Detoxification activity and energy cost is attenuated in the whiteﬂies feeding on Tomato yellow leaf curl China virus-infected tobacco plants
ABSTRACT: We compared the transcriptional profiles of female adult whiteflies of B. tabaci Middle East-Asia Minor 1 feeding on TYLCCNV-free and TYLCCNV-infected tobacco plants using the next-generation sequencing technique. Culture of B (MEAM1 cryptic species) whitefly was maintained on cotton plants. One thousand of newly emerged adults of whitefly on cotton were released onto the leaves of healthy and viruliferous tobacco plants. After 72 h of oviposition, all the adult whiteflies were discarded, and the progeny allowed to develop to adults. The cultures of MEAM1 on tobacco plants were maintained in climate chambers at 27 ± 1°C, a photoperiod of 14 h light/10 h darkness and 70 ± 10% relative humidity. Approximately 1,000 female adult whiteflies newly emerged from mock-inoculated tobacco plants and 1,000 female adult whiteflies newly emerged from virus-infected tobacco plants were collected and stored at -80°C. The RNA was extracted and sequenced using Illunima Analyzer II.
Project description:We investigated the transcriptional response of invasive B. tabaci B biotype to tomato yellow leaf curl China virus (TYLCCNV) using Illumina sequencing technology. We found that 1,606 genes involved in 157 biochemical pathways were differentially expressed in the viruliferous whiteflies. Culture of B biotype whitefly was maintained on cotton plants. Three thousands of newly emerged adults of whitefly on cotton were released onto the leaves of healthy and viruliferous tobacco plants. They were allowed to feed for 24 h. After that, non-viruliferous and viruliferous whiteflies were transferred respectively to cotton plants in different cages and allowed to feed for 120 h. Then approximately 1,000 non-viruliferous and viruliferous female adults of whitefly were collected, respectively. The RNA was extracted and sequenced using Illunima Analyzer II.
Project description:We investigated the transcriptional response of invasive Mediterranean (MED) species of the whitefly B. tabaci complex (commonly referred to as Q biotype) to entomopathogenic fungi Beauveria bassiana using Illumina sequencing technology. Nearly 1,000 of control whiteflies, 48h fungal-induced whiteflies and 72h fungal-induced whiteflies were collected, respectively.
Project description:The whitefly Bemisa tabaci is a species complex of more than 31 cryptic species which include some of the most destructive invasive pests of many ornamental and glasshouse crops worldwide. Among them, Middle East-Asia Minor 1 (herein MEAM1) and Mediterranean (herein MED) have invaded many countries around the world and displaced the native whitefly species. However, the molecular differences between invasive and indigenous whiteflies remain largely unknown. The global transcriptional difference between the two invasive whitefly Bemisia tabaci species (MEAM1, MED) and one indigenous whitefly species (Asia II 3) were analyzed using the Illumina sequencing technology.
Project description:The whitefly Bemisa tabaci is a species complex with global distribution and extensive genetic diversity. In this species complex, Middle East-Asia Minor 1 (MEAM1, previously referred to as the ‘B biotype’) species has been spreading rapidly in tropical and subtropical regions. we analyzed the transcriptional responses of the invasive MEAM1 and the indigenous Asia II 3 species of B. tabaci complex during host plant shift (from cotton to tobacco) using the Illumina sequencing technology.The different gene expression pattern of energy and carbonhydrate metabolism and detoxification metabolism between MEAM1 and Asia II 3 were the main reasons of their different capacity of adapation. The global transcriptional difference between the invasive whitefly Bemisia tabaci species (MEAM1) and the indigenous whitefly species (Asia II 3) on cotton and tobacco were analyzed using the Illumina sequencing technology.
Project description:Total poly(A) RNAs from wild-type adult stage worms were subjected to high-throughput sequencing using an Illumina platform to detect RNA cleavage fragment. Wild-type (N2) C. elegans
Project description:Here we performed a transcriptomic study on complete symptom development process of CMV-infected Nicotiana tabacum using Solexa/Illumina's high-throughput digital gene expression (DGE) system. 12 DGE libraries (from six virus-infected samples and six corresponding mock-inoculated samples) were constructed, and the gene expression variations between the virus-infected sample and the mock-inoculated sample in each symptom stage were compared. Thousands of differentially expressed genes were obtained by the comparison, and KEGG pathway analysis of these genes suggested that many biological processes (such as phtosynthesis, pigment metabolism and plant-pathogen interaction) were related to the symptom development. The authenticity of the DGE data was further confirmed by analyzing real-time RT-PCR using several random-selected genes. We sequenced a cDNA library constructed from mixture of total RNA from six virus-infected samples and six mock-inoculated samples to get gene information for tobacco leaves in different symptom stages, including vein clearing, mosaic, severe chlorosis, partial recovery, total recovery and re-mosaic, and 95,916 Unigenes were obtained Tobacco leaves were inoculated by CMV-infected leaf homogenate and healthy leaf homogenate, respectivley. Six time points with different symptom stage were selected, and one virus-infect sample and one mock-inoculated sample were collected at each time. In order to average out variation of different plants, five leaves from five different plants were mixed to prepare every RNA sample. Twelve individual tag libraries of samples (six infected samples and six mock-inoculated samples) were constructed in parallel. For the gene expression analysis, the twelve samples were grouped into six groups, and each group contained a virus-infected sample and a mock-inoculated sample collected at the same time. In each group, the DGE data of virus-infected sample were compared to that of mock-inoculated sample to obtain the gene expression variations. Illumina sequencing of transcripts from virus-infected and mock-inoculated samples to get gene information for tobacco leaves in different symptom stages. In order to get more gene information, the systemically infected leaves and mock-inoculated leaves were harvested at six time points and five leaves from five different plants were collected at each time point. The RNA-Seq analysis provided gene information for mock-inoculated and virus-infected tobacco leaves in different symptom stages.
Project description:Versatile roles of REVOLUTA (REV), a Class III homeodomain-leucine zipper (HD-ZIP III) transcription factor, have been mainly depicted in Arabidopsis and Populus. In this study, we investigated the functions of its tomato homolog, namely SlREV. Over-expression of a microRNA166-resistant version of SlREV (35S::REVRis) not only resulted in vegetative abnormities such as curly leaves and fasciated stems, but also caused dramatic reproductive alterations including continuous production of flowers at pedicel abscission zone (AZ) and ectopic fruit formation on receptacles. Microscopic analysis showed that meristem-like structures continuously emerged out from the exodermises of pedicel AZs and ectopic carpels formed between the first and the second whorl of floral buds in 35S::REVRis plants. Therefore, we performed Illumina’s digital gene expression (DGE) system, a tag-based transcriptome sequencing methodTranscriptional data to dicover differential expressed genes in early buds (1-2 mm floral buds at stage 6-8) of overexpression line SlREVRis-1. The result suggests that SlREV may regulate genes related to meristem maintenance and cell differentiation in the development of flower pedicel abscission zone, and modulate genes in homodomain and MADS-box families and hormone pathways during fruit formation. These results reveal important roles of SlREV in tomato. 1-2 mm floral buds at stage 6-8 were sampled from three individual plants of 35S::REVRis-1 and corresponding WT control. Three aliquots of RNA from transgenic or WT plants were pooled. Then, the digital expression profile were generated by Illumina Cluster Station and Illumina HiSeq™ 2000 System (BGI Inc.).
Project description:We used a digital gene expression (DGE) system to generate partial gene expression profiles for 12 selected samples. Significant differences in gene expression between early- and late-flowering samples were detected for 72 candidate genes for flowering time. Genes related to circadian rhythms were significantly overrepresented among the differentially expressed genes. Our data suggest that circadian clock genes play an important role in the evolution of flowering time, and C. bursa-pastoris plants exhibit expression differences for candidate genes likely to affect flowering time across the broad range of environments they face in China. Digital gene expression analysis was performed on RNA from 12 different flowering samples of C. bursa-pastoris. These samples are from different areas, 6 of which represent early-flowering ecotypes, and the other 6 samples represent late-flowering samples
Project description:Trichomes are the hair-like structures that are widely present on the surface of aerial organs and function in plant defense against biotic and abiotic stresses. Previous studies focus on the single cell trichomes in Arabidopsis and cotton, or multicellular glandular trichomes in tomato, but the developmental process and molecular mechanisms controlling multicellular non-glandular trichome development are largely neglected. Here, we extensively characterized the fruit trichome (spine) development in wild type cucumber and in a tiny branched hair (tbh) mutant that contains a spontaneous mutation and has hairless foliage and smooth fruit surface. Our data indicated that cucumber trichome was multicellular and non-glandular, with no branches or endoreduplication. Further, the major feature of cucumber trichome development was spine base expansion. Transcriptome profiling through Digital Gene Expression indicated that meristem-related genes and transcription factors were implicated in the fruit spine development, and polarity regulators were upregulated during spine base expansion. qRT-PCR verified the reliability of our RNA-SEQ data, and in situ hybridization confirmed the enriched expression of meristem regulators CUP-SHAPED COTYLEDON3 (CUC3) and STM (SHOOT MERISTEMLESS) , as well as the abaxial identity gene KANADI (KAN) in cucumber fruit spine. Together, our results suggest a distinct regulatory pathway involving meristem genes and polarity regulators in multicellular trichome development in cucumber. Using Digital Gene Expression technology to compare the genome-wide gene expression profiles in the fruit spines of wild type cucumber and the tbh mutant, as well as the fruit spines on fruits of 0.5cm and 1.6cm long, repectively. Two biological repelicates were generated for each tissue.
Project description:We report the application of single-molecule-based sequencing technology for high-throughput profiling of transcriptome in mazie plants. The ZmPIS gene coding PtdIns synthase from maize with a maize ubiquitin promoter was transferred into maize. The transgenic ZmPIS maize showed enhanced drought tolerance compared to non-transgenic maize. The differentially expressed genes between wide-type maize and transgenic ZmPIS maize were detected by the assay of digital gene expression profile and real time RT-PCR datas. The results displayed that the overexpression of ZmPIS resulted in the expression levels changes of a large number of genes including genes involved in the phosphatidylinositol (PI) metabolic pathway, photosynthesis metabolism, carbohydrate metabolism, aminoacid metabolism and genes coding transcription factors. Examination of The differences of the transcriptional profile between wide-type maize and transgenic ZmPIS maize and analysis of the network regulated by the ZmPIS gene