Table S1 (Ho et al.)

Table S1 Primers and their sequences used in this study.

Name Sequence

CDPK1-F1 5’-CTACAGATCTATGGGGAATCAGTGCCA-3'

CDPK1-R1 5'-TCATAGATCTCTAAACTCCATTTTCACAGAT-3'

CDPK1-F2 5'-TCTAGGGTACCGGATCCGATCGAGCGCGCGAGG-3'

CDPK1-R2 5'-CTCAACTGCAGATCCAACCCCCGCCTGAG-3'

GFP-F3 5'-TCTAGCTGCAGGTGAGCAAGGGCGAG-3'

GFP-R3.1 5'-TCTAGGTCGACTTACTTGTACAGCTCGTC-3'

GFP-R3.2 5'-TCTAGGTCGACACGCTGAACTTGTGGC-3'

GF14c-F4 5'-TAAGGATCCATGTCTCGGGAGGAGAAT-3'

GF14c-R4 5'-TCAGGATCCTTACTGGCCCTCGCAGGCGT-3'

GF14c-F5 5'-TCTAGGGTACCGGATCCACGAGACCACTCGAACCCGACCCGCCTC-3'

GF14c-R5 5'-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.

.

1 MGNQCQNGTL GSDYHNRFPR EHAVGYVQGD SYLDLKKFDD TWPEVNNFKP

51 TAASILRRGL DPTSINVLGR KTADLREHYI IGRKLGQAQF GTTYLCTEIN

101 TGCEYACKTI PKRKLITKED VEDVRREIQI MHHLSGHKNV VAIKDVYEDG

151 QAVHIVMELC AGGELFDRIQ EKGHYSERKA AELIRIIVSI VAMCHSLGVM

201 HRDLKPENFL LLDKDDDLSI KAIDFGLSVF FKPGQVFTEL VGSPYYVAPE

251 VLHKRYGPES DVWSAGVILY VLLSGVPPFW AETQQGIFDA VLKGHIDFQS

301 DPWPKISDSA KDLIRKMLSH CPSERLKAHE VLRHPWICEN GVATDQALDP

351 SVISRLKQFS AMNKLKKLAL RVIAERLSEE EIAGLREMFK AVDTKNRGVI

401 TFGELREGLR RFGAEFKDTE IGDIMEAAHN DNNVTIHYEE FIAATLPLNK

451 IEREEHLLAA FTYFDKDGSG YITVDKLQRA CGEHNMEDSL LEEIISEVDQ

501 NNDGQIDYAE FVAMMQGSNV GLGWQTMESS LNVALRDAPQ VH*

I

III IV

V VIa

VIb VII VIII

X

IX

II

Q

P R

R

E

W

P

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. S2Expression of OsCDPK1 in T4 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. S3Comparison of plant height between T1 generation of OsCDPK1(tr) (8 independent

lines) and OsCDPK1(Ri) (5 independent lines) and wild type (Wt) plants. (A)The independent T1 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 T1 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-1OEtr-1

60 daysA

A

Wt OEtr-1 Ri-1

Le

ngth

(cm

)

16.0

12.0

8.0

4.0

0.0

10 days +GA320.0

B

Wt OEtr-1 Ri-1

Fig. S4The semi-dwarf phenotype of the OEtr-1 line was recovered by GA3 treatment.Comparison of plant height between T4 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 GA3 for 10 days.

Fig. S4 (Ho et al.)

C

Wt OEtr-1 Ri-1

Le

ngth

(cm

)

16.0

12.0

8.0

4.0

0.0

10 days20.0

10 days -GA3

D

Wt OEtr-1 Ri-1

See

dlin

gs

len

gth

(cm

)

OsCDPK1

GA20ox1

GA20ox2

GA3ox2

Act1

Ri-1OEtr-1Wt

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 14-3-3 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-OxWt GF14c-Ri

Fig. S6 (Ho et al.)