Calcium : magnesium ratio affects environmental stress sensitivity in the serpentine-endemic Alyssum inflatum (Brassicaceae)
Rasoul Ghasemi A F , Zohreh Zare Chavoshi B , Robert S. Boyd C and Nishanta Rajakaruna D E
+ Author Affiliations
- Author Affiliations
A Department of Biology, Payame Noor University, 19395-4697, Tehran, I.R. of Iran.
B Department of Biology, Payame Noor University, 81395-671 Isfahan, I.R. of Iran.
C Department of Biological Sciences, Auburn University, Auburn, AL 36849, USA.
D College of the Atlantic, Bar Harbor, ME 04609, USA.
E Unit for Environmental Sciences and Management, North-West University, Potchefstroom, 2520, South Africa.
F Corresponding author. Email: rasoul.ghasemi@es.isfpnu.ac.ir
Australian Journal of Botany 63(2) 39-46 https://doi.org/10.1071/BT14235
Submitted: 8 September 2014 Accepted: 22 December 2014 Published: 8 April 2015
Abstract
Plants endemic to serpentine soils are adapted to harsh environmental conditions typical of those soils, particularly, low (<1) calcium (Ca) : magnesium (Mg) ratios. We compared survival of two perennial Alyssum species native to Iran under experimental manipulations of Ca : Mg ratio, including when Ca : Mg ratio was varied under conditions of high ammonium concentration and heat stress. Alyssum inflatum is a serpentine endemic capable of nickel (Ni) hyperaccumulation, whereas A. lanceolatum is found on non-serpentine soils and is not known to hyperaccumulate Ni. We grew plants of both species under four Ca : Mg ratios (0.4, 2, 20, 40) and tested survival when plants were exposed to elevated ammonium levels (0, 1 and 4 mM) and heat stress (control conditions vs a 5-h 36°C treatment daily for 5 days). Alyssum lanceolatum was more tolerant of Ca : Mg ratio variation (100% survival in all treatments), whereas A. inflatum survival was maximum at Ca : Mg = 2, reduced at Ca : Mg = 0.4, and very low for Ca : Mg ratios of 20 and 40. Alyssum lanceolatum also tolerated ammonium and heat stress, whereas survival of A. inflatum declined at higher Ca : Mg ratios when subjected to both stresses. We conclude that at higher Ca : Mg ratios, the serpentine endemic has reduced tolerance for these environmental stresses and may be more susceptible to human-driven climate change-associated stressors than the non-serpentine species.
Additional keywords: global climate change, heat stress, local adaptation, nitrogen deposition, serpentine endemism.
References
Alexander EB, Coleman RG, Keeler-Wolf T, Harrison S (2007) ‘Serpentine geoecology of western North America.’ (Oxford University Press: New York)Anacker BL (2011) Phylogenetic patterns of endemism and diversity. In ‘Serpentine: the evolution and ecology of a model system’. (Eds SP Harrison, N Rajakaruna) pp. 49–79. (University of California Press: Berkeley, CA)
Anacker BL (2014) The nature of serpentine endemism. American Journal of Botany 101, 219–224.
| The nature of serpentine endemism.Crossref | GoogleScholarGoogle Scholar | 24509800PubMed |
Anacker BL, Whittall J, Goldberg E, Harrison SP (2011) Origins and consequences of serpentine endemism in the California flora. Evolution 65, 365–376.
| Origins and consequences of serpentine endemism in the California flora.Crossref | GoogleScholarGoogle Scholar | 20812977PubMed |
Asemaneh T, Ghaderian SM, Baker AJM (2007) Responses to Mg/Ca balance in an Iranian serpentine endemic plant, Cleome heratensis (Capparaceae), and a related non-serpentine species, C. foliolosa. Plant and Soil 293, 49–59.
| Responses to Mg/Ca balance in an Iranian serpentine endemic plant, Cleome heratensis (Capparaceae), and a related non-serpentine species, C. foliolosa.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXltlaktLo%3D&md5=bca185ccc736925703c5c5e974b56b6fCAS |
Bradshaw HD (2005) Mutations in CAX1 produce phenotypes characteristic of plants tolerant to serpentine soils. New Phytologist 167, 81–88.
| Mutations in CAX1 produce phenotypes characteristic of plants tolerant to serpentine soils.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXmtl2nsbk%3D&md5=ff3a321b1b1d69e44199e8f63e37b18bCAS | 15948832PubMed |
Brady KU, Kruckeberg AR, Bradshaw HD (2005) Evolutionary ecology of plant adaptation to serpentine soils. Annual Review of Ecology Evolution and Systematics 36, 243–266.
| Evolutionary ecology of plant adaptation to serpentine soils.Crossref | GoogleScholarGoogle Scholar |
Brooks RR (1987) ‘Serpentine and its vegetation: a multidisciplinary approach.’ (Dioscorides: Portland, OR)
Damschen EI, Harrison S, Ackerly DD, Fernandez-Going BM, Anacker BL (2012) Endemic plant communities on special soils: early victims or hardy survivors of climate change? Journal of Ecology 100, 1122–1130.
| Endemic plant communities on special soils: early victims or hardy survivors of climate change?Crossref | GoogleScholarGoogle Scholar |
Demarty M, Morvan C, Thellier M (1984) Calcium and the cell wall. Plant, Cell & Environment 7, 441–448.
| Calcium and the cell wall.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL2cXlsVOktrc%3D&md5=8190ff42f971b72d62a662c17408abdcCAS |
Epstein E, Bloom AJ (2004) ‘Mineral nutrition of plants: principles and perspectives.’ 2nd edn. (Sinauer Associates: Sunderland, MA)
Eskelinen A, Harrison S (2014) Exotic plant invasions under enhanced rainfall are constrained by soil nutrients and competition. Ecology 95, 682–692.
| Exotic plant invasions under enhanced rainfall are constrained by soil nutrients and competition.Crossref | GoogleScholarGoogle Scholar | 24804452PubMed |
Fenn LB, Feagley S (1999) Review of beneficial uses of calcium and ammonium salts for stimulating plant growth and metabolite translocation. Communications in Soil Science and Plant Analysis 30, 2627–2641.
| Review of beneficial uses of calcium and ammonium salts for stimulating plant growth and metabolite translocation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXnsVaksrc%3D&md5=7b725d94b829bc5ea9aa10c997c52983CAS |
Fenn LB, Hasanein B, Burks CM (1995) Calcium-ammonium effects on growth and yield of small grains. Agronomy Journal 87, 1041–1046.
| Calcium-ammonium effects on growth and yield of small grains.Crossref | GoogleScholarGoogle Scholar |
Fernandez-Going BM (2014) Climate change and the future of edaphic floras. In ‘Plant ecology and evolution in harsh environments’. (Eds N Rajakaruna, RS Boyd, TB Harris) pp. 297–311. (Nova Science Publishers: New York)
Fernandez-Going BM, Harrison S (2013) Effects of experimental water addition depend on grassland community characteristics. Plant Ecology 214, 777–786.
| Effects of experimental water addition depend on grassland community characteristics.Crossref | GoogleScholarGoogle Scholar |
Fernandez-Going BM, Harrison SP, Anacker BL, Safford HD (2013) Climate interacts with soil to produce beta diversity in Californian plant communities. Ecology 94, 2007–2018.
| Climate interacts with soil to produce beta diversity in Californian plant communities.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC2c3ht12qug%3D%3D&md5=324918ffdebef43f4329b8b08f1166c2CAS | 24279272PubMed |
Fowler D, Coyle M, Skiba U, Sutton MA, Cape JN, Reis S, Sheppard LJ, Jenkins A, Grizzetti B, Galloway JN, Vitousek P, Leach A, Bouwman AF, Butterbach-Bahl K, Dentener F, Stevenson D, Amann M, Voss M (2013) The global nitrogen cycle in the twenty-first century. Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences 368,
| The global nitrogen cycle in the twenty-first century.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXht1Wkt77O&md5=8d2731fb6319dac2ba42001c3b59771dCAS | 23713128PubMed |
Galloway JN, Schlesinger WH, Levy H, Michaels A, Schnoor JL (1995) Nitrogen fixation: anthropogenic enhancement-environmental response. Global Biogeochemical Cycles 9, 235–252.
| Nitrogen fixation: anthropogenic enhancement-environmental response.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXmtlWlt7g%3D&md5=ef7dcab4a09459ab2e41bf2b9dff60a8CAS |
Ghasemi R, Ghaderian SM (2009) Responses of two populations of an Iranian nickel-hyperaccumulating serpentine plant, Alyssum inflatum Nyar., to substrate Ca/Mg quotient and nickel. Environmental and Experimental Botany 67, 260–268.
| Responses of two populations of an Iranian nickel-hyperaccumulating serpentine plant, Alyssum inflatum Nyar., to substrate Ca/Mg quotient and nickel.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhtV2qs7jM&md5=83866dafb6e62a4131fc1d9b8ddae0f6CAS |
Ghasemi R, Ghaderian SM, Krämer U (2009a) Accumulation of nickel in trichomes of a nickel hyperaccumulator plant, Alyssum inflatum. Northeastern Naturalist 16, 81–92.
| Accumulation of nickel in trichomes of a nickel hyperaccumulator plant, Alyssum inflatum.Crossref | GoogleScholarGoogle Scholar |
Ghasemi R, Ghaderian SM, Krämer U (2009b) Interference of nickel with copper and iron homeostasis contributes to metal toxicity symptoms in the nickel hyperaccumulator plant Alyssum inflatum. New Phytologist 184, 566–580.
| Interference of nickel with copper and iron homeostasis contributes to metal toxicity symptoms in the nickel hyperaccumulator plant Alyssum inflatum.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhsVOgs7jN&md5=e435886cc684c36a062ea35a005a7d20CAS | 19691676PubMed |
Ghasemi R, Chavoshi ZZ, Boyd RS, Rajakaruna N (2014) A preliminary study of the role of nickel in enhancing flowering of the nickel hyperaccumulating plant Alyssum inflatum Nyár. (Brassicaceae). South African Journal of Botany 92, 47–52.
| A preliminary study of the role of nickel in enhancing flowering of the nickel hyperaccumulating plant Alyssum inflatum Nyár. (Brassicaceae).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXmsVCjtLc%3D&md5=ad9bcc0684412b2463393bf2d34d4aa2CAS |
Going BM, Hillerislambers J, Levine JM (2009) Abiotic and biotic resistance to grass invasion in serpentine annual plant communities. Oecologia 159, 839–847.
| Abiotic and biotic resistance to grass invasion in serpentine annual plant communities.Crossref | GoogleScholarGoogle Scholar | 19139921PubMed |
Gong M, van der Luit AH, Knight MR, Trewavas AJ (1998) Heat-shock-induced changes in intracellular Ca2+ level in tobacco seedlings in relation to thermotolerance. Plant Physiology 116, 429–437.
| Heat-shock-induced changes in intracellular Ca2+ level in tobacco seedlings in relation to thermotolerance.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXkslGitQ%3D%3D&md5=499fe96398df1575a8099868bf588367CAS |
Gotelli NJ, Ellison EM (2002) Nitrogen deposition and extinction risk in northern pitcher plant, Sarracenia purpurea. Ecology 83, 2758–2765.
| Nitrogen deposition and extinction risk in northern pitcher plant, Sarracenia purpurea.Crossref | GoogleScholarGoogle Scholar |
Gryschko R, Kuhnle R, Terytze K, Breuer J, Stahr K (2005) Soil extraction of readily soluble heavy metals and As with 1 M NH4NO3-solution. Journal of Soils and Sediments 5, 101–106.
| Soil extraction of readily soluble heavy metals and As with 1 M NH4NO3-solution.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXpslSmu7s%3D&md5=48e60ba6d6a40d9e3e125033c1071888CAS |
Harrison SP, Rajakaruna N (Eds) (2011) ‘Serpentine: the evolution and ecology of a model system.’ (University of California Press: Berkeley, CA)
Harrison SP, Damschen E, Going BM (2009) Climate gradients, climate change, and special edaphic floras. Northeastern Naturalist 16, 121–130.
Hedge I, Rechinger KH (1968) Cruciferae. In ‘Flora Iranica’. (Ed. KH Rechinger) pp. 1–372. (Akademische Druck-u., Verlagsanstalt: Graz, Austria)
Hülber K, Dirnböck T, Kleinbauer I, Willner W, Dullinger S, Karrer G, Mirtl M (2008) Long-term impacts of nitrogen and sulphur deposition on forest floor vegetation in the northern limestone Alps, Austria. Applied Vegetation Science 11, 395–404.
| Long-term impacts of nitrogen and sulphur deposition on forest floor vegetation in the northern limestone Alps, Austria.Crossref | GoogleScholarGoogle Scholar |
Jarvis MC (1984) Structure and properties of pectin gels in plant cell walls. Plant, Cell & Environment 7, 153–164.
Jarvis MC, Briggs SPH, Knox JP (2003) Intercellular adhesion and cell separation in plants. Plant, Cell & Environment 7, 153–164.
Jiang Y, Huang B (2001) Effects of calcium on antioxidant activities and water relations associated with heat tolerance in two cool‐season grasses. Journal of Experimental Botany 52, 341–349.
| Effects of calcium on antioxidant activities and water relations associated with heat tolerance in two cool‐season grasses.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXjtVaju70%3D&md5=de0d4d954e7372783273bb857c65648dCAS | 11283179PubMed |
Kay KM, Ward KL, Watt LR, Schemske DW (2011) Plant speciation. In ‘Serpentine: the evolution and ecology of a model system’. (Eds SH Harrison, N Rajakaruna) pp. 181–199. (University of California Press: Berkeley, CA)
Kazakou E, Dimitrakopoulos PG, Baker AJM, Reeves RD, Troumbis AY (2008) Hypotheses, mechanisms and trade-offs of tolerance and adaptation to serpentine soils: From species to ecosystem level. Biological Reviews of the Cambridge Philosophical Society 83, 495–508.
Klein JD, Ferguson IB (1987) Effect of high temperature on calcium uptake by suspension-cultured pear fruit cells. Plant Physiology 84, 153–156.
| Effect of high temperature on calcium uptake by suspension-cultured pear fruit cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL2sXksFOltbk%3D&md5=6d04d2dedadca77c5ad9bb1283f33d09CAS | 16665390PubMed |
Kruckeberg AR (1984) ‘California serpentines: flora, vegetation, geology, soils, and management problems.’ (University of California Press: Berkeley, CA)
Lee JA (1998) The calcicole–calcifuge problem revisited. Advances in Botanical Research 29, 1–30.
| The calcicole–calcifuge problem revisited.Crossref | GoogleScholarGoogle Scholar |
Madhok OP (1965) Magnesium nutrition of Helianthus annuus L. and Helianthus bolanderi Gray subspecies exilis Heiser. PhD Dissertation. University of Washington, Seattle, WA.
Madhok OP, Walker RB (1969) Magnesium nutrition of two species of sunflower. Plant Physiology 44, 1016–1022.
| Magnesium nutrition of two species of sunflower.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaF1MXltVSqsro%3D&md5=4df248c256683534171760b6c77c0ebbCAS | 16657149PubMed |
Marschner H (2011) ‘Mineral nutrition of plants.’ 3rd edn. (Academic Press: San Diego, CA)
Mergemann H, Sauter M (2000) Ethylene induces epidermal cell death at the site of adventitious root emergence in rice. Plant Physiology 124, 609–614.
| Ethylene induces epidermal cell death at the site of adventitious root emergence in rice.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXnsF2rsL4%3D&md5=e45d97b26af3f0d97f8968868644f688CAS | 11027711PubMed |
Moore KA, Elmendorf SC (2011) Plant competition and facilitation in systems with strong environmental gradients. In ‘Serpentine: the evolution and ecology of a model system’. (Eds SP Harrison, N Rajakaruna) pp. 223–236. (University of California Press: Berkeley, CA)
Myers N, Mittermeier RA, Mittermeier CG, da Fonseca GAB, Kent J (2000) Biodiversity hotspots for conservation priorities. Nature 403, 853–858.
| Biodiversity hotspots for conservation priorities.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXhs1Olsr4%3D&md5=ad1dc1d3f067155f97ed4e9930487835CAS | 10706275PubMed |
O’Dell RE, Claassen VP (2006) Serpentine and nonserpentine Achillea millefolium accessions differ in serpentine substrate tolerance and response to organic and inorganic amendments. Plant and Soil 279, 253–269.
| Serpentine and nonserpentine Achillea millefolium accessions differ in serpentine substrate tolerance and response to organic and inorganic amendments.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XhsVaqsr8%3D&md5=2f3aa2492d14d86b3ddebecf861a8e7aCAS |
O’Dell RE, Rajakaruna N (2011) Intraspecific variation, adaptation, and evolution. In ‘Serpentine: the evolution and ecology of a model system’. (Eds SP Harrison, N Rajakaruna) pp. 97–137. (University of California Press: Berkeley, CA)
Palm ER, Van Volkenburgh E (2014) Physiological adaptations of plants to serpentine soils. In ‘Plant ecology and evolution in harsh environments’. (Eds N Rajakaruna, RS Boyd, TB Harris) pp. 129–147. (Nova Science Publishers Inc.: New York)
Palm ER, Brady K, Van Volkenburgh E (2012) Serpentine tolerance in Mimulus guttatus does not rely on exclusion of magnesium. Functional Plant Biology 39, 679–688.
| Serpentine tolerance in Mimulus guttatus does not rely on exclusion of magnesium.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38Xht1Smu7bL&md5=7a45652d32174ff5318afcda12ee17d8CAS |
Phoenix GK, Emmet BA, Britton AJ, Caporn SJM, Dise NB, Helliwell R, Jones L, Leake JR, Leith ID, Sheppard LJ, Sowerby A, Pilkington MG, Rowe EC, Ashmore MR, Power SA (2012) Impacts of atmospheric nitrogen deposition: responses of multiple plant and soil parameters across contrasting ecosystems in long-term field experiments. Global Change Biology 18, 1197–1215.
| Impacts of atmospheric nitrogen deposition: responses of multiple plant and soil parameters across contrasting ecosystems in long-term field experiments.Crossref | GoogleScholarGoogle Scholar |
Pievani T (2014) The sixth mass extinction: anthropocene and the human impact on biodiversity. Rendiconti Lincei 25, 85–93.
| The sixth mass extinction: anthropocene and the human impact on biodiversity.Crossref | GoogleScholarGoogle Scholar |
Rajakaruna N, Boyd RS (2008) The edaphic factor. In ‘Encyclopedia of ecology. Vol. 2’. (Eds SE Jorgensen, B Fath) pp. 1201–1207. (Elsevier: Oxford, UK)
Rajakaruna N, Siddiqi MY, Whitton J, Bohm BA, Glass ADM (2003) Differential responses to Na+/K+ and Ca2+/Mg2+ in two edaphic races of the Lasthenia californica (Asteraceae) complex: a case for parallel evolution of physiological traits. New Phytologist 157, 93–103.
| Differential responses to Na+/K+ and Ca2+/Mg2+ in two edaphic races of the Lasthenia californica (Asteraceae) complex: a case for parallel evolution of physiological traits.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXht1SiurY%3D&md5=3413142b42bee2a01b6540d156164e43CAS |
Rajakaruna N, Harris TB, Alexander EB (2009) Serpentine geoecology of eastern North America: a review. Rhodora 111, 21–108.
| Serpentine geoecology of eastern North America: a review.Crossref | GoogleScholarGoogle Scholar |
Reeves RD, Baker AJM, Borhidi A, Berazaín R (1999) Nickel hyperaccumulation in the serpentine flora of Cuba. Annals of Botany 83, 29–38.
| Nickel hyperaccumulation in the serpentine flora of Cuba.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXht1ajsL4%3D&md5=6f1477942217c49230dce50fdbfcb529CAS |
Safford HD, Viers JH, Harrison SP (2005) Serpentine endemism in the California flora: a database of serpentine affinity. Madrono 52, 222–257.
| Serpentine endemism in the California flora: a database of serpentine affinity.Crossref | GoogleScholarGoogle Scholar |
Sambatti JBM, Rice KJ (2007) Functional ecology of ecotypic differentiation in the Californian serpentine sunflower (Helianthus exilis). New Phytologist 175, 107–119.
| Functional ecology of ecotypic differentiation in the Californian serpentine sunflower (Helianthus exilis).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXot1Cks70%3D&md5=e03557f70f1c69d7cf3bb2df9812bd49CAS |
SAS Institute (2005) ‘StatView 5.0.’ (Thomson–Brooks/Cole: Belmont, CA)
Schneider HE, Allen EB (2012) Effects of elevated nitrogen and exotic plant invasion on soil seed bank composition in Joshua Tree National Park. Plant Ecology 213, 1277–1287.
| Effects of elevated nitrogen and exotic plant invasion on soil seed bank composition in Joshua Tree National Park.Crossref | GoogleScholarGoogle Scholar |
Stocker TF, Qin D, Plattner GK, Tignor M, Allen SK, Boschung J, Nauels A, Xia Y, Bex V, Midgley PM (2013) ‘Climate change 2013: the physical basis. Contribution of the Working Group I to the fifth assessment report of the Intergovernmental Panel on Climate Change.’ (Cambridge University Press: New York)
Tibbetts RA, Smith JAC (1993) Vacuolar accumulation of calcium and its interaction with magnesium availability. In ‘The vegetation of ultramafic (serpentine) soils: proceedings of the first international conference on serpentine ecology’. (Eds AJM Baker, J Proctor, RD Reeves) pp. 367–373. (Intercept: Andover, UK)
Vallano DM, Selmants PC, Zavaleta ES (2012) Simulated nitrogen deposition enhances performance of an exotic grass relative to native serpentine grassland competitors. Plant Ecology 213, 1015–1026.
| Simulated nitrogen deposition enhances performance of an exotic grass relative to native serpentine grassland competitors.Crossref | GoogleScholarGoogle Scholar |
Van der Ent A, Repin R, Sugau J, Wong KM (2014) ‘The ultramafic flora of Sabah: an introduction to the plant diversity on ultramafic soils.’ (Natural History Publications (Borneo): Kota Kinabalu, Borneo)
Wallace A, Jones MB, Alexander GV (1982) Mineral composition of native woody plants growing on serpentine soil in California. Soil Science 134, 42–44.
| Mineral composition of native woody plants growing on serpentine soil in California.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL38XlsVeks7c%3D&md5=fe4b5599978af2e415a6274fabf82464CAS |
Warren R, Price J, Fischlin A, de la Nava Santos S, Midgley G (2011) Increasing impacts of climate change upon ecosystems with increasing global mean temperature rise. Climatic Change 106, 141–177.
| Increasing impacts of climate change upon ecosystems with increasing global mean temperature rise.Crossref | GoogleScholarGoogle Scholar |
White PJ, Broadley MR (2003) Calcium in plants. Annals of Botany 92, 487–511.
| Calcium in plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXoslOmtLg%3D&md5=6a82ffe50050e2d57a53c777d71f085cCAS | 12933363PubMed |
Zar JH (1996) ‘Biostatistical analysis.’ (Prentice-Hall: Englewood Cliffs, NJ)
