Through transposon mutagenesis and DNA sequence analysis, the main disease determinant of the entomopathogenic bacterium Yersinia entomophaga MH96 was localized to an ϳ32-kb pathogenicity island (PAI) designated PAI Ye96 . Residing within PAI Ye96 are seven open reading frames that encode an insecticidal toxin complex (TC), comprising not only the readily recognized toxin complex A (TCA), TCB, and TCC components but also two chitinase proteins that form a composite TC molecule. The central TC gene-associated region (ϳ19 kb) of PAI Ye96 was deleted from the Y. entomophaga MH96 genome, and a subsequent bioassay of the ⌬TC derivative toward Costelytra zealandica larvae showed it to be innocuous. Virulence of the ⌬TC mutant strain could be restored by the introduction of a clone containing the entire PAI Ye96 TC gene region. As much as 0.5 mg of the TC is released per 100 ml of Luria-Bertani broth at 25°C, while at 30 or 37°C, no TC could be detected in the culture supernatant. Filter-sterilized culture supernatants derived from Y. entomophaga MH96, but not from the ⌬TC strain grown at temperatures of 25°C or less, were able to cause mortality. The 50% lethal doses (LD 50 s) of the TC toward diamondback moth Plutella xylostella and C. zealandica larvae were defined as 30 ng and 50 ng, respectively, at 5 days after ingestion. Histological analysis of the effect of the TC toward P. xylostella larva showed that within 48 h after ingestion of the TC, there was a general dissolution of the larval midgut.Toxin complexes (TCs) active on insects were first identified in the nematode-associated bacterium Photorhabdus luminescens and termed TCs, as three proteins combined to form a complex with insecticidal activity (5). TC toxins were subsequently identified in the genome of Serratia entomophila where they reside in the designated gene order sepA, sepB, and sepC (33); this toxin complex ABC designation defines the revised nomenclature of the TC proteins (25). The TC toxins derived from P. luminescens reside as multiple but dissimilar orthologues throughout the P. luminescens T011 genome (22), and different insecticidal activities may be attributed to a different TC cluster (32). The S. entomophila sepABC genes are plasmid borne, and their translated products are host specific, only causing amber disease in larvae of the New Zealand grass grub Costelytra zealandica (Coleoptera: Scarabaeidae) (33). TC-like toxins have since been identified in the genome of Xenorhabdus nematophilus (60), Pseudomonas syringae pv. tomato DC3000 (9), and some Yersinia species. The toxin complex A (TCA)-like (tcaB) gene of Yersinia pestis CO92 contains a frameshift mutation, and the toxin complex B (TCB)-like (tcaC) gene contains an internal deletion (51), indicative of a loss of function, while the corresponding TCA-like and TCBlike orthologues in Y. pestis KIM and 91001 do not (18, 62). Tennant et al. (66) showed that mutations in each of the Yersinia enterocolitica biotype 1A T83 genes, TCA-like (tcbA), TCB-like (tcaC), and TCC-like (tccC) gen...

A highly virulent strain, AGR96X, exhibiting specific pathogenicity against larvae of the New Zealand grass grub (; Coleoptera: Scarabaeidae) and the New Zealand manuka beetle ( and ; Coleoptera: Scarabaeidae), was isolated from a diseased grass grub larva. A 12-day median lethal dose of 4.89 × 10 ± 0.92 × 10 cells per grass grub larva was defined for AGR96X, and death occurred within 5 to 12 days following the ingestion of a high bacterial dose. During the infection period, the bacterium rapidly multiplied within the insect host and invaded the hemocoel, leading to a mean bacterial load of 8.2 × 10 cells per larva at 6 days postingestion. Genome sequencing of strain AGR96X revealed the presence of a variant of the antifeeding prophage (Afp), a tailocin designated AfpX. Unlike Afp, AfpX contains two Afp16 tail-length termination protein orthologs and two putative toxin components. A 37-kb DNA fragment encoding the AfpX-associated region was cloned, transformed into, and fed to and larvae, causing mortality. In addition, the deletion of the putative chaperone component abolished the virulence of AGR96X. Unlike Afp, the AfpX tailocin could be induced by mitomycin C. Transmission electron microscopy analysis revealed the presence of Afp-like particles of various lengths, and when the purified AfpX tailocin was fed to grass grub or manuka beetle larvae, they underwent phenotypic changes similar to those of larvae fed AGR96X. strain AGR96X shows dual activity against larvae of endemic New Zealand pasture pests, the grass grub () and the manuka beetle ( spp.). Unlike , the causal agent of amber disease, which takes 3 to 4 months to kill grass grub larvae, AGR96X causes mortality within 5 to 12 days of ingestion and invades the insect hemocoel. AGR96X produces a unique variant of the antifeeding prophage (Afp), a cell-free phage-like entity that is proposed to deliver protein toxins to the grass grub target site, causing a cessation of feeding activity. Unlike other Afp variants, AGR96X Afp, named AfpX, contains two tail-length termination proteins, resulting in greater variability in the AfpX length. AfpX shows dual activity against both grass grub and manuka beetle larvae. AGR96X is a viable alternative to for pest control in New Zealand pasture systems.

Azole resistance is uncommon in Australian clinical and environmental A. fumigatus isolates; further surveillance is indicated.

In a collection of 735 Botrytis isolates sampled from Australian wine grape‐growing regions, a single isolate from clade I and group I (based on Bc‐hch RFLP analysis) was found. As many Botrytis species are known to live sympatrically, it was hypothesized that this isolate might be a new Botrytis species. After phenotypic and molecular assays supported this hypothesis, the species was designated B. medusae. Phylogenetic analyses using the nuclear genes G3PDH, HSP60, RPB2, NEP1 and NEP2 consistently placed B. medusae in an early‐diverging clade I Botrytis spp. lineage. Botrytis medusae produced white aerial mycelium, grew faster at 30 °C and produced long‐branched conidiophore extensions, compared with B. cinerea and B. pseudocinerea. Botrytis medusae was only able to infect wounded grape leaves and was significantly less virulent on wounded leaves and berries than B. cinerea. Botrytis medusae also lacked villiform appendages on the conidial surface and long conidiophores, which are defining features of B. sinoviticola and B. californica, respectively. Identification and characterization of new cryptic Botrytis species living in sympatry on grapevines could potentially provide information to assist disease management strategies for B. cinerea.

Here we report the draft genome of Yersinia entomophaga type strain MH96T. The genome shows 93.8% nucleotide sequence identity to that of Yersinia nurmii type strain APN3a-cT, and comprises a single chromosome of approximately 4,275,531 bp. In silico analysis identified that, in addition to the previously documented Y. entomophaga Yen-TC gene cluster, the genome encodes a diverse array of toxins, including two type III secretion systems, and five rhs-associated gene clusters. As well as these multicomponent systems, several orthologs of known insect toxins, such as VIP2 toxin and the binary toxin PirAB, and distant orthologs of some mammalian toxins, including repeats-in-toxin, a cytolethal distending toxin, hemolysin-like genes and an adenylate cyclase were identified. The genome also contains a large number of hypothetical proteins and orthologs of known effector proteins, such as LopT, as well as genes encoding a wide range of proteolytic determinants, including metalloproteases and pathogen fitness determinants, such as genes involved in iron metabolism. The bioinformatic data derived from the current in silico analysis, along with previous information on the pathobiology of Y. entomophaga against its insect hosts, suggests that a number of these virulence systems are required for survival in the hemocoel and incapacitation of the insect host.