Kit for rapidly diagnosing peach bacterial shot hole disease

Abstract

The invention relates to a kit for rapidly diagnosing peach bacterial shot hole, and belongs to the technical field of plant protection. The kit comprises an RPA primer pair, a specific crRNA and a single-stranded DNA probe; the sequence of the RPA primer pair is as shown in SEQ ID NO.1-2, the sequence of the specific crRNA is as shown in SEQ ID NO.3, and the sequence of the single-stranded DNA probe is as shown in SEQ ID NO.4 and / or SEQ ID NO.5. The kit is used for detecting the main pathogenic bacteria Xap of the peach bacterial shot-hole disease on the basis of an RPA technology and a Crispr / Cas system, visual detection is realized through a transverse flow detection test strip and a miniature ultraviolet flashlight, the kit is simple and flexible, low in cost, high in sensitivity and high in specificity, a user can freely select the kit, the problem that the kit cannot be used in some scenes is effectively solved, and the method has a good application prospect in the field of peach disease diagnosis and monitoring.

Description

technical field [0001] The invention relates to a kit for rapidly diagnosing peach bacterial perforation disease, which belongs to the field of combined use of plant protection and RPA-Crispr / Cas technology. Background technique [0002] In recent years, peach bacterial perforation disease caused by Xanthomonas arboricola pv.pruni (also known as Xap) is one of the main diseases with high incidence and serious damage on peach trees. In the early stage of the disease, small water-soaked gray-brown spots formed on the fruit surface and leaves, which continued to extend and aggravate in the later stage, forming large-scale perforation or cracking. Anthracnose and other symptoms are confused, making it difficult for grassroots agricultural technology extension personnel and fruit farmers to distinguish, and bringing technical difficulties to accurate diagnosis. In addition, the pathogen will quickly transfer and colonize new tissues after causing leaf necrosis, and the survival ...

AI Technical Summary

Problems solved by technology

The traditional morphological identification method requires practitioners to have solid professional theoretical knowledge of plant pathology and rich experience in disease identification. The identification cycle is long and the operation is cumbersome, and it is easy to be affected by different varieties, different tissues, different growth stages, disease symptoms or subjective symptoms. Cognition and other influences cause diagnostic errors, which are not applicable in field testing and production practice

Most of the existing molecular identification methods, such as conventional PCR, real-time fluorescence quantitative PCR, Bio-PCR, LAMP, etc., require cyclers or water baths, which cannot truly realize real-time detection in the field. Application space is limited

At present, the detection methods related to PCR technology based on the above-mentioned conservative marker fragments still have the difficulty that the detection sensitivity and specificity cannot meet the requirements of rapid and accurate diagnosis in the field

This is because, because the peach hole disease lesion tissue is often also infected by other pathogenic bacteria at the same time, such as peach brown spot hole bacteria (Wilsonomyces carpophilus), peach scab bacteria (Carpophila Venturia carpophila), and a small amount of disease The Xap strain DNA extracted from plaque tissue is often very limited, which brings technical obstacles to the detection based on PCR technology, making the detection sensitivity not high enough, and the results need to be judged under laboratory conditions, which cannot meet the needs of rapid and accurate detection in the field. Urgent need for testing

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