In situ Detection of MLOs for Solanaceae Stolbur and Faba Bean Phyllody by Indirect Immunofluorescence

Abstract

MLOs of faba bean phyllody and stolbur diseases were detected by a simple sensitive indirect immunofluorescence technique. Antisera prepared against membrane fractions of diseased plants were used with both 40 ,urn-thick sections obtained with a freezing microtome and with 7 ,urn-thick sections prepared from aldehyde-fixed paraffin-embedded plant materials. The technique is highly sensitive, simple to practice and utilizes small quantities of reagents. Specific MLO labeling was obtained using antisera with low titers and containing high proportions of anti-plant antibodies. The ease of using sections from fresh plant material suggests the potential use of the technique for routine indexing and screening of MLO infections. Immunofluorescence on sections from paraffin-embedded plant tissues improved the visualization of labeled MLOs, factilitated the accessibility of antigens, reduced non-specific labeling and allowed more precise localization and interpretation of immunofluorescent reactions. ZusammenfassungIn situ-Nachweis von Mycoplasrna-ahnlichen Organismen (MLOs)-Erreger von Stolbur bei Nachtschattengewachsen und Phyllodie bei Feldbohnen -durch indirekte Immunfluoreszenz

Global Status of Phytoplasma Diseases in Vegetable Crops

“…Microscopy coupled with serology enhances the sensitivity of their detection. In tomato plants infected with “stolbur” and faba bean phyllody, phytoplasmas were confirmed by in situ immunofluorescence technique (Cousin et al, 1989). The presence of phytoplasma was also established by electron microscopy in ultrathin sections of sieve tubes of shoots and roots of potato showing witches’ broom (Harrison and Roberts, 1969), as well as in cauliflower and bottle gourd exhibiting symptoms of phyllody and witches’ broom (Chou et al, 1976; Bertaccini et al, 1983) where pleomorphic structures lacking the cell wall were observed.…”

Occurrence and distribution of ‘Candidatus Phytoplasma trifolii’ associated with diseases of solanaceous crops in Lebanon

“…Infection of tomato, pepper and potato by phytoplasma have been reported from several areas in the Mediterranean basin (Avinent and Llacer 1995;Cousin et al 1968;Cousin et al 1989;Fos et al 1992;Marcone and Ragozzino 1995;ZimmermanGries and Klein 1978). In Italy, phytoplasmas from phylogenetic groups 16SrI, III, V and XII were identified by PCR and RFLP analysis of 16S-rDNA amplicons (Del Serrone et al 2001).…”

Influence du milieu d'extraction sur la détection du bois noir et de la flavescence dorée de la vigne, par des anticorps poly- et monoclonaux dirigés contre les phytoplasmes du stolbur et de la flavescence dorée

“…Parallèlement aux approches de la biologie moléculaire, la sérologie avec des anticorps polyclonaux ou monoclonaux (AcMc) dirigés contre les phytoplasmes de plusieurs jaunisses (Caudwell et al, 1982 ;Clark et al, 1988 ;Boudon-Padieu et al, 1989 ;Cousin et al, 1989 ;Schwartz et al, 1989 ; (Daire et al, 1993a ;Kuszala et al, 1993 (Cousin et al, 1989). Puis la production d'un AcMc dirigé contre le phytoplasme du stolbur (Garnier et al, 1990) (Maixner, 1994 (Gugerli, 1984 (Kirkpatrick et al, 1987 ; (Deng et Hiruki, 1990 ;Daire et al, 1992 ;Jaraush et al, 1994 ;Saeed et al, 1994).…”