Abstract
The tea plant (Camellia sinensis L.) is vulnerable to the geometrid Ectropis oblique; although microRNAs (miRNAs) are important for plant growth, development and stress response, the function of miRNAs in the response of C. sinensis to stress from E. oblique is unclear. To identify E. oblique stress-responsive miRNAs and their target genes in tea plant, three small RNA libraries were constructed from leaves subjected to mechanical wounding (MW), geometrid attack (GA) and from healthy control (CK) leaves. Using high-throughput sequencing, 130 known miRNAs and 512 novel miRNAs were identified; of these, differential expression under GA stress was observed for 36 known and 139 novel miRNAs. Furthermore, 169 GA-responsive and 173 MW-responsive miRNAs were detected by miRNA microarray. The expression patterns of six GA-responsive miRNAs were validated by qRT-PCR. Several target genes for these miRNAs encode various transcription factors, including ethylene-responsive transcription factors and squamosa promoter-binding-like proteins, which suggests that these miRNAs may regulate stress-responsive transcriptional processes in tea plant. The present findings provide novel insights into miRNA-mediated regulatory mechanisms underlying the response to GA stress, and also offer valuable information for development of pest resistance using RNA interference-based strategies in tea plants.
Highlights
Small non-protein-coding RNAs have emerged as key eukaryotic regulatory molecules, and are involved in diverse biological processes including transcription, oxidation-reduction, transport, and stress response
This study aims to identify the differential conserved and species-specific novel miRNAs responsive to insect feeding in tea plant by high-throughput small RNA sequencing; to achieve this, we constructed three small RNA libraries from tea plant leaves subjected to mechanical wounding (MW), geometrid attack (GA), and healthy control (CK) leaves
Each sRNA library was generated from an equal mixture of small RNAs obtained at different time points from GA, MW and CK tea leaves, respectively
Summary
Small non-protein-coding RNAs (sRNAs) have emerged as key eukaryotic regulatory molecules, and are involved in diverse biological processes including transcription, oxidation-reduction, transport, and stress response. Several conserved and novel miRNAs were identified in response to drought and cold stress in the tea plant by high-throughput sequencing[26,27,28]. This study aims to identify the differential conserved and species-specific novel miRNAs responsive to insect feeding in tea plant by high-throughput small RNA sequencing; to achieve this, we constructed three small RNA (sRNA) libraries from tea plant leaves subjected to mechanical wounding (MW), geometrid attack (GA), and healthy control (CK) leaves. The potential target genes of these miRNAs were involved in diverse biological processes including transcription, signal transduction, stress response, and plant growth and maintenance These results lay the foundation for understanding miRNA-based regulation in the response to the infestation of tea plants with E. oblique
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