Abstract

IRACHETA CARDENAS, María Magdalena; SALAZAR MARTINEZ, Sanjuanita Lyzzeth; CARDENAS AVILA, María Luisa  and  ROCHA PENA, Mario A.. In Situ Localization of Citrus tristeza virus Inclusion Bodies with Antibodies Developed to the Recombinant p20 Non-Structural Protein. Rev. mex. fitopatol [online]. 2012, vol.30, n.2, pp.161-166. ISSN 2007-8080.

An in situ immunoassay is described for localization of Citrus tristeza virus (CTV) inclusion bodies by light microscopy with antibodies developed to the recombinant p20 non-structural protein. Free hand bark sections of ca. 100 µm tick from healthy or CTV infected plants were fixed with 70 % ethanol and subsequently incubated with specific polyclonal antiserum anti-p20 either from rat or rabbit at dilutions 1:5,000 y 1:10,000 y 1:1,000 and 1:3,000 respectively and IgG anti species at 1:30,000 dilution; at the end, the chromogenic solution NBT/BCIP was added. Viral inclusion bodies were visualized by light microscopy at 10 and 40X magnifications as dark blue irregular areas in phloem associated tissues in CTV infected plants; no dark blue color was observed in the phloem tissues of healthy plants.

Keywords : DIBA; serology; recombinant protein; light microscopy.