Method for rapidly detecting dendrobe virus

Abstract

The invention discloses a method for rapidly detecting dendrobe virus. The invention provides a method for detecting whether dendrobe plants carry dendrobe virus or not. The method comprises the following steps of obtaining juice from tissues of to-be-detected dendrobe plants; carrying out reverse transcription by taking the obtained stock solution or diluent of the juice as a template; and carrying out PCR amplification by using the reverse transcription product as a template and the dendrobium virus universal primer, and determining whether the to-be-detected dendrobium plant carries the dendrobium virus according to a result. The method is an effective method for rapidly, simply and conveniently detecting and cloning the dendrobe virus, plays a considerable role in quarantine and control of DVd under natural conditions, and can also provide a method reference for detection of other quasi-virus and even virus.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for rapidly detecting dendrobium viroids, which is mainly used for rapid detection of dendrobium viroids (Dendrobium viroid, DVd) by rapidly extracting juice from dendrobium stem / leaf tissues combined with RT-PCR. Background technique [0002] Dendrobium, which belongs to the genus Dendrobium of the family Orchidaceae, is a monocotyledonous plant widely distributed in many countries in Asia. It is an important traditional Chinese medicine, and its stems and leaves are both succulent. Dendrobium has high ornamental value in addition to its wide range of medicinal effects Khentry, Y., Paradornuwat, A., Tantiwiwat, S., Phansiri, S., Thaveechai, N., 2006. andodontoglossum ringspot virus in Dendrobium spp.in Thailand. Crop Prot.25,926-932.; Bulpitt,C.J.,Li,Y.,Bulpitt,P.F.,Wang,J.,2007.The use of orchids in Chinese medicine.J.R.Soc.Med.100,558 -563.). Dendrobium plants are suscept...

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Problems solved by technology

However, extraction of nucleic acids by these traditional methods generally requires more plant samples (at least 0.1-0.2 g fresh weight leaves per sample), these methods require complicated steps to remove polyphenols and polysaccharides in nucleic acids, and the cost is high (Hadidi, A., Candresse, T., 2003. Polymerase chain reaction. In: Hadidi, A., Flores, R., Randles, J.W., Semanick, J.S. (Eds.), Viroids. CSIRO, Collingwood, Australia, pp .115-122.)

There are also many biological companies on the market that have developed kits for the extraction of viroid nucleic acid, but most of the kits are expensive, and ordinary laboratories cannot afford the detection of large-scale samples

In addition, the effect of nucleic acid extraction with kits varies greatly, which is mainly related to the polyphenol polysaccharide content of plants.

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