Orchids are distributed around the world, however, the factors shaping their specific distribution and habitat preferences are largely unknown. Moreover, many orchids are at risk of becoming threatened as landscapes change, sometimes declining without apparent reason. One important factor affecting plant distribution is nutrient levels in the environment. Nitrates can inhibit not only orchid growth and persistence, but also seed germination. We used in vitro axenic cultures to exactly determine the germination sensitivity of seven orchid species to nitrates and correlated this with soil properties of the natural sites and with the species' habitat preferences. We found high variation in response to nitrate between species. Orchids from oligotrophic habitats were highly sensitive, while orchids from more eutrophic habitats were almost insensitive. Sensitivity to nitrate was also associated with soil parameters that indicated a higher nitrification rate. Our results indicate that nitrate can affect orchid distribution via direct inhibition of seed germination. Nitrate levels in soils are increasing rapidly due to intensification of agricultural processes and concurrent soil pollution, and we propose this increase could cause a decline in some orchid species.

Department of Botany Faculty of Science Charles University Prague Czech Republic

Department of Experimental Plant Biology Faculty of Science Charles University Prague Czech Republic

Institut de Systématique Évolution Biodiversité Muséum national d'Histoire naturelle CNRS Sorbonne Université Paris France

Prague Botanical Garden Prague Czech Republic

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Aber J., McDowell W., Nadelhoffer K., Magill A., Berntson G., Kamakea M., McNulty S., Currie W., Rustad L., Fernandez I. (1998) Nitrogen saturation in temperate forest ecosystems. BioScience, 48, 921-934.

Alboresi A., Gestin C., Leydecker M.T., Bedu M., Meyer C., Truong H.N. (2005) Nitrate, a signal relieving seed dormancy in Arabidopsis. Plant, Cell and Environment, 28, 500-512.

Antonopoulos Z., Tsiftsis S. (2017) ATLAS of the Greek orchids, Vol. I. Mediterraneo Editions, Rethymno, Greece, p 509.

Apg I.V. (2016) An update of the Angiosperm Phylogeny Group classification for the orders and families of flowering plants: APG IV. Botanical Journal of the Linnean Society, 181, 1-20.

Arditti J. (1967) Factors affecting the germination of orchid seeds. The Botanical Review, 33, 1-97.

Arditti J., Ghani A. (2000) Numerical and physical properties of orchid seeds and their biological implications. New Phytologist, 145, 367-421.

Beyrle H., Penningsfeld F., Hockf B. (1991) The role of nitrogen concentration in determining the outcome of the interaction between Dactylorhiza incarnata (L.) Soó and Rhizoctonia sp. New Phytologist, 117, 665-672.

Beyrle H.F., Smith S.E., Franco C.M.M., Peterson R.L. (1995) Colonization of Orchis morio protocorms by a mycorrhizal fungus: effects of nitrogen nutrition and glyphosate in modifying the responses. Canadian Journal of Botany, 73, 1128-1140.

Borenstein A., Linker R., Shmulevich I., Shaviv A. (2006) Determination of soil nitrate and water content using attenuated total reflectance spectroscopy. Applied Spectroscopy, 60, 1267-1272.

Borken W., Matzner E. (2009) Reappraisal of drying and wetting effects on C and N mineralization and fluxes in soils. Global Change Biology, 15, 808-824.

Bowles M., Zettler L., Bell T., Kelsey P. (2005) Relationships between soil characteristics, distribution and restoration potential of the federal threatened eastern prairie fringed orchid, Platanthera leucophaea (Nutt.) Lindl. The American Midland Naturalist, 154, 273-285.

Callesen I., Raulund-Rasmussen K., Gundersen P., Stryhn H. (1999) Nitrate concentrations in soil solutions below Danish forests. Forest Ecology and Management, 114, 71-82.

Chase M.W., Cameron K.M., Freudenstein J.V., Pridgeon A.M., Salazar G., van den Berg C., Schuiteman A. (2015) An updated classification of Orchidaceae. Botanical Journal of the Linnean Society, 177, 151-174.

R Core Team (2014) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. http://www.R-project.org/.

Corrêa M.M., Ker J.C., Mendonça E.S., Ruiz H.A., Bastos R.S. (2003) Atributos físicos, químicos e mineralógicos de solos da região de várzeas de Souza (PB). Revista Brasileira de Ciência do Solo, 27, 311-324.

Crawford N.M. (1995) Nitrate: nutrient and signal for plant growth. The Plant Cell, 7, 859-868.

Davis B.J., Phillips R.D., Wright M., Linde C.C., Dixon K.W. (2015) Continent-wide distribution in mycorrhizal fungi: implications for the biogeography of specialized orchids. Annals of Botany, 116, 413-421.

Dechorgnat J., Nguyen C.T., Armengaud P., Jossier M., Diatloff E., Filleur S., Daniel-Vedele F. (2011) From the soil to the seeds: the long journey of nitrate in plants. Journal of Experimental Botany, 62, 1349-1359.

Delforge P. (2006) Orchids of Europe, North Africa and the Middle East, 3rd edn. A&C Black Publishers, London, UK, p 544.

Dijk E., Eck N. (1995a) Axenic in vitro nitrogen and phosphorus responses of some Dutch marsh orchids. New Phytologist, 131, 353-359.

Dijk E., Eck N. (1995b) Ammonium toxicity and nitrate response of axenically grown Dactylorhiza incarnata seedlings. New Phytologist, 131, 361-367.

Dijk E., Eck N. (1995c) Effects of mycorrhizal fungi on in vitro nitrogen response of some Dutch indigenous orchid species. Canadian Journal of Botany, 73, 1203-1211.

Dijk E., Olff H. (1994) Effects of nitrogen, phosphorus and potassium fertilization on field performance of Dactylorhiza majalis. Acta Botanica Neerlandica, 43, 383-392.

Djordjević V., Tsiftsis S., Lakušić D., Jovanović S., Stevanović V. (2016a) Factors affecting the distribution and abundance of orchids in grasslands and herbaceous wetlands. Systematics and Biodiversity, 4, 355-370.

Djordjević V., Tsiftsis S., Lakušić D., Stevanović V. (2016b) Niche analysis of orchids of serpentine and non-serpentine areas: Implications for conservation. Plant Biosystems, 150, 710-719.

Duchaufour P. (1976) Dynamics of organic matter in soils of temperate regions: its action on pedogenesis. Geoderma, 15, 31-40.

Duermeyer L., Khodapanahi E., Yan D., Krapp A. (2018) Regulation of seed dormancy and germination by nitrate. Seed Science Research, 28, 150-157.

Ellenberg H., Weber H.E., Düll R., Wirth V., Werner W., Paulißen D. (1991) Zeigerwerte von Pflanzen in Mitteleuropa. Scripta Geobotanica, 18, 1-248.

Engardt M., Simpson D., Schwikowski M., Granat L. (2017) Deposition of sulphur and nitrogen in Europe 1900-2050. Model calculations and comparison to historical observations. Tellus B: Chemical and Physical Meteorology, 69, 1328945.

Fetter J.C., Brown R.N., Görres J.H., Lee C., Amador J.A. (2012) Nitrate and phosphate leaching under turfgrass fertilized with a squid-based organic fertilizer. Water, Air, & Soil Pollution, 223, 1531-1541.

Greenwell B.M., Kabban C.M.S. (2014) Investr: an R package for inverse estimation. The R Journal, 6, 90-100. Available from http://journal.r-project.org/archive/2014-1/greenwell-kabban.pdf

Guarino R., Domina G., Pignatti S. (2012) Ellenberg’s Indicator values for the Flora of Italy: first update: Pteridophyta, Gymnospermae, and Monocotyledonae. Flora Mediterranea, 22, 197-209.

Hejcman M., Schellberg J., Pavlů V. (2010) Dactylorhiza maculata, Platanthera bifolia and Listera ovata survive N application under P limitation. Acta Oecologica, 36, 684-688.

Hew C.S., Lim L.Y., Low C.M. (1993) Nitrogen uptake by tropical orchids. Environmental and Experimental Botany, 33, 273-281.

Janssen B.H. (1996) Nitrogen mineralization in relation to C: N ratio and decomposability of organic materials. Plant and Soil, 181, 39-45.

Jeřábková K. (2006) Ecological demands and optimal management of Pseudorchis albida. Master thesis, Czech University of Life Sciences, Prague, Czech Republic.

Jersáková J., Malinová T., Jeřábková K., Dötterl S. (2011) Biological flora of the British Isles: Pseudorchis albida (L.) Á. & D. Löve. Journal of Ecology, 99, 1282-1298.

Kaiser D.R., Reinert D.J., Reichert J.M., Streck C.A., Pellegrini A. (2010) Nitrate and ammonium in soil solution in tobacco management systems. Revista Brasileira de Ciência do Solo, 34, 379-387.

Kabala C., Karczewska A., Gałka B., Cuske M., Sowiński J. (2017) Seasonal dynamics of nitrate and ammonium ion concentrations in soil solutions collected using MacroRhizon suction cups. Environmental Monitoring and Assessment, 189, 304.

Kauth P.J., Vendrame W.A., Kane M.E. (2006) In vitro seed culture and seedling development of Calopogon tuberosus. Plant Cell, Tissue and Organ Culture, 85, 91-102.

Keiluweit M., Wanzek T., Kleber M., Nico P., Fendorf S. (2017) Anaerobic microsites have an unaccounted role in soil carbon stabilization. Nature Communications, 8, 1771.

Knudson L. (1922) Nonsymbiotic germination of orchid seeds. Botanical Gazette, 73, 1-25.

Krehan M. (2009) Nährstoffansprüche Winterblatt bildender Orchideenarten in Mittel- und Südeuropa. Berichte aus den Arbeitskreisen Heimische Orchideen, 26, 88-97.

Kretzschmar H. (2013) Die Orchideen Deutschlands und angrenzender Länder, 2nd edn. Quelle & Meyer, Wiebelsheim, Germany, p 285.

Kubát K. (2010) Orchis L. - vstavač. In: Chrtek J., Kaplan Z., Štěpánková J. (Eds), Květena České republiky 8. Czech Republic, Academia, Praha, pp 524-541.

Lange O.L., Nobel P.S., Osmond C.B., Ziegler H. (2013) Physiological plant ecology III: responses to the chemical and biological environment. Springer Science & Business Media, Berlin, Germany, p 710.

Leake J. (1994) The biology of myco-heterotrophic (“saprophytic”) plants. New Phytologist, 127, 171-216.

Luna B., Moreno J.M. (2009) Light and nitrate effects on seed germination of Mediterranean plant species of several functional groups. Plant Ecology, 203, 123-135.

Manns H.R., Parkin G.W., Martin R.C. (2016) Evidence of a union between organic carbon and water content in soil. Canadian Journal of Soil Science, 96, 305-316.

Masclaux-Daubresse C., Daniel-Vedele F., Dechorgnat J., Chardon F., Gaufichon L., Suzuki A. (2010) Nitrogen uptake, assimilation and remobilization in plants: challenges for sustainable and productive agriculture. Annals of Botany, 105, 1141-1157.

Mattila E., Kuitunen M.T. (2000) Nutrient versus pollination limitation in Platanthera bifolia and Dactylorhiza incarnata (Orchidaceae). Oikos, 89, 360-366.

McCormick M.K., Jacquemyn H. (2014) What constrains the distribution of orchid populations? New Phytologist, 202, 392-400.

McCormick M.K., Taylor D.L., Juhaszova K., Burnett R.K., Whigham D.F., O’Neill J.P. (2012) Limitations on orchid recruitment: not a simple picture. Molecular Ecology, 21, 1511-1523.

McCormick M.K., Taylor D.L., Whigham D.F., Burnett R.K. (2016) Germination patterns in three terrestrial orchids relate to abundance of mycorrhizal fungi. Journal of Ecology, 104, 744-754.

Miller A.J., Fan X., Orsel M., Smith S.J., Wells D.M. (2007) Nitrate transport and signalling. Journal of Experimental Botany, 58, 2297-2306.

Moran-Zuloaga D., Dippold M., Glaser B., Kuzyakov Y. (2015) Organic nitrogen uptake by plants: reevaluation by position-specific labeling of amino acids. Biogeochemistry, 125, 359.

Mujica M.I., Saez N., Cisternas M., Manzano M., Armesto J.J., Pérez F. (2016) Relationship between soil nutrients and mycorrhizal associations of two Bipinnula species (Orchidaceae) from central Chile. Annals of Botany, 118, 149-158.

Murashige T., Skoog F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15, 473-497.

Musolff A., Schmidt C., Rode M., Lischeid G., Weise S.M., Fleckenstein J.H. (2016) Groundwater head controls nitrate export from an agricultural lowland catchment. Advances in Water Resources, 96, 95-107.

Nadarajan J., Wood S., Marks T.R., Seaton P.T., Pritchard H.W. (2011) Nutritional requirements for in vitro seed germination of 12 terrestrial, lithophytic and epiphytic orchids. Journal of Tropical Forest Science, 23, 204-212.

Näsholm T., Kielland K., Ganeteg U. (2009) Uptake of organic nitrogen by plants. New Phytologist, 182, 31-48.

Oksanen J., Blanchet F.G., Kindt R., Legendre P., Minchin P.R., O'Hara R.B., Simpson G.L., Solymos P., Stevens M.H.H., Wagner H. (2015) Vegan: Community Ecology Package. R package, version, 2.3-1. Available from: http://CRAN.R-project.org/package=vegan (accessed 5 July 2018).

Oyonarte C., Pérez-Pujalte A., Delgado G., Delgado R., Almendros G. (1994) Factors affecting soil organic matter turnover in a Mediterranean ecosystem from Sierra de Gador (Spain): an analytical approach. Communications in Soil Science and Plant Analysis, 25, 1929-1945.

Park S., Kim H.K., Kim M.S., Lee G.M., Song D.H., Jeon S.H., Kim T.S. (2016) Monitoring nitrate natural attenuation and analysis of indigenous micro-organism community in groundwater. Desalination and Water Treatment, 57, 24096-24108.

Pedersen A., Petersen B.M., Eriksen J., Hansen S., Jensen L.S. (2007) A model simulation analysis of soil nitrate concentrations - Does soil organic matter pool structure or catch crop growth parameters matter most? Ecological Modelling, 205, 209-220.

Pereira M.G., Schiavo J.A., Fontana A., Neto A.H.D., Miranda L.P.M. (2013) Caracterização e classificação de solos em uma topossequência sobre calcário na Serra da Bodoquena, MS. Revista Brasileira de Ciência do Solo, 37, 25-36.

Petrou N., Petrou M., Giannakoulias M. (2011) Orchids of Greece. Hellenic Society for the Protection of Nature, Koan Publishing House, Athens, Greece, p 320.

Pignatti S., Menegoni P., Pietrosanti S. (2005) Bioindicazione attraverso le piante vascolari. Valori di indicazione secondo Ellenberg (Zeigerwerte) per le specie della Flora d’Italia. Braun-Blanquetia, 39, 1-97.

Ponert J., Vosolsobě S., Kmecová K., Lipavská H. (2011) European orchid cultivation - from seed to mature plant. European Journal of Environmental Sciences, 1, 95-107.

Ponert J., Figura T., Vosolsobě S., Lipavská H., Vohník M., Jersáková J. (2013) Asymbiotic germination of mature seeds and protocorm development of Pseudorchis albida (Orchidaceae) are inhibited by nitrates even at extremely low concentrations. Botany-Botanique, 91, 662-670.

Procházka F., Jongepierová I., Marhold K. (2010) Gymnadenia R. Br. - pětiprstka. In: Chrtek J., Kaplan Z., Štěpánková J. (Eds), Květena České republiky 8. Czech Republic, Academia, Praha, pp 492-497.

Rasmussen H.N. (1995) Terrestrial orchids: from seed to mycotrophic plant. Cambridge University Press, Cambridge, UK, p 444.

Rasmussen H.N., Pedersen H.AE. (2011) Cypripedium calceolus germination in situ: seed longevity, and dormancy breakage by long incubation and cold winters. American Journal of Botany, 100, 2066-2073.

Rasmussen H.N., Dixon K.W., Jersáková J., Těšitelová T. (2015) Germination and seedling establishment in orchids: a complex of requirements. Annals of Botany, 116, 391-402.

Rivett M.O., Buss S.R., Morgan P., Smith J.W., Bemment C.D. (2008) Nitrate attenuation in groundwater: a review of biogeochemical controlling processes. Water Research, 42, 4215-4232.

Selosse M.A. (2014) The latest news from biological interactions in orchids: in love, head to toe. New Phytologist, 202, 337-340.

Sgarbi E., Grimaudo M., del Prete C. (2009) In vitro asymbiotic germination and seedling development of Limodorum abortivum (Orchidaceae). Plant Biosystems, 143, 114-119.

Silvertown J., Wells D.A., Gillman M., Dodd M.E., Robertson H., Lakhani K.H. (1994) Short-term effects and long-term after-effects of fertilizer application on the flowering population of green-winged orchid Orchis morio. Biological Conservation, 69, 191-197.

Singer S.F. (2012) The changing global environment. Springer Science & Business Media, Berlin, Germany, p 399.

Skopp J., Jawson M.D., Doran J.W. (1990) Steady-state aerobic microbial activity as a function of soil water content. Soil Science Society of America Journal, 54, 1619-1625.

Swarts N.D., Dixon K.W. (2009) Terrestrial orchid conservation in the age of extinction. Annals of Botany, 104, 543-556.

Swarts N.D., Dixon K.W. (2017) Conservation methods for terrestrial orchids. J. Ross Publishing, Plantation, FL, USA, p 235.

Trépanier M., Lamy M.P., Dansereau B. (2009) Phalaenopsis can absorb urea directly through their roots. Plant and Soil, 319, 95-100.

Vaněk V., Šilha J., Němeček R. (2003) The level of soil nitrate content at different management of organic fertilizers application. Plant, Soil and Environment, 49, 197-202.

van Waes J.M., Debergh P.C. (1986) In vitro germination of some Western European orchids. Physiologia Plantarum, 67, 253-261.

Wakelin S., Maclean P., Cave V., Zhou J., Grelet G.A., Whitehead D. (2019) Characterising the soil ecosystem phenotype associated with relatively low nitrate-N concentrations. Applied Soil Ecology, 142, 189-198.

Wang H.H., Wonkka C.L., Treglia M.L., Grant W.E., Smeins F.E., Rogers W.E. (2015) Species distribution modelling for conservation of an endangered endemic orchid. AoB Plants, 7, 039.

Waterman R.J., Bidartondo M.I., Stofberg J., Combs J.K., Gebauer G., Savolainen V., Pauw A. (2011) The effects of above- and belowground mutualisms on orchid speciation and coexistence. The American Naturalist, 177, E54-E68.

Weil R.R., Brady N.C. (2016) The nature and properties of soils. 15th edn. Pearson Education, Essex, UK, p 1104.

Whigham D.F., O’Neill J.P., Rasmussen H.N., Caldwell B.A., McCormick M.K. (2006) Seed longevity in terrestrial orchids - Potential for persistent in situ seed banks. Biological Conservation, 129, 24-30.

World Health Organisation (2011) Nitrate and nitrite in drinking water. Background document for development of WHO Guidelines for Drinking water Quality. http://www.who.int/water_sanitation_health/dwq/chemicals/nitratenitrite2ndadd.pdf.

Wotavová K., Balounová Z., Kindlmann P. (2004) Factors affecting persistence of terrestrial orchids in wet meadows and implications for their conservation in a changing agricultural landscape. Biological Conservation, 118, 271-279.

Yukawa T., Ogura-Tsujita Y., Shefferson R.P., Yokoyama J. (2009) Mycorrhizal diversity in Apostasia (Orchidaceae) indicates the origin and evolution of orchid mycorrhiza. American Journal of Botany, 96, 1997-2009.

Zajac M., Ujházy K., Škodová I., Kuzemko A., Borsukevych L., Danylyuk K., Duchoň M., Figura T., Kish R., Smatanová J., Turis P., Turisová I., Uhliarová E., Janišová M. (2016) Classification of semi-natural mesic grasslands in the Ukrainian Carpathians. Phytocoenologia, 46, 257-293.

Fungal symbionts may modulate nitrate inhibitory effect on orchid seed germination