Tissue Print Hybridization for Detection and Characterization of Viroids. 2022

Maria S Kaponi, and Teruo Sano, and Panayota E Kyriakopoulou
Department of Heraklion, Hellenic Agricultural Insurance Organization (ELGA), Maiandrou & Petrakogiorgi, Crete, Greece.

Simplifying sample preparation by transferring plant sap (from plant sections) directly onto a membrane is advantageous for any routine viroid detection technique, such as tissue printing. After fixing the samples on the membrane, hybridization steps similar to Northern or dot blot can be successfully implemented as long as factors like stringency and low viroid titer are properly adjusted to enable enhancement of the detection limit. The protocol described allows for indexing hundreds of field samples as a phytosanitary control measure.

UI MeSH Term Description Entries
D009693 Nucleic Acid Hybridization Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503) Genomic Hybridization,Acid Hybridization, Nucleic,Acid Hybridizations, Nucleic,Genomic Hybridizations,Hybridization, Genomic,Hybridization, Nucleic Acid,Hybridizations, Genomic,Hybridizations, Nucleic Acid,Nucleic Acid Hybridizations
D010935 Plant Diseases Diseases of plants. Disease, Plant,Diseases, Plant,Plant Disease
D010944 Plants Multicellular, eukaryotic life forms of kingdom Plantae. Plants acquired chloroplasts by direct endosymbiosis of CYANOBACTERIA. They are characterized by a mainly photosynthetic mode of nutrition; essentially unlimited growth at localized regions of cell divisions (MERISTEMS); cellulose within cells providing rigidity; the absence of organs of locomotion; absence of nervous and sensory systems; and an alternation of haploid and diploid generations. It is a non-taxonomical term most often referring to LAND PLANTS. In broad sense it includes RHODOPHYTA and GLAUCOPHYTA along with VIRIDIPLANTAE. Plant
D006824 Hybridization, Genetic The genetic process of crossbreeding between genetically dissimilar parents to produce a hybrid. Crossbreeding,Hybridization, Intraspecies,Crossbreedings,Genetic Hybridization,Genetic Hybridizations,Hybridizations, Genetic,Hybridizations, Intraspecies,Intraspecies Hybridization,Intraspecies Hybridizations
D012367 RNA, Viral Ribonucleic acid that makes up the genetic material of viruses. Viral RNA
D014772 Viroids A group of pathogens comprising the smallest known agents of infectious disease. They are unencapsulated and are capable of replicating autonomously in susceptible cells. Positively identified viroids composed of single-stranded RNA have been isolated from higher plants, but the existence of DNA viroids pathogenic to animals is suspected. Viroid
D015151 Immunoblotting Immunologic method used for detecting or quantifying immunoreactive substances. The substance is identified by first immobilizing it by blotting onto a membrane and then tagging it with labeled antibodies. Dot Immunoblotting,Electroimmunoblotting,Immunoelectroblotting,Reverse Immunoblotting,Immunoblotting, Dot,Immunoblotting, Reverse,Dot Immunoblottings,Electroimmunoblottings,Immunoblottings,Immunoblottings, Dot,Immunoblottings, Reverse,Immunoelectroblottings,Reverse Immunoblottings

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