Identification of miRNAs associated with Aspergillus flavus infection and their targets in groundnut (Arachis hypogaea L.).

Abstract

Background: The quality of groundnut produce is adversely impacted due to aflatoxin contamination by the fungus Aspergillus flavus. Although the transcriptomic control is not fully understood, the interaction between long non-coding RNAs and microRNAs in regulating A. flavus and aflatoxin contamination remains unclear. This study was carried out to identify microRNAs (miRNAs) to enhance the understanding of in vitro seed colonization (IVSC) resistance mechanism in groundnut.

Result: In this study, resistant (J 11) and susceptible (JL 24) varieties of groundnut were treated with toxigenic A. flavus (strain AF-11-4), and total RNA was extracted at 1 day after inoculation (1 DAI), 2 DAI, 3 DAI and 7 DAI. Seeds of JL 24 showed higher mycelial growth than J 11 at successive days after inoculation. A total of 208 known miRNAs belonging to 36 miRNA families, with length varying from 20-24 nucleotides, were identified, along with 27 novel miRNAs, with length varying from 20-22 nucleotides. Using psRNATarget server, 952 targets were identified for all the miRNAs. The targeted genes function as disease resistant proteins encoding, auxin responsive proteins, squamosa promoter binding like proteins, transcription factors, pentatricopeptide repeat-containing proteins and growth regulating factors. Through differential expression analysis, seven miRNAs (aly-miR156d-3p, csi-miR1515a, gma-miR396e, mtr-miR2118, novo-miR-n27, ptc-miR482d-3p and ppe-miR396a) were found common among 1 DAI, 2 DAI, 3 DAI and 7 DAI in J 11, whereas ten miRNAs (csi-miR159a-5p, csi-miR164a-3p, novo-miR-n17, novo-miR-n2, osa-miR162b, mtr-miR2118, ptc-miR482d-3p, ptc-miR167f-3p, stu-miR319-3p and zma-miR396b-3p) were found common among 1 DAI, 2 DAI, 3 DAI and 7 DAI in JL 24. Two miRNAs, ptc-miR482d-3p and mtr-miR2118, showed contrasting expression at different time intervals between J 11 and JL 24. These two miRNAs were found to target those genes with NBS-LRR function, making them potential candidates for marker development in groundnut breeding programs aimed at enhancing resistance against A. flavus infection.

Conclusion: This study enhances our understanding of the involvement of two miRNAs namely, ptc-miR482d-3p and mtr-miR2118, along with their NBS-LRR targets, in conferring resistance against A. flavus-induced aflatoxin contamination in groundnut under in vitro conditions.