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Bias in leaf dry mass estimation after oven-drying isoprenoid-storing leaves

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Abstract

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Air-drying, cooling and freezing leaf samples rich in isoprenoid content evaporates isoprenoids and consequently causes an underestimation of the leaf dry matter content.

Abstract

Foliage isoprenoid content or essential oil content is a key plant characteristic important in plant science, but also for food, cosmetic and pharmaceutical industry. Commonly, the amount of foliage chemicals is normalized with respect to leaf dry mass (content per dry mass). However, the protocols for foliage drying have received little attention. In particular, volatile and semi-volatile isoprenoids and other compounds with low boiling point may partly volatilize during drying, reducing both isoprenoid and leaf dry matter contents, thereby potentially underestimating the content of volatiles and overestimating that of non-volatile constituents. Three leaf preservation (flash freezing in liquid nitrogen, freezing at −82 and at −20 °C, and cooling at 4 °C) and two leaf desiccation (freeze-drying at −54 °C and 1.2 mbar, and air-drying at 30, 40 60, 70, 80, 100, 110, 125, 150, 200, and 300 °C) procedures were applied to replicate leaves, and the leaf dry matter content as well as the leaf isoprenoid content was measured afterwards. The results of this experiment suggested that freeze-drying method (FD) performed the best leaf isoprenoid preservation followed by FF method, whilst air-drying methods performed a dramatic isoprenoid loss in the leaf samples, as well as preserving the leaves at freezing and cooling temperatures. Leaf isoprenoid content and leaf dry to fresh mass ratio were correlated and there were evidences of leaf dry matter content (LDMC) miscalculation in isoprenoid-storing species when using traditional air-drying methods and preservation of samples, possibly related to the isoprenoid loss as well as other volatile compounds. In addition, losses of high C content volatiles and respiration of sugars and carbohydrate pyrolysis with low carbon content may differently affect the bulk leaf C content.

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Acknowledgments

This work was supported by the Internationalization Program DoRa for doctoral studies through the European Social Fund, and the Estonian Ministry of Science and Education (institutional grant IUT8-3), the European Commission through the European Regional Fund: the Estonian Center of Excellence in Environmental Adaptation (project F11100PKTF), and the Environmental Conservation and Environmental Technology R&D Programme: BioAtmos (project 8-2/T13006PKTF). We thank Dana Copolovici, Milvi Purgas and Ülle Püttsepp for helping with their lab facilities.

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Authors and Affiliations

  1. Department of Plant Physiology, Institute of Agricultural and Environmental Sciences, Estonian University of Life Sciences, Kreutzwaldi 1, 51014, Tartu, Estonia

    Miguel Portillo-Estrada, Lucian Copolovici & Ülo Niinemets

  2. Department of Biology, Centre of Excellence PLECO (Plant and Vegetation Ecology), University of Antwerp, Universiteitsplein 1, B-2610, Wilrijk, Belgium

    Miguel Portillo-Estrada

  3. Institute of Technical and Natural Sciences Research-Development of Aurel Vlaicu University, Elena Dragoi 2, 310330, Arad, Romania

    Lucian Copolovici

  4. Estonian Academy of Sciences, Kohtu 6, 10130, Tallinn, Estonia

    Ülo Niinemets

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  1. Miguel Portillo-Estrada
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  3. Ülo Niinemets
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Correspondence to Miguel Portillo-Estrada.

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Communicated by H. Rennenberg.

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Portillo-Estrada, M., Copolovici, L. & Niinemets, Ü. Bias in leaf dry mass estimation after oven-drying isoprenoid-storing leaves. Trees 29, 1805–1816 (2015). https://doi.org/10.1007/s00468-015-1262-8

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