Chinese Wild-Growing Vitis amurensis ICE1 and ICE2 Encode MYC-Type bHLH Transcription Activators that Regulate Cold Tolerance in Arabidopsis
Figure 2
Phylogenetic tree, alignment and expression profiles of VaICE1,2.
(A) Phylogenetic tree based on the deduced amino acid sequences of ICE from a range of plant species. A neighbor-joining tree, with the following predicted ICE protein sequences, with GenBank accession numbers listed in parentheses: V. amurensis (VaICE1, AGP04217; VaICE2, AGP04218; VaICE14, ADY17816), V. vinifera (VvICE1, AFI49627; VvICE1a, AGQ03810; VvICE1b, AGQ03811), V. riparia (VrICE1, AGG34704; VrICE2, AIA58705; VrICE3, AIA58706; VrICE4, AIA58707), Arabidopsis thaliana (AtICE1, NP_189309; AtICE2, NP_172746), Brassica napus (BnICE1, AEL33687), Brassica rapa subsp. Chinensis (BrICE1, ACB70963), Capsella bursa-pastoris (CbICE1, AAS79350), Camellia sinensis (CsICE, ACT90640), Eucalyptus camaldulensis (EcICE1,ADY68776), Eucalyptus globules (EgICE1, AEF33833), Eutrema salsugineum (EsICE, ACT68317), Glycine max (GmICE1, ACJ39211), Hordeum vulgare (HvICE2, ABA25896), Malus x domestica (MdbHLH1, ABS50251), Oryza sativa (OsICE1, Os11g0523700; OsICE2, Os01g0928000), Populus suaveolens (PsICE1, ABF48720), Populus trichocarpa (PtrICE1, ABN58427), Raphanus sativus (RsICE1, ADY68771), Triticum aestivum (TaICE41, ACB69501; TaICE87, ACB69502), Zea mays (ZmICE2, ACG46593), was produced by ClustalX 2.0 alignment followed by tree construction using MEGA 5.0 with 100 bootstrap tests. The branch support values are indicated. The length of the scale bar corresponds to 5 substitutions per site. (B) Comparison of ICE amino acid sequences from V. amurensis (VaICE1,2) and Arabidopsis thaliana (AtICE1,2). Deduced amino acid sequences were aligned using ClustalW. Sequences and accession numbers are shown for the following: Vitis amurensis (VaICE1, KC815984; VaICE2, KC815985) and A. thaliana (AtICE1, AAP14668; AtICE2, AAO63441). Residues in black and gray regions indicate identical and similar residues, respectively, between isoforms. Four predicted domains are labeled: a S-rich motif, a basic-helix-loop-helix-leucine zipper (bHLH-ZIP) region, ICE-specific domain [38] and a ACT-UUR-ACR-like domain. The red triangles indicate the position of the mutation isolated by Chinnusamy et al. [32] and Kanaoka et al. [54], and the residue targeted for sumoylation by SIZ1[53]. Green triangle indicates E-box/N-box specificity site [69], blue asterisks indicate core residues for DNA binding sites [70], and red diamonds indicate dimerization interface/polypeptide binding sites [70]. (C) Expression profiles of VaICE1 and VaICE2 during a cold stress time-course experiment. Semi-quantitative RT-PCR was used to determine VaICE1 and VaICE2 transcript levels in cold treated grapevine leaves at indicted times. Grapevine GAPDH was used as a loading control.