pressing the P1/HC-ProTu gene (P1/HCProTu plant) was previously performed by means of high-throughput (HTP) RNA sequencing (RNASeq) [1]. The transcriptomic profiles had been subsequently analyzed by means of a comparative network making use of the ContigViews system. The network highlighted many critical gene silencing components, which includes AGO1, AGO2, and AGO3, as well as many miRNA targets, calcium signaling components, hormone signaling components, and defense responserelated genes [1]. Hu et al. (2020) demonstrated that ethylene signaling genes inside the P1/HC-ProTu plants are substantially highlighted within the gene-to-gene network and that endogenous ethylene is also hugely accumulated inside the P1/HC-ProTu plants [1]. Moreover, Pasin et al. (2020) showed that the P1 (P1Pp ) of plum pox virus (PPV) triggers endogenous abscisic acid (ABA) accumulation in PPV-infected plants [5]. HTP RNA-Seq gives deep bioinformation; however, the abundant info obtained by RNA-Seq increases the analysis threshold for data mining and also the troubles in excluding the false-positive results generated with the low abundance gene profile. HTP RNA-Seq also has a greater price for the deep sequencing. One example is, the cost and sample determination for HTP RNA-Seq may possibly limit the experimental design and style of a preliminary transcriptome study. Here, we propose the use of low-throughput (LTP) RNA-Seq in a preliminary study. The LTP RNA-Seq profiles had been generated from P1/HC-ProTu -related transgenic plants and compared together with the connected P1/HC-ProTu -related profiles obtained previously by means of HTP RNA-Seq by Hu et al. [1]. Within this study, we performed the P1/HC-ProTu -related transcriptomic profiling making use of diverse logic analysis approaches to investigate the suppression mechanism further. We also performed LTP RNA-Seq of these P1/HC-ProTu -related components and compared the networks obtained in the LTP datasets and previously published HTP profiles. The outcomes indicate that LTP RNA-Seq has the prospective to decrease the sequencing budgets and exclude genes with low expressions, which may well yield a false-positive, and as a result, this approach could enable researchers swiftly CYP2 Activator drug identify significant pathways for ATM Inhibitor custom synthesis additional study. 2. Materials and Approaches two.1. Plant Supplies and Transgenic Plants Arabidopsis thaliana ecotype Col-0, three P1/HC-ProTu -related transgenic plants (P1Tu , HC-ProTu , and P1/HC-ProTu ), and ago1-27 mutant have been employed in this study [1,6]. The Arabidopsis seeds have been surface-sterilized, chilled at four C for two days, after which sown on Murashige and Skoog (MS) medium with/without appropriate antibiotics. All the plants had been grown at 24 C within a development room with 16 h of light/8 h of dark. 2.2. cDNA Library Building and RNA Sequencing Ten-day-old and 14-day-old seedlings in the wild-type Col-0, P1Tu , HC-ProTu , and P1/HC-ProTu plants were used for the collecting samples for HTP and LTP wholetranscriptome deep sequencing, respectively. Three biological replicates of each of the wild-type Col-0, P1Tu , HC-ProTu , and P1/HC-ProTu samples have been integrated in this study, and every biological replicate consisted of 250 seedlings. Total RNA was extracted in the seedlings working with a silica-gel membrane method (Viogene, New Taipei City, Taiwan). The mRNAs for LTP sequencing were isolated applying the poly(A) mRNA magnetic isolation module (New England Biolabs, San Diego, CA, USA). All RNA sequencing libraries were constructed by using the cDNA library kit (Invitrogen Thermo Fisher Scientific, Waltham,