Using optical fibers to measure absorption in intact conifer leaves, relative numbers of chloroplasts, and pigment content

  • Ksenija RadotićEmail author
  • Thor B. Melø
Original Paper


For investigations of ongoing processes in plants, such as photosynthesis in conifer leaves, nondestructive and noninvasive measuring techniques are needed. In this paper, a novel approach has been developed for the measurement of chloroplasts’ numbers and pigment contents in conifer leaves based on the measurements of leaf absorption spectra using optical fibers and an array spectrophotometer. To eliminate the effect of scattering on the measured absorption spectra, a strategy has been applied taking advantage of the combined use of thin optical fibers normal to the needle's longitudinal axis and the phenomenon that scattering is largest in the forward direction. The optical path in the leaf is nearly the distance between the fiber tips; thus, we were able to obtain the absorption spectrum of the pigments in situ. A effect of the measured absorption spectra, occurring due to the organization of pigments in the leaf and interaction between light and leaf interior, can be accounted for by using the so-called Duysens transformation. Using this transformation, pigment contents and the relative number of chloroplasts can be obtained from the measured absorption spectra. We applied the method to observe pigment concentrations in different stages of the greening process in the leaves of two conifer species, Taxus baccata and Picea abies. The presented method may be used to estimate changes in chloroplast number and pigment content during various phases of greening of a species and to observe differences among various species.


Taxus baccata and Norwegian spruce Optical fibers Leaf absorption spectra Duysens transformation Relative chloroplasts’ number Pigment content 



We are thankful to Prof Kalbe Razi Naqvi, who inspired this work, and to The Institute of Physics, Norwegian University of Science and Technology (NTNU), where these measurements were done. The authors also acknowledge Ministry of the Education, Science and Technology of the Republic of Serbia (Project No. 173017).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Jensen, P.E., Leister, D.: Chloroplast evolution, structure and functions. F1000Prime Rep 6, 40 (2014). CrossRefGoogle Scholar
  2. 2.
    Moss, R.A., Loomis, W.E.: Absorption spectra of leaves. i. The visible spectrum. Plant Physiol. 27, 370–391 (1952)CrossRefGoogle Scholar
  3. 3.
    Cerovic, Z.G., Masdoumier, G., Ben Ghozlen, N., Latouche, G.: A new optical leaf-clip meter for simultaneous non-destructive assessment of leaf chlorophyll and epidermal flavonoids. Physiol. Plant. 146, 251–260 (2012)CrossRefGoogle Scholar
  4. 4.
    Louis, J., Meyer, S.D., Maunoury-Danger, F., Fresneau, C., Meudec, E., Cerovic, Z.G.: Seasonal changes in optically assessed epidermal phenolic compounds and chlorophyll contents in leaves of sessile oak (Quercus petraea): towards signatures of phenological stage. Funct. Plant Biol. 36, 732–741 (2009)CrossRefGoogle Scholar
  5. 5.
    Sumanta, N., Haque, C.I., Nishika, J., Suprakash, R.: Spectrophotometric analysis of chlorophylls and carotenoids from commonly grown fern species by using various extracting solvents. Res. J. Chem. Sci. 4, 63–69 (2014)Google Scholar
  6. 6.
    Gerber, F., Marion, R., Olioso, A., Jacquemoud, S., da Luz, R., Fabre, S.: Modeling directional-hemispherical reflectance and transmittance of fresh and dry leaves from 0.4 μm to 5.7 μm with the PROSPECT-VISIR model. Remote Sens. Environ. 115, 404–414 (2011)ADSCrossRefGoogle Scholar
  7. 7.
    Haardt, H., Maske, H.: Specific in vivo absorption coefficient of chlorophyll a at 675 nm. Limnol. Oceanogr. 32(3), 608–619 (1987)ADSCrossRefGoogle Scholar
  8. 8.
    Menke, W.: Structure and chemistry of plastids. Annu. Rev. Plant Physiol. 13, 27–44 (1962)CrossRefGoogle Scholar
  9. 9.
    Merzlyak, M.N., Chikunova, O.B., Zhigalova, T.V., Naqvi, K.R.: Light absorption by isolated chloroplasts and leaves: effects of scattering and ‘packing’. Photosynth. Res. 102, 31–41 (2009)CrossRefGoogle Scholar
  10. 10.
    Duysens, L.N.M.: The flattening of the absorption spectrum of suspensions, as compared to that of solutions. Biochim. Biophys. Acta 19, 1–12 (1956)CrossRefGoogle Scholar
  11. 11.
    Naqvi, K.R.: Screening hypochromism (sieve effect) in red blood cells: a quantitative analysis. Biomed. Opt. Express 5, 1290–1295 (2014)CrossRefGoogle Scholar
  12. 12.
    Bengtson, C., Klockare, B., Larsson, S., Sundqvist, C.: The effect of phytohormones on chlorophyll (ide), protochlorophyll (ide) and carotenoid formation in greening dark grown wheat leaves. Physiol. Plant. 40, 198–204 (1977)CrossRefGoogle Scholar
  13. 13.
    Czech, A.S., Strzałka, K., Schurr, U., Matsubara, S.: Developmental stages of delayed-greening leaves inferred from measurements of chlorophyll content and leaf growth. Funct. Plant Biol. 36, 654–664 (2009)CrossRefGoogle Scholar

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© Springer Nature B.V. 2020

Authors and Affiliations

  1. 1.Institute for multidisciplinary researchUniversity of BelgradeBelgradeSerbia
  2. 2.Department of PhysicsNorwegian University of Science and Technology (NTNU)TrondheimNorway

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