Hostname: page-component-cd9895bd7-mkpzs Total loading time: 0 Render date: 2024-12-25T05:21:09.916Z Has data issue: false hasContentIssue false

Corrections for interferences and extraction conditions make a difference: use of the TBARS assay for lipid peroxidation of orthodox Spartina pectinata and recalcitrant Spartina alterniflora seeds during desiccation

Published online by Cambridge University Press:  01 February 2011

James H. Chappell Jr
Affiliation:
Department of Plant Pathology and Crop Physiology, 302 Life Sciences Building, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA
Marc Alan Cohn*
Affiliation:
Department of Plant Pathology and Crop Physiology, 302 Life Sciences Building, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA
*
*Correspondence Fax: +1 225 578 1415 Email: [email protected]

Abstract

Lipid peroxidation and membrane damage are often proposed as causes of recalcitrant seed death, and the thiobarbituric acid reactive substances (TBARS) assay is commonly used to measure lipid peroxidation. However, several artefacts can cause an overestimation of TBARS values, and these have not been routinely addressed in experiments with recalcitrant seeds. In the present report, TBARS was assayed as recalcitrant Spartina alterniflora and orthodox S. pectinata seeds were dried rapidly. Using the traditional Heath and Packer (1968) protocol with tissue extraction at 4°C, S. alterniflora had higher overall TBARS values than S. pectinata, and TBARS products increased when recalcitrant S. alterniflora and orthodox S. pectinata seeds were dried. However, when corrections for interfering substances, such as sugars and anthocyanins, were made, the TBARS values between the two species were almost identical. When seeds were freeze-clamped in liquid nitrogen prior to extraction, TBARS did not increase during desiccation for either species. These findings may indicate that lipid peroxidation is not the cause of desiccation-induced death in S. alterniflora. Therefore, freeze-clamping during tissue extraction and corrections for TBARS interfering substances must be applied to avoid overestimation of lipid peroxidation values.

Type
Short Communication
Copyright
Copyright © Cambridge University Press 2011

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Berjak, P. and Pammenter, N.W. (2001) Seed recalcitrance – current perspectives. South African Journal of Botany 67, 7989.CrossRefGoogle Scholar
Chandel, K.P.S., Chaudhury, R., Radhamani, J. and Malik, S.K. (1995) Desiccation and freezing sensitivity in recalcitrant seeds of tea, cocoa and jackfruit. Annals of Botany 76, 443450.CrossRefGoogle Scholar
Chappell, J.H. (2008) Is oxidative stress the cause of death when recalcitrant Spartina alterniflora seeds are dried? PhD dissertation, Louisiana State University, Baton Rouge.Google Scholar
Cheng, H.-Y. and Song, S.-Q. (2008) Possible involvement of reactive oxygen species scavenging enzymes in desiccation sensitivity of Antiaris toxicaria seeds and axes. Journal of Integrative Plant Biology 50, 15491556.CrossRefGoogle ScholarPubMed
Du, Z. and Bramlage, W.J. (1992) Modified thiobarbituric acid assay for measuring lipid oxidation in sugar-rich plant tissue extracts. Journal of Agricultural and Food Chemistry 40, 15661570.CrossRefGoogle Scholar
Finch-Savage, W.E., Blake, P.S. and Clay, H.A. (1996) Desiccation stress in recalcitrant Quercus robur L. seeds results in lipid peroxidation and increased synthesis of jasmonates and abscisic acid. Journal of Experimental Botany 47, 661667.CrossRefGoogle Scholar
Greggains, V., Finch-Savage, W.E., Quick, W.P. and Atherton, N.M. (2000) Putative desiccation tolerance mechanisms in orthodox and recalcitrant seeds of the genus Acer. Seed Science Research 10, 317327.CrossRefGoogle Scholar
Greggains, V., Finch-Savage, W.E., Atherton, N.M. and Berjak, P. (2001) Viability loss and free radical processes during desiccation of recalcitrant Avicennia marina seeds. Seed Science Research 11, 235242.Google Scholar
Halliwell, B. and Gutteridge, J.M.C. (2007) Free radicals in biology and medicine (4th edition). Oxford, Oxford University Press.Google Scholar
Heath, R.L. and Packer, L. (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Archives of Biochemistry and Biophysics 125, 189198.CrossRefGoogle ScholarPubMed
Hendry, G.A.F., Finch-Savage, W.E., Thorpe, P.C., Atherton, N.M., Buckland, S.M., Nilsson, K.A. and Seel, W.E. (1992) Free radical processes and loss of seed viability during desiccation in the recalcitrant species Quercus robur L. New Phytologist 122, 273279.CrossRefGoogle ScholarPubMed
Hodges, D.M., DeLong, J.M., Forney, C.F. and Prange, R.K. (1999) Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta 207, 604611.CrossRefGoogle Scholar
Kermode, A.R. and Finch-Savage, B.E. (2002) Desiccation sensitivity in orthodox and recalcitrant seeds in relation to development. pp. 149184 in Black, M.; Pritchard, H.W. (Eds) Desiccation and survival in plants: drying without dying. Wallingford, CABI Publishing.CrossRefGoogle Scholar
Leprince, O. and Golovina, E.A. (2002) Biochemical and biophysical methods for quantifying desiccation phenomena in seeds and vegetative tissues. pp. 111146 in Black, M.; Pritchard, H.W. (Eds) Desiccation and survival in plants: drying without dying. Wallingford, CABI Publishing.CrossRefGoogle Scholar
Li, C. and Sun, W.Q. (1999) Desiccation sensitivity and activities of free radical-scavenging enzymes in recalcitrant Theobroma cacao seeds. Seed Science Research 9, 209217.CrossRefGoogle Scholar
Shuter, S.L., Davies, M.J., Garlick, P.B., Hearse, D.J. and Slater, T.F. (1990) Myocardial tissue preparation for ESR spectroscopy: some methods may cause artifactual generation of signals. Free Radical Research Communications 9, 5563.CrossRefGoogle ScholarPubMed
Wesley-Smith, J., Pammenter, N.W., Berjak, P. and Walters, C. (2001) The effects of two drying rates on the desiccation tolerance of embryonic axes of recalcitrant jackfruit (Artocarpus heterophyllus Lamk.) seeds. Annals of Botany 88, 653664.CrossRefGoogle Scholar
Xin, X., Jing, X.-M., Liu, Y. and Song, S.-Q. (2010) Viability loss pattern under rapid dehydration of Antiaris toxicaria axes and its relation to oxidative damage. Journal of Integrative Plant Biology 52, 434441.CrossRefGoogle ScholarPubMed