MicroRNAs (miRNAs) are noncoding RNAs, ~21–24 nucleotides in length, that play a pivotal role in post-transcriptional gene regulation by inducing cleavage or translational repression of target mRNAs with complementary sequences. In this study, we identified miRNAs expressed during the early developmental stage of mung bean (Vigna radiata), a major legume crop, using small RNA sequencing (sRNA-seq), and analyzed their expression profiles across various mung bean tissues. Mung bean-specific miRNAs were found to be highly expressed in the aerial parts of seedlings, particularly in the leaves. Furthermore, the expression of these miRNAs was effectively validated using Tailed-Hoogsteen triplex DNA-encapsulated silver nanocluster (DNA/AgNC) sensors. The nanosensor enables rapid detection of target miRNAs within 30 min and is easy to apply for field-based assessments. The predicted target mRNAs of the identified miRNAs were associated with a range of biological processes relevant to early-stage development. This study highlights the potential of nanosensor-based approaches for the efficient identification of novel miRNAs in staple crops, offering a promising strategy to reduce the cost, time, and labor required during the transition from laboratory research to field applications.

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