sensitivity assay. (A) Determination on the endogenous ABA amounts. The imply values SD have been obtained from three biological repeats. Comparisons amongst two groups were performed using a Student’s t test. p 0.001. (B) ABA sensitivity assay. The germination phenotypes were classified into four phases: I, rupture on the seed coat; II, radicle protrusion; III, fully-opened cotyledons; and IV, correct leaf development.3.4. P1/HC-ProTu Triggers Immune Responses within a Calcium-Dependent Manner We identified that the genes in the P1/HC-ProTu -only section encode numerous Ca2+ transporters, like Ca2+ channels [CMCU (AT5G66650), CNGC2 (AT5G15410), and CNGC14 (AT2G24610)], Ca2+ co-transporter [CCX2 (AT5G17850)], and Ca2+ pump [ACA1 (AT1G27770)] (Figure 4A(panel i) and Table 3). We also identified a number of Ca2+ sensors, such as Ca2+ /calmodulin (CaM), CaM-like (CaML) (Figure 4A(panel ii)), calcium binding proteins (Figure 4A(panel iii)), calmodulin-binding proteins (Figure 4A(panel iv)), and CDPKs/ CIPKs (Figure 4A(panel v) and Table 3). The members of your various families of Ca2+ sensors identified in the P1/HC-ProTu -only section were anticipated to contribute for the con-Viruses 2021, 13,9 ofversion of Ca2+ signals into either cellular strain responses or developmental processes (Figure 4A(panel vi and vii)). For example, the expressions of genes encoded calmodulinbinding proteins, which includes CAMBP25 (AT2G41010) and IQM4 (AT2G26190), whose gene expressions are induced by drought and salt/osmotic pressure. CPK32 (AT3G57530) and CPK28 (AT5G66210) encode calcium-dependent Bax Inhibitor site protein kinases that regulate plant H2 Receptor Modulator review development in addition to resetting pathogen-associated molecular pattern (PAMP)-induced defense signaling. The majority of genes involved in the Ca2+ signaling pathway have been expressed at greater levels inside the P1/HC-ProTu plants than within the Col-0, P1Tu , and HC-ProTu plants (Figure 4B) except for CNGC2 (AT5G15410), which suggests that the induction from the calcium signaling pathway will depend on ectopic-expressing P1/HC-ProTu . In summary, the outcomes indicate that P1/HC-ProTu may trigger many strain responses and developmental processes through the calcium signaling pathway.Figure 4. Calcium signaling pathway in response to P1/HC-ProTu . (A) Calcium signaling pathway inside the P1/HC-ProTu -only section of the HTP profiles within the Col-0 vs. P1/HC-ProTu , Col-0 vs. P1Tu , and Col-0 vs. HC-ProTu comparison sets obtained in the HTP profiles. (B) Heatmap showing the expression patterns from the P1/HC-ProTu -only genes involved within the calcium signaling pathway. Table 3. List of genes within the calcium signaling pathway found within the P1/HC-ProTu -only section of your HTP profiles. Categories Subcategories AGI Locus Code AT1G27770 AT2G24610 Transporter Uniporter AT5G15410 AT5G17850 AT5G66650 AT1G66400 AT1G76650 AT2G43290 AT3G22930 AT5G37770 AT5G39670 Gene Name ACA1 CNGC14 CNGC2/DND1 CCX2 CMCU CML23 CML38 CML5/MSS3 CML11 CML24/TCH2 CML46 Annotations Autoinhibited Ca2+ -ATPase 1 Cyclic nucleotide-gated channel 14 Cyclic nucleotide-regulated ion channel family members protein Sodium/calcium exchanger household protein Protein of unknown function (DUF607) Calmodulin like 23 Calmodulin-like 38 Calcium-binding EF-hand family protein Calmodulin-like 11 EF hand calcium-binding protein household Calcium-binding EF-hand family members proteinSensorCaMs/CaMLsViruses 2021, 13,ten ofTable 3. Cont. Categories Subcategories AGI Locus Code AT3G01830 Other calcium-binding proteins AT3G16510 AT3G25600 AT4G34150 AT2