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Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 115, Issue 1, pp 55–68 | Cite as

Proteomic analysis of cotyledonary explants during shoot organogenesis in Vigna radiata

  • Suparna Ghosh
  • Amita PalEmail author
Original Paper

Abstract

Vigna radiata or mungbean belongs to the legume family of plants. Mature mungbean seeds are rich source of dietary proteins for human nutrition. The present study was aimed to analyze the comparative protein profiles of two cotyledon types, Cot and Cot E, prior to and during early time points of shoot morphogenic induction to understand the unique differential regeneration response in these two explant types which was reported earlier. These explants were grown separately in vitro on the shoot induction medium (SIM) containing Gamborg’s B5 basal nutrient composition supplemented with 15 μM N6-benzyladenine. Isolation and characterization of the proteomes from Cot and Cot E explants at different time points, during early events of shoot differentiation, were performed using two dimensional gel electrophoresis following matrix assisted laser desorption-ionization tandem mass spectrometry. A total of 112 differentially identified proteins were classified according to their putative biological function. The differential control of protein synthesis between these explants under control condition, i.e. before in vitro culture, was also noted. In Cot E explants SIM induced prompt acquisition of competence for direct shoot morphogenesis probably through fast phytohormone signaling. Over accumulated proteins in Cot E indicated stimulation of several metabolic and associated pathways earlier than Cot explants. Abundance of stress and defense related proteins in Cot E explants was presumably to cope up with stressful cultural condition. Enhanced accumulation of folding-assisted proteins involved in organogenesis mediated cellular reprogramming in Cot E explants contributed further in rapid and efficient regeneration responsiveness.

Keywords

Cytokinin MALDI-TOF-TOF Proteomics Shoot regeneration Vigna radiata 

Abbreviations

SIM

Shoot induction medium

2-D GE

Two-dimensional gel electrophoresis

BA

N6-benzyladenine

BSA

Bovine serum albumin

Cv

Cultivar

CV

Coefficient of variation

DTT

Dithiothreitol

IEF

Isoelectric focusing

MALDI TOF MS

Matrix assisted laser desorption-ionization time-of-flight mass spectrometry

MEV

MultiExperiment viewer

MOWSE

Molecular weight search

PMSF

Phenylmethylsulfonyl fluoride

RT-PCR

Reverse transcription polymerase chain reaction

SDS PAGE

Sodium dodecyl sulphate polyacrylamide gel electrophoresis

Notes

Acknowledgments

Authors are thankful to the Department of Science and Technology, Government of India (DST Sanction no. SR/SO/PS-58/05) for constant financial support in this area of research; and to the Director, Bose Institute for providing all infrastructural facilities and a Senior Research Fellowship to SG. The proteomic facilities provided by DST through IRHPA project (IR/SO/LF02/2002) are thankfully acknowledged.

Supplementary material

11240_2013_340_MOESM1_ESM.jpg (418 kb)
Supplementary Fig. 1. Involvement of the identified proteins in the Glycolysis/Gluconeogenesis pathway assigned by KEGG database in Blast2go software. (JPEG 417 kb)
11240_2013_340_MOESM2_ESM.jpg (355 kb)
Supplementary Fig. 2. Involvement of the identified protein in the tricarboxylic acid cycle pathway assigned by KEGG database in Blast2go software. (JPEG 354 kb)
11240_2013_340_MOESM3_ESM.doc (200 kb)
Supplementary material 3 (DOC 199 kb)
11240_2013_340_MOESM4_ESM.doc (34 kb)
Supplementary material 4 (DOC 34 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  1. 1.Division of Plant BiologyBose InstituteKolkataIndia

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