Assessment of factual measurement times for chlorophyll-a fluorescence in rubber (Hevea brasiliensis) clones




Abstract. Cahyo AN, Murti RH, Putra ETS, Nuringtyas TR, Fabre D, Montoro P. 2021. Assessment of factual measurement times for chlorophyll-a fluorescence in rubber (Hevea brasiliensis) clones. Biodiversitas 22: 3470-3477. Chlorophyll-a fluorescence is widely used to determine the stress tolerance levels of some plant species. Measurement of chlorophyll-a fluorescence is accurate if the duration of dark adaptation is well defined and optimal Fv/Fm (maximum quantum yield of primary photochemistry/photosynthesis) is achieved. Leaf clips are usually used to darken the leaf prior to measurement. This procedure takes time and limits the use of chlorophyll-a fluorescence parameter in high-throughput screening of genetic populations. This study aimed to determine the most suitable time for the chlorophyll-a fluorescence measurement. This study was carried out on several rubbers (Hevea brasiliensis Müll. Arg.) clones and consisted of two steps. The first step was conducting the measurements at five different times at night: at 7.30, 8.00, 8.30, 9.00, and 9.30 p.m. The second step was conducting the measurements at daytime, which consisted of two factors. The first factor was the measurement time, which was divided into two categories: 7.30 a.m. and 1.30 p.m. The second factor was the duration of dark adaptation using leaf clips, which consisted of nine levels: 0, 15, 30, 45, 60, 75, 90, 105, and 120 min. Additional treatment (measurement at 9.00 p.m. without using leaf clips to darken the leaf) was used as a control. This study revealed that a dark adaptation time of two hours after the sunset was long enough for the rubber leaves chlorophyll-a fluorescence transient to be measured without using leaf clips for the dark adaptation. If the measurement is conducted by 7.30 a.m., the clone RRIM 600, GT1, and SP 217 required 15 min of dark adaptation, whereas clone PB 260 required 60 min of dark adaptation. Furthermore, measurement of chlorophyll-a fluorescence in the afternoon is not recommended due to the potentially high microclimate fluctuation.


An F, Kong L, Gong L, Wang Z, Lin W. 2011. Involvement of rootstocks and their hydraulic conductance in the drought resistance of grafted rubber trees. Afr J Biotechnol 10 (51): 10393–10404.
Aoki S, Nagaoka M. 1989. Measurement of Heat Sensitivity in Cucumber Leaves by Chlorophyll Fluorescence. Jpn Agric Res Q 22 (4): 297–301.
Azhar A, Sathornkich J, Rattanawong R, Kasemsap P. 2013. Responses of Chlorophyll Fluorescence, Stomatal Conductance, and Net Photosynthesis Rates of Four Rubber (Hevea brasiliensis) Genotypes to Drought. Adv Mater Res 844: 11–14.
Boureima S, Oukarroum A, Diouf M, Cisse N, Van Damme P. 2012. Screening for drought tolerance in mutant germplasm of sesame (Sesamum indicum) probing by chlorophyll a fluorescence. Environ Exp Bot 81: 37–43.
Cahyo AN, Murti RH, Putra ETS, Nuringtyas TR, Fabre D, Montoro P. 2020. SPAD-502 and atLEAF CHL PLUS values provide good estimation of the chlorophyll content for Hevea brasiliensis Müll. Arg. leaves. Menara Perkeb 88 (1): 1–8.
Çiçek N, Arslan Ö. 2015. Are The Photosynthetic Performance Indexes and The Drought Factor Index Satisfactory Selection Criterion for Stress? Fresenius Environ Bull 24 (11): 4190–4198.
Falqueto AR, da Silva Júnior RA, Gomes MTG, Martins JPR, Silva DM, Partelli FL. 2017. Effects of drought stress on chlorophyll a fluorescence in two rubber tree clones. Sci Hortic 224: 238–243.
FT GREEN LLC. 2019. atLEAF CHL PLUS Chlorophyll Meter User Manual 0131-50 Ver 1.1.
Gholamin R, Khayatnezhad M. 2011. The effect of end season drought stress on the chlorophyll content, chlorophyll fluorescence parameters and yield in maize cultivars. Sci Res Essays 6 (25): 5351–5357.
Govindjee. 1995. Sixty-Three Years Since Kautsky: Chlorophyll a Fluorescence. Funct Plant Biol 22 (2): 131–160.
Hansatech Instrument Ltd. 2018. Handy PEA+ and Pocket PEA System Manual.
Inonu I, Budianta D, Umar M, Yakup, Wiralag AYA. 2011. Rubber clone response on irrigation frequency in sand tailing medium post thin mining. J Agron Indones 39 (2): 131–136.
Jedmowski C, Ashoub A, Momtaz O, Brüggemann W. 2015. Impact of Drought, Heat, and Their Combination on Chlorophyll Fluorescence and Yield of Wild Barley (Hordeum spontaneum). J Bot 2015: 1–9.
Kalaji HM, Jajoo A, Oukarroum A, Brestic M, Zivcak M, Samborska IA, Cetner MD, ?ukasik I, Goltsev V, Ladle RJ. 2016. Chlorophyll a fluorescence as a tool to monitor physiological status of plants under abiotic stress conditions. Acta Physiol Plant 38 (102): 1–11.
Kalaji HM, Schansker G, Ladle RJ, Goltsev V, Bosa K, Allakhverdiev SI, Brestic M, Bussotti F, Calatayud A, D?browski P, Elsheery NI, Ferroni L, Guidi L, Hogewoning SW, Jajoo A, Misra AN, Nebauer SG, Pancaldi S, Penella C, Poli D, Pollastrini M, Romanowska-Duda ZB, Rutkowska B, Serôdio J, Suresh K, Szulc W, Tambussi E, Yanniccari M, Zivcak M. 2014. Frequently asked questions about in vivo chlorophyll fluorescence: practical issues. Photosynth Res 122 (2): 121–158.
Krishan B. 2017. Assessment of drought tolerance in few clones of natural rubber (Hevea brasiliensis) under dry hot climate of Odisha, India. J Exp Biol Agric Sci 5 (1): 106–110.
LI-COR Inc. 1990. LI-189 Quantum/Radiometer/Photometer Operating Manual.
Maxwell K, Johnson GN. 2000. Chlorophyll fluorescence—a practical guide. J Exp Bot 51 (345): 659–668.
Ögren E, Sjöström M. 1990. Estimation of the effect of photoinhibition on the carbon gain in leaves of a willow canopy. Planta 181: 560–557.
Oukarroum A, Madidi SE, Schansker G, Strasser RJ. 2007. Probing the responses of barley cultivars (Hordeum vulgare L.) by chlorophyll a fluorescence OLKJIP under drought stress and re-watering. Environ Exp Bot 60 (3): 438–446.
Oukarroum A, Schansker G, Strasser RJ. 2009. Drought stress effects on photosystem I content and photosystem II thermotolerance analyzed using Chl a fluorescence kinetics in barley varieties differing in their drought tolerance. Physiol Plant 137 (2): 188–199.
SAS Institute Inc. 2002. The SAS System for Windows. SAS Institute Inc., Cary, NC, USA.
Sasaki H, Li Z, Tsuji K, Oda M. 1994. Factors Affecting the Measurement of Chlorophyll a Fluorescence in Cucumber Leaves. Jpn Agric Res Q 28 (4): 242–246.
Schansker G, Tóth SZ, Strasser RJ. 2006. Dark recovery of the Chl a fluorescence transient (OJIP) after light adaptation: The qT-component of non-photochemical quenching is related to an activated photosystem I acceptor side. Biochim Biophys Acta 1757: 787–797.
Schansker G, Yuan Y, Strasser RJ. 2008. Chl a Fluorescence and 820 nm Transmission Changes Occurring During a Dark-to-Light Transition in Pine Needles and Pea Leaves: A Comparison. In: Allen JF, Gantt E, Golbeck JH, Osmond B. (eds.) Energy from the Sun. Springer, Dordrecht.
Stirbet A, Govindjee. 2011. On the relation between the Kautsky effect (chlorophyll a fluorescence induction) and Photosystem II: Basics and applications of the OJIP fluorescence transient. J Photochem Photobiol B 104 (1–2): 236–257.
Stirbet A, Lazár D, Kromdijk J, Govindjee. 2018. Chlorophyll a fluorescence induction: Can just a one-second measurement be used to quantify abiotic stress responses? Photosynthetica 56 (1): 86–104.
Strasser RJ, Tsimilli-Michael M, Srivastava A. 2004. Analysis of the Chlorophyll a Fluorescence Transient. In: Papageorgiou GC, Govindjee (eds.) Chlorophyll a Fluorescence. Springer Netherlands, Dordrecht.
Strauss AJ, Krüger GHJ, Strasser RJ, Heerden PDRV. 2006. Ranking of dark chilling tolerance in soybean genotypes probed by the chlorophyll a fluorescence transient O-J-I-P. Environ Exp Bot 56 (2): 147–157.
Thomas, Lasminingsih M. 1994. Respons of some rubber clones on drought. War Perkaretan 12 (3): 1–4.
Weng JH. 2006. Underestimate of PS2 efficiency in the field due to high leaf temperature resulting from leaf clipping and its amendment. Photosynthetica 44 (3): 467–470.
Zushi K, Matsuzoe N. 2017. Using of chlorophyll a fluorescence OJIP transients for sensing salt stress in the leaves and fruits of tomato. Sci Hortic 219: 216–221.

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