Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

Non-destructive measurement of chlorophyll b : a ratios and identification of photosynthetic pathways in grasses by reflectance spectroscopy

Katharina Siebke A B and Marilyn C. Ball A C
+ Author Affiliations
- Author Affiliations

A Division of Plant Sciences, Research School of Biology, Australian National University, Canberra, ACT 0200, Australia.

B Present address: Heinz Walz GmbH, Eichenring 6, 91090 Effeltrich, Germany.

C Corresponding author. Email: marilyn.ball@anu.edu.au

This paper originates from a presentation at the 1st International Plant Phenomics Symposium, Canberra, Australia, April 2009.

Functional Plant Biology 36(11) 857-866 https://doi.org/10.1071/FP09201
Submitted: 29 July 2009  Accepted: 28 September 2009   Published: 5 November 2009

Abstract

Equations for non-destructive determination of chlorophyll b : a ratios in grasses were developed from reflectance spectra of intact leaves of barley (Hordeum vulgare L.) and two barley mutants: clorina f2, which lacks chlorophyll b and clorina f104, which has a low chlorophyll b content. These plants enabled separation of effects of chlorophyll composition on reflectance spectra due to differential light absorption by chlorophylls a and b and to measure the effects of chlorophyll b on the contribution of fluorescence emitted by chlorophyll a to the reflectance spectra. Indices developed from these data were then tested on growth chamber-grown leaves from six C3 and 17 C4 grass species (7 NAD-ME and 10 NADP-ME subtypes). We used the chlorophyll b : a ratio because the data were less skewed than the chlorophyll a : b ratio. The best index for determination of the chlorophyll b : a ratio utilised wavelengths affected by chlorophyll absorbance: [R626 – 0.5 (R603 + R647)]/[R552– R626]. The chlorophyll b : a ratio was significantly lower in the C4 than C3 grasses, but was not sufficient in itself to separate these two functional groups. However, because of differences in fluorescence characteristics, C3 and C4 species could be distinguished by an index based on wavelengths affected by chlorophyll fluorescence: [R696 to 709/R545 to 567].

Additional keywords: C3-photosynthesis, C4-photosynthesis, leaf pigments, remote sensing.


Acknowledgements

We thank Professors Terry Caelli, Paul Kriedemann and Jochen Zeil and Drs Adam Gilmore and Adrienne Nicotra for critical discussion of previous versions of the manuscript, Dr Oula Ghannoum for providing C4 grass seeds, Dr David Barker for pigment determinations, Drs Luigi Ranzullo, Glenn Newnham and Susanne Thulin for inspiring discussions.


References


Boardman NK (1977) Comparative photosynthesis of sun and shade plants. Annual Review of Plant Physiology 28, 355–377.
CrossRef | CAS |

Bossman B, Knoetzel J, Jansson S (1997) Screening of clorina mutants of barley (Hordeum vulgare L.) with antibodies against light-harvesting proteins of PSI and PSII: absence of specific antenna proteins. Photosynthesis Research 40, 287–294.

Clark RN, Roush TL (1984) Reflectance spectroscopy: quantitative analysis techniques for remote sensing applications. Journal of Geophysical Research 89, 6329–6340.
CrossRef | CAS |

Curran PJ, Dungan JL, Peterson DL (2001) Estimating the foliar biochemical concentration of leaves with reflectance spectrometry. Testing the Kokaly and Clark methodologies. Remote Sensing of Environment 76, 349–359.
CrossRef |

Davies AMC, Fearn T (2002) Doing it faster and smarter (Lesson 6 of matrix algebra). Spectroscopy Europe 14, 24–26.

Edwards G , Walker D (1983) ‘C3, C4: mechanisms, and cellular and environmental regulation of photosynthesis.’ (Blackwell Scientific Publications: Oxford, UK)

Ghannoum O, Evans JR, Chow WS, Andrews TJ, Conroy JP, von Caemmerer S (2005) Faster Rubisco is the key to superior nitrogen-use efficiency in NADP-malic enzyme relative to NAD-malic enzyme C4 grasses. Plant Physiology 137, 638–650.
CrossRef | CAS | PubMed |

Gilmore AM, Yamamoto HY (1991) Resolution of lutein and zeaxanthin using a lightly carbon-loaded C18 high performance liquid chromatographic column. Journal of Chromatography. A 543, 137–145.
CrossRef | CAS |

Gilmore AM, Itoh S, Govindjee (2000) Global spectral-kinetic analysis of room temperature chlorophyll a fluorescence from light-harvesting antenna mutants of barley. Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences 355, 1371–1384.
CrossRef | CAS | PubMed |

Gitelson AA, Merzlyak MN (1994) Quantitative estimation of chlorophyll-a using reflectance spectra: experiments with autumn chestnut and maple leaves. Journal of Photochemistry and Photobiology. B, Biology 22, 247–252.
CrossRef | CAS |

Gitelson AA, Merzlyak MN (1997) Remote estimation of chlorophyll content in higher plants. International Journal of Remote Sensing 18(12), 2691–2697.
CrossRef |

Gitelson AA, Zur Y, Chivkunova OB, Merzlyak MN (2002) Assessing carotenoid content in plant leaves with reflectance spectroscopy. Photochemistry and Photobiology 75, 272–281.
CrossRef | CAS | PubMed |

Grant L (1987) Diffuse and specular characteristics of leaf reflectance. Remote Sensing of Environment 22, 309–322.
CrossRef |

Green BR, Durnford DG (1996) The chlorophyll–carotenoid proteins of oxygenic photosynthesis. Annual Review of Plant Physiology and Plant Molecular Biology 47, 685–714.
CrossRef | CAS | PubMed |

Harrison MA, Nemson JA, Melis A (1993) Assembly and composition of the chlorophyll ab light harvesting complex of barley (Hordeum vulgare L.): immunochemical analysis of chlorophyll b-less and chlorophyll b-deficient mutants. Photosynthesis Research 38, 141–151.
CrossRef | CAS |

Hatch MD (1987) C4 photosynthesis: a unique blend of modified biochemistry, anatomy and ultrastructure. Biochimica et Biophysica Acta 895, 81–106.
CAS |


Heber U (1969) Conformational changes of chloroplasts induced by illumination of leaves in vivo. Biochimica et Biophysica Acta 180, 302–319.
CrossRef | CAS | PubMed |

Heinze I, Pfündel E, Hühn M, Dau H (1997) Assembly of light harvesting complexes II (LHC-II) in the absence of lutein – a study on the a-carotenoid-free mutant C-2A′-34 of the green alga Scenedesmus obliquus. Biochimica et Biophysica Acta 1320, 188–194.
CrossRef | CAS |

Kitajima K, Hogan KP (2003) Increases of chlorophyll a/b ratios during acclimation of tropical woody seedlings to nitrogen limitation and high light. Plant, Cell & Environment 26(6), 857–865.
CrossRef | PubMed |

Knoetzel J, Simpson D (1991) Expression and organisation of antenna proteins in the light- and temperature-sensitive barley mutant clorina f104. Planta 185, 111–123.
CrossRef | CAS |

Kokaly RF, Clark RN (1999) Spectroscopic determination of leaf biochemistry using band-depth analysis of absorption features and stepwise multiple linear regression. Remote Sensing of Environment 67, 267–287.
CrossRef |

Kühlbrandt W, Wang DA, Fujiyoshi Y (1994) Atomic model of plant light-harvesting complex. Nature 367, 614–621.
CrossRef | PubMed |

Lee AL-C, Thornber JP (1995) Analysis of the pigment stoichiometry of pigment-protein complexes from barley (Hordeum vulgare). Plant Physiology 107, 565–574.
CrossRef | CAS | PubMed |

Lichtenthaler H (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods in Enzymology 148, 350–382.
CrossRef | CAS |

Morosinotto T, Caffarri S, Dall’Osto L, Bassi R (2003) Mechanistic aspects of the xanthophyll dynamics in higher plant thylakoids. Physiologia Plantarum 119, 347–354.
CrossRef | CAS |

Nicotra AB, Hofmann M, Siebke K, Ball MC (2003) Spatial patterning of pigmentation in evergreen leaves in response to freezing stress. Plant, Cell & Environment 26, 1893–1904.
CrossRef | CAS |

Paulsen H (1995) Chlorophyll a/b-binding proteins. Photochemistry and Photobiology 62(3), 367–382.
CrossRef | CAS |

Pfündel E, Pfeffer M (1997) Modification of photosystem I light harvesting of bundle-sheath chloroplasts occurred during the evolution of NADP-malic enzyme C4 photosynthesis. Plant Physiology 114, 145–152.
PubMed |


Pfündel E, Nagel E, Meister A (1996) Analyzing the light energy distribution in the photosynthetic apparatus of C4 plants using highly purified mesophyll and bundle sheath thylakoids. Plant Physiology 112, 1055–1070.
PubMed |


Porra RJ, Thompson WA, Kriedemann PE (1989) Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochimica et Biophysica Acta 975, 384–394.
CrossRef | CAS |

Rühle W, Wild A (1979) The intensification of absorbance changes in leaves by light-dispersion. Planta 146, 551–557.
CrossRef |

Sage RF, Kubien DS (2007) The temperature response of C3 and C4 photosynthesis. Plant, Cell & Environment 30, 1086–1106.
CrossRef | CAS | PubMed |

von Caemmerer S (2000) ‘Biochemical models of leaf photosynthesis.’ (CSIRO Publishing: Collingwood, Australia)

Yoder BJ, Daley LS (1989) Development of a visible spectroscopic method for determining chlorophyll a and b in vivo in leaf samples. Spectroscopy 5, 44–50.

Zarco-Tejada PJ, Miller JR, Mohammed GH, Noland TL (2000) Chlorophyll fluorescence effects on vegetation apparent reflectance: I. Leaf-level measurements and model simulation. Remote Sensing of Environment 74(3), 582–595.
CrossRef |








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