Foliar Carbon and Nitrogen Content and Stable Isotopic Composition of Selected Philippine Flora
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Philippine Journal of Science 150 (S1): 539-550, Special Issue on Biodiversity ISSN 0031 - 7683 Date Received: 04 Oct 2020 Foliar Carbon and Nitrogen Content and Stable Isotopic Composition of Selected Philippine Flora Roland V. Rallos*, Gerald P. Dicen, Andrea Luz G. Nery, and John Leonard R. Labides Agriculture Research Section Philippine Nuclear Research Institute Department of Science and Technology (DOST-PNRI) Commonwealth Ave., Diliman, Quezon City National Capital Region 1101 Philippines The elemental carbon and nitrogen content, as well as the stable isotopic composition of plants, can provide information on their nutrient dynamics and physiological characteristics. With applications ranging from paleoecology to ecological modeling, this information is critical in biodiversity conservation and management especially for highly dynamic ecosystems such as the Philippines’. Here, we determined the carbon and nitrogen content and their stable isotopic ratios in selected Philippine flora across different classifications and photosynthetic pathways. Fully developed leaves from different species of grasses, succulents, shrubs, and trees were collected and analyzed using isotope ratio mass spectrometry for carbon and nitrogen concentration and stable isotopic composition. Our results showed that trees had the highest carbon and nitrogen content, while grasses that utilized the C4 photosynthetic pathway were most efficient in terms of nutrient utilization as evidenced by their high C/N ratios. Foliar stable carbon isotopic composition of the surveyed Philippine flora was an excellent measure for distinguishing among photosynthetic pathways. The stable nitrogen isotopic composition was not distinct across plant classifications and showed indications of being sensitive to environmental factors, thus limiting its use for phylogenetic tracing. Our findings thus indicate that foliar carbon and nitrogen content and stable isotopic composition provide insights that cannot be easily achieved with other measurements. Keywords: carbon, isotope ratio mass spectrometry, nitrogen, photosynthesis, Philippine flora, stable isotopes INTRODUCTION development, and biochemical functioning. In addition, plant C and N are also critical for ecological processes Carbon (C) and nitrogen (N) are essential elements in all such as energy flow and nutrient cycling, while the C/N living organisms, and their metabolism is closely linked ratio is an indicator of N use efficiency (NUE) and an (Raven et al. 2004). For plants, C and N are crucial for input variable for many ecological and ecosystem models fundamental cellular activities (Coruzi and Bush 2001; (Zhang et al. 2020). Zheng 2009), which make them essential for plant growth, Information on plant isotopic compositions is essential in *Corresponding Author: [email protected] predicting future patterns of many ecosystem processes 539 Philippine Journal of Science Rallos et al.: Stable Isotopic Composition Vol. 150 No. S1, Special Issue on Biodiversity of Philippine Flora and functions (e.g. primary productivity, C and N characterizing plant functional groups and photosynthetic sequestration, nutrient fluxes). Likewise, it is also used pathways (Descolas-Gros and Schölzel 2007; Cernusak to trace the plant's geographic origin and is an important et al. 2013), stable isotopes analysis can also be used to tool in forensic settings from an ecological perspective determine the diet of animals (Dalerum and Angerbjörn (Chesson et al. 2018). The stable isotopic composition 2005; Hopkins and Ferguson 2012) to make sense of their of plants depends on the source of their nutrition and movement and habitat (Hobson 1999, 2007; Bowen et al. the conditions involved in their uptake. To some extent, 2005) and is, therefore, useful in biodiversity conservation information on the processes involved in the transport and management (Walter et al. 2014). In the soil, being a and transformation of plant-derived materials is preserved huge repository of dead organic materials, stable isotopes in their stable isotope ratios (Marshall et al. 2007). For are a useful proxy for studying both long-term and short- this reason, the stable isotopes in organic materials term biogeochemical dynamics in the environment. For – in conjunction with their corresponding elemental example, the stable C and N isotope ratios can be used composition – can be considered as natural environmental to provide a mechanistic understanding of the long- tracers (Michener and Lajtha 2007). Thus, stable isotope term C storage in mangroves (Dicen et al. 2019) and in measurement has become one of the indispensable tools elucidating the effect of burning on soil organic matter used in ecological studies. biodegradability (Dicen et al. 2020). However, in all the applications, prior data on the isotopic composition of Leaf stable C isotope ratios vary based on how different relevant flora must be available as they are central to the photosynthetic pathways discriminate against heavier inter-trophic cycling of elements, especially C and N. C atoms (Farquhar et al. 1989; Cernusak et al. 2013). The C3 photosynthetic pathway tends to discriminate Studying tropical ecosystems, which are extremely diverse heavier CO2 molecules as compared with the C4 and dynamic, such as the Philippines’ can greatly benefit photosynthetic pathway – as a factor of the ratio of its from stable isotopes measurement. Consolidated stable intracellular and ambient CO2 concentration, water use isotope data from this study and other similar reports can efficiency, coordination between stomatal conductance then be used to generate isotope landscape, or isoscapes, and photosynthesis, and leaf area adjustment (Cernusak which is a very useful tool for data interpretation in et al. 2013). ecological and forensic studies. However, no study has been done on the stable isotope composition of diverse On the other hand, stable N isotope ratios in plants are Philippine flora to date. In this study, we determined reflective of the short-term dynamics in N cycling within the C and N content and their stable isotopic ratios in ecosystems (Ometto et al. 2006; Craine et al. 2015). For selected Philippine flora, across different classifications instance, microbial activities spell the difference in leaf and photosynthetic pathways. In addition, we explored stable isotope composition between temperate and tropical patterns that emerged from the relationships among plants (Martinelli et al. 1999), as well as the difference measured parameters. between leguminous and non-leguminous species (Boddey et al. 2000). Spatial, temporal, and climatic factors can also have subtle but otherwise significant effects on the stable C and N isotopic composition of plants (Liu et al. MATERIALS AND METHODS 2008; Ma et al. 2012; Xu et al. 2017). Earlier studies indicated that the measurement of C and N stable isotopic About 10–15 fully developed leaf samples of selected compositions in plants would provide an understanding Philippine flora – classified into grass (n = 17), shrub of the cycling of these nutrients. For example, Garten et (n = 22), succulent (n = 24), and tree (n = 25) species – al. (2008) and Craine et al. (2009) have shown that with were obtained around the Greater Manila Area (GMA), increasing N availability and potential N mineralization Philippines. GMA is the contiguous developed zones through natural N supply or N availability gradients, plant surrounding Metro Manila, including the adjacent stable N isotope ratios have increased. Moreover, stable C provinces of Cavite, Laguna, Batangas, Rizal, Bulacan, and N ratios have also been used to investigate the rate of and Pampanga (Figure 1). Grass species were further C sequestration and N cycling under different vegetations classified according to their photosynthetic pathways, and land use (Baldos and Rallos 2019). with n = 4 for C4 grasses and n = 13 for C3 grasses. Soils in the study area were classified under different series Variation in environmental conditions and other with varying fertility levels. The soil in Pampanga area biogeochemical processes can also have a strong influence is under Angeles series with low fertility level, Quingua over the natural abundances of stable C and N isotopes series in Bulacan area, Novaliches series and San Manuel in plants. Knowledge of the different factors that cause series in Metro Manila area, Tagaytay series in Cavite area, variations in stable isotope ratios can, thus, be explored and Calumpang series in Laguna area – which all have for several ecological research applications. Aside from 540 Philippine Journal of Science Rallos et al.: Stable Isotopic Composition Vol. 150 No. S1, Special Issue on Biodiversity of Philippine Flora Figure 1. Map of the sampled area. moderate fertility levels (PhilRice 2010, 2013; Carating by the presence of the catalyst Cr2O3, which was used as et al. 2014). The study area’s mean annual precipitation packing material in the combustion reactor. (MAP) and mean annual temperature (MAT) based on the WorldClim database (Fick and Hijmans 2017) were 2047 Following combustion, the resulting gaseous products mm and 27.3 ºC, respectively. Leaf samples were collected were allowed to pass through a reduction furnace from the middle part of tree and shrub canopies. Sampled maintained at 650 °C. The reduction column contains trees and shrubs have fully good vigor while sampled copper where the excess oxygen was scavenged by succulents and grasses were not foraged obviously by the reduced