Abstract
BackgroundMicroRNAs (miRNAs) are a class of 21–24 nucleotide endogenous non-coding small RNAs that play important roles in plant development and defense responses to biotic and abiotic stresses. Tobacco curly shoot virus (TbCSV) is a monopartite begomovirus, cause leaf curling and plant stunting symptoms in many Solanaceae plants. The betasatellite of TbCSV (TbCSB) induces more severe symptoms and enhances virus accumulation when co-infect the plants with TbCSV.MethodsIn this study, miRNAs regulated by TbCSV and TbCSB co-infection in Nicotiana benthamiana were characterized using high-throughput sequencing technology.ResultsSmall RNA sequencing analysis revealed that a total of 13 known miRNAs and 42 novel miRNAs were differentially expressed in TbCSV and TbCSB co-infected N. benthamiana plants. Several potential miRNA-targeted genes were identified through data mining and were involved in both catalytic and metabolic processes, in addition to plant defense mechanisms against virus infections according to Gene Ontology (GO) analyses. In addition, the expressions of several differentially expressed miRNAs and their miRNA-targeted gene were validated through quantitative real time polymerase chain reaction (qRT-PCR) approach.ConclusionsA large number of miRNAs are identified, and their target genes, functional annotations also have been explored. Our results provide the information on N. benthamiana miRNAs and would be useful to further understand miRNA regulatory mechanisms after TbCSV and TbCSB co-infection.
Highlights
MicroRNAs are a class of 21–24 nucleotide endogenous non-coding small RNAs that play important roles in plant development and defense responses to biotic and abiotic stresses
A miRNA-sequencing approach was used to identify N. benthamiana miRNAs regulated by Tobacco curly shoot virus (TbCSV)/ TbCSV isolate Y35 and its betasatellite (TbCSB) co-infection
A total of 16,753,586 and 17, 822,708 raw reads were generated via Illumina sequencing to represent these samples
Summary
MicroRNAs (miRNAs) are a class of 21–24 nucleotide endogenous non-coding small RNAs that play important roles in plant development and defense responses to biotic and abiotic stresses. Since the first report of plant miRNA in Arabidopsis thaliana [5, 6], numerous studies have shown that plant miRNAs play critical roles in regulating plant development [7, 8], gene translational repression [9], flowering and sex determination [10], phytohormone signaling [11], and plant responses to abiotic and/or biotic stresses [12] such as drought stress [13] and infections with viruses or other pathogens [14,15,16].
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