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Table 1 Oligonucleotide primers used in this study

From: Overexpression of PtrABF gene, a bZIP transcription factor isolated from Poncirus trifoliata, enhances dehydration and drought tolerance in tobacco via scavenging ROS and modulating expression of stress-responsive genes

Genes

Primers

Annealing temperature (°C)

Product size (bp)

Sequences (5'-3')

 
    

Forward

Reverse

Used for

PtrABF

GSP1

60

1511

ATTGGAGCAAGCTGTTGCCACCGC

CGCAGCTGCCTACACTCCATGG

Amplification of full-length cDNA

Actin

Actin

58

107

ATTGTAAGCAACTGGGATGATA

AGAGGTGCCTCAGTGAGAAG

Quantitative real-time PCR

PtrABF

GSP2

57

113

CCATCACCAGTTCCTTATGTGTT

GATCTTGCAGCTGACTCTCTGTT

 

PtrABF

GSP3

58

1363

GGCTCGAGATGGGATCTCAAATGAACTTCAAG (XhoI site is underlined)

AAGGATCCCGTCAGTGTCCTCCTCAAGCACAG (BamHI site is underlined)

Subcellular localization

GFP

GFP-GSP1

60

732

CGGGATCCATGAGTAAAGGAGAAG (BamHI site is underlined)

CGGTACCTTATTTGTATAGTTCATCC (KpnI site is underlined)

 

PtrABF

GSP4

60

1366

GGCCATGGCGATGGGATCTCAAATGAACTTCAAG (NcoI site is underlined)

ATAGGATCCCTACCAAGGGCCCGTCAGTGTCC (BamHI site is underlined)

Transcriptional activation assay

PtrABF

GSP5

58

160

ATTGGAGCAAGCTGTTGCCACCGC

CGCAGCTGCCTACACTCCATGG

Transgene overexpression

CaMV35S-PtrABF

35S-PtrABF

60

2013

CGCCGTAAAGACTGGCGAACAGTTCATACAGAGT

CGCAGCTGCCTACACTCCATGG

Confirmation of transgenic plants

NPTII

NPTII

56

742

AGACAATCGGCTGCTCTGAT

TCATTTCGAACCCCAGAGTC

 

NtADC1

 

58

306

CTTGCTGATTACCGCAATTTATC

CCTTACTGCAGGCTTTTCATCTA

Analysis of expression levels

NtADC2

 

58

350

GCCGGCCCTAGGTTGTTGTGTAGATG

AGCGAACAACAAGAGGCAGCTGAAGCC

 

NtAREB

 

58

224

TCTTCACAGCAAAAGCCTCA

GTGACCCCATTATGCAATCC

 

NtCAT

 

58

151

AGGTACCGCTCATTCACACC

AAGCAAGCTTTTGACCCAGA

 

NtCDPK2

 

58

267

AGGTGAGCTTTTCGATAGGATTATT

ACTTCTGGTGCAACATAGTAAGGAC

 

NtDREB

 

58

503

GGACCCACTTGCTGATTCTT

GCGCCTCCTCATCCATATAA

 

NtERD10C

 

58

366

ACGTGGAGGCTACAGATCGTGGTTTG

TCTCCACTGGTACAGCCGTGTCCTCAC

 

NtERF

 

58

220

ACACTTCATTTTCACATTCCAACTT

TCTTCTAATTCTTGACCATGGCTAC

 

NtLEA5

 

58

350

TTGTTAGCAGGCGTGGGTAT

CTCTCGCTCTTGTTGGGTTC

 

NtAPX

 

58

262

CAAATGTAAGAGGAAACTCAGAGGA

AGCAACAACTCCAGCTAATTGATAG

 

NtSAMDC

 

58

313

ATTGGTTTTGAAGGTTTTGAGAAG

TCACGTCTTGTACTTTGAGAGACAG

 

NtSOD

 

58

238

AGCTACATGACGCCATTTCC

CCCTGTAAAGCAGCACCTTC

 

Tubulin

 

58

163

TCCAGGACAAGGAGGGTAT

CATCAACAACAGGCAACCTAG

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