Abstract
Increasing incidence of viral infections in crop plants adversely affects their growth and yield. Tomato (Solanum lycopersicum) is considered to be a favorite host for viruses with over 50 species of begomoviruses naturally infecting this crop. Tomato leaf curl virus (ToLCV) is among the most widespread and devastating begomoviruses affecting tomato production. microRNAs (miRs) have been established as key regulators of gene expression and plant development. The miR pathways are disturbed during infection by viruses. Thus, comprehension of regulatory miR networks is crucial in understanding the effect of viral pathogenicity. To identify key miRs involved in ToLCV infection, a high throughput approach involving next generation sequencing was employed. Healthy and infected leaf tissues of two tomato varieties, differing in their susceptibility to ToLCV infection were analyzed. NGS data analysis followed by computational predictions, led to identification of 91 known miRs, 15 novel homologs and 53 novel miRs covering two different varieties of tomato, susceptible (Pusa Ruby) and tolerant (LA1777) to ToLCV infection. The cleaved targets of these miRs were identified using online available degradome libraries from leaf, flower and fruit of tomato and showed their involvement in various biological pathways through KEGG Orthology. With detailed comparative profiling of expression pattern of these miRs, we could associate the specific miRs with the resistant and infected genotypes. This study depicted that in depth analysis of miR expression patterns and their functions will help in identification of molecules that can be used for manipulation of gene expression to increase crop production and developing resistance against diseases.
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This research was supported by grants from Department of Biotechnology, New Delhi, India. The authors acknowledge the efforts of Dr. Afsar Raza Naqvi in preparing the tomato sRNA libraries.
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12298_2017_482_MOESM1_ESM.xlsx
Supplementary File 1: Lists all the predicted novel miRs, their precursors, genomic loci, associated family and isoforms
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Supplementary File 2: Secondary structure of all predicted miRs and their isoforms. miR is highlighted with blue and miR* is highlighted with red color and the circle at the last base represents 5′ end of the precursor
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Supplementary File 4: Validation of miRs by stem-loop RT-PCR in tissues infiltrated with agrobacterium clone. Lane 1: Low range DNA ladder (25 bp), 7 s: shoot tissue 7 days post infiltration, 7 r: root tissue 7 days post infiltration, 28 s: shoot tissue 28 days post infiltration, 28r: root tissue 28 days post infiltration
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Supplementary File 6: Target-plots (t-plots) and cleavage sites of all the degradome validated targets. Red dot indicates signatures consistent with miR-directed cleavage site. The abundance of each signature is plotted as a function of its position in the transcript
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Tripathi, A., Goswami, K., Tiwari, M. et al. Identification and comparative analysis of microRNAs from tomato varieties showing contrasting response to ToLCV infections. Physiol Mol Biol Plants 24, 185–202 (2018). https://doi.org/10.1007/s12298-017-0482-3
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DOI: https://doi.org/10.1007/s12298-017-0482-3