Isolation of intact and pure chloroplasts from leaves of Arabidopsis thaliana plants acclimated to low irradiance for studies on Rubisco regulation

Magda Grabsztunowicz, Grzegorz Jackowski

Abstract


A protocol is presented for low-cost and fast isolation of intact and pure chloroplasts from leaves of plants acclimated to low irradiance. The protocol is based on a differential centrifugation of cleared leaf homogenate and omits a centrifugation on Percoll gradient step. The intactness and purity of the chloroplasts isolated from leaves of low irradiance-acclimated plants by using this protocol (confirmed by phase contrast microscopy as well as enzymatic and immunological approaches) allows plausible studies on low irradiance-dependent Rubisco regulation.


Keywords


acclimation; chloroplast isolation; differential centrifugation; low irradiance; Arabidopsis thaliana; Rubisco

Full Text:

PDF

References


Parry MAJ. Rubisco activity: effects of drought stress. Ann Bot. 2002;89(7):833–839. http://dx.doi.org/10.1093/aob/mcf103

Chaitanya KV, Jutur PP, Sundar D, Reddy RA. Water stress effect on photosynthesis in different mulberry cultivars. Plant Growth Regul. 2003;40:75–80. http://dx.doi.org/10.1023/A:1023064328384

Krapp A, Hofmann B, Schäfer C, Stitt M. Regulation of the expression of rbcS and other photosynthetic genes by carbohydrates: a mechanism for the “sink regulation” of photosynthesis? Plant J. 1993;3(6):817–828. http://dx.doi.org/10.1111/j.1365-313X.1993.00817.x

Cohen I. A proposed mechanism for the inhibitory effects of oxidative stress on Rubisco assembly and its subunit expression. Plant Physiol. 2005;137(2):738–746. http://dx.doi.org/10.1104/pp.104.056341

Knopf JA, Shapira M. Degradation of Rubisco SSU during oxidative stress triggers aggregation of Rubisco particles in Chlamydomonas reinhardtii. Planta. 2005;222(5):787–793. http://dx.doi.org/10.1007/s00425-005-0023-0

Mehta RA, Fawcett TW, Porath D, Mattoo AK. Oxidative stress causes rapid membrane translocation and in vivo degradation of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem. 1992;267(4):2810–2816.

Dionisio-Sese ML, Tobita S. Antioxidant responses of rice seedlings to salinity stress. Plant Sci. 1998;135(1):1–9. http://dx.doi.org/10.1016/S0168-9452(98)00025-9

Smirnoff N. Plant resistance to environmental stress. Curr Opin Biotechnol. 1998;9(2):214–219. http://dx.doi.org/10.1016/S0958-1669(98)80118-3

Orozco-Cardenas ML. Hydrogen peroxide acts as a second messenger for the induction of defense genes in tomato plants in response to wounding, systemin, and methyl jasmonate. Plant Cell. 2001;13(1):179–191. http://dx.doi.org/10.1105/tpc.13.1.179

Moreno J, Garcia-Murria MJ, Marin-Navarro J. Redox modulation of Rubisco conformation and activity through its cysteine residues. J Exp Bot. 2007;59(7):1605–1614. http://dx.doi.org/10.1093/jxb/erm310

Marin-Navarro J, Moreno J. Cysteines 449 and 459 modulate the reduction-oxidation conformational changes of ribulose-1,5-bisphosphate carboxylase/oxygenase and the translocation of the enzyme to membranes during stress. Plant Cell Environ. 2006;29(5):898–908. http://dx.doi.org/10.1111/j.1365-3040.2005.01469.x

Albuquerque JA, Esquıvel MG, Teixeira AR, Ferreira RB. The catabolism of ribulose bisphosphate carboxylase from higher plants. A hypothesis. Plant Sci. 2001;161(1):55–65. http://dx.doi.org/10.1016/S0168-9452(01)00389-2

Landry LG, Pell EJ. Modification of Rubisco and altered proteolytic activity in O3-stressed hybrid poplar (Populus maximowizii × trichocarpa). Plant Physiol. 1993;101(4):1355–1362.

Garcia-Ferris C, Moreno J. Oxidative modification and breakdown of ribulose-1,5-bisphosphate carboxylase/oxygenase induced in Euglena gracitis by nitrogen starvation. Planta. 1994;193(2). http://dx.doi.org/10.1007/BF00192532

Prioul JL, Reyss A. Acclimation of ribulose bisphosphate carboxylase and mRNAs to changing irradiance in adult tobacco leaves: differential expression in LSU and SSU mRNA. Plant Physiol. 1987;84(4):1238–1243. http://dx.doi.org/10.1104/pp.84.4.1238

Luciński R, Misztal L, Samardakiewicz S, Jackowski G. The thylakoid protease Deg2 is involved in stress-related degradation of the photosystem II light-harvesting protein Lhcb6 in Arabidopsis thaliana. New Phytol. 2011;192(1):74–86. http://dx.doi.org/10.1111/j.1469-8137.2011.03782.x

Adamiec M, Drath M, Jackowski G. Redox state of plastoquinone pool regulates expression of Arabidopsis thaliana genes in response to elevated irradiance. Acta Biochim Pol. 2008;55(1):161–173.

Urantowka A, Knorpp C, Olczak T, Kolodziejczak M, Janska H. Plant mitochondria contain at least two i-AAA-like complexes. Plant Mol Biol. 2005;59(2):239–252. http://dx.doi.org/10.1007/s11103-005-8766-3

Watanabe M, Nakayama H, Watanabe Y, Shimada N. Induction of a specific isoenzyme of glutamate dehydrogenase during isolation and the first 48 h of culture of Brassica napus leaf protoplasts. Physiol Plant. 1992;86(2):231–235. http://dx.doi.org/10.1034/j.1399-3054.1992.860207.x

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72(1-2):248–254. http://dx.doi.org/10.1016/0003-2697(76)90527-3

Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680–685. http://dx.doi.org/10.1038/227680a0

Aronsson H, Jarvis P. The chloroplast protein import apparatus, its components, and their roles. FEBS Lett. 2002;529(2–3):215–220. http://dx.doi.org/10.1007/978-3-540-68696-5_3

Belknap WR. Partial purification of intact chloroplasts from Chlamydomonas reinhardtii. Plant Physiol. 1983;72(4):1130–1132. http://dx.doi.org/10.1104/pp.72.4.1130

Schulz A, Knoetzel J, Scheller HV, Mant A. Uptake of a fluorescent dye as a swift and simple indicator of organelle intactness: import-competent chloroplasts from soil-grown arabidopsis. J Histochem Cytochem. 2004;52(5):701–704. http://dx.doi.org/10.1177/002215540405200514

Lang EGE, Mueller SJ, Hoernstein SNW, Porankiewicz-Asplund J, Vervliet-Scheebaum M, Reski R. Simultaneous isolation of pure and intact chloroplasts and mitochondria from moss as the basis for sub-cellular proteomics. Plant Cell Rep. 2010;30(2):205–215. http://dx.doi.org/10.1007/s00299-010-0935-4

Peltier JB. The oligomeric stromal proteome of Arabidopsis thaliana chloroplasts. Mol Cell Proteomics. 2005;5(1):114–133. http://dx.doi.org/10.1074/mcp.M500180-MCP200

Miyadai K, Mae T, Makino A, Ojima K. Characteristics of ribulose-1,5-bisphosphate carboxylase/oxygenase degradation by lysates of mechanically isolated chloroplasts from wheat leaves. Plant Physiol. 1990;92(4):1215–1219. http://dx.doi.org/10.1104/pp.92.4.1215




DOI: https://doi.org/10.5586/asbp.2012.043

Journal ISSN:
  • 2083-9480 (online)
  • 0001-6977 (print; ceased since 2016)
This is an Open Access journal, which distributes its content under the terms of the Creative Commons Attribution License, which permits redistribution, commercial and non-commercial, provided that the content is properly cited.
The journal is a member of the Committee on Publication Ethics (COPE) and aims to follow the COPE’s principles.
The journal publisher is a member of the Open Access Scholarly Publishers Association.
The journal content is indexed in Similarity Check, the Crossref initiative to prevent scholarly and professional plagiarism.
Publisher
Polish Botanical Society