Sample preparation method and application for detecting cucumber green mottle mosaic virus carried by Cucurbitaceae seeds

Abstract

The invention discloses a preparing method and an application of a sample for detecting cucumber green mottle mosaic viruses (CGMMV) carried by cucurbitaceae seeds. The preparing method includes the steps that starting from treating seed samples, shell-kernel separating treatment is carried out on the cucurbitaceae seeds, seed kernels with the low virus content are removed, seed shells with the high virus content are left to serve as to-be-tested samples, the seed samples obtained after treatment are sufficiently grinded through a full-automatic rapid sample grinding instrument, and the CGMMV in the large quantity of cucurbitaceae seeds can be rapidly detected through the sufficiently-grinded samples with a RT-PCR method and can also be rapidly detected through the sufficiently-grinded samples with a dot enzyme-linked immunosorbent assay (dot-ELISA) method. The preparing method and the application are suitable for virus detection of the common cucurbitaceae seeds such as calabash seeds, cucumber seeds, lagenaria sicerariae seeds, pumpkin seeds, long gourd seeds and watermelon seeds. A detecting method is high in sensitivity, good in specificity, short in consumed time and low in cost, technical supports are provided for large-scale detection and scientific prevention and control of the CGMMV carried by the cucurbitaceae seeds, and references are provided for detection of plant viruses transmitted by other seeds.

Description

technical field [0001] The field of the invention belongs to the field of biotechnology, and relates to the treatment of a Cucurbitaceae seed sample carrying cucumber green mottled mosaic virus, as well as serological and molecular biological detection methods and applications. Background technique [0002] Cucumber green mottle mosaic virus (CGMMV) was first reported in the United Kingdom in 1935. With the increasingly frequent exchange and introduction of cucurbit crops among countries, it was subsequently introduced in Germany, Finland, Denmark, Russia, Greece, etc. in Europe. Many countries and regions have reported the harm of the virus one after another. [0003] Cucumber green mottle mosaic virus is a member of the genus Tobamovirus in the family Tymoviridae. It mainly infects Cucurbitaceae crops, and can cause yellowing and deformity of leaves of gourds, cucumbers, watermelons, pumpkins, etc., slow growth, delayed fruiting, most of the fruits turn yellow and white, ...

AI Technical Summary

Problems solved by technology

However, due to the high fat content of the poisonous Cucurbitaceae seeds, it is difficult to extract high-quality RNA or the virus content of CGMMV is too low to produce enough templates, so the conventional liquid nitrogen grinding method is used to extract the RNA of the poisonous Cucurbitaceae seeds. , and detected by RT-PCR method, the target band is often not amplified

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