Open AccessDNA probes in detection of spiroplasmas and mycoplasma‐like organisms in plants and insects
Author(s) -
Nur I.,
Bove J.M.,
Saillard C.,
Rottem S.,
Whitcomb R.M.,
Razin S.
Publication year - 1986
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1986.tb01519.x
Subject(s) - spiroplasma , biology , plasmid , mollicutes , mycoplasma , gene , ribosomal rna , hybridization probe , dot blot , dna–dna hybridization , suppression subtractive hybridization , dna , homology (biology) , nucleic acid thermodynamics , southern blot , 16s ribosomal rna , microbiology and biotechnology , genetics , complementary dna , rna , cdna library
DNA probes were applied to detect spiroplasmas and uncultivable mycoplasma‐like organisms (MLOs) in infected plants and insects. The probes consisted of pMC5, a plasmid carrying the RNA genes of Mycoplasma capricolum and pRA1, a plasmid recovered from Spiroplasma citri . Southern blot hybridization of pMC5 with digested DNAs of periwinkle plants infected with S. citri , or with various MLOs, yielded 2 heavy and several weaker bands. The heavy hybridization bands were shown to represent rRNA genes of the plant chloroplasts, indicating significant nucleotide sequence homology between the mycoplasmal rRNA genes and those of plant chloroplasts. Some of the weaker hybridization bands, not revealed in DNA of healthy plants, appeared to represent rRNA gene sequences of the infectious agent. Use of the spiroplasma plasmid as a probe enabled the detection of S. citri in infected plant material and in hemolymph of infected leafhoppers at a high sensitivity level.