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Identification of differential proteins of mungbean cotyledons during seed germination: a proteomic approach

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Abstract

Proteomic analyses of seed germination, a complex physiological process is not yet explored in leguminous plants. Such study has been undertaken to reveal the variations that occur in cotyledonary proteins during seed gemination of mungbean (Vigna radiata L. Wilczek). Cotyledons were harvested for protein extraction and two dimensional gel electrophoreses at two developmental stages (I and II) of seed germination. Comparative qualitative and quantitative analyses between these two stages revealed differential expressions and de novo appearance or disappearance of proteins. Seventy-four proteins were identified by MALDI-TOF-MS and MS/MS and categorized according to their functions including metabolism and energy production, stress related, protein processing, transcription, storage etc. Majority of these proteins are associated with metabolisms indicating that cotyledons acquire an active metabolic state during seed germination. Many of the identified proteins were enzymes involved in starch and sucrose metabolism and those of glycolysis, tricarboxylic acid cycle etc. A pathway illustrating starch synthesis/breakdown, degradation of sucrose and the fates of reducing sugars in the germinating mungbean cotyledons has been elucidated. Predominance of antioxidant enzymes can be attributed to combat with reactive oxygen species generated due to active metabolism. Thus, the study comprehensively dealt with mechanisms of stored reserve mobilization and the roles of additional cotyledonary proteins of mungbean essential for successful seed germination and seedling development.

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References

  • Alexander R, Alamillo JM, Salamini F, Bartels D (1994) A novel embryo-specific barley cDNA clone encodes a protein with homologies to bacterial glucose and ribitol dehydrogenase. Planta 192:519–525

    Article  CAS  PubMed  Google Scholar 

  • Alpert P, Oliver MJ (2002) Drying without dying. In: Black M, Pritchard HW (eds) Desiccation and survival in plants, drying without dying. CABI Publishing, New York, pp 3–44

    Chapter  Google Scholar 

  • Angeles-Nunez JG, Tiessen A (2010) Arabidopsis sucrose synthase 2 and 3 modulate metabolic homeostasis and direct carbon towards starch synthesis in developing seeds. Planta 232:701–718

    Article  CAS  PubMed  Google Scholar 

  • Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399

    Article  CAS  PubMed  Google Scholar 

  • Arai M, Mori H, Imasek H (1991) Roles of sucrose-metabolizing enzymes in growth of seedlings: purification of acid invertase from growing hypocotyls of mung bean seedlings. Plant Cell Physiol 32:1291–1298

    CAS  Google Scholar 

  • Bewley JD (1997) Seed germination and dormancy. Plant Cell 9:1055–1066

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Bewley JD, Black M (1994) Seeds: physiology of development and germination. Plenum Press, New York

    Book  Google Scholar 

  • Borghetti F, Noda FN, De Sá CM (2002) Possible involvement of proteasome activity in ethylene induced germination of dormant sunflower embryos. Braz J Plant Physiol 14:125–131

    Article  CAS  Google Scholar 

  • Boudet J, Buitink J, Hoekstra FA, Rogniaux H, Larre C, Satour P, Leprince O (2006) Comparative analysis of the heat stable proteome of radicles of Medicago truncatula seeds during germination identifies late embryogenesis abundant proteins associated with desiccation tolerance. Plant Physiol 140:1418–1436

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Cheng L, Gao X, Li S, Shi M, Javeed H, Jing X, Yang G, He G (2010) Proteomic analysis of soybean [Glycine max (L.) Meer.] seeds during imbibition at chilling temperature. Mol Breeding 26:1–17

    Article  CAS  Google Scholar 

  • Chrispeels MJ, Boulter D (1975) Control of storage protein metabolism in the cotyledons of germinating mung beans: role of endopeptidase. Plant Physiol 55:1031–1037

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Das S, Sengupta DN, Pal A (2006) Differential protein pattern of two cotyledon explants of Vigna radiata during induced in vitro differentiation: probable implication in the conundrum of differential regeneration response. J Plant Biochem Biotechnol 15:123–129

    Article  CAS  Google Scholar 

  • Dey PM (1986) Changes in the forms of invertase during germination of mung bean seeds. Phytochem 25:51–53

    Article  CAS  Google Scholar 

  • Dure L (1993) A repeat 11-mer amino acid motif and plant desiccation. Plant J 3:363–369

    Article  CAS  PubMed  Google Scholar 

  • Finnie C, Maeda K, Østergaard O, Bak-Jensen KS, Larsen J, Svensson B (2004) Aspects of the barley seed proteome during development and germination. Prot Plant Proteins 32:517–519

    CAS  Google Scholar 

  • Fu X, Richards DE, Ait-ali T, Hynes LW, Ougham H, Peng J, Harberd NP (2002) Gibberellin-mediated proteasome-dependent degradation of the barley DELLA protein SLN1 repressor. Plant Cell 14:3191–3200

    Google Scholar 

  • Gallardo K, Job C, Groot SPC, Puype M, Demol H, Vandekerckhove J, Job D (2001) Proteomic analysis of Arabidopsis seed germination and priming. Plant Physiol 126:835–848

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Garnczarska M, Lukasz W (2008) Differential response of antioxidative enzymes in embryonic axes and cotyledons of germinating lupine seeds. Acta Physiol Plant 30:427–432

    Article  CAS  Google Scholar 

  • Geigenberger P, Stitt MA (1991) “Futile” cycle of sucrose synthesis and degradation is involved in regulating partitioning between sucrose, starch and respiration in cotyledons of germinating Ricinus communis L. seedlings when phloem transport is inhibited. Planta 185:81–90

    CAS  PubMed  Google Scholar 

  • Han B, Kermode AR (1996) Dehydrin-like proteins in castor bean seeds and seedlings are differentially produced in response to ABA and water-deficit-related stresses. J Exp Bot 47:933–939

    Article  CAS  Google Scholar 

  • Harris N (1976) Starch grain breakdown in cotyledon cells of germinating mung bean seeds. Planta (Berl) 129:271–272

    Article  CAS  Google Scholar 

  • Hebert DN, Foelimer B, Helenius A (1996) Calnexin and calreticulin promote folding, delay oligomerization and suppress degradation of influenza hemagglutinin microsomes. EMBO J 15:2961–2968

    CAS  PubMed  Google Scholar 

  • Hedley CL (2001) Grain legume carbohydrates. In: Hedley CL (ed) Carbohydrates in grain legume seeds: improving nutritional quality and agronomic characteristics. CABI Publishing, New York, pp 2–14

  • Herrmann KM (1999) The shikimate pathway. Annu Rev Plant Physiol Plant Mol Biol 50:473–503

    Article  CAS  PubMed  Google Scholar 

  • Job C, Rajjou L, Lovigny Y, Belghazi M, Job D (2005) Patterns of protein oxidation in Arabidopsis seeds and during germination. Plant Physiol 138:790–802

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Juliano BO, Varner JE (1969) Enzymic degradation of starch granules in the cotyledons of germinating peas. Plant Physiol 44:886–892

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Kim SY, Chung HJ, Thomas TL (1997) Isolation of a novel class of bZIP transcription factors that interact with ABA-responsive and embryo-specification elements in the Dc3 promoter using a modified yeast one-hybrid system. Plant J 11:1237–1251

    Article  CAS  PubMed  Google Scholar 

  • Kundu S, Chakraborty D, Pal A (2010) Proteomic analysis of salicylic acid induced resistance to Mungbean Yellow Mosaic India Virus in Vigna mungo. J Prot 74:337–349

    Article  Google Scholar 

  • Linthorst HJM (1991) Pathogenesis-related proteins in plants. Crit Rev Plant Sci 10:123–150

    Article  CAS  Google Scholar 

  • Maiti S, Chakraborty D, Kundu S, Paul S, Sengupta S, Pal A (2011) Developmentally regulated temporal expression and differential acid invertase activity in differentiating cotyledonary explants of mung bean [Vigna radiata (L.) Wilczek]. Plant Cell Tissue Organ Culture 107:417–425. doi:10.1007/s11240-011-9992-9

    Article  CAS  Google Scholar 

  • Mak Y, Willows RD, Roberts TH, Wrigley CW, Sharp PJ, Copeland L (2009) Germination of wheat: a functional proteomics analysis of the embryo. Cereal Chem 86:281–289

    Article  CAS  Google Scholar 

  • Manickam A, Carlier AR (1980) Isolation and function of a low molecular weight protein of mung bean embryonic axes. Planta 149:234–240

    Article  CAS  PubMed  Google Scholar 

  • Martinez M, Somoza IR, Carbonero P, Diaz I (2003) A cathepsin B-like cysteine protease gene from Hordeum vulgare (gene CatB) induced by GA in aleurone cells is under circadian control in leaves. J Exp Bot 54:951–959

    Article  CAS  PubMed  Google Scholar 

  • Mauseth JD (1998) Botany: an introduction to plant biology. Jones and Bartlett publishers, Sudbury

    Google Scholar 

  • Mitsuhashi W, Sasaki S, Kanazawa A, Yang YY, Kamiya Y, Toyomasu T (2004) Differential expression of acid invertase genes during seed germination in Arabidopsis thaliana. Biosci Biotechnol Biochem 68:602–608

    Article  CAS  PubMed  Google Scholar 

  • Molina LL, Mongrand S, Chua NH (2001) A postgermination developmental arrest checkpoint is mediated by abscisic acid and requires the ABI5 transcription factor in Arabidopsis. PNAS 98:4782–4787

    Article  Google Scholar 

  • Muntz K, Belozersky MA, Dunaevsky YE, Schlereth A, Tidemann J (2001) Stored proteinases and the initiation of storage protein mobilization in the seeds during germination and seedling growth. J Exp Bot 52:1741–1752

    Article  CAS  PubMed  Google Scholar 

  • Murray DRA (1979) Storage role for albumins in pea cotyledons. Plant Cell Env 2:221–226

    Article  Google Scholar 

  • Obroucheva NV, Lityagina SV (2009) Acid vacuolar invertase in dormant and germinating seeds of the horse chestnut. Rus J Dev Biol 40:339–344

    Article  CAS  Google Scholar 

  • Quettier AL, Eastmond PJ (2009) Storage oil hydrolysis during early seedling growth. Plant Physiol Biochem 47:485–490

    Article  CAS  PubMed  Google Scholar 

  • Roncarati R, Salamini F, Bartels D (1995) An aldose reductase homologous gene from barley: regulation and function. Plant J 7:809–822

    Article  CAS  PubMed  Google Scholar 

  • Ross HA, McRae D, Davies HV (1996) Sucrolytic enzyme activities in cotyledons of the faba bean. Plant Physiol 11:329–338

    Google Scholar 

  • Russel SD (1993) The egg cell: development and role in fertilization and early embryogenesis. Plant Cell 5:1349–1359

    Google Scholar 

  • Tsai YC, Wu MT, Hsieh JS, Hsing YIC, Shih MD (2008) Expression of Glycine max physiologically mature genes in soybean (Glycine max L.) tissues. J Genet Mol Biol 19:168–181

    Google Scholar 

  • Tunnacliffe A, Wise MJ (2007) The continuing conundrum of the LEA proteins. Naturwissenschaften 94:791–812

    Article  CAS  PubMed  Google Scholar 

  • Vasconcelos EAR, Nogueira FCS, Abreu EFM, Goncalves EF, Souza PAS, Campos FAP (2005) Protein extraction from cowpea tissues for 2-D gel electrophoresis and MS analysis. Chromatographia 62:447–450

    Article  CAS  Google Scholar 

  • Waters ER, Lee GJ, Vierling E (1996) Evolution, structure and function of the small heat shock proteins in plants. J Exp Bot 47:325–338

    Article  CAS  Google Scholar 

  • Wehmeyer N, Vierling E (2000) The expression of small heat shock proteins in seeds responds to discrete developmental signals and suggests a general protective role in desiccation tolerance. Plant Physiol 122:1099–1108

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Wu X, Liu H, Wang W, Chen S, Hu X, Li C (2011) Proteomic analysis of seed viability in maize. Acta Physiol Plant 33:181–191

    Article  CAS  Google Scholar 

  • Xu DP, Sung SJS, Black CC (1989) Sucrose metabolism in lima bean seeds. Plant Physiol 89:1106–1116

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Yang P, Li X, Wang X, Chen H, Chen F, Shen S (2007) Proteomic analysis of rice (Oryza sativa) seeds during germination. Proteomics 7:3358–3368

    Article  CAS  PubMed  Google Scholar 

  • Zhao PS, Liu F, Zheng GC, Liu H (2011) Group 3 late embryogenesis abundant protein in Arabidopsis: structure, regulation, and function. Acta Physiol Plant 33:1063–1073

    Article  CAS  Google Scholar 

  • Zou M, Guan Y, Ren H, Zhang F, Chen F (2008) A bZIP transcription factor, OsABI5, is involved in rice fertility and stress tolerance. Plant Mol Biol 66:675–683

    Article  CAS  PubMed  Google Scholar 

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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.

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  1. Division of Plant Biology, Bose Institute, Kolkata, 700054, India

    Suparna Ghosh & Amita Pal

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  1. Suparna Ghosh
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  2. Amita Pal
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Correspondence to Amita Pal.

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Communicated by M. Stobiecki.

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Ghosh, S., Pal, A. Identification of differential proteins of mungbean cotyledons during seed germination: a proteomic approach. Acta Physiol Plant 34, 2379–2391 (2012). https://doi.org/10.1007/s11738-012-1042-7

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  • DOI: https://doi.org/10.1007/s11738-012-1042-7

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