Electronic supplementary material

Supplementary Table S1. Primers used for amplification of Vitis amurensis cDNAs in PCR and real-time PCRs

Application / Primer, 5'-3'
Full-length cDNA of VaCPK1 (KC488321) / VaCPK1F
5'ATG GGG AAT AAC TGT GTG GGA;
VaCPK1R
5'TCA ATA GAC TGG TAG CGG CT
Full-length cDNA of VaCPK26 (KC488323) / VaCPK26F
5'ATG GGC AAC ACA TGC CGG GGA
VaCPK26R
5'CTA AAA AGC TCC TGG TGC ATC TC
Transfer of VaCPK1 cDNA to the pSAT1 vector / VaCPK1-pSAT-F
5'GCT CAG ATC TAT GGG GAA TAA CTG TGT GGG A;
VaCPK1-pSAT-R
5'TCG AGG TAC CTC AAT AGA CTG GTA GCG GCT
Transfer of VaCPK26 cDNA to the pSAT1 vector / VaCPK26-pSAT-F
5'GCT CGA GCT CAT GGG CAA CAC ATG CCG GGG A
VaCPK26-pSAT-R
5'TCG AGG TAC CCT AAA AAG CTC CTG GTG CAT CTC
Analyzing the total VaCPK1 expression / VaCPK1-S
5'CAA TGA TGG CCG CAT AGA TTA T;
VaCPK1-A1
5'TCA ATA GAC TGG TAG CGG CT
Analyzing the exogenous VaCPK1 expression / VaCPK1-S;
pSAT-A
5'GAG AGA CTG GTG ATT TTT GCG
Analyzing the endogenous VaCPK1 expression / VaCPK1-S;
VaCPK1-A3
5'GCT CTT GGC TGA GTG ACT AT
Analyzing the total VaCPK26 expression / VaCPK26-S
5'TAT GAT GCA AAA GGG CAA TGC;
VaCPK26-A1
5'CTC CTG GTG CAT CTC TCA A
Analyzing the exogenous VaCPK26 expression / VaCPK26-S;
pSAT-A
Analyzing the endogenous VaCPK26 expression / VaCPK26-S;
VaCPK26-A3
5'CGA TTC TCC TTT CTA GCT CA

The forward VaCPK1-pSAT-F primer contained a BglII restriction site and the reverse VaCPK1-pSAT-R primer contained a KpnI restriction site, which are underlined. The forward VaCPK26-pSAT-F primer contained a SacI restriction site and the reverse VaCPK26-pSAT-R primer contained a KpnI restriction site.


Table S2. Comparison of the deduced amino acid sequences of VaCPK1, 20, 26, and 3a genes and some known CDPKs from Arabidopsis thaliana (AtCPK1), Theobroma cacao (TcCDPK1), and Citrus clementina (CcCDPK1)

VaCPK1
(KC488321) / VaCPK20
(KC488322) / VaCPK26
(KC488323) / VaCDPK3a
(JQ793892) / AtCPK1
(L14771) / TcCDPK1
(EOY19314) / CcCDPK1
(XP_006422398)
VaCPK1 / - / 69 (81) / 63 (75) / 57 (68) / 70 (78) / 77 (85) / 64 (76)
VaCPK20 / 69 (81) / - / 62 (73) / 56 (66) / 70 (79) / 73 (83) / 63 (73)
VaCPK26 / 63 (75) / 62 (73) / - / 62 (72) / 61 (71) / 66 (77) / 90 (94)
VaCDPK3a / 57 (68) / 56 (66) / 62 (72) / - / 54 (64) / 59 (69) / 60 (72)

The amino acid sequence identities and positives in brackets (in %) were determined using the NCBI BLAST program (http://blast.nbi.nlm.nih.gov) using the algorithm blastp (protein-protein BLAST).


Fig. S1. Comparison of the deduced amino acid sequences of the VaCPK1, 20, 26, and 3a genes. Dashes indicate absent residues. Dark grey shading indicates identical amino acid residues. Grey shading indicates similar amino acid residues. I-IX - catalytic subdomains of the CDPK kinase domain. The junction (autoinhibitory) domain and the calmodulin-like domain containing the putative EF-hands are indicated in the figure