.-J.; Lin, S.-S. Investigation of P1/HC-Pro-Mediated ABA/Calcium Signaling Responses through Gene Silencing through High- and Low-Throughput RNA-seq Approaches. Viruses 2021, 13, 2349. doi.org/10.3390/v13122349 Academic Editor: Yau-Heiu Hsu Received: 4 August 2021 Accepted: 19 November 2021 Published: 23 NovemberAbstract: The P1/HC-Pro viral suppressor of potyvirus suppresses posttranscriptional gene silencing (PTGS). The fusion protein of P1/HC-Pro can be cleaved into P1 and HC-Pro by way of the P1 self-cleavage activity, and P1 is required and enough to improve PTGS suppression of HCPro. To address the modulation of gene regulatory relationships induced by turnip mosaic virus (TuMV) P1/HC-Pro (P1/HC-ProTu ), a comparative transcriptome analysis of 3 types of transgenic plants (P1Tu , HC-ProTu , and P1/HC-ProTu ) had been carried out using both high-throughput (HTP) and low-throughput (LTP) RNA-Seq methods. The outcomes showed that P1/HC-ProTu disturbed the endogenous abscisic acid (ABA) accumulation and genes in the signaling pathway. Moreover, the integrated responses of stress-related genes, in distinct to drought stress, cold tension, senescence, and stomatal dynamics, altered the expressions by the ABA/calcium signaling. Crosstalk amongst the ABA, jasmonic acid, and salicylic acid CD30 Inhibitor custom synthesis pathways might simultaneously modulate the anxiety responses triggered by P1/HC-ProTu . Furthermore, the LTP network evaluation revealed important genes in frequent with these identified by the HTP network in this study, demonstrating the effectiveness of the miniaturization in the HTP profile. Overall, our Bcl-2 Modulator Synonyms findings indicate that P1/HC-ProTu -mediated suppression in RNA silencing altered the ABA/calcium signaling as well as a wide range of tension responses. Keywords and phrases: ABA signaling; calcium signaling; HTP-Seq; LTP-Seq; P1/HC-ProTu ; anxiety responsePublisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.1. Introduction P1/HC-Pro would be the initially identified viral suppressor of potyvirus and can trigger the suppression of RNA silencing within the microRNA (miRNA) and short-interfering RNA (siRNA) regulatory pathways [1]. Our prior research indicate that the FRNK motif of HC-Pro plays an essential part within the suppression on the miRNA pathway but still suppresses 40 from the siRNA pathway [2]. In addition, Hu et al. (2020) demonstrated that many potyviral species of P1/HC-Pro, i.e., turnip mosaic virus (TuMV), zucchini yellow mosaic virus (ZYMV), and tobacco etch virus (TEV), possess the exact same function in RNA silencing suppression [1]. However, the P1/HC-Pro of TuMV (P1/HC-ProTu ) triggers ARGONAUTE1 (AGO1) degradation, whereas those of ZYMV (P1/HC-ProZy ) and TEV (P1/HC-ProTe ) do not trigger AGO1 degradation, which suggests that viral P1/HC-Pros exhibit functional diversity. Additionally, Sanobar et al. (2021) demonstrated that HC-ProTuCopyright: 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access write-up distributed under the terms and conditions in the Creative Commons Attribution (CC BY) license ( creativecommons.org/licenses/by/ 4.0/).Viruses 2021, 13, 2349. doi.org/10.3390/vmdpi/journal/virusesViruses 2021, 13,2 ofinhibits HEN1 activity in miRNA three -end 2 -O-methylation in vitro and in vivo by means of the binding activity of HC-ProTu FRNK motif with HEN1 [4]. To know P1/HC-Pro-mediated RNA silencing suppression further, a transcriptomic analysis according to transgenic Arabidopsis ex