Bacillus popilliae and Bacillus lentimorbus, bacteria causing milky disease in Japanese beetles and related scarab larvae

Abstract

Bacillus popilliae and Bacillus lentimorbus, causative agents of milky disease in Japanese beetle and related scarab larvae, have hitherto been differentiated based upon a small number of phenotypic characteristics, but they have not previously been examined at the molecular level. In this study 34 isolates of these bacteria were examined for DNA similarity and by random amplified polymorphic DNA (RAPD) analysis. Two distinct but related similarity groups were identified: the f i r s t contained strains of B. popilliae and the second contained strains of B. lentimorbus. Two strains distinct from but related t o B. popilliae may represent a subspecies. Some strains received as B. popilliae were found to be most closely related to B. lentimorbus and some received as B. lentimorbus were found to be most closely related t o B. popilliae. RAPD analysis confirmed the DNA similarity results. Paraspore formation, previously believed to be a characteristic unique to B. popilliae, was found to occur among a sub-group of B. lentimorbus strains. Growth in media supplemented with 2% NaCl was found to be a somewhat less reliable characteristic in distinguishing the species than vancomycin resistance, the latter being present only in B. popilliae.

A new strain of Paenibacillus lentimorbus isolated from larvae of the oriental beetle, Blitopertha orientalis (Coleoptera: Scarabaeidae), in Chiba Prefecture, Japan

“…rhopaea isolated from Rhopaea verreauxi had a parasporal body, but was incapable of growth in 2% NaCl. Rippere et al (1998) and Harrison et al (2000) also reported that some strains identified as P. popilliae were unable to grow in MYPGP medium containing 2% NaCl. Further, Rippere et al (1998) and Harrison et al (2000) reported that parasporal body formation was present in isolates of both species as defined by DNA similarity rather than being restricted to P. popilliae, as originally described.…”

“…Rippere et al (1998) and Harrison et al (2000) also reported that some strains identified as P. popilliae were unable to grow in MYPGP medium containing 2% NaCl. Further, Rippere et al (1998) and Harrison et al (2000) reported that parasporal body formation was present in isolates of both species as defined by DNA similarity rather than being restricted to P. popilliae, as originally described. Stahly et al (1992) reported that most B. popilliae strains originally isolated from Japanese and oriental beetles exhibited vancomycin resistance, while four of five Cyclocephala strains were vancomycin sensitive.…”

“…Three characteristics, growth in 2% NaCl, presence of a parasporal body (Claus and Berkeley, 1986), and resistance to vancomycin (Stahly et al, 1992;Rippere et al, 1998), have been described as phenotypic traits to distinguish B. popilliae from B. lentimorbus. Semadara strain was unable to grow in 2% NaCl, it was susceptible to vancomycin, and had a parasporal body.…”

Susceptibility of the Japanese beetle, Popillia japonica (Newman) (Coleoptera: Scarabaeidae), to entomopathogenic Hypocreales fungi

“…galleriae SDS-502 (Yamaguchi et al 2013) and Paenibacillus popilliae (formely Bacillus popilliae) (Bulla et al 1975;Klein 1992;Redmond and Potter 1995;Matsuki et al 1997;Rippere et al 1998), the nematodes Steinernema glaseri Steiner and Heterorhabditis bacteriophora Poinar, and the fungi Metarhizium anisopliae (Lacey et al 1994;Villani et al 1994;Lacey et al 1995b;Abalos et al 2001;Cappaert and Smitley 2002;Petty et al 2012) and Beauveria bassiana (Hanula and Andreadis 1988;Hanula et al 1991;Klein 1992;Lacey et al 1995a, b;Bixby et al 2007). …”

Evaluation of the hypervariable region in the 16S rDNA sequence as an index for rapid species identification in the genus Paenibacillus.

“…1) because these informative positions were deleted by gap removal to create the final data set to be used for subsequent phylogenetic calculations. On the other hand, insect pathogens P. lentimorbus and P. popilliae are closely related species (Klein and Kaya, 1995;Rippere et al, 1998;Pettersson et al, 1999) From the above observations, these three species may be in subspecific relation to each other, and further studies with more strains and under same experimental conditions are necessary to clarify the appropriate taxonomic positions of these species and intraspecific group. Strains of P. polymyxa were subdivided into two groups, A-1 and A-2, based on a single nucleotide difference at position 212 (group A-1: G and group A-2: A), and the sequences of group A-1 were identical to that of P. polymyxa IAM 13419…”