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Acta Prataculturae Sinica ›› 2022, Vol. 31 ›› Issue (1): 164-180.DOI: 10.11686/cyxb2020501

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Identification and functional analysis of lncRNA167 and its cleavage product miR167c in Medicago truncatula

Jia-ju ZHANG(), Jie YU, Ming-na LI, Jun-mei KANG, Qing-chuan YANG, Rui-cai LONG()   

  1. Institute of Animal Science,the Chinese Academy of Agricultural Sciences,Beijing 100193,China
  • Received:2020-11-10 Revised:2021-01-04 Online:2021-12-01 Published:2021-12-01
  • Contact: Rui-cai LONG


Long non-coding RNA (lncRNA) is a class of functional RNA molecules with a length of more than 200 nucleotides, which plays important roles in various biological regulation processes and response to biological and abiotic stresses. lncRNAs often interacts with microRNAs (miRNAs). In this study, a salt stress induced lncRNA (Mt-lncRNA167) obtained by high-throughput sequencing was identified to be a precursor of miRNA. The chromosome location of Mt-lncRNA167 coincided with that of miR167c, a member of miR167 family in Medicago truncatula. Mt-lncRNA167 was predicted to play a role as a cleavage precursor of Mt-miR167c. The online software analysis showed that auxin response factor 6/8ARF6/8) were two target genes of Mt-miR167c. The expression pattern analysis showed that Mt-lncRNA167 mainly expressed in leaves. Mt-lncRNA167 and Mt-miR167c responded to 100 mmol·L-1 NaCl, 5% PEG 6000, and 4 ℃ cold stress, which was consistent with the prediction of their promoter cis-acting elements. Under salt, drought and low temperature stresses, the expression patterns of Mt-lncRNA167 and Mt-miR167c were almost the same, which were contrary to the expression patterns of target genes ARF6/8. Mt-lncRNA167 and Mt-miR167c overexpressing Arabidopsis thaliana showed fertility defect phenotypes, such as short fruit pod and filament, anther deformity, and low pollen activity. The results of germination rate under salt and drought stress showed that the resistance of Mt-lncRNA167 and Mt-miR167c overexpressing A. thaliana plants was better than that of wild type plants. In addition, the hypocotyls of Mt-lncRNA167 and Mt-miR167c overexpressing A. thaliana plants were significantly longer than that of the wild type plants. In conclusion, Mt-lncRNA167 was predicted to be the precursor of Mt-miR167c, while Mt-miR167c inhibited the expression of ARF6/8. The regulatory network of Mt-lncRNA167-Mt-miR167c-ARF6/8 may play an important role in stress tolerance and developmental regulation of M. truncatula.

Key words: alfalfa, long non-coding RNAs, microRNAs, abiotic stress, growth and development