fluorescence quantitative, downy mildew technology, applied in the field of bioengineering

Inactive Publication Date: 2020-06-19
NINGXIA UNIVERSITY
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Problems solved by technology

At present, there is no report on the quantitative method of overwintering amount of grape downy mildew in soil in China

Method used

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  • Method for detecting overwintering amount of plasmopara viticola in soil by real-time fluorescence quantitative PCR
  • Method for detecting overwintering amount of plasmopara viticola in soil by real-time fluorescence quantitative PCR
  • Method for detecting overwintering amount of plasmopara viticola in soil by real-time fluorescence quantitative PCR

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Embodiment 1

[0054] 1. Materials and methods

[0055] 1.1 Source of soil samples

[0056] Soil samples were collected from April to July 2018, from Pushang Chateau Vineyard, Zhihuiyuanshi Chateau Vineyard, Changyu Chateau Vineyard and Qingtongxia City in Yinchuan City, the eastern foot of Helan Mountain in Ningxia. Vineyards of Yuhuang Chateau (Table 1). The grid method was used for the entire field, and soil samples were collected by the "ten"-shaped 5-point method for each soil sample point, and the soil samples were taken as one sample point after mixing. The specific sampling method is as follows: 4 points are determined in four directions at a distance of 3-5m from the center of the sample point (determined according to the size of the field block). The point is 5-10cm away from the ground, and a 2cm thick soil sample is taken. After mixing the 5 soil samples, pass through a 20-mesh sieve, take about 20g of soil samples, seal them in a plastic bag, put them in an ice box and bring ...

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Abstract

The invention provides a method for detecting overwintering amount of plasmopara viticola in soil by real-time fluorescence quantitative PCR. The method of the invention is of great significance for accurately estimating the field disease condition in an early stage, determining the disease place, and formulating timely and effective control measures; a specific primer pair Pv5 / Pv6 is designed according to a full-length ITS gene (2337 bp) of plasmopara viticola, and a molecular detection system of plasmopara viticola real-time PCR is constructed, to quantitatively detect the overwintering amount of plasmopara viticola in soil, and to provide technical support for early monitoring, early warning and accurate prevention and control of plasmopara viticola in the field.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for detecting the overwintering amount of grape peronospora in soil by real-time fluorescent quantitative PCR. Background technique [0002] Grape downy mildew caused by Plasmopara viticola spreads all over the world's grape producing areas, and is one of the most serious fungal diseases that harm grapes, causing serious economic losses to grape production. The fungus overwinters in the soil mainly as oospores. In the following year, under suitable conditions, the oospores germinate to produce sporangia, and then zoospores are produced from the sporangia. It spreads by wind and rain, invades from the stomata on the back of leaves, and conducts initial infection. After a incubation period of 7-12 days, sporangia and sporangia were produced in the diseased part, and the sporangia germinated to produce zoospores for re-infection. Therefore, it is of gr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6851C12Q1/06C12R1/645
CPCC12Q1/6895C12Q1/6851C12Q2531/113C12Q2545/114C12Q2563/107
Inventor 顾沛雯闫思远肖瑞刚李文学张游李嘉泓
Owner NINGXIA UNIVERSITY
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