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. 2023 Aug 29;14(9):1727.
doi: 10.3390/genes14091727.

mRNA-Seq and miRNA-Seq Analyses Provide Insights into the Mechanism of Pinellia ternata Bulbil Initiation Induced by Phytohormones

Wenxin Xu  1 , Haoyu Fan  1 , Xiaomin Pei  1 , Xuejun Hua  1 , Tao Xu  1 , Qiuling He  2
Affiliations

mRNA-Seq and miRNA-Seq Analyses Provide Insights into the Mechanism of Pinellia ternata Bulbil Initiation Induced by Phytohormones

Wenxin Xu et al. Genes (Basel). .

Abstract

Pinellia ternata (Thunb.) Breit (abbreviated as P. ternata) is a plant with an important medicinal value whose yield is restricted by many factors, such as low reproductive efficiency and continuous cropping obstacles. As an essential breeding material for P. ternata growth and production, the bulbils have significant advantages such as a high survival rate and short breeding cycles. However, the location effect, influencing factors, and molecular mechanism of bulbil occurrence and formation have not been fully explored. In this study, exogenously applied phytohormones were used to induce in vitro petiole of P. ternata to produce bulbil structure. Transcriptome sequencing of mRNA and miRNA were performed in the induced petiole (TCp) and the induced bulbil (TCb). Gene Ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed for the identification of key genes and pathways involved in bulbil development. A total of 58,019 differentially expressed genes (DEGs) were identified. The GO and KEGG analysis indicated that DEGs were mainly enriched in plant hormone signal transduction and the starch and sucrose metabolism pathway. The expression profiles of miR167a, miR171a, and miR156a during bulbil induction were verified by qRT-PCR, indicating that these three miRNAs and their target genes may be involved in the process of bulbil induction and play an important role. However, further molecular biological experiments are required to confirm the functions of the identified bulbil development-related miRNAs and targets.

Keywords: P. ternata; RNA-seq; bulbil; induction; microRNA; phytohormone.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
The process of bulbil induction in P. ternata. 3 days (A), 6 days (B), 9 days (C), and 12 days (D) show the early stage of bulbil induction, and 36 days (E) and 108 days (F) show the mature induced bulbil.
Figure 2
Figure 2
The GO enrichment analysis of TCb and TCp differentially expressed genes (DEGs). The top 20 GO terms with significant enrichment; rich factor indicates the enrichment degree of differential genes, and the higher the value, the higher the enrichment degree.
Figure 3
Figure 3
The KEGG pathway enrichment analysis of TCb and TCp differentially expressed genes (DEGs). The top 20 KEGG pathways with significant enrichment; rich factor indicates the enrichment degree of differential genes, and the higher the value, the higher the enrichment degree.
Figure 4
Figure 4
The KEGG pathways enrichment analysis target genes of differentially expressed miRNAs (DEmiRNAs). The top 20 KEGG pathways with significant enrichment; rich factor indicates the enrichment degree of DEmiRNAs, and the higher the value, the higher the enrichment degree.
Figure 5
Figure 5
Verification of expression profiles of key genes, miRNAs, and their target genes via qRT-PCR. P0 (red) was an uninduced petiole and served as a blank control, P1 = 3 d, P2 = 6 d, P3 = 9 d, P4 = 12 d. There was no significant difference with the same letter (p > 0.05), while there was a significant difference without the same letter (p < 0.05).
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