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. 2021 Jan 28;22:90. doi: 10.1186/s12864-021-07396-6

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© The Author(s) 2021

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 5 — Characteristics of RcDREB2B. a Phylogenetic tree of RcDREB2B and other plant DREB proteins. The phylogenetic tree was constructed using Evolview and MEGA 7.0 with bootstrap values of 1000 replicates. The accession numbers of the protein sequences that have been retrieved from NCBI and species designations are listed below: Arabidopsis thaliana: AtDREB2A (AB007790), AtDREB2B (NM_111939.2), AtABI4(A0MES8), AtTINY2 (AY940160.1), AtTINY(Q39127), AtRAP2.1 (Q8LC30), AtRAP2.9 (NM_179009.1), AtRAP2 (AAP04063.1); Oryza sativa: OsDREB1A (AF300970), OsDREB3 (NP_001048142); Glycine max: GmDREB2 (ABB36645), GmDREBb (AAQ57226); Gossypium hirsutum: GhDBP1(AAO43165.1); Zea mays: ZmDREB1A (AF450481), ZmDREB2A (NP_001105876), ZmABI4 (AY125490), ZmDBF2 (AF493799); Hordeum vulgare: HvDRF1 (AY223807); Triticum aestivum: TaDREB1 (DQ195068), TaCBF1 (AF376136); Secale cereal: ScCBF1 (AF370730); Brassica napus: BnCBF17 (AF499034); Morus notabilis: MnDREB4D (AHJ25980.1). Different color lines indicate different subgroups of the DREB subfamily. b Transcript levels of RcDREB2B. The expression levels of RcDREB2B in leaves and roots during no drought (ND), mild drought (MD), and severe drought (SD) treatments. The mean fold changes of RcDREB2B in MD and SD were compared with ND. We present results as means ± SD. n = 3. Different letters denote significant differences at P < 0.05 with one-way ANOVA analysis. c Subcellular localization of RcDREB2B in Arabidopsis protoplast cells. RcDREB2B-GFP fusion protein or GFP alone is expressed under the control of pCAMBIA 1300 in Arabidopsis protoplast cells. The photographs were taken in the green channel, bright channel, and merge channel. Bars = 20 μm. d Transcriptional activity analysis of RcDREB2B. The full-length ORF of RcDREB2B was fused to the GAL4 DNA-binding domain in the vector pGBKT7 to generate pGBKT7-RcDREB2B, and the construct was converted into yeast strain Y2HGold. Yeast dilutions were grown on SD medium lacking Trp, and the same medium containing with or without X-gal but lacking with Trp, His, Ade. pGBKT7 and pGAL4 plasmids were respectively used as negative and positive control