Reagent kit for detecting tomato yellow leaf curl virus carried by single-head whiteflies

Abstract

The invention discloses a reagent kit for detecting tomato yellow leaf curl virus carried by single-head whiteflies. A primer group includes a primer 1, a primer 2, a primer 3 and a primer 4, wherein nucleotide sequences of the primer 1, the primer 2, the primer 3 and the primer 4 are respectively a sequence 1, a sequence 2, a sequence 3 and a sequence 4 in a sequence table. An experiment of the reagent kit proves that the reagent kit aims for the single-head whiteflies, a LAMP primer is designed according to genome sequences of TYLCV preponderant strains occurred in our country, toothpicks or suckers are directly utilized to puncture the single-head whiteflies, deoxyribonucleic acid (DNA) is not required to be extracted, the LAMP reaction is directly performed, and the reagent kit has the advantages of being simple, quick, sensitive and the like and overcomes the shortcomings including time consumption, nucleic acid extraction, high instrument requirements, complicated steps and the like in traditional technologies.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for detecting tomato yellow leaf curl virus carried by single bemisia tabaci. Background technique [0002] Tomato yellow leaf curl virus (Tomato yellow leaf curl virus, TYLCV) is an important source of virus that causes a devastating disease in tomato production, "Tomato yellow leaf curl virus disease", which has caused serious economic damage to tomato production in my country. loss. The carrier of the virus is Bemisia tobaci Gennadius. Bemisia tobaci Gennadius is very common in crops, vegetables and weeds, and it harms plants such as Brassicaceae, Cucurbitaceae, Leguminosae, Solanaceae, Malvaceae, etc. , the main economic crops that are damaged are cotton, tobacco, tomato, sweet potato and so on. Bemisia tabaci carries the poison for life after being infected by sucking plants. Under protected cultivation conditions, Bemisia tabaci can safely overwinter in the north and oc...

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Problems solved by technology

[0003] In the integrated control of tomato yellow leaf curl virus disease, the use of non-toxic tomato seedlings and the monitoring of bemisia tabaci are the key to successful integrated control. A single infected bemisia tabaci can cause disease by sucking tomato plants , immediately causing disease epidemics and large-scale outbreaks, causing devastating disasters to tomato production. In view of this, it is necessary to carry out virus detection on Bemisia tabaci in a timely, fast and simple manner, which requires the development of a simple, fast and sensitive detection method Whether a single whitefly carries TYLCV

[0004] At present, there are two common methods to detect whether Bemisia tabaci carry TYLCV. One is molecular biology detection method, mainly using PCR method, but PCR detection is time-consuming, the product is inconvenient to detect, and the operation is cumbersome. The reaction requires a PCR machine and special reagents. Not suitable for production line use

The second is the serological detection method, mainly ELISA method, but the detection sensitivity and specificity of ELISA are not high, so it is not suitable for rapid and accurate detection

The deficiencies of the existing technology mainly include: first, the existing primers are not suitable for the detection of TYLCV that occurs in my country, which makes the detection efficiency low and the false negative rate is high; second, the current technology needs to extract DNA, the process is complicated, and it is not easy to popularize and apply; The existing technology is not suitable for the simple and fast detection of single whitefly

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