The complete mitochondrial genome of a jujube geometrid, Sucra jujuba (Lepidoptera: Geometridae) and its phylogenetic analysis.

Sucra jujuba Chu, 1979 (Lepidoptera: Geometridae) is a major insect pest in jujube plantation. In this study, we have sequenced the complete mitochondrial genome of S. jujuba. The circular genome was 15,557 bp in length and contained 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), and one AT-rich region (GenBank accession no. MZ507574). The nucleotide composition was significantly biased (A, T, C, and G were 41.85%, 39.65%, 10.97%, and 7.53%, respectively) with A + T contents of 81.50%. The Bayesian phylogenetic analysis of the concatenated nucleotide sequences of 13 PCGs from 30 species in the subfamily Ennominae and two outgroup species was performed. The results indicated that S. jujuba was closely related to Amraica recursaria in the subfamily Ennominae.

The Sequence Characteristics and Binding Properties of the Odorant-Binding Protein SvelOBP1 from Sympiezomias velatus (Coleoptera: Curculionidae) to Jujube Volatiles.

Sympiezomias velatus (Chevrolat) (Coleoptera: Curculionidae) has caused serious damage on jujube trees (Ziziphus jujuba Mill) in northern China. Semiochemicals emerging from the host are essential in the process of insects identifying and localizing the host. The highly expressed odorant-binding protein 1 of S. velatus (SvelOBP1) was assumed to play a possible role in the recognition of host volatiles. In this study, SvelOBP1 was cloned based on the antennal transcriptome of S. velatus. The recombinant SvelOBP1 protein was expressed in Escherichia coli and purified by Ni-NTA resin. The predicted protein SvelOBP1 belonged to a classic OBP subfamily. The expression patterns revealed that SvelOBP1 was mainly expressed in the antennae of both males and females, whereas the expression of SvelOBP1 in other body parts could be neglected. The fluorescence binding assay indicated that SvelOBP1 displayed very strong binding affinities to dibutyl benzene-1,2-dicarboxylate and (Z)-hex-3-en-1-ol (Ki = 6.66 ± 0.03 and 7.98 ± 0.06 µM). The molecular docking results showed that residues Trp114, Phe115 and Asp110 may be involved in binding to both dibutyl benzene-1,2-dicarboxylate and (Z)-hex-3-en-1-ol and may have a great impact on odorant recognition of S. velatus. Our results provide evidence that SvelOBP1 might participate in the olfactory molecular perception of S. velatus and would promote the development of pest attractants for S. velatus control.

Identification and sex-specific expression of chemosensory genes in the antennal transcriptomes of Pachyrhinus yasumatsui (Coleoptera: Curculionidae).

Pachyrhinus yasumatsui Kono et Morimoto is a major pest of Chinese jujube, which is widespread in northern China and causes severe economic losses in the jujube industry. Chemosensory genes play crucial roles in insect behaviors. Currently, little is known about chemosensory genes in P. yasumatsui. In the present study, antennal transcriptomes of female and male adult P. yasumatsui were annotated. In total, 113 genes involved in chemosensory functions were identified, including 41 odorant receptors, 28 odorant-binding proteins, 16 ionotropic receptors, 15 chemosensory proteins, 9 gustatory receptors, and 4 sensory neuron membrane proteins. Subsequently, the phylogenetic analyses of these olfactory-related proteins in P. yasumatsui were conducted using multiple sequence alignment. Furthermore, sex-specific expression levels of 113 genes were analyzed based on fragments per kilobase of transcript per million mapped reads (FPKM). Then, the quantitative real-time PCR (RT-qPCR) was used to quantify gene expression profiles of 28 P. yasumatsui OBPs (PyasOBPs) and 15 CSPs (PyasCSPs). The results revealed that 20 PyasOBPs and 13 PyasCSPs exhibited significantly higher expression in the antennae than in the bodies, suggesting that they might have functions in olfaction. Moreover, some OBPs and CSPs (PyasOBP6, PyasOBP7, PyasOBP16, PyasOBP21, and PyasCSP4) exhibited female-biased expression, indicating that they might take part in several female-specific behaviors. This study will promote the understanding of olfactory mechanism in P. yasumatsui, and our findings lay the groundwork for developing environmentally friendly pest management measures.

Functional Characterization of Odorant Binding Protein PyasOBP2 From the Jujube Bud Weevil, Pachyrhinus yasumatsui (Coleoptera: Curculionidae).

Odorant binding proteins (OBPs) play an important role in insect olfaction. The jujube bud weevil Pachyrhinus yasumatsui (Coleoptera: Curculionidae) is a major pest of Zizyphus jujuba in northern China. In the present study, based on the antennal transcriptome, an OBP gene of P. yasumatsui (PyasOBP2) was cloned by reverse transcription PCR (RT-PCR). Expression profile analyses by quantitative real-time PCR (qRT-PCR) revealed that PyasOBP2 was highly expressed in the antennae of both male and female P. yasumatsui adults, while its expression was negligible in other tissues. PyasOBP2 was prokaryotically expressed, and purified by Ni-NTA resin. The fluorescence competitive binding assays with 38 plant volatiles from Z. jujuba showed that PyasOBP2 could bind with a broad range of plant volatiles, and had strongest binding capacities to host-plant volatiles like ethyl butyrate (Ki = 3.02 µM), 2-methyl-1-phenylpropene (Ki = 4.61 µM) and dipentene (Ki = 5.99 µM). The three dimensional structure of PyasOBP2 was predicted by homology modeling, and the crystal structure of AgamOBP1 (PDB ID: 2erb) was used as a template. The molecular docking results indicated that the amino acid residue Phe114 of PyasOBP2 could form hydrogen bonds or hydrophobic interactions with some specific ligands, so this residue might play a key role in perception of host plant volatiles. Our results provide a basis for further investigation of potential functions of PyasOBP2, and development of efficient monitoring and integrated pest management strategies of P. yasumatsui.

Characterization of Erwinia tracheiphila Bacteriophage FBB1 Isolated from Spotted Cucumber Beetles that Vector E. tracheiphila.

Erwinia tracheiphila, the causal pathogen of bacterial wilt of cucurbit crops, is disseminated by cucumber beetles. A bacteriophage, designated FBB1 (Fu-Beattie-Beetle-1), was isolated from spotted cucumber beetles (Diabrotica undecimpunctata) that were collected from a field in which E. tracheiphila is endemic. FBB1 was classified into the Myoviridae family based on its morphology, which includes an elongated icosahedral head (106 × 82 nm) and a putatively contractile tail (120 nm). FBB1 infected all 62 E. tracheiphila strains examined and three Pantoea spp. strains. FBB1 virions were stable at 55°C for 1 h and tolerated a pH range from 3 to 12. FBB1 has a genome of 175,994 bp with 316 predicted coding sequences and a GC content of 36.5%. The genome contains genes for a major bacterial outer-membrane protein, a putative exopolysaccharide depolymerase, and 22 predicted transfer RNAs. The morphology and genome indicate that FBB1 is a T4-like virus and thus in the Tevenvirinae subfamily. FBB1 is the first virulent phage of E. tracheiphila to be reported and, to date, is one of only two bacteriophages to be isolated from insect vectors of phytopathogens. Collectively, the results support FBB1 as a promising candidate for biocontrol of E. tracheiphila based on its virulent (lytic) rather than lysogenic lifestyle, its infection of all E. tracheiphila strains examined to date, and its infection of a few nonpathogenic bacteria that could be used to support phage populations when pathogen numbers are low.

Antifungal Effect of Magnolol and Honokiol from Magnolia officinalis on Alternaria alternata Causing Tobacco Brown Spot.

In this study, two phenol compounds, magnolol and honokiol, were extracted from Magnolia officinalis and identified by LC-MS, 1H- and 13C-NMR. The magnolol and honokiol were shown to be effective against seven pathogenic fungi, including Alternaria alternata (Fr.) Keissl, Penicillium expansum (Link) Thom, Alternaria dauci f.sp. solani, Fusarium moniliforme J. Sheld, Fusarium oxysporum Schltdl., Valsa mali Miyabe & G. Yamada, and Rhizoctonia solani J.G. Kühn, with growth inhibition of more than 57%. We also investigated the mechanisms underlying the potential antifungal activity of magnolol and honokiol. The results showed that they inhibited the growth of A. alternata in a dose-dependent manner. Moreover, magnolol and honokiol treatment resulted in distorted mycelia and increased the cell membrane permeability of A. alternata, as determined by conductivity measurements. These results suggest that magnolol and honokiol are potential antifungal agents for application against plant fungal diseases.