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Srijani Ghanta, Dipto Bhattacharyya, Ragini Sinha, Anindita Banerjee, Sharmila Chattopadhyay


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Nicotiana tabacum overexpressing γ-ECS exhibits biotic stress tolerance likely through NPR1-dependent salicylic acid mediated pathway


Srijani Ghanta, Dipto Bhattacharyya, Ragini Sinha, Anindita Banerjee, Sharmila Chattopadhyay*


Plant Biotechnology Laboratory, Drug Development/Diagnostics & Biotechnology Division, Indian Institute of Chemical Biology (A unit of Council of Scientific & Industrial Research), 4, Raja S.C.Mullick Road, Kolkata 700 032, India
*Corresponding author: e-mail- sharmila@iicb.res.in

Tel: 033-2499 5868

Fax: 033-2473 5197



Supplementary Fig. S1 Analysis of NtGB lines. a Southern blot indicating the insertion of T-DNA containing LeECS introduced into tobacco genome of different NtGB lines. Genomic DNA from NtGB lines were digested with EcoRI (E) and HindIII (H) separately and blot hybridized to LeECS radiolabeled probe

GSH and SA status in NtGB lines as compared to wild-type plants. b The amount of total glutathione. c GSH: GSSG ratio. d SA content. Data are the mean ± SD for three individual experiments (n = 3) using plants grown independently





Supplementary Fig. S2 Time-dependent expression analysis of NtPR1, NtPR2, NtPR4 and NtPR5 in NtGB lines and wild-type plants infected with P. syringae pv. tabaci (OD 0.05). Samples were collected at indicated time intervals after pathogen infection for quantitative RT-PCR analysis. Actin was used as an internal control. Data are the mean ± SD for three individual experiments (n = 3) using plants grown independently


Supplementary Fig. S3 Time-dependent expression analysis of NtPR1, NtPR2, NtPR4 and NtPR5 in NtGB lines and wild-type plants infected with P. syringae pv. tabaci grown upto an OD of 0.7. Samples were collected at indicated time intervals after pathogen infection for quantitative RT-PCR analysis. Actin was used as an internal control. Data are the mean ± SD for three individual experiments (n = 3) using plants grown independently

Supplementary Table S1. List of primers used in semiquantitative and quantitative RT-PCR

Gene name

Forward primer (5' to 3')

Reverse primer (5' to 3')

Accession no.

NptII

GAGGCTATTCGGCTATGACTG

ATCGGGAGGGGCGATACCGTA

AF485783

ECS700bp

AATATTTCTTCACGGAATTCCTC

AAGAGAGCTGTAGCAATAGGC

AF017983

PR1a

ATGGGATTTGTTCTCTTTTCACA

TTAGTATGGACTTTCGCCTCT

X12737

GLS

ATGGCTCATCTTTCAGATCTC

ATGAGACCACAACTCCAGCA

X95932

MAPKK

ATGAAGAAAGGATCTTTAGCTCCT

ATAGTTGTAAGTGCCGACAAAGG

AF165186

NPR1

ATGGATAATAGTAGGACTGCGTT

AATGTCATGAGGCAAGGCTTTATC

AF480488

TRXh

ATGGCTGCTAACGATGCTACTTC

AGCAGTGACAGTAGCAGGAGC

X58527

GSNOR1

TGCTGTGGCATGGGAGGCTG

ATTCACAAACTCTGTCACACCAAA

EU652335

ICS1

CAGGTTGAGTTTGATGAGCT

ACTCATCATCTTCAGTCTGGA

AY740529

PR2

ATGACTTTATGCATTAAAAATGG

CTGCAGGATTTGCTTCTTGTT

M60460

PR5

ATGAGAAAGACCCACGTC

ATGCCTTCTTTGCAGCAG

AF154636

SDRLP

ATGGCCAGCTATTCTCTTCCA

CAGATTCATACCACTCACATATT

AJ223177

AOS

ATGGCAGTAGCAACAGCAACA

CTGGGTTCGCTCCGTACAAT

AJ295274

ACCOx

ATGGAGTTGCTTAACACTGAACAA

TCTTTCTCCAACCATCAGGCTGA

X83229

PR4

TGCTTTCTGCGCTACTTGGG

GCCCCTTCTTATTCTAAACGGC

X60281

PR10

GTTCGTATAGGGGGCAAGTT

TTAAGCATAGGCTTCAGGATT

AY055111

Hsp70

ATGGCCGGAAAAGGAGAAGG

AGTAAGGAGGGACACATCGAAAGT

AY253326

CA

ATGTCAACTGCTTCCATTAACAG

AATGACAACAATGTTCTCTACCTT

M94135

ADH

ATAGAAGAAGTGGAGGTTGCACC

ATTGAACCTGCTGGGATTCAAATC

X81853

ADC

ATGCCGGCCCTAGGTTGTTG

GAAATGTACTCAGCGTCCTTG

AF321137

CBP

ATGGAAGAAACTTGCAGCTTCAG

CCTAAGGAAGTCTTCAACAGTAAA

U58971

Actin

ATGGCAGACGGTGAGGATATTCA

TGGCGCAACACGAAGTTCGTT

EU938079


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