Open AccessDetection of Pasteuria penetrans infection in Meloidogyne arenaria race 1 in planta by polymerase chain reaction
Author(s) -
Schmidt L.M.,
Preston J.F.,
g G.,
Dickson D.W.,
Aldrich H.C.
Publication year - 2004
Publication title -
fems microbiology ecology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.377
H-Index - 155
eISSN - 1574-6941
pISSN - 0168-6496
DOI - 10.1016/j.femsec.2004.03.011
Subject(s) - biology , endospore , gene , polymerase chain reaction , 16s ribosomal rna , microbiology and biotechnology , bacillus subtilis , bacteria , ribosomal dna , primer (cosmetics) , spore , phylogenetic tree , genetics , chemistry , organic chemistry
We report on the development of a PCR‐based assay to detect Pasteuria penetrans infection of Meloidogyne arenaria in planta using specific primers for recently sequenced sigE , spoIIAB and atpF genes of P. penetrans biotype P20. Amplification of these genes in crude DNA extracts of ground tomato root galls using real‐time kinetic PCR distinguished infected from uninfected M. arenaria race 1 by analysis of consensus thresholds for single copy genes. Fluorescent in situ hybridization (FISH) using the sigE primer sequence as a probe shows hybridization to P. penetrans cells in various stages of vegetative (pre‐endospore) development. Ratios of gene copies for sigE and 16S rDNA were obtained for P. penetrans and compared to Bacillus subtilis as a genomic paradigm of endospore‐forming bacteria. Phylogenetic analysis of the sigE gene from Gram‐positive, endospore‐forming bacteria finds P. penetrans most closely related Paenbacillus polymyxa . The sporulation genes ( spo genes), particularly si gE , have sequence diversity that recommends them for species and biotype differentiation of the numerous Pasteuria isolates that infect a large number of plant‐parasitic nematodes.