Leaf Xanthophyll content and composition in sun and shade determined by HPLC
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As a part of our investigations to test the hypothesis that zeaxanthin formed by reversible de-epoxidation of violaxanthin serves to dissipate any excessive and potentially harmful excitation energy we determined the influence of light climate on the size of the xanthophyll cycle pool (violaxanthin + antheraxanthin + zeaxanthin) in leaves of a number of species of higher plants. The maximum amount of zeaxanthin that can be formed by de-epoxidation of violaxanthin and antheraxanthin is determined by the pool size of the xanthophyll cycle. To quantitate the individual leaf carotenoids a rapid, sensitive and accurate HPLC method was developed using a non-endcapped Zorbax ODS column, giving baseline separation of lutein and zeaxanthin as well as of other carotenoids and Chl a and b.
The size of the xanthophyll cycle pool, both on a basis of light-intercepting leaf area and of light-harvesting chlorophyll, was ca. four times greater in sun-grown leaves of a group of ten sun tolerant species than in shade-grown leaves in a group of nine shade tolerant species. In contrast there were no marked or consistent differences between the two groups in the content of the other major leaf xanthophylls, lutein and neoxanthin. Also, in each of four species examined the xanthophyll pool size increased with an increase in the amount of light available during leaf development whereas there was little change in the content of the other xanthophylls. However, the α-carotene/β-carotene ratio decreased and little or no α-carotene was detected in sun-grown leaves. Among shade-grown leaves the α-carotene/β-carotene ratio was considerably higher in species deemed to be umbrophilic than in species deemed to be heliophilic.
The percentage of the xanthophyll cycle pool present as violaxanthin (di-epoxy-zeaxanthin) at solar noon was 96–100% for shade-grown plants and 4–53% for sun-grown plants with zeaxanthin accounting for most of the balance. The percentage of zeaxanthin in leaves exposed to midday solar radiation was higher in those with low than in those with high photosynthetic capacity.
The results are consistent with the hypothesis that the xanthophyll cycle is involved in the regulation of energy dissipation in the pigment bed, thereby preventing a buildup of excessive excitation energy at the reaction centers.
Key wordsHPLC shade plants sun plants violaxanthin xanthophyll cycle zeaxanthin
epoxidation state (V+0.5A)/(V+A+Z)
photon flux density
- Bilger W, Björkman O and Thayer SS (1989) Light-induced spectral absorbance changes in relation to photosynthesis and the epoxidation state of xanthophyll cycle components in cotton leaves. Plant Physiol 91: 542–551Google Scholar
- Braumann TH and Grimme LH (1981) Reversed-phase high performance liquid chromatography of chlorophylls and carotenoids. Biochim Biophys Acta 637: 8–17Google Scholar
- Braumann TH, Weber G and Grimme LH (1982) Carotenoid and chlorophyll composition of light harvesting and reaction center proteins of the thylakoid membrane. Photobiochem Photobiophys 4: 1–8Google Scholar
- Davies BH (1976) Carotenoids. In: Goodwin TW (ed.) Chemistry and Biochemistry of Plant Pigments Vol 2, pp 38–165 London: Academic Press Inc.Google Scholar
- Demmig B and Björkman O (1987) Comparison of the effect of excessive light on chlorophyll fluorescence (77 K) and photon yield of O2 evolution in leaves of higher plants. Planta 171: 171–184Google Scholar
- Demmig B, Winter K, Krüger A and Czygan F-C (1987) Photoinhibition and zeaxanthin formation in intact leaves. Plant Physiol 84: 218–224Google Scholar
- Demmig B, Winter K, Krüger A and Czygan F-C (1988) Zeaxanthin and the heat dissipation of excess light energy in Nerium oleander exposed to a combination of high light and water stress. Plant Physiol 87: 17–24Google Scholar
- Demmig-Adams B, Winter K, Krüger A and Czygan F-C (1989) Light response of CO2 assimilation, dissipation of excess excitation energy, and zeaxanthin content of sun and shade leaves. Plant Physiol 90: 881–886Google Scholar
- Grill D and Pfeifhofer HW (1985) Carotinoide in Fichtennadeln II. Quantitative Untersuchungen. Phyton 25: 1–15Google Scholar
- Krinsky NI and Welankiwar S (1964) Assay of carotenoids. Meth Enzymol 105: 155–162Google Scholar
- Lémoine Y, Zabulon G and Cornu A (1987) Chlorophyll-protein complexes; changes associated with chloroplast development in a virescent Petunia hybrida mutant. In Biggins J (ed.) Progress in Photosynthesis Research Vol II, pp. 371–374 Dordrecht: Martinus Nijhoff PublishersGoogle Scholar
- Lichtenthaler HK, Burgstahler R, Buschmann C, Meier D, Prenzel U and Schönthal A (1982) Effect of high light and high light stress on composition, function and structure of the photosynthetic apparatus. In: Marcelle R (ed.) Stress Effects on Photosynthesis, pp 353–370. The Hague: Dr W. Junk PublishersGoogle Scholar
- Mantoura RFC and Llewellyn CA (1983) Rapid determination of algal chlorophyll and carotenoid pigments and their break-down products in natural waters by reverse-phase high-performance liquid chromatography. Anal Chim Acta 151: 297–314Google Scholar
- Powles S (1984) Photoinhibition of photosynthesis induced by visible light. Ann Rev Plant Physiol 35: 15–44Google Scholar
- Prenzel U and Lichtenthaler HK (1982) Localization of β-carotene in chlorophyll a-proteins and changes in its levels during short-term high light exposure of plants. In: Wintermans JFGM and Kuiper PJC (eds.) Biochemistry and Metabolism of Plant Lipids, pp 565–572 Amsterdam: Elsevier Biomedical Press B.V.Google Scholar
- Rau W (1988) Functions of carotenoids other than in photosynthesis. In: Goodwin TW (ed.) Plant Pigments, pp 231–255 San Diego, CA: Academic Press Inc.Google Scholar
- Schäfer C and Björkman O (1989) Relationship between efficiency of photosynthetic energy conversion and chlorophyll fluorescence quenching in upland cotton (Gossypium hirsutum L.). Plants 178: 367–376Google Scholar
- Siefermann-Harms D (1988) High performance liquid chromatography of chloroplast pigments. One-step separation of carotene and xanthophyll isomers, chlorophylls and pheophytins. J. Chromatog 448: 411–416Google Scholar
- Strain HH (1958) Chloroplast pigments and chromatographic analysis. 32nd Annual Priestley Lectures, Penn State University, University Park, PA, USAGoogle Scholar
- Wieckowski S and Goodwin TW (1966) Assimilatory pigments in cotyledons of four species of pine seedlings grown in darkness and in light. Phytochem 5: 1345–1348Google Scholar
- Willstätter R and Stoll A (1913) Untersuchungen über Chlorophyll. Julius Springer, BerlinGoogle Scholar
- Wright SW and Shearer JD (1984) Rapid extraction and high-performance liquid chromatography of chlorophylls and carotenoids from marine phytoplankton. J Chromatog 294: 281–295Google Scholar