Skip to main content
SpringerLink
Log in

Protein metabolism during natural ageing in desiccating recalcitrant seeds of Shorea robusta

Acta Physiologiae Plantarum volume 36, pages 1649–1659 (2014)Cite this article

Abstract

Seeds of Shorea robusta (sal) are recalcitrant owing to its high desiccation sensitivity. Germinability in sal seed was lost rapidly from 100 to 0 % within 8 days. Protein oxidation examined separately in axis and cotyledon of ageing sal seeds by monitoring the levels of carbonyls, hydroperoxide, malondialdehyde and 4-hydroxy-2-nonenal adducts with protein, Amadori and Maillard reaction products. Changes in protease and proteasome activity were also estimated. The levels of all the modified proteins and activities of protease and proteasome were similar in axis and cotyledons. The amounts of carbonyls (5.5 fold in axis and 3.9 fold in cotyledons) and hydroperoxides (13.5 fold in axis and 12 fold in cotyledons) increased significantly as the seeds became non-viable. Similarly, the levels of malondialdehyde and 4-hydroxy-2-nonenal adducts promoted as the storage period advanced and reached tenfold both in the axis and cotyledons in non-viable seeds. The ageing also promoted levels of reducing sugar along with rapid enhancement in the levels of Amadori and Maillard reaction products, respectively, by 4.4 and 1.8 fold in 5 days sal seeds. Substantial promotion in protease activity both in the axis (sevenfold) and cotyledons (tenfold) of absolutely aged seeds was discernible. The activity of proteasome exhibited steady decline from 0.767 to 0.170 nmol min−1 g−1 DM in axis and 0.20–0.086 nmol min−1 g−1 DM in cotyledons of ageing sal seeds. Changes in the ROS and protein catabolism/oxidation have been discussed to establish loss of germinability in desiccating recalcitrant sal seeds.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2

Abbreviations

AMC:

7-Amido-4-methylcoumarin

DAH:

Days after harvest

DNPH:

2,4-Dinitrophenyl hydrazine

EA:

Embryonic axis

G:

Germination

HNE:

4-Hydroxy-2-nonenal

LN2 :

Liquid nitrogen

MC:

Moisture content

MDA:

Malondialdehyde

NBT:

Nitroblue tetrazolium

PCO:

Protein carbonyl

PrOOH:

Protein hydroperoxide

ROS:

Reactive oxygen species

SDS:

Sodium dodecyl sulphate

TCA:

Trichloroacetic acid

References

  • Berjak P, Pammenter NW (1997) Progress in understanding and manipulation of desiccation sensitive (recalcitrant) seeds. In: Ellis RH, Black M, Murdoch AJ, Hong TD (eds) Basic and applied aspects of seed biology. Proc Fifth Int Workshop on Seeds. Reading, Kluwer Academic Publishers, The Netherlands, pp 689–703

    Chapter  Google Scholar 

  • Berjak P, Pammenter NW (2008) From Avicennia to Ziziana: seed recalcitrance in perspective. Ann Bot 101:213–228

    Article  PubMed Central  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Bruni F, Leopold AC (1991) Glass transitions in soybean seed: relevance to anhydrous biology. Plant Physiol 96:660–663

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Carrard G, Friguet B (2003) The proteasome in aging. In: von Zglinicki T (ed) Aging at the molecular level. Kluwer Academic Publisher, The Netherlands, pp 213–231

    Chapter  Google Scholar 

  • Castellion M, Matiacevich S, Buera P, Maldonado S (2010) Protein deterioration and longevity of quinoa seeds during long term storage. Food Chem 121:952–958

    Article  CAS  Google Scholar 

  • Chairpotto E, Scavazza A, Leonarduzzi G, Camandola S, Biasi F, Teggia PM, Garavogilia M, Robecchi A, Roncari A, Poli G (1997) Oxidative damage and transforming growth factor β 1 expression in pretumorial and tumoral lesions of human intestine. Free Radic Bio Med 22:889–894

    Article  Google Scholar 

  • Firth CA, Crone EM, Flavall EA, Roake JA, Gieseg SP (2008) Macrophage mediated protein hydroperoxide formation and lipid oxidation in low density lipoprotein are inhibited by the inflammation marker 7,8-dihydroneopterin. Biochimia et Biophysica Acta 1783:1095–1101

    Article  CAS  Google Scholar 

  • Galleschi L, Capocchi A, Ghiringhelli S, Saviozzi F, Calucci L, Pinzino C, Zandomeneghi M (2002) Antioxidants, free radicals, storage proteins and proteolytic activities in wheat (Triticum durum) seeds during accelerated aging. J Agric Food Chem 50:5450–5457

    Article  CAS  PubMed  Google Scholar 

  • Gay C, Collins J, Gebicki JM (1999) Hydroperoxide assay with the ferric-xylenol orange complex. Anal Biochem 273:149–155

    Article  CAS  PubMed  Google Scholar 

  • Gebicki S, Gebicki JM (1993) Formation of peroxides in amino acids and proteins exposed to oxygen free radicals. Biochem J 289:743–749

    CAS  PubMed Central  PubMed  Google Scholar 

  • Gebicki S, Gebicki JM (1999) Crosslinking of DNA and proteins induced by protein hydroperoxide. J Biochem 338:629–636

    Article  CAS  Google Scholar 

  • Hammond CJ (2008) Is oxidative stress the cause of death when recalcitrant Spartina alterniflora seeds are dried? Ph. D. Thesis, Louisiana State University and Agriculture and Mechanical College, Louisiana, pp 1–150

  • Hendry GAF, Finch-Savage WE, Thorpe PC, Antherton NM, Buckland SM, Nilsson KA, Seel WE (1992) Free radical activity and loss of viability during desiccation in the recalcitrant Quercus robur L. New Phytol 122:273–279

    Article  CAS  Google Scholar 

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

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Kehinde TO, Ajala MO, Daniel IO, Oyelakin OO (2013) Physiological and genetic integrity of Amaranth (Amaranthus spp.) seeds during storage. Int J Plant Breed Genet 7:35–46

    Article  Google Scholar 

  • Keshavkant S, Padhan J, Parkhey S, Naithani SC (2012) Physiological and antioxidant responses of germinating Cicer arietinum seeds to salt stress. Russ J Plant Physiol 59:206–211

    Article  CAS  Google Scholar 

  • Khan MF, Wu X, Tipins UR, Ansari GA, Boor PJ (2002) Protein adducts of malondialdehyde and 4-hydroxynonenal in livers of iron loaded rats: quantitation and localization. Toxicology 173:193–201

    CAS  PubMed  Google Scholar 

  • Kumar GNM, Knowles NR (1993) Age of potato seed tubers influences protein synthesis during sprouting. Physiol Plant 89:262–270

    Article  CAS  Google Scholar 

  • Kumar GNM, Houtz RL, Knowles RN (1999) Age induced protein modification and increased proteolysis in potato seed tubers. Plant Physiol 119:89–99

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Kumar M, Kak A, Singh S (2011) Analysis of biochemical and physiological changes during ultra-desiccation in radish (Raphanus sativus). Asian J Plant Sci Res 1:5–21

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of head bacteriophage T4. Nature 227:680–685

    Article  CAS  PubMed  Google Scholar 

  • Levine RL, Willians JA, Stadtman ER, Schater E (1994) Carbonyl assay for determination of oxidatively modified proteins. Method Enzymol 233:346–363

    Article  CAS  Google Scholar 

  • Martinez-Vincente M, Sovak G, Cuervo AM (2005) Protein degradation and ageing. Exp Gerontol 40:622–633

    Article  Google Scholar 

  • Merheb CW, Cabral H, Gomes E, Da-Silva R (2007) Partial characterization of protease from a thermophilic fungus, Thermoascus aurantiacus, and its hydrolytic activity on bovine casein. Food Chem 104:127–131

    Article  CAS  Google Scholar 

  • Murthy UMN, Sun WQ (2000) Protein modification by Amadori and Millard reaction during seed storage: roles of sugar hydrolysis and lipid peroxidation. J Exp Bot 51:1221–1228

    Article  CAS  PubMed  Google Scholar 

  • Murthy UMN, Liang Y, Kumar PP, Sun WQ (2002) Non-enzymatic protein modification by the Millard reaction reduces the activities of scavenging enzymes in Vigna radiata. Physiol Plant 115:213–220

    Article  CAS  PubMed  Google Scholar 

  • Murthy UMN, Kumar PP, Sun WQ (2003) Mechanisms of seed ageing under different storage conditions for Vigna radiata (L.) Wilczek: lipid peroxidation, sugar hydrolysis, Maillard reactions and their relationship to glass state transition. J Exp Bot 54:1057–1067

    Article  CAS  PubMed  Google Scholar 

  • Ogé L, Bourdais G, Bove J, Collet B, Godin B, Granier F, Boutin JP, Job D, Jullien M, Grappin P (2008) Protein repair l-isoaspartyl methyltransferase 1 is involved in both seed longevity and germination vigor in Arabidopsis. Plant Cell 20:3022–3037

    Article  PubMed Central  PubMed  Google Scholar 

  • Oracz K, El-Maarouf Bouteau H, Farrant JM, Cooper K, Belghazi M, Job C, Job D, Corbineau F, Bailly C (2007) ROS production and protein oxidation as a novel mechanism for seed dormancy. Plant J 50:452–465

    Article  CAS  PubMed  Google Scholar 

  • Parkhey S, Naithani SC, Keshavkant S (2012) ROS production and lipid catabolism in desiccating Shorea robusta seeds during ageing. Plant Physiol Biochem 57:261–267

    Article  CAS  PubMed  Google Scholar 

  • Ponist S, Mihalova D, Jancinova V, Snir V, Ondrejickova O, Mascia C, Poli G, Stancikova M, Nosal R, Bauerov K (2010) Reduction of oxidative stress in adjuvant arthritis. Comparison of efficacy of two pyridoindoles: stobadine dipalmitate and SMe1∙2HCl. Acta Biochim Polo 57:223–228

    CAS  Google Scholar 

  • Pukacka S, Ratajczak E (2006) Antioxidative response of ascorbate-glutathione pathway enzymes and metabolites to desiccation of recalcitrant Acer saccharinum seeds. J Plant Physiol 163:1259–1266

    Article  CAS  PubMed  Google Scholar 

  • Rajjou L, Lovigny Y, Groot PC, Steven Belghaz M, Job C, Job D (2008) Proteome-wide characterization of seed aging in Arabidopsis: a comparison between artificial and natural aging protocols. Plant Physiol 148:620–641

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Roach T, Beckett RP, Minibayeva FV, Colville L, Whitaker C, Chen H, Bailly C, Kranner I (2010) Extracellular superoxide production, viability and redox poise in response to desiccation in recalcitrant Castanea sativa seeds. Plant Cell Environ 33:59–75

    CAS  PubMed  Google Scholar 

  • Sitte N, Huber M, Grune T, Ladhoff A, Doecke WD, Von Zglinicki T, Davies KJ (2000) Proteasome inhibition by lipofuscin during postmitotic aging of fibroblasts. FASEB J 14:1490–1498

    Article  CAS  PubMed  Google Scholar 

  • Spiro RG (1966) Analysis of sugars found in glycoproteins. Methods Enzymol 8:3–52

    Article  CAS  Google Scholar 

  • Strelec I, Ugarcic-Hardi Z, Mario H (2008) Accumulation of Amadori and Maillard products in wheat seeds aged under different storage conditions. Croatica Chemi Acta 81:131–137

    CAS  Google Scholar 

  • Sun WQ, Leopold AC (1995) The Maillard reaction and oxidative stress during ageing of soybean seeds. Physiol Plant 94:94–105

    Article  CAS  Google Scholar 

  • Szweda PA, Friguet B, Szweda LI (2002) Proteolysis, free radicals and aging. Free Radic Bio Med 33:29–36

    Article  CAS  Google Scholar 

  • Tanito M, Michael HE, Kotake Y, Anderson RE (2005) Protein modifications by 4-hydroxynonenal and 4-hydroxyhexanal in light-exposed rat retina. Invest Ophthal Vis Sci 46:3859–3868

    Article  PubMed  Google Scholar 

  • Vinayachandra Chandrashekar KR (2012) Effect of desiccation on viability and biochemical changes in Knema attenuata seeds. J For Res 23:703–706

    Article  CAS  Google Scholar 

  • Wang W, Vignani R, Scali M, Cresti M (2006) A universal and rapid protocol for protein extraction from recalcitrant plant tissues for proteomic analysis. Electrophoresis 27:2782–2786

    Article  CAS  PubMed  Google Scholar 

  • Wettlaufer SH, Leopold AC (1991) Relevance of Amadori and Maillard products to seed deterioration. Plant Physiol 97:165–169

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Yang P, Fu H, Walker J, Papa CM, Smalle J, Yu-Ming JJ, Vierstra RD (2004) Purification of the Arabidopsis 26 S proteasome biochemical and molecular analysis revealed the presence of multiple isoforms. J Biol Chem 279:6401–6413

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors gratefully acknowledge financial assistance awarded to S. Parkhey by University Grants Commission, New Delhi (F.4-1/2006(BSR)/7-145/2007(BSR), dated 24.06.2011).

Author information

Authors and Affiliations

  1. School of Life Sciences, Pt. Ravishankar Shukla University, Raipur, 492 010, India

    Suruchi Parkhey & S. C. Naithani

  2. School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur, 492 010, India

    S. Keshavkant

Authors
  1. Suruchi Parkhey
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S. C. Naithani
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. Keshavkant
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to S. Keshavkant.

Additional information

Communicated by S. Weidner.

Electronic supplementary material

Below is the link to the electronic supplementary material.

11738_2014_1540_MOESM1_ESM.doc

Figs. S1 and S3 Showing peak density graph in EA and cotyledon respectively during natural storage of Shorea robusta seeds at laboratory conditions (Recorded by Quantity One software of Gel Doc System, Bio-Rad, USA). Figs. S2 and S4 showing relative intensity values of EA and cotyledon of Shorea robusta seed in different lanes during natural storage at laboratory conditions (Recorded by Quantity One software of Gel Doc System, Bio-Rad, USA). (DOC 165 kb)

Rights and permissions

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Parkhey, S., Naithani, S.C. & Keshavkant, S. Protein metabolism during natural ageing in desiccating recalcitrant seeds of Shorea robusta . Acta Physiol Plant 36, 1649–1659 (2014). https://doi.org/10.1007/s11738-014-1540-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11738-014-1540-x

Keywords

search

Navigation