Studies on the isolation of plant pigments from Spinach leaves

ravikumar velpula

Abstract


The leaves of plants contain a number of colored pigments generally falling into two categories, chlorophylls and carotenoids. The green chlorophylls a and b, which are highly conjugated compounds capture the light energy used in photosynthesis. Carotenoids are part of a larger collection of plant-derived compounds called terpenes. These naturally occurring compounds contain 10, 15, 20, 25, 30 and 40 carbon atoms, which suggest that there is a compound with five carbon atoms that serves as their building block. Their structures are consistent with the assumption that they were made by joining together isoprene units, usually in a "head to tail" fashion. Isoprene is the common name for 2-methyl-1,3-butadiene. The branched end is the "head" and the unbranched is the "tail". That isoprene units are linked in a head to tail fashion to form terpenes is known as the isoprene rule. Carotenoids are tetraterpenes. Lycopene, the compound responsible for the red coloring of tomatoes and watermelon, and β-carotene, the compound that causes carrots and apricots to be orange, are examples of carotenoids. β-Carotene which cleaves to form two molecules of vitamin A when it is ingested, is the major dietary source of this vitamin.

Spinach leaves, which you will use in this assignment, contain chlorophyll a and b and β- carotene as major pigments as well as smaller amounts of other pigments such as xanthophylls. The xanthophylls, which are oxidized versions of carotenes and pheophytins, look like chlorophyll except that the, Mg+2 is replaced by two hydrogen ions. In this assignment you will isolate and separate the plant pigments using differences in polarity to effect the separation. Since the different components are colored differently, the separation is easily followed visually.


Keywords


Isolation, Extraction, Column Chromatography, Thin-layer chromatography

Full Text:

PDF

References


AL-BABILI S. and BEYER P. 2005. Golden Rice – five years on the road – five years to go? Trends in Plant Science 12: 565–573.

BEALE SI. 2003. Photosynthetic pigments: Perplexing persistent prevalence of "Superfluous " pigment production. Current Biology 8: R342.

BEALE SI. 2005. Green genes gleaned. Trends in Plant Science 10: 309–312.

BRIDLE P, and GARCIA VIGUERA C. 1997. Analysis of anthocyanins in strawberries and elderberries. A comparison of capillary zone electrophoresis and HPLC. Food Chemistry 59: 299–304.

BRITTON G. 1995. Structure and properties of carotenoids in relation to function. The Federation of American Societies for Experimental Biology Journal 9:1551–1558.

DAVIES KM. 2004. Plant Pigments and their Manipulation. Annual Review of Plant Biology 14. Blackwell Publishing Ltd, Oxford UK.

DŻUGAN M. 2006. Czynniki wpływające na stabilność zielonych barwników roślin. Zeszyty Naukowe Polskiego Towarzystwa Inżynierii Ekologicznej 7: 26–33.

ESPINEDA CE, LINFORD AS, DEVINE D, BRUSSLAN JA. 1999. The AtCAO gene, encoding chlorophyll a oxygenase, is required for chlorophyll b synthesis in Arabidopsis thaliana. Proceedings of the National Academy of Sciences of the United States og America 96: 10507–10511.

GANDÍA-HERRERO F, GARCÍA-CARMONA F, and ESCRIBANO J. 2005. Floral fluorescence effect. Nature 437: 334–336. GIULIANO G, AQUILANI R, and DHARMAPURI S. 2000. Metabolic engineering of plant carotenoids. Trends in Plants Science 10: 406–409.

GRIESBACH R. 2005. Biochemistry and genetics of flower colour. Plant Breeding Reviews 25: 89–114.

GROTEWOLD E. 2006. The genetics and biochemistry of floral pigments. Annual Review of Plant Biology 57: 761–780.

HAMILTON WD, and BROWN SP. 2001. Autumn tree colours as a handicap signal. Proceedings of the Royal Society of London. Biology Letters 268: 1489–1493.

HATIER JHB, and GOULD KS 2007. Black coloration in leaves of Ophiopogon planiscapus "Nigrescens". Leaf optics, chromaticity, and internal light gradients. Functional Plant Biology 34: 130–138.

HELDT HW. 2005. Plant Biochemistry. Third Edition. Elsevier Academic Press, Burlington, USA.

HOCH WA, SINGSAAS EL, and MCCOWN BH. 2003. Resorption protection. Anthocyanins facilitate nutrient recovery in autumn by shielding leaves from potentially damaging light levels. Plant Physiology 133 : 1296–1305.

HOLTON TA, BRUGLIERA F, LESTER DR, TANAKA Y, HYLAND CD, MENTING JG, LU CY, FARCY E, STEVENSON TW, and CORNISH EC 1993. Cloning and expression of cytochrome P450 genes controlling flower colour. Nature 366: 276–279.

HORTON P, and RUBAN A. 2005. Molecular design of the photosystem II light-harvesting antenna: photosynthesis and photoprotection. Journal Experimental of Botany 56: 365–373.

HU G, YALPANI N, BRIGGS SP, and JOHAL GS 1998. A porphyrin pathway impairment is responsible for the phenotype of a dominant disease lesion mimic mutant of maize. Plant Cell 10:1095–1105.

KATSUMOTO Y. et al. 2007. Engineering of the rose flavonoid biosynthetic pathway successfully generated blue-hued flowers accumulating delphinidin. Plant and Cell Physiology 48: 1589–1600.

KEVAN P, GIURFA M, and CHITTKA L. 1996. Why are there so many and so few white flowers? Trends in Plant Science 1: 280–284.

LEBEDEV N, VAN CLEVE B, ARMSTRONG G, and APEL K.1995. Chlorophyll synthesis in a Deetiolated (det340) Mutant of Arabidopsis without NADPH-Protochlorophyllide (PChlide) Oxidoreductase (POR) A and PhotoactivePChlide-F655. Plant Cell 7(12): 2081–2090.

LEE DW, and GOULD KS. 2002. Why leaves turn red. American Scientist 90: 524–531.

MARKHAM KR, BLOOR SJ, NICHOLSON R, RIVERA R, SHEMLUCK M, KEVAN PG, and MICHENER C. 2004. Black flower coloration in wild Lisianthius nigrescens: Its chemistry and ecological consequences. Zeitschrift für Naturforschung 59c: 625–630.

MARKHAM KR, GOULD KS, WINEFIELD CS, MITCHELL KA, BLOOR SJ, and BOASE MR. 2000. Anthocyanic vacuolar inclusions – their nature and significance in flower colouration. Phytochemistry 55: 327–336.

MARSAC NT. 2003. Phycobiliproteins and phycobilisomes: the early observations. Photosynthesis Reaearch 76: 197–205.

MCCLINTOCK B. 1983. The significance of the genome to challenge. In: Frangsmyr T and Lindsten J [eds.], NobelLectures Physiology or Medicine 1981– 990, 180–199. World Scientific Pub. Co., Singapore, for the Nobel Foundation.

MIMURO M. 2002. Visualization of excitation energy transfer processes in plants and algae. Photosynthesis Research 73: 133–138.

NICHOLSON R. 1999. The blackest flower in the world. Natural History 108: 60–63.

NISHIO JN. 2000. Why are higher plants green? Evolution of the higher plant photosynthetic pigment complement. Plant Cell and Environment 23: 539–548.

NIYOGI K. 2000. Safety values for photosynthesis. Current Opinion in Plant Biology 3: 455–560.

PIGA A, DEL CARO A, PINNA I, and AGABBIO M. 2005. Anthocyanin and colour evolution in naturally black table olives during anaerobic processing. Food Science and Technology38: 425–429.

RAVEN PH. 2005. Biology of Plants. Chapter 20, 452–465. Seventh Edition. W.H. Freeman Publishing, New York,USA.

SHIONO M. et al. 2005. Structure of the blue cornflower pigment. Nature 436: 791.

TANAKA A and TANAKA R. 2006. Chlorophyll metabolism. Current Opinion in Plant Biology 9: 248–255.

VAN DER KROL AR, MUR LA, BELD M, MOL JNM, and STUITJE AR. 1990. Flavonoid genes in petunia: addition of a limited Plant pigments 15 number of gene copies may lead to suppression of gene expression. Plant Cell 2: 291–299.

VERWEIJ W, SPELT C, DI SEBASTIANO GP, VERMEER J, REALE L, FERRANTI F, KOES R, and QUATTROCCHIO F. 2008. An H+P-ATPase on the tonoplast determines vacuolar pH and flower colour. Nature Cell Biology 10: 1456–1462.

VON WETTSTEIN D. 2000. Chlorophyll biosynthesis I: from analysis of the mutants to the genetic engineering of the pathway. Discoveries in Plant Biology 3: 75–93.

WINKEL-SHIRLEY B. 2001. Flavonoid biosynthesis. a colorfulmodel for genetics, biochemistry, cell biology, and biotechnology. Plant Physiology 126: 485–493.

WINKEL-SHIRLEY B. 2002. Molecular genetics and control of anthocyanin expression. Advances in Botanical Research 37: 75–88.

WINKEL-SHIRLEY B. 2006. The Biosynthesis of flavonoids. In: The Regulation of Flavonoid Biosynthesis. Springer, New York, USA.

TAIZ LZ, and EIGER E. 2006. Photosynthesis; the light reaction. In: Plant Physiology. Fourth Edition. Sinauer Associates Inc. Publishers, Sunderland, MA USA.

YAKUSHIJI H, KOBAYASHI S, GOTO-YAMAMOTO N, TAE JEONGT S, SUETA T, MITANI N, and CZUMA A. 2006. A skin color mutation of grapevine, from black-skinned pinot noir to white-skinned pinot blanc, is caused by deletion of the functional VvmybA1 allele. Bioscience, Biotechnology, and Biochemistry 70(6): 1506–1508.

YOUNG AJ, PHILIP D, RUBAN AV, HORTON P, and FRANK H. 1997. The xanthophyll cycle and carotenoid-mediated dissipation of excess excitation energy in photosynthesis. Pure and Applied Chemistry 10: 2125–2130.

ZHANG H, WANG L, DEROLES S, BENNETT R, and DAVIES K. 2006. New insight into the structures and formation of anthocyanic vacuolar inclusions in flower petals. Bio Med Central Plant Biology 6: 29–43.


Refbacks

  • There are currently no refbacks.


Creative Commons License
This work is licensed under a Creative Commons Attribution 3.0 License.