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 tomato SlWRKY3 transcription factor was overexpressed in cultivated tomato (Solanum lycopersicum)and transgenic plants transcriptome was compared to that of wild-type plants.
Project description:Small RNA (sRNA)-guided RNA silencing is a critical antiviral defense mechanism employed by a variety of eukaryotic organisms. Although the induction of RNA silencing by bipartite and monopartite begomoviruses has been described in plants, the nature of begomovirus/betasatellite complexes remains undefined. We profiled Tomato yellow leaf curl China virus (TYLCCNV) and its associated betasatellite (TYLCCNB)-derived small RNAs (V-sRNAs and S-sRNAs) using Solexa-based deep sequencing to evaluate the role of betasatellites in V-sRNA modulation. Both sense and anti-sense V-sRNAs and S-sRNAs accumulated preferentially as 22 nucleotide species in infected Solanum lycopersicum and Nicotiana benthamiana plants, indicating that secondary siRNAs were triggered. High resolution mapping of V-sRNA and S-sRNA revealed heterogeneous distribution of V-sRNA and S-sRNA sequences across the TYLCCNV and TYLCCNB genomes. In TYLCCNV-infected S. lycopersicum or N. benthamiana and TYLCCNV and betaC1-mutant TYLCCNB co-infected N. benthamiana plants, the primary TYLCCNV targets were AV2 and the 5’ terminus of AV1. In TYLCCNV and betasatellite-infected plants, the number of V-sRNAs targeting this region decreased and the production of V-sRNAs increased corresponding to the overlapping regions of AC2 and AC3, as well as the 3’ terminal of AC1. betaC1 is the primary determinant mediating symptom induction and also the primary silencing target of the TYLCCNB genome even in its mutated form. In addition, the betasatellite affected the amount of V-sRNAs detected in S. lycopersicum and N. benthamiana plants. characterization of Tomato yellow leaf curl China virus and Tomato yellow leaf curl China betasatellite-derived small interfering RNAs from five cDNA libraries of two plant species
Project description:The tomato SlWRKY3 transcription factor was overexpressed in cultivated tomato (Solanum lycopersicum)and transgenic plants transcriptome was compared to that of wild-type plants. At least 4 plants were collected for RNA extraction. The aim of the experiment was to compare transcriptomes of 35::SlWRKY3 plants and wild-type plants grown together and on MS (Murashige and Skoog) medium in vitro for 4 weeks. A technical replicate (dye swap) was conducted.
Project description:Plants represent the nutritional basis of virtually all life on earth and protein-rich foods from crop plants are a global megatrend essential for sustaining an increasing human population and counteracting climate change. While the genomes of crops are increasingly elucidated, little is known about crop proteomes – the entirety of proteins that execute and control nearly every aspect of life. To address this shortcoming we optimized a protocol for mapping the proteome of different crops such as Solanum lycopersicum (tomato) fruit and included four technical replicates and three biological replicates from different tomato plants to demonstrate the robustness of the workflow.
Project description:RNA sequencing in tomato for detect mRNA expression of Solanum lycopersicum Axillary bud.The two cultivars (monomaker, raceme) at Axillary bud for transcriptome sequencing
Project description:We sequenced mRNA from immature green (15 days after anthesis) and red (Breaker+10 days) tomato (Solanum lycopersicum) fruit tissues from plants over-expressing SlGLK1 and SlGLK2 and from control plants 'M82' to compare gene expression levels between transgenic fruit and the control. Note: Samples in SRA were assigned the same sample accession. This is incorrect as there are different samples, hence “Source Name” was replaced with new values. Comment[ENA_SAMPLE] contains the original SRA sample accessions.
Project description:RNA sequencing in tomato for detect mRNA expression of Solanum lycopersicum flower.The two cultivars (monomaker, raceme) had three different flowering stages (budlet, Flower bud, Full bloom) for transcriptome sequencing
Project description:Lines nearly isogenic for fw3.2 in the cultivated background Solanum lycopersicum c.v. Yellow Stuffer were grown in the greenhouse in a completely randomized design. fw3.2 (ys) and fw3.2 (wt) are NILs carrying cultivated and wild alleles for fw3.2 locus. Young flower buds were harvested. For each sample, three replicates were used. RNA was extracted using Trizol and Stand-specific libraries were prepared from total RNA and sequences of 51 bp were generated on an Illumina HiSeq2000.
