Yu-Di Liu

In this study, a vitellogenin like1 gene (SfVg-like1) in Sogatella furcifera was identified. The open reading frame (ORF) encoded 1,321 amino acid sequence. Structure analysis reveals that the amino acid sequence of SfVg-like1 has 3 conserved LPD_N, DUF1943 and VWFD domains. Phylogenetic analyses showed that SfVg-like1 was clustered in the same branch with the Vg-like1 of Nilaparvata lugens (100% bootstrap value) compared with other Hemiptera insects Vgs associated with vitellogenesis. Temporo-spatial expression analyses showed that SfVg-like1 expressed during all stages, and in both genders. The relative expression levels of SfVg-like1 mRNA were higher in adults than in nymph developmental stages. The knockdown of SfVg-like1 gene resulted in the inhibition of the ovarian development in female adults, whereas the morphology of the testis in male adults was not been affected. The silence of SfVg-like1 could decrease the relative expression levels of target of rapamycin (SfTOR, GenBank MW193765) and vitellogenin (SfVg, GenBank MH271114) genes significantly in female adults. However, the knockdown of SfTOR or SfVg genes in female adults did not affect the transcript level of SfVg-like1. Therefore, it demonstrated that SfVg-like1 might locate on the upstream signaling pathways of SfTOR and SfVg. These results demonstrate that SfVg-like1 is essential for S. furcifera reproduction, and it could be the potential target for the control of this pest.

The white-backed planthopper (WBPH), Sogatella furcifera, is one of the most important piercing-sucking pests of rice (Oryza sativa) in Asia. Mucin-like salivary protein (SFMLP) is highly expressed in the salivary glands of WBPH, which plays an important role in WBPH feeding. In this study, WBPH injected with dsSFMLP had difficulty in sucking phloem sap from rice plants, which significantly reduced their food intake, weight, and survival. In contrast, the knockdown of the SFMLP gene had only a marginal effect on the survival of WBPH fed an artificial diet. Further studies showed that silencing SFMLP resulted in the short and single-branched salivary sheaths secretion and less formation of salivary flanges in rice. These data suggest that SFMLP is involved in the formation of the salivary sheath and is essential for feeding in WBPH. Overexpression of the SFMLP gene in rice plants promoted the feeding of WBPH, whereas silencing the gene in rice plants significantly decreased WBPH performance. Additionally, it was found that overexpression of SFMLP in rice plants elicited the signalling pathway of SA (salicylic acid) while suppressing JA (jasmonic acid); in contrast, silencing of the SFMLP gene in rice plants showed the opposite results. This study clarified the function of SFMLP in WBPH feeding as well as mediating rice defences.

Vitellogenin (Vg) and the target of rapamycin (TOR) are important genes involved in insect reproduction regulation. In this work, the full lengths of the Vg (SfVg) and TOR (SfTOR) genes of the white-backed planthopper Sogatella furcifera were cloned. The expression pattern in females showed that SfVg was highly expressed in fat bodies, and SfTOR was highly expressed in Malpighian tubules. After silencing SfVg or SfTOR, female adults did not deposit eggs. Their ovarian development was delayed, and yolk protein deposition in the oocytes was reduced. However, wild-type females mated with SfTOR-silenced males could lay eggs and produce offsprings normally. The dissections of testes and accessory glands of males with SfTOR knockdown showed that their development was not affected. Therefore, the silencing of the SfVg or SfTOR genes can effectively inhibit female reproduction, but SfTOR knockdown has no significant effect on male reproductive capacity. Furthermore, silencing SfTOR can cause SfVg expression to decrease significantly. All of the above results revealed that SfVg and SfTOR are essential for white-backed planthopper reproduction and may provide a potential target for pest control.

The brown planthopper (BPH), Nilaparvata lugens Stal (Hemiptera: Delphacidae), is a serious rice pest because of its destructive feeding. The salivary glands of the BPH play a key role in successful feeding. In this study, we explored the salivary gland transcriptome (sialotranscriptome) of adult BPHs using Illumina sequencing and a total of 55,913 transcripts and 45,421 unigenes were obtained. We identified one reference gene RPL9 (Ribosomal protein L9) and 19 salivary protein genes from the BPH sialotranscripome, which were categorized as those involved in sugar metabolism, extra-oral digestion of cell wall components, detoxification, and suppression of plant defenses. Tissue expression profiles of 19 salivary protein genes analysis revealed that the expression level of alpha-glucosidase family 31 had no difference in five tissues, suggesting that it may have functions in the whole-body parts. Glucose dehydrogenase (flavine adenine dinucleotide, quinone)-like was expressed highly in the salivary gland, which might play putative role in insect feeding. Glucose dehydrogenase (acceptor) was expressed the highest level in head without salivary gland. Other salivary protein genes were highly expressed in gut or malpighian tubule, suggesting that they may play roles in digestive and detoxification mechanism. Moreover, we detected RPL9 is one of the genes that is most consistently present for acquisition of gene expression in different tissues. Thus, RPL9 can be a new reference for expression studies of BPH. The obtained BPH sialotranscripome provides a list of genes that have potential roles in feeding and interaction between BPHs and rice plants.

The white-backed planthopper, Sogatella furcifera, is a phloem sap feeder that secretes watery and gelling saliva during feeding. In this study, we identified the major proteins in watery saliva of S. furcifera by shotgun LC-MS/MS analysis combined with transcriptomic analysis. A total of 161 proteins were identified, which were divided into 8 function categories, including enzymes, transporter, calcium ion binding protein, salivary sheath protein, cytoskeleton protein, DNA-, RNA-, and protein-binding or regulating proteins, other non-enzyme proteins and unknown proteins. Gene expression pattern of 11 secretory proteins were analyzed by real time quantitative-PCR. We detected the mucin-like protein, which had a unique expression level in salivary gland, most likely to be a candidate effector involved in regulation of plant defense. This study identified the watery saliva component of S. furcifera and it provided a list of proteins which may play a role in interaction between S. furcifera and rice.

Trichogramma wasps were collected from the parasitized eggs of lepidopteran pests from 21 sampling sites in East Asia and South-East Asia. Six Trichogramma species were identified based on the molecular identificationmethod using the internal transcribed spacer 2 (ITS2) region of the rDNAof Trichogramma chilonis, T. evanescens, T. ostriniae, T. embryophagum, T. dendrolimi and T. japonicum. The results of molecular identification were confirmed by morphological identification. Additionally, natural populations were screened for the prevalence of Wolbachia. Five out of 21 populations were infected by the same Wolbachia strain, which was identified by using Wolbachia wsp gene and multilocus sequencing approach. The phylogenetic analysis of Wolbachia wsp sequences revealed that the Wolbachia strain was classified in the strain wEvaA in the group of EvA of the supergroup A.

The white-backed planthopper (WBPH), Sogatella furcifera (Horváth), is one of the serious rice pests because of its destructive feeding. The salivary glands of the WBPH play an important role in the feeding behaviour. Currently, however, very little is known about the salivary glands at the molecular level. We sequenced the salivary gland transcriptome (sialotranscripome) of adult WBPHs using the Illumina sequencing. A total of 65,595 transcripts and 51,842 unigenes were obtained from salivary glands. According to annotations against the Nr database, many of the unigenes identified were associated with the most studied enzymes in hemipteran saliva. In the present study, we identified 32 salivary protein genes from the WBPH sialotranscripome, which were categorized as those involved in sugar metabolism, detoxification, suppression of plant defense responses, immunity-related responses, general digestion, and other phytophagy processes. Tissue expression profiles analysis revealed that four of 32 salivary protein genes (multicopper oxidase 4, multicopper oxidase 6, carboxylesterase and uridine phosphorylase 1 isform X2) were primarily expressed in the salivary gland, suggesting that they played putative role in insect-rice interactions. 13 of 32 salivary protein genes were primarily expressed in gut, which might play putative role in digestive and detoxify mechanism. Development expression profiles analysis revealed that the expression level of 26 of 32 salivary protein genes had no significant difference, suggesting that they may play roles in every developmental stages of salivary gland of WBPH. The other six genes have a high expression level in the salivary gland of adult. 31 of 32 genes (except putative acetylcholinesterase 1) have no significant difference in male and female adult, suggesting that their expression level have no difference between sexes. This report analysis of the sialotranscripome for the WBPH, and the transcriptome provides a foundational list of the genes involved in feeding. Our data will be useful to investigate the mechanisms of interaction between the WBPH and the host plant.

The white-backed planthopper, Sogatella furcifera (Hemiptera, Delphacidae), is one of the most devastating rice pests. For a better control strategy, various genetic studies have been conducted using reverse-transcription quantitative real-time polymerase chain reaction (qRT-PCR). The appropriate application of qRT-PCR requires reliable endogenous controls; however, studies on this aspect of the white-backed planthopper are lacking. In the present study, nine commonly used reference genes, elongation factor 1-α (EF1-α), polyubiquitin (UB), ribosomal protein S18 (RPS18), actin 1 (ACT), α-1 tubulin (TUB), glyceraldehyde-3-phosphate (GAPDH), ribosomal protein L9 (RPL9), ribosomal protein L10 (RPL10), and 18S ribosomal RNA (18S), were evaluated by qRT-PCR for their expression stability under four different experimental conditions (different developmental stages, acquisition of Southern rice black-streaked dwarf virus (SRBSDV), different tissues, and different temperature stress). These results were analyzed using four software programs (geNorm, NormFinder, BestKeeper, and the delta Ct method) and a Web-based comprehensive tool RefFinder to compare and rank candidate reference genes. According to the results of RefFinder analysis, which integrates the abovementioned four software programs, TUB was ranked as the most suitable reference gene at different developmental stages and under different temperature stress, and GAPDH and RPL9 showed the highest stability for acquisition of SRBSDV and different tissues, respectively. These results will provide a solid foundation for future gene expression study on the white-backed planthopper, and also will give aids in establishing a standardized qRT-PCR procedure for other related insects.

The white-backed planthopper, Sogatella furcifera (Horvath), is currently the only confirmed vector of Southern rice black-streaked dwarf virus (SRBSDV), which causes severe rice production losses in China. In this study, an absolute quantification qPCR method was used to detect viral gene mRNA expression levels at different developmental stages of white-backed planthoppers fed SRBSDV-infected rice plants. A comparison of viral copy numbers of the SRBSDV S10 gene at the same developmental stage indicated that the white-backed planthopper had higher viral copy numbers when the virus was acquired at the earlier developmental stages. The adult-stage white-backed planthoppers that had acquired the virus at the first–second nymphal stage displayed significantly higher viral titers than white-backed planthoppers that acquired the virus at the third–fourth nymphal stage, at the fifth nymphal stage, and at the adult stage. The fifth nymphal stage white-backed planthoppers that acquired the virus at the first–second nymphal stage displayed higher viral copy numbers than fifth nymphal stage white-backed planthoppers that acquired the virus at the third–fourth nymphal stage and at the fifth nymphal stage. The highest viral load value appeared in the middle adult stage. The annual immigration characteristics of white-backed planthoppers would be beneficial for the dispersal of SRBSDV because this virus could be transmitted far away following the migration of vigorous planthoppers. Therefore, investigating the change in the viral load at different life stages of SRBSDV-positive individuals is required to develop more effective control of the spread of SRBSDV in the field.

The properties of red soil may change over time when paddy fields are developed on what was previously dry land. The effect of rice cultivation duration on soil invertebrates is unknown. Five chronosequences of rice cultivation (1, 10, 20, 50 and 100 years) were selected to investigate the temporal changes of soil microbiological properties and nematode assemblages. The results showed that soil microbial properties (microbial biomass C, microbial biomass N, basal respiration), nutrient availability (mineral N and P), nematode abundance and richness of nematode genera generally increased with the duration of rice cultivation. Notably, most soil measurements peaked after 50 years of cultivation (P<0.05) and decreased slightly after 100 years. As the period of rice cultivation increased, the proportion of nematode herbivores rose significantly (P < 0.05), while that of predators/omnivores slightly declined. Nematode Channel Ratio (NCR) also increased with cultivation duration (P < 0.05), indicating that the bacterial energy channel was more dominant in old rice fields when compared to those cultivated for a shorter period. Other ecological indices of nematode assemblage such as maturity index and structure index, did not reveal consistent trends with an increased period of rice cultivation. In summary, a change in land use from dry land to paddy fields promotes soil microbial properties and nematode assemblages in the first few decades, which then become stable after 50 years of cultivation