Abstract
Horsegram (Macrotyloma uniflorum (Lam.) Verdc.) is a drought hardy food and fodder legume of Indo-African continents with diverse germplasm sources demonstrating alternating mechanisms depicting contrasting adaptations to different climatic zones. Tissue specific expression of genes contributes substantially to location specific adaptations. Regulatory networks of such adaptive genes are elucidated for downstream translational research. MicroRNAs are small endogenous regulatory RNAs which alters the gene expression profiles at a particular time and type of tissue. Identification of such small regulatory RNAs in low moisture stress hardy crops can help in cross species transfer and validation confirming stress tolerance ability. This study outlined prediction of conserved miRNAs from transcriptome shotgun assembled sequences and EST sequences of horsegram. We could validate eight out of 15 of the identified miRNAs to demonstrate their role in deficit moisture stress tolerance mechanism of horsegram variety Paiyur1 with their target networks. The putative mumiRs were related to other food legumes indicating the presence of gene regulatory networks. Differential miRNA expression among drought specific tissues indicted the probable energy conservation mechanism. Targets were identified for functional characterization and regulatory network was constructed to find out the probable pathways of post-transcriptional regulation. The functional network revealed mechanism of biotic and abiotic stress tolerance, energy conservation and photoperiod responsiveness.
Highlights
Investigation over decades has enriched the miRNA’s knowhow in biogenesis and explicit regulatory machinery
Significant miRNA homologs within reported 8496 miRNAs were identified by executing nucleotide blast (BLASTn) with 27,997 Transcriptome Shotgun Assembly (TSA) contigs, SSH-Mu library sequences of moisture stressed horsegram cDNA (NCBI dbEST ID 75866463) and 1008 Expressed Sequence Tags (EST) sequences as query resulted in 16 ESTs and 6303 TSA hits (E value 0.1) for further analysis
The predicted miRNAs were analyzed for various structural features to distinguish from other small RNAs such as tRNAs, rRNAs and mRNAs (Table 1 and Fig. 2)
Summary
Investigation over decades has enriched the miRNA’s knowhow in biogenesis and explicit regulatory machinery. Being a diploid (2n = 20, 22 or 24], short duration (120–180 days to maturity) plant species and adapted to grow on wide range of agro-climatic conditions, horsegram can be weighed as appropriate model for moisture stress tolerance genes/QTLs investigation challenging undernourishment in drought prone r egions[6,7]. It can be envisaged as nutraceuticals, forage c rop[8] and anti-calcifying inhibitors[9]. We report differential expression of identified miRs through a stringent in-silico schema elucidating conserved miRNAs, their characterization, validation with their target annotation and networking
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