Characterization and analysis of the transcriptome in Opisina arenosella from different developmental stages using single-molecule real-time transcript sequencing and RNA-seq.

Opisina arenosella is one of the main pests harming coconut trees. To date, there have been few studies on the molecular genetics, biochemistry and physiology of O. arenosella at the transcriptional level, and there are no available reference genomes. Here, Illumina RNA sequencing combined with PacBio single-molecule real-time analysis was applied to study the transcriptome of this pest at different developmental stages, providing reference data for transcript expression analysis. Twelve samples of O. arenosella from different stages of development were sequenced using Illumina RNA sequencing, and the pooled RNA samples were sequenced with PacBio technology (Iso-Seq). A full-length transcriptome with 41,938 transcripts was captured, and the N50 and N90 lengths were 3543 bp and 1646 bp, respectively. A total of 36,925 transcripts were annotated in public databases, 6493 of which were long noncoding RNAs, while 2510 represented alternative splicing events. There were significant differences in the gene expression profiles at different developmental stages, with high levels of differential gene expression associated with growth, development, carbohydrate metabolism and immunity. This work provides resources and information for the study of the transcriptome and gene function of O. arenosella and provides a valuable foundation for understanding the changes in gene expression during development.

Transcriptome and gene expression analysis of Rhynchophorus ferrugineus (Coleoptera: Curculionidae) during developmental stages.

BACKGROUND: Red palm weevil, Rhynchophorus ferrugineus Olivier, is one of the most destructive pests harming palm trees. However, genomic resources for R. ferrugineus are still lacking, limiting the ability to discover molecular and genetic means of pest control. METHODS: In this study, PacBio Iso-Seq and Illumina RNA-seq were used to generate transcriptome from three developmental stages of R. ferrugineus (pupa, 7th-instar larva, adult) to increase the understanding of the life cycle and molecular characteristics of the pest. RESULTS: Sequencing generated 625,983,256 clean reads, from which 63,801 full-length transcripts were assembled with N50 of 3,547 bp. Expression analyses revealed 8,583 differentially expressed genes (DEGs). Moreover, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that these DEGs were mainly related to the peroxisome pathway which associated with metabolic pathways, material transportation and organ tissue formation. In summary, this work provides a valuable basis for further research on the growth and development, gene expression and gene prediction, and pest control of R. ferrugineus.

SMRT sequencing of the full-length transcriptome of the Rhynchophorus ferrugineus (Coleoptera: Curculionidae).

BACKGROUND: Red palm weevil Rhynchophorus ferrugineus (Coleoptera: Curculionidae) is one of the most destructive insects for palm trees in the world. However, its genome resources are still in the blank stage, which limits the study of molecular and growth development analysis. METHODS: In this study, we used PacBio Iso-Seq and Illumina RNA-seq to first generate transcriptome from three developmental stages of R. ferrugineus (pupa, 7th larva, female and male) to increase our understanding of the life cycle and molecular characteristics of R. ferrugineus. RESULTS: A total of 63,801 nonredundant full-length transcripts were generated with an average length of 2,964 bp from three developmental stages, including the 7th instar larva, pupa, female adult and male adult. These transcripts showed a high annotation rate in seven public databases, with 54,999 (86.20%) successfully annotated. Meanwhile, 2,184 alternative splicing (AS) events, 2,084 transcription factors (TFs), 66,230 simple sequence repeats (SSR) and 9,618 Long noncoding RNAs (lncRNAs) were identified. In summary, our results provide a new source of full-length transcriptional data and information for the further study of gene expression and genetics in R. ferrugineus.

Inter-country trade, genetic diversity and bio-ecological parameters upgrade pest risk maps for the coconut hispid Brontispa longissima.

BACKGROUND: Invasions of a number of tree-feeding beetles have increased globally and pose a mounting threat to the world's trees, production forests and natural habitats. An in-depth understanding of the determinants of invasion potential of a given species and invasibility of novel environments can help forecast future invasions and avert undesirable socio-economic impacts. Here, we quantitatively assess the (multivariate) drivers of historic invasions of the coconut hispid Brontispa longissima (Coleoptera: Chrysomelidae) across the Asia-Pacific region and critically assess its invasion potential for other key coconut-growing regions. RESULTS: Genetic variation of B. longissima in its invaded range indicated multiple incursions, likely associated with (short-range) natural dispersal and (long-range) trade in ornamental palms and coconut plantlets. Interception records at China's ports of entry accentuate the role of traded planting material. The high fecundity and prolonged, yet adaptable, oviposition period of B. longissima further enhance the invasiveness of this species and aid its successful establishment. Coconut-growing areas are identified with high climatic suitability for B. longissima, and where strengthened biosecurity protocols can prevent future invasions. CONCLUSION: A combined assessment of inter-country trade patterns, population genetics and species bio-ecology (e.g. climate-related development) illuminates the dispersal pathways of invasive species, assesses invasibility of particular geographies, guides quarantine interventions and thus can effectively avert future invasions. © 2019 Society of Chemical Industry.

Potential distribution of an invasive pest, Euplatypus parallelus, in China as predicted by Maxent.

BACKGROUND: Euplatypus parallelus is a highly polyphagous invasive pest native to Central and South America. In recent years it has invaded many countries in Africa and Asia and resulted in considerable economic loss. In China it has been reported to have invaded Taiwan, and been also recorded in Hainan Province. Until now there has been no invasion into the mainland. In order to better manage this invasive pest, here we predicted the suitable area of E. parallelus in China by the Maxent model. RESULTS: The Maxent model predicted the potential distribution of E. parallelus with a test AUC value of 0.962 and a training AUC value of 0.978. Temperature seasonality (Bio_04), annual temperature range (Bio_07), annual precipitation (Bio_12) and mean temperature of the coldest quarter (Bio_11) were the strongest predictors of E. parallelus distribution with 32.1%, 19.8%, 15% and 10.4% contributions, respectively. The potential suitable area for E. parallelus was mainly distributed in the southeastern coast, the southwestern border, and Taiwan and Hainan provinces in China, and the highly suitable areas were located in the northern coast of Hainan Province and the southwestern coast of Taiwan Province. This pest prefers a stable, warm and rainy climate, which indicates that tropics and subtropics would be its ideal area. CONCLUSION: Euplatypus parallelus has invaded Hainan and Taiwan in China. Measures should be taken to prevent it from spreading on these two islands. Moreover, strict quarantine, biological study and control measures are necessary to block its spread, invasion and damage, especially in these climate-suitable areas. © 2018 Society of Chemical Industry.

Nutrient addition increases the capacity for division of labor and the benefits of clonal integration in an invasive plant.

Many of the most invasive plants are clonal, and clonal integration has been proposed as an important mechanism promoting invasiveness. When the availabilities of two essential resources are negatively correlated in space, clonal integration may benefit clonal plants through division of labor. We hypothesized that environments with reciprocal patchiness of light and soil water may induce division of labor, and nutrient addition may increase both the division of labor and the benefits of clonal integration. To test this, we grew pairs of connected and disconnected ramets of the clonal invader Mikania micrantha under negative spatial covariance of light and soil water such that the proximal ramets were grown under high light and low soil water conditions and the distal ramets were grown under low light and high soil water conditions. In half of the ramet pairs, both ramets of a pair received a nutrient addition treatment. The results showed that connection decreased the root to shoot ratio in proximal ramets and increased it in distal ramets, indicating that division of labor was induced. In addition, connection increased the root to shoot ratio of distal ramets more under high soil nutrient conditions than under low soil nutrient conditions, indicating that nutrient addition increased the division of labor. Connection increased plant biomass at the whole clonal fragment level, and this increase was larger under high soil nutrient conditions than under low soil nutrient conditions. This study showed, for the first time, that in environments with reciprocal patchiness of two essential resources, the capacity for division of labor and its influence on plant performance may depend on the availability of a third essential resource. Because invasive plants often can acquire a larger amount of soil resources than native plants, our study may also contribute to the understanding why clonality is related to invasiveness.

Identification of putative odorant binding protein genes in Asecodes hispinarum, a parasitoid of coconut leaf beetle (Brontispa longissima) by antennal RNA-Seq analysis.

Asecodes hispinarum (Hymenoptera: Eulophidae) is an endoparasitoid and an efficient biological control agent which attacks larvae of Brontispa longissima, a serious insect pest of Palmae plants in China. Odorant binding proteins (OBPs) are believed to be important for transporting semiochemicals through the aqueous sensillar lymph to the olfactory receptor cells within the insect antennal sensilla. No previous study has reported on OBPs in A. hispinarum. In this study, we conducted the large-scale identification of OBP genes from the antennae of A. hispinarum by using transcriptome sequencing. Approximately 28.4 million total raw reads and about 27.3 million total clean reads were obtained, and then 46,363 unigenes were assembled. Of these unigenes, a total of 21,263 can be annotated in the NCBI non-redundant database. Among the annotated unigenes, 16,623 of them can be assigned to GO (Gene Ontology). Furthermore, we identified 8 putative OBP genes, and a phylogenetic tree analysis was performed to characterize the 8 OBP genes. In addition, the expression of the 8 OBP genes in different A. hispinarum body tissues was analyzed by real-time quantitative polymerase chain reaction (qRT-PCR). The results indicated that the 8 OBP genes were expressed accordingly to sexes and tissues, but all highly expressed in antennae. The finding of this study will lay the foundation for unraveling molecular mechanisms of A. hispinarum chemoperception.

Effect of ozone on respiration of adult Sitophilus oryzae (L.), Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.).

The effect of ozone on the respiration of three species of adult stored-product Coleoptera was tested in an air-tight flask. Sitophilus oryzae (L.), Rhyzopertha dominica (F.) and Tribolium castaneum (Herbst) adults were exposed to atmosphere containing 0.1, 0.2 or 0.4microg/ml initial ozone at 23-25 degrees C and 50% r.h. Carbon dioxide (CO(2)) production reflected the respiration rates of insects and was determined with a gas chromatograph (GC). The experiments showed that the effects of ozone on respiration had two distinct phases. Phase 1 involved a lower respiration rate of the adult stored-product Coleoptera under ozone atmosphere and reflected the need for insects to reduce ozone toxicity. After 1h, CO(2) production of S. oryzae was 3.19, 2.63, 2.27 and 1.99microl/mg for the ozone concentration of 0, 0.1, 0.2 and 0.4microg/ml, respectively. The results also showed that there were decreases in the rate of respiration in R. dominica and T. castaneum with an increase in ozone concentration. During phase 2, respiration of S. oryzae, R. dominica, and T. castaneum adults treated with ozone increased as the ozone degraded to oxygen. After 7h, the effect of ozone on CO(2) production, relative to the control, changed from a decrease to an increase. The findings in relation to control strategies were discussed.