Barthová, V.

Nauš, J.

Vacek, K.

Zrůst, J.

Institute of Mathematics AS CR, v. v. i.

Acta Universitatis Carolinae. Mathematica et PhysicaKarel Vacek; J. Nauš; V. Barthová; J. ZrůstFluorescence, absorption and photosynthetic rate measurements of potato leaves(Solanum tuberosum L.) during their vegetationsActa Universitatis Carolinae. Mathematica et Physica, Vol. 20 (1979), No. 2, 3--9Persistent URL: http://dml.cz/dmlcz/142436Terms of use:© Univerzita Karlova v Praze, 1979Institute of Mathematics of the Academy of Sciences of the Czech Republic provides access todigitized documents strictly for personal use. Each copy of any part of this document mustcontain these Terms of use.This paper has been digitized, optimized for electronic delivery and stampedwith digital signature within the project DML-CZ: The Czech DigitalMathematics Library http://project.dml.cz1979 ACTA UNIVERSITATTS CAROLINAE - MATHEMATICA ET PHYSICA VOL. 20. NO. 2. Pag. 3—9 Fluorescence, Absorption and Photosynthetic Rate Measurements of Potato Leaves (Solanum tuberosum L.) During Their Vegetation K. VACEK, J. N A U S and V. BARTHOVA Department of Chemical Physics, Charles University, Prague*) J. Z R t i S T Institut for Potato Research, Havlickuv Brod**) Received 27 October 1978 A certain correlation between ontogenetic dependences of spectral properties (absorption, fluorescence and polarized fluorescence) and the photosynthetic rate of in vivo potato leaves has been found. The spectral properties reflect the state of the leaf pigment system which can be slightly different for different potato varieties. Bwjia o6HapyaceHa B3aHM0CBH3b Meac/Ty OHToreHera-iecKOH 3aBHCHMOCTbK> cneKTpajn>m>rx CBOHCTB (norJiomeHHe, (JmyopecueHiuw H nojiapH3amui (JmyopecuemniH) H CKOpocrwo <j>OTOCHHTe3a jmcTbeB KapTo4>ejiH. B cneKTpajibHbix CBoňCTBax OTpaacaeTCH cocroHHHe nurMeHTHOH CHcreMbi jiHCTbeB, KOTOpoe MO»ceT oTjnnaTbCH fljiui pa3Hbix pa3H0BHAH0CTeH KapTo4)ejn>Horo pacreHHi!. Byla zjištěna určitá korelace mezi ontogenetickou závislostí spektrálních vlastností (absorp­ce, fluorescence a polarizace fluorescence) a rychlostí fotosyntézy listů brambor in vivo. Ve spek­trálních vlastnostech se odráží stav pigmentové soustavy listů, který může být poněkud odlišný u různých odrůd. 1. Introduct ion Relatively many papers have been devoted to the absorption and luminescence measurements of plant pigments in the material in vivo (e.g. [1], [2], [3], [4]), but only few of them have studied the change of the mentioned properties during the ontogenesis (e.g. recently [5], [6]). Although it is difficult to gain quantitative con­clusions from such measurements, mainly due to a number of disturbing effects (scattering, non-homogenous pigment distribution in the tissue in vivo), these measurements can provide some qualitative information concerning, for example, relative amounts of different pigments, the changes of average pigment concentration or its aggregation during the ontogenesis of the leaf etc. The information about the * 121 16 Praha 2, Ke Karlovu 3, Czechoslovakia. ** 580 01 Havlíčkův Brod, Czechoslovakia. correlation between physical and photosynthetic properties of the leaves and the final products of photosynthesis (i.e. tubers) can contribute to the application aspects of these experiments. 2. Material and Methods Four potato varieties Blanik, Cira, Nora and Radka were chosen for optical studies during a vegetation period of 4 weeks in the greenhouse. All spectroscopic measurements were carried out on the terminal leaflet of the fourth leaf (counted from the top of the plant) at room temperature. Fluorescence was excited and detected at the upper side of the leaf. The experiments were three times repeated during the years 1975 — 7. The relative absorption was measured by modified spectrophotometer Specord UV-VIS (neutral filters), the fluorescence was measured by means of laboratory spectrofluorometer (excitation prism monochromator SPM 1 and measuring grating monochromator SPM 2) with a cooled photomultiplier M12FD35 connected to registration potentiometer. Monochromators and photomultiplier are products of Carl Zeiss, Jena. The experimental arrangement is shown in Fig. 1. The absorption and fluorescence spectra are not corrected for the scattering and reabsorption. 7 6 8 вґ Fig. 1. Design of the laboratory-made spectrofluorometer. 1 — Excitation high pressure mercury lamp HBO 500; 2 — monochromator SPM1 (Carl Zeiss, Jena); 3 — interference filter; 4 — sample holder; 5 — Schott filter RG 2; 6 — monochromator SPM 2 (Carl Zeiss, Jena); 7 — photomultiplier M 12 FD 35; 8 — recorder EZ 4 (Laboratorní přístroje, Praha) The rate of photosynthesis was measured by the method of Bartos et al. [7] adapted for potato plants by Zriist [8]. Two series of experiments were performed. In the first series all varieties were planted simultaneously, in the second one the plants were grown in such a way that the beginning of the bud growth was approximately the same for all varieties. The results of the first and second series are shown in Fig. 4 - 5 and in Fig. 6, respectively. The basic character of dependences and all conclusions are the same for both series of experiments. The first measurement was performed at the beginning of bud formation. 3. Results Fig. 2 shows a typical spectrum of the relative absorption of green potato leaf at room temperature. The main absorption maxima are numbered from the short-to the longwavelength side of the spectrum. The spectrum reflects the presence of main photosynthetic pigments - chlorophylls and carotenoids. Both the optical density ratio of the 2nd and the 7th bands (D2/.D7) and also the position of the 7th maximum A7 decreased during the studied period of ontogenesis. There was found no unambiguous difference among the varieties in these dependences. 600 650 700 XCntтü 25 23 21 19 17 15 13 ЪСЮ3ст~Ь Fig. 2. Relative absorption spectrum of the terminal leaflet of the fourth potato leaf from the top (variety Blanik, two weeks after commencement of bud formation) at room temperature Fig. 3 presents the fluorescence spectra of leaf at room tempetature excited by the light of different wavelengths. The fluorescence band intensity ratio ljl2 de­creases with the change of the excitation wavelength from 435, 465 nm to 578 nm. For Aex = 578 nm the first fluorescence maximum is shifted to longer wavelengths in comparison with that excited by Aex = 435 and 465 nm. These changes were proved for all measurements. For example the ljl2 ontogenetic dependence for .4ex = 435 nm and for four potato varieties is shown in Fig. 4. The ljl2 ratio for all three excitation wavelengths slightly increases in the followed time interval. In mutual comparison of different varieties Blanik and Radka show an extreme behaviour. For AeY = 435 and 465 nm \ex-Jt35nm \ex-5F8r.m \ex-^65nm 7A0 A EnmJ Fig. 3. Fluorescence spectrum of the terminal leaflet of the fourth potato leaf from the top (variety Blanik, three weeks after the full plant emergence) at room temperature for three different excitation wavelengths the band ratio ljl2 is, in the average, the greatest one and the smallest one for the variety Blanik and Radka, respectively (see Fig. 4). For Xtx = 578 nm the opposite situation was found. Fig. 5 illustrated the ontogenetic dependence of fluorescence polarization degree P of the first fluorescence band (684 nm) excited by Aex = 435 nm at room temperature. The decrease is not monotonous but "sinusoidally modulated". The average absolute value of P is the greatest one and the smallest one for the variety Blanik and Radka, respectively. In the Fig. 6 the corresponding ontogenetics dependence of photosynthetic rate is shown. In the course of ontogenesis the photosynthetic rate decreases. Again the varieties Blanik and Radka behave extremely (maximum mean rate for Blanik and minimum for Radka). A greet natural dispersity of plant material and hence a great statistical error is demonstrated in all measured values. Furthermore, the influence of a systematic error (the influence of weather, illumination etc.) cannot also" be excluded. Due to 4 t (weeks) to the Fig. 4. The change of the fluorescence intensity ratio of the shortwavelength (684 nm) ._ . longwavelength (735 nm) band Ix/I2 during the studied ontogenesis interval. Fourth leaf from the top, room temperature. Curves: 1 — variety Blanik, 2 — variety Cira, 3 — variety Nora, d — vnrifitv RaHk-a 4  a iety adk  ,M 1 2 3 4 t(wcch») Fig. 5. The change of the fluorescence polarization degree P during the ontogenesis of potato plant. Fourth leaf from the top, room temperature. Curves: 1 — variety Blanik, 2 — variety Cira, 3 — variety Nora, 4 — variety Radka the above reasons, the accuracy of measurements is lowered, and only qualitative conclusions can be drawn from our data. t (weeks) Fig. 6. The change of the photosynthetic rate of the potato leaf during the studied ontogenesis interval. Curves: 1 — variety Blanik, 2 — variety Cira, 3 — variety Nora, 4 — variety Radka 4. Discussion The relative increase of the second fluorescence band intensity and simultaneous shift of the first band to the longer wavelengths with increasing excitation wavelength (Fig. 3) were observed also in a model system (chlorophyll a in polymer matrix [9]). These facts belong to the general properties of higly concentrated pigment systems, and they reflect a series of phenomena due to low distance of pigment molecules (different pigment forms, aggregated states, electron excitation energy transfer, reabsorption and, in leaves, also number of chloroplasts in a cell). During the studied part of potato plant ontogenesis the local concentration of chlorophyll a (fluorescent species) decreases and hence the ratio ljl2 and the absolute value of polarization degree P increase. Simultaneously the relative amount of other pigments in comparison with chlorophyll a decreases (decrease of D2lD7). The auxiliary pigments (chlorophyll b and carotenoids) as well as chlorophyll a contribute to the second absorption band, the 7th band represents mainly the absorption of chlorophyll a. A systematic study of spectral parameters and photosynthetic rate also revealed the distinct behaviour of the potato varieties Blanik and Radka. In our opinion the variety Blanik shows some features of a more effective photosynthetic system (a greater photosynthetic rate, lower local chlorophyll a concentration) than other varieties, the variety Radka represents the opposite case. The found correlation of the spectral and photosynthetic parameters of the leaf indicates the possibility of the use of nondestructive optical and luminescence methods for the determination of the photosynthetic leaf activity in different plants. 8 References [1] GULJAJEV B. A., LITVIN F. F. : Biofizika 22, 845 (1967). [2] GURINOVIČ G. P., SEVČENKO A. N. , SOLOVJEV K. N.: Spektroskopija chlorofilla i rodstvennych sojedin nij, izd. „Nauka i těchnika", Minsk, 1968. [3] FRENCH C S., BROWN J. S., LAWRENCE M. C : Plant Physiol. 49, 421 (1972). [4] ŠESTÁК Z.: Photosynthetica 11, 367 (1977). [5] GOEDHEER J. C , GUUAJEV B. A.: Proceedings of Eur. Biophys. Congress 4, 43 (1971). [6] VACEК K., VAVŘINEC E., BARTHOVÁ V., N A U Š J., ZRŮST J.: Abstracts of the Tagung der Sektion Physiologie deг E A P R (7.—12. 6. 1976) Göttingen. [7] BARTOŠ J., КUBÍN Š., ŠETLÍК L: Biologia Plantarum (Praha) 2, 201 (1960). [8] ZRŮST J.: Rostlinná výгoba (Praha) 19, 243 (1973). [9] VACEК K., VAVŘINEC E., ČERNÁ V., VINDUŠКOVÁ O., NAUŠ J., LOКAJ P.: Studia biophysica 44 155 (1974). 

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Fluorescence, absorption and photosynthetic rate measurements of potato leaves (Solanum tuberosum L.) during their vegetations

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Fluorescence, absorption and photosynthetic rate measurements of potato leaves (Solanum tuberosum L.) during their vegetations

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