Colorado potato beetle, Leptinotarsa decemlineata (Say), is a devastating pest of potatoes in North America and Europe. L. decemlineata has developed resistance to insecticides used for its control. In this study, in order to find a more effective potential biological control agent against L. decemlineata, we investigated its microbiota and tested their insecticidal effects. According to morphological, physiological and biochemical tests as well as 16S rDNA sequences, microbiota was identified as Leclercia adecarboxylata (Ld1), Acinetobacter sp. (Ld2), Acinetobacter sp. (Ld3), Pseudomonas putida (Ld4), Acinetobacter sp. (Ld5) and Acinetobacter haemolyticus (Ld6). The insecticidal activities of isolates at 1.8×109 bacteria/mL dose within five days were 100%, 100%, 35%, 100%, 47% and 100%, respectively, against the L. decemlineata larvae. The results indicate that Leclercia adecarboxylata (Ld1) and Pseudomonas putida (Ld4) isolates may be valuable potential biological control agents for biological control of L. decemlineata.

Thaumetopoea pityocampa (pine processionary moth) is one of the most important pine pests in the forests of Mediterranean countries, Central Europe, the Middle East and North Africa. Apart from causing significant damage to pinewoods, T. pityocampa occurrence is also an issue for public and animal health, as it is responsible for dermatological reactions in humans and animals by contact with its irritating hairs. High throughput sequencing technologies have allowed the fast and cost-effective generation of genetic information of interest to understand different biological aspects of non-model organisms as well as the identification of potential pathogens. Using these technologies, we have obtained and characterized the transcriptome of T. pityocampa larvae collected in 12 different geographical locations in Turkey. cDNA libraries for Illumina sequencing were prepared from four larval tissues, head, gut, fat body and integument. By pooling the sequences from Illumina platform with those previously published using the Roche 454-FLX and Sanger methods we generated the largest reference transcriptome of T. pityocampa. In addition, this study has also allowed identification of possible viral pathogens with potential application in future biocontrol strategies.

In this study, we have isolated and identified 5 bacterial isolates from European shot-hole borer, Xyleborus dispar (Coleoptera: Scolytidae), an important pest of hazelnut. After various morphological, physiological, biochemical and molecular characteristics were determined in detail, bacterial isolates were identified as Pseudomonas fluorescens (Xd1), Bacillus megaterium (Xd2), Bacillus thuringiensis (Xd3), Pseudomonas rhizosphaerae (Xd4) and Pantoea cedenensis (Xd5). Especially, the B. thuringiensis strain was identified in more detail as Bacillus thuringiensis subsp. tenebrionis (H8ab). The insecticidal activities were determined against the larvae of Agelastica alni L. (Col.: Chrysomelidae), Amphimallon solstitiale L. (Col.: Scarabaeidae), Leptinotarsa decemlineata (Say) (Col.: Chrysomelidae), and Melolontha melolontha L. (Col.: Scarabaeidae) larvae at 1.8 x 10 9 CFU/ml dose, within five days. The highest insecticidal activities are 100, 100, 80 and 100% for Bacillus thuringiensis subsp. tenebrionis (Xd3) on A. alni, A. solstitiale, L. decemlineata, and M. melolontha larvae, respectively. Our results indicate that especially B. thuringiensis subsp. tenebrionis (Xd3) may be valuable as a microbial control agent for coleopteran pests, respectively.

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The Amsacta moorei entomopoxvirus (AMEV) genome has 279 open reading frames (ORFs) among which is the AMV197, composed of 900 nt and potentially encoding a protein of 299 amino acids. Sequence-derived amino acid analysis suggested it to be a serine/threonine protein kinase (PK) having conserved PK and serine/ threonine PK domains. For transcriptional analysis of the AMV197 pk gene, Ld652 cells were infected with AMEV and mRNA was isolated at different times thereafter. RT-PCR analysis indicated that the transcription of the AMV197 pk gene started at 4 h post infection (h p.i.) and continued to be expressed through 24 h p.i. Infection of Ld cells in the presence of Ara-C (inhibits DNA replication), followed by RT-PCR showed that AMV197 pk is transcribed as an early gene. Transcription was initiated at 54 nt upstream of the translation start site. The vaccinia virus early promoter element G was also found at the correct position (−21) in the AMV197 pk gene. Rapid amplification of the 3′ ends of the AMV197 pk transcript showed that there are two polyadenylation start points. They are located at 22 and 32 nucleotides downstream of translation stop site. Also, the translational stop site and poly (A) signal of AMV197 pk are overlapped. The termination signal TTTTTGT sequence of vaccinia virus early genes was found just upstream of the 3′ end of AMV197 pk gene. Conserved amino acid subdomains of the AMV197 PK were found by sequence comparisons with PK's from other organisms. Analysis of the protein sequence of AMV197 pk gene reveals close identity with PK genes of other organisms.