biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 55:75-82, 2011 | DOI: 10.1007/s10535-011-0010-1

Characterization of Mg-dechelating substance in senescent and pre-senescent Arabidopsis thaliana leaves

A. M. Büchert1, P. M. Civello1,2, G. A. Martínez1,2,*
1 Instituto de Investigaciones Biotecnológicas, CONICET-UNSAM, Chascomús, Argentina
2 Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Argentina

The removal of Mg2+ is an important step in the chlorophyll degradation pathway and extracts from senescent and presenescent Arabidopsis thaliana leaves were analyzed for Mg-dechelatase activity, using chlorophyllin, an artificial derivative of the natural substrate, chlorophyllide. The optimum temperature and pH for this reaction were determined to be at approximately 50 °C and 7.2, respectively. Mg-dechelatase activity was enhanced by addition of EDTA and inhibited by MgCl2, HgCl2 and reduced glutathione, indicating phenomenons such as retroinhibition by reaction products and dependence on the redox state of the mixture. Size exclusion chromatography was performed on Arabidopsis leaf extracts, and Mg-dechelatase activity was found in the fraction corresponding to molecular mass of about 42 kDa, which indicates that the Mg-dechelating compound in Arabidopsis is considerably larger than in other systems. During dark-induced senescence, the activity increased over time until reaching a maximum at day 4, and then decreased. The addition of plant growth regulators indicated that the accumulation of Mg-dechelatase was activated by ethylene and delayed by 6-benzylaminopurine.

Keywords: chlorophyll degradation; chlorophyllin; pheophorbide

Received: June 19, 2009; Accepted: February 17, 2010; Published: March 1, 2011  Show citation

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Büchert, A.M., Civello, P.M., & Martínez, G.A. (2011). Characterization of Mg-dechelating substance in senescent and pre-senescent Arabidopsis thaliana leaves. Biologia plantarum, 55(1), 75-82. doi: 10.1007/s10535-011-0010-1
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    References

    1. Benedetti, C.E., Arruda, P.: Altering the expression of the chlorophyllase gene ATHCOR1 in transgenic Arabidopsis caused changes in the chlorophyll-to-chlorophyllide ratio. - Plant Physiol. 128: 1255-1263, 2002. Go to original source...
    2. Bertani, G.: Studies on lysogenesis I.: The mode of phage liberation by lysogenic Escherichia coli. - J. Bacteriol. 62: 293, 1951. Go to original source...
    3. Cho, C., Chung, E., Kim, K., Soh, H., Jeong, Y., Lee, S., Lee, Y., Kim, K., Chung, Y., Lee, J.: Plasma membrane localization of soybean matrix metalloproteinase differentially induced by senescence and abiotic stress. - Biol. Plant. 53: 461-467, 2009. Go to original source...
    4. Costa, M.L., Civello, P.M., Chaves, A.R., MartÍnez, G.A.: Characterization of Mg-dechelatase activity obtained from Fragaria × ananassa fruit. - Plant Physiol. Biochem. 40: 111-118, 2002. Go to original source...
    5. Costa, M.L., Civello, P.M., Chaves, A.R., MartÍnez, G.A.: Effect of ethephon and 6-benzylaminopurine on chlorophyll degrading enzymes and a peroxidase-linked chlorophyll bleaching during post-harvest senescence of broccoli (Brassica oleracea L.) at 20 °C. - Postharvest Biol. Technol. 35: 191-199, 2005. Go to original source...
    6. Costa, M.L., Civello, P.M., Chaves, A.R., MartÍnez, G.A.: Hot air treatment decreases chlorophyll catabolism during postharvest senescence of broccoli (Brassica oleracea L. var. italica) heads. - J. Sci. Food Agr. 86: 1125-1131, 2006a. Go to original source...
    7. Costa, M.L., Vicente, A.R., Civello, P.M., Chaves, A.R., MartÍnez, G.A.: UV-C treatment delays postharvest senescence in broccoli florets. - Postharvest Biol. Technol. 39: 204-210, 2006b. Go to original source...
    8. Hörtensteiner, S.: Chlorophyll degradation during senescence. - Annu. Rev. Plant Biol. 57: 55-77, 2006. Go to original source...
    9. Inskeep, W.P., Bloom, P.R.: Extinction coefficients of chlorophyll a and b in N,N-dimethylformamide and 80 % acetone. - Plant Physiol. 77: 483-485, 1985. Go to original source...
    10. Li, J.R., Yu, K., Wei, J.R., Ma, Q., Wang, B.Q., Yu, D.: Gibberellin retards chlorophyll degradation during senescence of Paris polyphylla. - Biol. Plant. 54: 395-399, 2010. Go to original source...
    11. Matile, P., Hörtensteiner, S., Thomas, H.: Chlorophyll degradation. - Annu. Rev. Plant Biol. 50: 67-95, 1999. Go to original source...
    12. Matile, P., Hörtensteiner, S., Thomas, H., Kräutler, B.: Chlorophyll breakdown in senescent leaves. - Plant Physiol. 112: 1403-1409, 1996. Go to original source...
    13. Park, S.Y., Yu, J.W., Park, J.S., Li, J., Yoo, S.C., Lee, N.Y., Lee, S.K., Jeong, S.W., Hak, S.S., Koh, H.J., Jeon, J.S., Park, Y.I., Paek, N.C.: The senescence-induced staygreen protein regulates chlorophyll degradation. - Plant Cell 19: 1649-1664, 2007. Go to original source...
    14. Procházková, A., Wilhelmová, N.: Antioxidant protection during ageing and senescence in transgenic tobacco with enhanced activity of cytokinin oxidase/dehydrogenase. - Biol. Plant. 53: 691-696, 2009. Go to original source...
    15. Pružinská, A., Tanner, G., Anders, I., Roca, M., Hörtensteiner, S.: Chlorophyll breakdown: pheophorbide a oxygenase is a Rieske-type iron-sulfur protein, encoded by the accelerated cell death 1 gene. - Proc. nat. Acad. Sci. USA 100: 15259-15264, 2003. Go to original source...
    16. Rodoni, S., Nakayama, K., Anderl, M., Kräutler, B., Moser, D., Thomas, H., Matile, P., Hörtensteiner, S.: Chlorophyll breakdown in senescent chloroplasts: cleavage of pheophorbide a in two enzymic steps. - Plant Physiol. 115: 669-676, 1997. Go to original source...
    17. Schelbert, S., Aubry, S., Burla, B., Agne, B., Kessler, F., Krupinska, K., Hortensteiner, S.: Pheophytin pheophorbide hydrolase (pheophytinase) is involved in chlorophyll breakdown during leaf senescence in Arabidopsis. - Plant Cell 21: 767-785, 2009. Go to original source...
    18. Schenk, N., Schelbert, S., Kanwischer, M., Goldschmidt, E.E., Dörmann, P., Hörtensteiner, S.: The chlorophyllases AtCLH1 and AtCLH2 are not essential for senescencerelated chlorophyll breakdown in Arabidopsis thaliana. - FEBS Lett. 581: 5517-5525, 2007. Go to original source...
    19. Shioi, Y., Tomita, N., Tsuchiya, T., Takamiya, K.I.: Conversion of chlorophyllide to pheophorbide by Mg-dechelating substance in extracts of Chenopodium album. - Plant Physiol. Biochem. 34: 41-47, 1996.
    20. Suzuki, T., Kunieda, T., Murai, F., Morioka, S., Shioi, Y.: Mg-dechelation activity in radish cotyledons with artificial and native substrates, Mg-chlorophyllin a and chlorophyllide a. - Plant Physiol. Biochem. 43: 459-464, 2005. Go to original source...
    21. Suzuki, T., Shioi, Y.: Re-examination of Mg-dechelation reaction in the degradation of chlorophylls using chlorophyllin a as a substrate. - Photosynth. Res. 74: 217-223, 2002. Go to original source...
    22. Tanaka, A., Tanaka, R.: Chlorophyll metabolism. - Curr. Opin. Plant Biol. 9: 248-255, 2006. Go to original source...
    23. Tang, L., Okazawa, A., Fukusaki, E., Kobayashi, A.: Removal of magnesium by Mg-dechelatase is a major step in the chlorophyll-degrading pathway in Ginkgo biloba in the process of autumnal tints. - Z. Naturforsch. Sect. C 55: 923-926, 2000. Go to original source...
    24. Tsuchiya, T., Ohta, H., Okawa, K., Iwamatsu, A., Shimada, H., Masuda, T., Takamiya, K.I.: Cloning of chlorophyllase, the key enzyme in chlorophyll degradation: finding of a lipase motif and the induction by methyl jasmonate. - Proc. nat. Acad. Sci. USA 96: 15362-15367, 1999. Go to original source...
    25. Vicente, A.R., Costa, L., Covatta, F., MartÍnez, G.A., Chaves, A.R., Civello, P.M., Sozzi, G.O.: Physiological changes in boysenberry fruit during growth and ripening. - J. hort. Sci. Biotechnol. 81: 525-531, 2006. Go to original source...
    26. Vicentini, F., Iten, F., Matile, P.: Development of an assay for Mg-dechelatase of oilseed rape cotyledons, using chlorophyllin as the substrate. - Physiol. Plant. 94: 57-63, 1995. Go to original source...
    27. Wüthrich, K.L., Bovet, L., Hunziker, P.E., Donnison, I.S., Hörtensteiner, S.: Molecular cloning, functional expression and characterisation of RCC reductase involved in chlorophyll catabolism. - Plant J. 21: 189-198, 2000. Go to original source...

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