ABSTRACT: Expression analysis of tomato plants TYLCV resistant, susceptible and resistant line silenced in the hexose transporter (LeHT1) gene before and 7 days after inoculation of tomato with Tomato yellow leaf curl virus (TYLCV).
Project description:Investigation of whole genome gene expression level changes in a resistance susceptible and LeHT1-silenced resistance tomato plants, infected with TYLCV and compared to the non infected control. Two inbred tomato lines (named R and S) were used issued from a breeding program describe by Vidavsky and Czosnek, 1998, and Silencing of the hexose transporter gene LeHT1 and the properties of these plants have been described by Eybishtz et al., 2010. The array was designed to include all the known tomato genes retrieved from TIGR and Sol Genomics public databases. Following filtration for redundancies, 25,591 known genes and 7,335 uncertain or unknown genes were retrieved The data were transferred to Roche NimblGen company (http://www.nimblegen.com/) who has generated a 60-mer oligonucleotide array where each of the ~30,000 tomato genes is represented by at least two specific probes, and unrelated controls. Each slide contains four arrays of ~70,000 oligos. TYLCV resistant and susceptible lines infection with TYLCV and LeHT1 silenced TYLCV resistant line infected with TYLCV
Project description:Investigation of whole genome gene expression level changes in a resistance susceptible and LeHT1-silenced resistance tomato plants, infected with TYLCV and compared to the non infected control. Two inbred tomato lines (named R and S) were used issued from a breeding program describe by Vidavsky and Czosnek, 1998, and Silencing of the hexose transporter gene LeHT1 and the properties of these plants have been described by Eybishtz et al., 2010.
Project description:A comparative study to determine the pepper leaf curl virus resistance machanism between resistant and susceptible genotypes at three leaf stage. To study the molecular mechanism of pepper leaf curl virus (PepLCV) resistance, pepper plants were exposed to PepLCV through artificial inoculation and hybridization on Agilent tomato microarrays. The expression analysis of PepLCV resistant and susceptible genotypes after artificial inoculation at three leaf stage showed that the resistance against PepLCV is due to sum of expression of hundreds of genes at a particular stage. Tomato microarrays consisting of 43,803 probes were used for whole genome expression analysis of chilli peppers for resistance against PepLCV. Transcripts from the leaves of resistant (BS-35) and susceptible plants (IVPBC-535) were compared in response to PepLCV inoculation at three leaf stage.
Project description:A comparative study to determine the pepper leaf curl virus resistance machanism between resistant and susceptible genotypes at three leaf stage. To study the molecular mechanism of pepper leaf curl virus (PepLCV) resistance, pepper plants were exposed to PepLCV through artificial inoculation and hybridization on Agilent tomato microarrays. The expression analysis of PepLCV resistant and susceptible genotypes after artificial inoculation at three leaf stage showed that the resistance against PepLCV is due to sum of expression of hundreds of genes at a particular stage.
Project description:In this study, an GFP-based affinity purification followed by mass spectrometry (AP-MS) approach is used in order to identify new tomato proteins that interact with the Replication initiator protein (Rep, AL1, C1) from Tomato yellow leaf curl virus (TYLCV).
Project description:Transcriptional overlap between transgenic Arabidopsis plants expressing C4G2A from the Tomato yellow leaf curl virus (TYLCV) and a cas-1 mutant upon activation of plant immunity by treatment with the bacterial peptide elicitor flg22 (1 µM, 12 h).
Project description:RNA interference (RNAi) is a widely-used approach to generate virus-resistant transgenic crops. However, durability of RNAi-mediated resistance under extreme field conditions and side-effects of stable RNAi expression have not been thoroughly investigated. Here we performed field trials and molecular characterization of two RNAi-transgenic Solanum lycopersicum lines resistant to Tomato yellow leaf curl virus (TYLCV) disease, the major constraint for tomato cultivation in Cuba and worldwide. In order to determine potential impact of the hairpin RNA transgene expression on tomato genome expression and development, differences in the phenotypes and the transcriptome profiles between the transgenic and non-transgenic plants were examined. Transcriptome profiling revealed a common set of up- and down-regulated tomato genes, which correlated with slight developmental abnormalities in both transgenic lines.
Project description:The whitefly, Bemisia tabaci MEAM1 is a devastating vector capable of transmitting hundreds of plant viruses, including Tomato yellow leaf curl virus (TYLCV), to important food and fiber crops. Here we performed genome-wide profiling of micro RNAs (miRNAs) and piwi-interacting RNAs (piRNAs) in whiteflies after feeding on TYLCV-infected tomato or uninfected tomato for 24, 48 and 72 h. Overall, 160 miRNAs were discovered, 68 of which were conserved and 92 were B. tabaci-specific miRNAs. Majority of the genes that were predicted to be targeted by miRNAs had gene ontologies related to metabolic processes. We identified two miRNAs that were differentially expressed in whiteflies when fed on TYLCV-infected tomato compared to whiteflies that fed on uninfected tomato. The identified piRNAs were expressed as clusters throughout the whitefly genome. A total of 53 piRNA clusters were expressed across all time points and treatments, while 5 piRNA clusters were exclusively expressed in whiteflies that fed on TYLCV-infected tomato, and 24 clusters were exclusively expressed in whiteflies that fed on uninfected tomato. Approximately 62% of all identified piRNAs were derived from non-coding sequences that included intergenic regions, introns, and UTRs with unknown functions. The remaining 38% of piRNAs were derived from coding sequences (CDS) and repeat elements. Transposable elements targeted by piRNA clusters included both class I retrotransposons such as Gypsy, Copia, and LINEs and class II DNA transposons such as MITE, hAT, and TcMar. Lastly, six protein coding genes were targeted in whiteflies that fed on TYLCV-infected tomato. Information on how TYLCV influences miRNA and piRNA expression in whiteflies provides a greater understanding of regulatory pathways involved in mediating whitefly-virus interactions, and will facilitate the identification of novel targets for RNAi control.
Project description:Female whiteflies are used to inoculate the susceptible and resistant tomato genotypes as follows. Whiteflies are caged with TYLCV-infected tomato plants for a 48 h acquisition access period. Then, the viruliferous insects are collected and used to inoculate 20 plants of each genotype. The plants at their 4 leaf-stage are inoculated by caging ten viruliferous whiteflies with the youngest true leaf (more than 1 cm) of each plant, using leaf cages. Mock inoculations are conducted using the same number of non-viruliferous insects. After a 12 h inoculation access period (which ensures 100% infection), the insect cages are removed and the plants are treated with insecticide (imidacloprid). The plants are kept for 3 or 7 days in an insect-proof greenhouse, in natural daylight (12 h light/12 h dark). Keywords: Direct comparison, loop design
Project description:Tomato curly stunt virus (ToCSV) is a monopartite begomovirus infecting tomatoes in South Africa, with sequence similarity to tomato yellow leaf curl virus (TYLCV). While there are numerous reports on the mechanism of TYLCV resistance in tomato, the underlying mechanisms in the tomato-ToCSV pathosystem is still relatively unknown. The main aim of this study was to investigate and compare the global methylation profile of ToCSV in two near-isogenic tomato lines, one with a tolerant phenotype (T, NIL396) and one with a susceptible phenotype (S, NIL395). Bisulfite conversion and PCR amplification, coupled with a next-generation sequencing approach, were used to elucidate the global pattern of methylation of ToCSV cytosine residues in T and S leave tissue at 35 days post-infection (dpi). The extent of methylation was more pronounced in tolerant plants compared to susceptible plants in all sequence (CG, CHG and CHH) contexts, however, the overall methylation levels were relatively low (<3%). Notably, a significant interaction (p < 0.05) was observed between the viral genomic region and susceptible vs. tolerant status for CG methylated regions where it was observed that the 3'IR CG methylation was significantly (p < 0.05) higher than CG methylation of other genomic regions in tolerant and susceptible plants. Additionally, statistically significant (EdgeR p < 0.05) differentially methylated cytosines were located primarily in the genomic regions V2/V1 and C4/C1 of ToCSV. The relative expression, using RT-qPCR, was also employed in order to quantify the expression of various key methylation-related genes, MET1, CMT2, KYP4/SUVH4, DML2, RDM1, AGO4 and AGO6 in T vs. S plants at 35dpi. The differential expression between T and S was significant for MET1, KYP4/SUVH4 and RDM1 at p<0.05 which further supports more pronounced methylation observed in ToCSV from T plants vs. S plants. While this study provides new insights into the differences in methylation profiles of ToCSV in S vs. T tomato plants, further research is required to link tolerance and susceptibility to ToCSV.
