Presentation on theme: "Table S1 (Ho et al.) Table S1 Primers and their sequences used in this study. Name Sequence CDPK1-F1 5-CTACAGATCTATGGGGAATCAGTGCCA-3' CDPK1-R15'-TCATAGATCTCTAAACTCCATTTTCACAGAT-3'"— Presentation transcript:
Table S1 (Ho et al.) Table S1 Primers and their sequences used in this study. Name Sequence CDPK1-F1 5-CTACAGATCTATGGGGAATCAGTGCCA-3' CDPK1-R15'-TCATAGATCTCTAAACTCCATTTTCACAGAT-3' CDPK1-F25'-TCTAGGGTACCGGATCCGATCGAGCGCGCGAGG-3' CDPK1-R25'-CTCAACTGCAGATCCAACCCCCGCCTGAG-3' GFP-F35'-TCTAGCTGCAGGTGAGCAAGGGCGAG-3' GFP-R3.15'-TCTAGGTCGACTTACTTGTACAGCTCGTC-3' GFP-R3.25'-TCTAGGTCGACACGCTGAACTTGTGGC-3' GF14c-F45'-TAAGGATCCATGTCTCGGGAGGAGAAT-3' GF14c-R45'-TCAGGATCCTTACTGGCCCTCGCAGGCGT-3' GF14c-F55'-TCTAGGGTACCGGATCCACGAGACCACTCGAACCCGACCCGCCTC-3' GF14c-R55'-ACCTTAGATTCTGCAGCAGTAGATGAG-3' CDPK1-F6 5-ATGGGGAATCAGTGCCAGAAC-3 CDPK1-R6 5-AAGTTGTGCCAAACTGGCCCT-3 Gene accession number:OsCDPK1 (AY158077),GFP (U43284),OsGF14c (U65957). Restriction enzyme recognition sites are underlined..
Fig. S1 Structure and amino acids sequence of OsCDPK1. Subdomains conserved in protein kinases are indicated by Roman numerals and underlined. Four EF-hand structures of calcium binding regions are indicated by dash underlines. Double dash underline indicates autoinhibitory regions of kinase activity. Green and red letters represent changes of amino acid residue from OsCDPK1 to OsCDPK12 and to OsCDPK13, respectively. Fig. S1 (Ho et al.)
Fig. S2 Expression of OsCDPK1 in T 4 transgenic rice. Total RNA was isolated from 14- day-old seedlings and subjected to Northern blot analysis using the OsCDPK1- specific probe. Ethidium bromide staining detected 25S and 18S rRNA bands. Lane 1, OsCDPK1 knockdown transgenic line (Ri-1); lane 2, OsCDPK1(tr) overexpressing transgenic line (OEtr-1); lane 3, wild-type (Wt). Fig. S2 (Ho et al.)
Fig. S3 Comparison of plant height between T 1 generation of OsCDPK1(tr) (8 independent lines) and OsCDPK1(Ri) (5 independent lines) and wild type (Wt) plants. (A)The independent T 1 transgenic plants were corresponding to the transformed cell lines as showed in Figure 2, respectively, were grown in the isolated field for 60 days. (B) The T 1 transgenic plants of OEtr-1 and Ri-1, and wild type plants were planted in an another isolated field and grown for 60 days. Wt Ri- 1, 2, 3, 4, 5 OEtr- 1, 3, 4, 6, 7, 8, 9,15 Ri-1 Ri-2 Ri-3 Ri-4 Ri-5 OEtr-1 OEtr-3 OEtr-4 OEtr-6 OEtr-7 OEtr-8 OEtr-9 OEtr-15 Fig. S3 (Ho et al.) B Wt Ri-1 OEtr-1 60 days A
A Wt OEtr-1 Ri-1 Length (cm) days +GA B Wt OEtr-1 Ri-1 Fig. S4 The semi-dwarf phenotype of the OEtr-1 line was recovered by GA 3 treatment. Comparison of plant height between T 4 generation of OEtr-1 and Ri-1 and wild type (Wt) plants. Seeds were germinating on medium containing1/2 MS salt and supplemented with (A and B) or without (C and D) 10 μM GA 3 for 10 days. Fig. S4 (Ho et al.) C Wt OEtr-1 Ri-1 Length (cm) days days -GA 3 D Wt OEtr-1 Ri-1 Seedlings length (cm)
OsCDPK1 GA20ox1 GA20ox2 GA3ox2 Act1 Ri-1 OEtr-1 Wt Fig. S5 The expression of GA biosynthesis-related genes in OEtr-1 and Ri-1 plants. Total RNA was isolated from 14-day-old seedlings and subjected to RT-PCR analysis using primers specific for OsCDPK1, GA20ox1, GA20ox2, and GA3ox2; the rice Act1 gene was used as an internal control. Fig. S5 (Ho et al.)
Fig. S6 Drought tolerance in transgenic rice is enhanced by overexpression of the protein GF14c. Rice seedlings were grown on 1/2 MS agar medium for 14 days, transferred to the pot and grown for another 7 days, withheld water for 5 days, and re-watered for 4 days. GF14c-OxWtGF14c-Ri Fig. S6 (Ho et al.)