Table 1.
Key differentially expressed genes in HvNAT2-OX and GP under the control and Cd.
| Gene ID |
Fold change (OX vs GP) |
Predicted molecular function | |||
|---|---|---|---|---|---|
|
RNA-seq |
qRT-PCR |
||||
| Control | Cd | Control | Cd | ||
| HORVU3Hr1G073180 | 4.19 | 4.54 | 5.81 | 6.44 | Nucleobase-ascorbate transporter 2 (NAT2) contributes to Cd tolerance in barley in this study |
| HORVU1Hr1G057630 | −0.18 | 1.38 | −0.07 | 1.55 | CSC1-like protein (ERD4) for hyperosmolarity-gated non-selective cation channel that permeates Ca2+ and potentially Cd2+ ions |
| HORVU0Hr1G020790 | −0.31 | 1.04 | 0.33 | 1.05 | Calcium-transporting ATPase 9 (ACA9) permeates Ca2+ and potentially Cd2+ ions |
| HORVU7Hr1G097210 | −0.14 | 1.00 | −0.33 | 1.20 | Cadmium/zinc-transporting ATPase 3 (HMA3) for Cd homeostasis |
| HORVU1Hr1G043470 | 3.56 | 2.91 | 8.29 | 7.56 | Glutathione S-transferase T3 (GST3) for oxidative stress response |
| HORVU2Hr1G018440 | −0.39 | 1.14 | −0.02 | 1.10 | Glutathione Peroxidase 2 (GPX2) protects against oxidative stresses |
| HORVU2Hr1G044360 | −0.3 | 1.03 | −0.07 | 1.36 | Glutathione Peroxidase 2 (GPX2) protects against oxidative stresses |
| HORVU3Hr1G024510 | −0.19 | 1.29 | −0.33 | 1.19 | Glutathione Peroxidase 25 (GPX25) protects against oxidative stresses |
| HORVU6Hr1G090560 | −0.09 | 1.29 | −0.66 | 1.22 | Glutathione S-transferase isoform X2 (GSTU6) for toxin catabolic process |
| HORVU3Hr1G099080 | −0.45 | 1.14 | −0.82 | 1.38 | Glutaredoxin-C2 (GRXC6) for cellular response to oxidative stress |
| HORVU7Hr1G087250 | −0.53 | 1.39 | 0.41 | 1.04 | L-ascorbate oxidase-like (AAO) in redox system of oxidative stress |
| HORVU6Hr1G026820 | 3.85 | 3.63 | 2.58 | 2.34 | Thioredoxin-like protein (Trx) participates as a hydrogen donor in redox reactions |
| HORVU6Hr1G065430 | 0.99 | 1.67 | 0.25 | 1.81 | Ethylene-responsive transcription factor 27-like (ERF027) acts as the components of stress signal transduction pathways |
| HORVU2Hr1G004200 | −0.34 | 1.6 | −0.51 | 1.59 | 1-aminocyclopropane-1-carboxylate oxidase homolog 1-like (ACO1) for Ethylene biosynthesis, Plant defense |
| HORVU2Hr1G108180 | 0.19 | 1.19 | 0.22 | 1.39 | Anthocyanidin reductase ((2S)-flavan-3-ol-forming)-like isoform X2 (ANRX2) with oxidoreductase activity |
| HORVU0Hr1G005420 | 1.36 | −1.03 | 1.24 | −0.68 | Alternative oxidase (AOX) controls the synthesis of ROS and NO |
| HORVU2Hr1G101990 | 1.83 | −0.88 | 1.37 | −0.70 | Alternative oxidase 1a (AOX1a) controls the synthesis of ROS and NO |
| BGI_novel_G000483 | 2.1 | −1.93 | 2.11 | −1.15 | Phenylalanine ammonia-lyase (PAL) in response to oxidative stress |
| HORVU5Hr1G093700 | 0.6 | 1.08 | −0.68 | 1.07 | Linoleate 9S-lipoxygenase 3 (LOX3) in the pathway of fatty acid metabolism and in Lipid metabolism |
| HORVU4Hr1G066270 | 0.49 | 1.16 | 0.25 | 1.59 | Allene oxide synthase 2 (AOS3) in the pathway of fatty acid metabolism and in lipid metabolism |
| HORVU5Hr1G059310 | 4.16 | 3.53 | 7.5 | 6.75 | Glutathione gamma-glutamylcysteinyltransferase 1 (PCS1) for phytochelatins and homophytochelatins for heavy metal binding |
| HORVU6Hr1G036920 | 0.31 | 1.39 | 0.21 | 1.25 | Cytokinin-O-glucosyltransferase 3 for cellular detoxification |
| HORVU7Hr1G113830 | 0.79 | 1.04 | 0.15 | 2.09 | WRKY transcription factor 2 in the pathway of MAPK signaling |
| HORVU7Hr1G057410 | −0.75 | 1.09 | −0.43 | 1.27 | R2R3 MYB transcriptional factor (MYB) may involve in stress regulation |
| HORVU2Hr1G010030 | 2.65 | 2.41 | 3.12 | 1.41 | Arginine decarboxylase 2 (ADC2) in the pathway of arginine metabolism |
| HORVU4Hr1G085450 | 4.67 | 5.04 | 7.31 | 8.56 | MADS-box transcription factor 51 (MADS51) may involve in stress regulation |
List of redox related genes in ROS regulation, and signaling, transport related genes differentially expressed in HvNAT2-OX vs GP plants. Fold change (OX vs GP) is log2N, log2N ≥ 1 are up-regulated, between 0 < |log2N| < 1 are unchanged and log2N ≤ -1 are down-regulated, Q-value ≤ 0.001. The Predicted Molecular Function is obtained from https://www.uniprot.org/. The qRT-PCR of genes: an additional hydroponic experiment was carried out again using GP and HvNAT2-OX lines under control and 10 μM Cd treatment with three replicates. Total RNA was isolated from leaves of barley plants after 24 h of Cd treatment using TRIzol reagent (Invitrogen, Karlsruhe, Germany). qRT–PCR reaction was performed on LightCycler 480 System (Roche, Germany). All primers used are listed in Tables S2. GAPDH was used as internal control.