Advertisement

Plant Molecular Biology

, Volume 57, Issue 5, pp 709–730 | Cite as

Temporal evolution of the Arabidopsis oxidative stress response

  • Ramamurthy MahalingamEmail author
  • Nigam Shah
  • Alexandra Scrymgeour
  • Nina Fedoroff
Article

Abstract

We have carried out a detailed analysis of the changes in gene expression levels in Arabidopsis thaliana ecotype Columbia (Col-0) plants during and for 6 h after exposure to ozone (O3) at 350 parts per billion (ppb) for 6 h. This O3 exposure is sufficient to induce a marked transcriptional response and an oxidative burst, but not to cause substantial tissue damage in Col-0 wild-type plants and is within the range encountered in some major metropolitan areas. We have developed analytical and visualization tools to automate the identification of expression profile groups with common gene ontology (GO) annotations based on the sub-cellular localization and function of the proteins encoded by the genes, as well as to automate promoter analysis for such gene groups. We describe application of these methods to identify stress-induced genes whose transcript abundance is likely to be controlled by common regulatory mechanisms and summarized our findings in a temporal model of the stress response.

Keywords

Arabidopsis CLENCH gene ontology ozone microarrays stress response 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Supplementary material

supp.pdf (14.4 mb)
Supplementary material

References

  1. Aro, E.M., Ohad, I. 2003Redox regulation of thylakoid protein phosphorylationAntioxid. Redox Signal.55567CrossRefPubMedGoogle Scholar
  2. Azuaje, F. 2002A cluster validity framework for genome expression dataBioinformatics18319320CrossRefPubMedGoogle Scholar
  3. Baker, B., Zambryski, P., Staskawicz, B., Dinesh-Kumar, S.P. 1997Signaling in plant-microbe interactionsScience276726733CrossRefPubMedGoogle Scholar
  4. Baker, C.J., Orlandi, E.W. 1995Active oxygen in plant pathogenesisAnnu. Rev. Phytopathol.33299321CrossRefGoogle Scholar
  5. Baker, C.J., Orlandi, E.W. 1999Sources and effect of reactive oxygen species in plantsGilbert, D.L.Colton, C.A. eds. Reactive Oxygen Species in Biological Systems: An Interdisciplinary ApproachKluwerNew York481501Google Scholar
  6. Baldasano, J.M., Valera, E., Jimenez, P. 2003Air quality data from large citiesSci. Total Environ.307141165CrossRefPubMedGoogle Scholar
  7. Barnes, D., Mayfield, S.P. 2003Redox control of posttranscriptional processes in the chloroplastAntioxid. Redox. Signal.58994CrossRefPubMedGoogle Scholar
  8. Beier, M., Hoheisel, J.D. 1999Versatile derivatisation of solid support media for covalent bonding on DNA-microchipsNucl. Acids Res.2719701977CrossRefPubMedGoogle Scholar
  9. Bell, E., Creelman, R.A., Mullet, J.E. 1995A chloroplast lipoxygenase is required for wound-induced jasmonic acid accumulation in ArabidopsisProc Natl Acad Sci USA9286758679PubMedGoogle Scholar
  10. Ben-Hur, A., Horn, D., Siegelmann, H.T., Vapnik, V. 2002Support vector clusteringJ. Mach. Learn. Res.2125137CrossRefGoogle Scholar
  11. Bowler, C., Fluhr, R. 2000The role of calcium and activated oxygens as signals for controlling cross-toleranceTrends Plant Sci.5241246CrossRefGoogle Scholar
  12. Brazma, A., Hingamp, P., Quackenbush, J., Sherlock, G., Spellman, P., Stoeckert, C., Aach, J., Ansorge, W., Ball, C.A., Causton, H.C., Gaasterland, T., Glenisson, P., Holstege, F.C.P., Kim, I.F., Markowitz, V., Matese, J.C., Parkinson, H., Robinson, A., Sarkans, U., Schulze-Kremer, S., Stewart, J., Taylor, R., Vilo, J., Vingron, M. 2001Minimum information about a microarray experiment (MIAME) – toward standards for microarray dataNat. Genet.29365371Google Scholar
  13. Campbell, P., Braam, J. 1998Co- and/or post-translational modifications are critical for TCH4 XET activityPlant J.15553561CrossRefPubMedGoogle Scholar
  14. Carlberg, I., Hansson, M., Kieselbach, T., Schroder, W.P., Andersson, B., Vener, A.V. 2003A novel plant protein undergoing light-induced phosphorylation and release from the photosynthetic thylakoid membranesProc. Natl. Acad. Sci. USA100757762CrossRefPubMedGoogle Scholar
  15. Chan, C.S., Guo, L., Shih, M.C. 2001Promoter analysis of the nuclear gene encoding the chloroplast glyceraldehyde-3-phosphate dehydrogenase B subunit of Arabidopsis thalianaPlant Mol. Biol.46131141CrossRefPubMedGoogle Scholar
  16. Conklin, P.L., Last, R.L. 1995Differential accumulation of antioxidant mRNAs in Arabidopsis thaliana exposed to ozonePlant Physiol.109203212CrossRefPubMedGoogle Scholar
  17. Creelman, R.A., Mullet, J.E. 1997Biosynthesis and action of jasmonates in plantsAnnu. Rev. Plant Physiol. Plant Mol. Biol.48355381CrossRefPubMedGoogle Scholar
  18. Cruz Castillo, M., Martinez, C., Buchala, A., Metraux, J.P., Leon, J. 2004Gene-specific involvement of beta-oxidation in wound-activated responses in ArabidopsisPlant Physiol.1358594CrossRefPubMedGoogle Scholar
  19. Desikan, R., Mackerness, S.A.H., Hancock, J.T., Neill, S.J. 2001Regulation of the Arabidopsis transcriptome by oxidative stressPlant Physiol.127159172CrossRefPubMedGoogle Scholar
  20. Despres, C., Chubak, C., Rochon, A., Clark, R., Bethune, T., Desveaux, D., Fobert, P.R. 2003The Arabidopsis NPR1 disease resistance protein is a novel cofactor that confers redox regulation of DNA binding activity to the basic domain/leucine zipper transcription factor TGA1Plant Cell1521812191CrossRefPubMedGoogle Scholar
  21. Dievart, A., Clark, S.E. 2004LRR-containing receptors regulating plant development and defenseDevelopment131251261CrossRefPubMedGoogle Scholar
  22. Duval, M., Hsieh, T.F., Kim, S.Y., Thomas, T.L. 2002Molecular characterization of AtNAM: a member of the Arabidopsis NAC domain superfamilyPlant Mol. Biol.50237248CrossRefPubMedGoogle Scholar
  23. Eisen, M.B., Spellman, P.T., Brown, P.O., Botstein, D. 1998Cluster analysis and display of genome-wide expression patternsProc Natl. Acad. Sci. USA951486314868CrossRefPubMedGoogle Scholar
  24. Escoubas, J.M., Lomas, M., LaRoche, J., Falkowski, P.G. 1995Light intensity regulation of cab gene transcription is signaled by the redox state of the plastoquinone poolProc. Natl. Acad. Sci. USA921023710241PubMedGoogle Scholar
  25. Eulgem, T., Rushton, P.J., Robatzek, S., Somssich, I.E. 2000The WRKY superfamily of plant transcription factorsTrends Plant Sci.5199206CrossRefPubMedGoogle Scholar
  26. Fan, W., Dong, X. 2002In vivo interaction between NPR1 and transcription factor TGA2 leads to salicylic acid-mediated gene activation in ArabidopsisPlant Cell1413771379CrossRefPubMedGoogle Scholar
  27. Fotopoulos, V., Gilbert, M.J., Pittman, J.K., Marvier, A.C., Buchanan, A.J., Sauer, N., Hall, J.L., Williams, L.E. 2003The monosaccharide transporter gene, AtSTP4, and the cell-wall invertase, Atbetafruct1, are induced in Arabidopsis during infection with the fungal biotroph Erysiphe cichoracearumPlant Physiol.132821829CrossRefPubMedGoogle Scholar
  28. Gillissen, B., Burkle, L., Andre, B., Kuhn, C., Rentsch, D., Brandl, B., Frommer, W.B. 2000A new family of high-affinity transporters for adenine, cytosine, and purine derivatives in ArabidopsisPlant Cell12291300CrossRefPubMedGoogle Scholar
  29. Gray, J.C., Sullivan, J.A., Wang, J.H., Jerome, C.A., MacLean, D. 2003Coordination of plastid and nuclear gene expressionPhilos. Trans. R Soc. Lond. B Biol. Sci.358135144; discussion 144–145CrossRefPubMedGoogle Scholar
  30. Greenberg, J.T. 1996Programmed cell death: a way of life for plantsProc. Natl. Acad. Sci. USA931209412097CrossRefPubMedGoogle Scholar
  31. Higo, K., Ugawa, Y., Iwamoto, M., Higo, H. 1998PLACE: a database of plant cis-acting regulatory DNA elementsNucl. Acids Res.26358359CrossRefPubMedGoogle Scholar
  32. Hull, A.K., Celenza, J.L. 2000Bacterial expression and purification of the Arabidopsis NADPH-cytochrome P450 reductase ATR2Protein Expr. Purif.18310315CrossRefPubMedGoogle Scholar
  33. Ikura, M., Osawa, M., Ames, J.B. 2002The role of calcium-binding proteins in the control of transcription: structure to functionBioessays24625636CrossRefPubMedGoogle Scholar
  34. Jarvis, P. 2003Intracellular signalling: the language of the chloroplastCurr. Biol.13R314R316CrossRefPubMedGoogle Scholar
  35. Jones-Rhoades, M.W., Bartel, D.P. 2004Computational identification of plant microRNAs and their targets, including a stress-induced miRNAMol. Cell14787799CrossRefPubMedGoogle Scholar
  36. Joo, J.H., Bae, Y.S., Lee, J.S. 2001Role of auxin-induced reactive oxygen species in root gravitropismPlant Physiol.12610551060CrossRefPubMedGoogle Scholar
  37. Joo, J.H., Wang, S., Chen, J.G., Jones, A.M. and Fedoroff, N.V. 2005. Different signaling and cell-death roles of heterotrimeric G protein α and β subunits in the Arabidopsis oxidative stress response to ozone. Plant Cell. 17: 957--970Google Scholar
  38. Karpinski, S., Gabrys, H., Mateo, A., Karpinska, B., Mullineaux, P.M. 2003Light perception in plant disease defence signallingCurr. Opin. Plant Biol.6390396CrossRefPubMedGoogle Scholar
  39. Kiyosue, T., Abe, H., Yamaguchi-Shinozaki, K., Shinozaki, K. 1998ERD6, a cDNA clone for an early dehydration-induced gene of Arabidopsis, encodes a putative sugar transporterBiochim. Biophys. Acta.1370187191PubMedGoogle Scholar
  40. Lamb, C.J., Dixon, R.A. 1994Molecular mechanisms underlying induction of plant defence gene transcriptionBiochem. Soc. Symp.60241248PubMedGoogle Scholar
  41. Lamb, C.J., Dixon, R.A. 1997The oxidative burst in plant disease resistanceAnnu. Rev. Plant Physiol. Plant Mol. Biol.48251275Google Scholar
  42. Lee, K., Lee, J., Kim, Y., Bae, D., Kang, K.Y., Yoon, S.C., Lim, D. 2004Defining the plant disulfide proteomeElectrophoresis25532541CrossRefPubMedGoogle Scholar
  43. Louerat-Oriou, B., Perret, A., Pompon, D. 1998Differential redox and electron-transfer properties of purified yeast, plant and human NADPH-cytochrome P-450 reductases highly modulate cytochrome P-450 activitiesEur. J. Biochem.25810401049CrossRefPubMedGoogle Scholar
  44. Low, P.S., Merida, J.R. 1996The oxidative burst in plant defense: function and signal transductionPhysiol. Plant.96533542CrossRefGoogle Scholar
  45. Mahalingam, R., Gomez-Buitrago, A., Eckardt, N., Shah, N., Guevara-Garcia, A., Day, P., Raina, R., Fedoroff, N.V. 2003Characterizing the stress/defense transcriptome of ArabidopsisGen. Biol.4R20CrossRefGoogle Scholar
  46. Mahalingam, R., Fedoroff, N.V. 2003Stress response, cell death and signaling: the many faces of reactive oxygen speciesPhysiol. Plant.1195668CrossRefGoogle Scholar
  47. Meyers, B.C., Dickerman, A.W., Michelmore, R.W., Sivaramakrishnan, S., Sobral, B.W., Young, N.D. 1999Plant disease resistance genes encode members of an ancient and diverse protein family within the nucleotide-binding superfamily [In Process Citation]Plant J.20317332PubMedGoogle Scholar
  48. Meyers, B.C., Kozik, A., Griego, A., Kuang, H., Michelmore, R.W. 2003Genome-wide analysis of NBS-LRR-encoding genes in ArabidopsisPlant Cell15809834CrossRefPubMedGoogle Scholar
  49. Moeder, W., Barry, C.S., Tauriainen, A.A., Betz, C., Tuomainen, J., Utriainen, M., Grierson, D., Sandermann, H., Langebartels, C., Kangasjarvi, J. 2002Ethylene synthesis regulated by biphasic induction of 1-aminocyclopropane-1-carboxylic acid synthase and 1-aminocyclopropane-1-carboxylic acid oxidase genes is required for hydrogen peroxide accumulation and cell death in ozone-exposed tomatoPlant Physiol.13019181926CrossRefPubMedGoogle Scholar
  50. Montane, M.H., Tardy, F., Kloppstech, K., Havaux, M. 1998Differential control of xanthophylls and light-induced stress proteins, as opposed to light-harvesting chlorophyll a/b proteins, during photosynthetic acclimation of barley leaves to light irradiancePlant Physiol.118227235CrossRefPubMedGoogle Scholar
  51. Mou, Z., Fan, W., Dong, X. 2003Inducers of plant systemic acquired resistance regulate NPR1 function through redox changesCell113120CrossRefPubMedGoogle Scholar
  52. Mullineaux, P., Ball, L., Escobar, C., Karpinska, B., Creissen, G., Karpinski, S. 2000Are diverse signalling pathways integrated in the regulation of Arabidopsis antioxidant defence gene expression in response to excess excitation energy?Philos. Trans. R Soc. Lond. B Biol. Sci.355 15311540CrossRefPubMedGoogle Scholar
  53. Mullineaux, P., Karpinski, S. 2002Signal transduction in response to excess light: getting out of the chloroplastCurr. Opin. Plant Biol.54348CrossRefPubMedGoogle Scholar
  54. Nawrath, C., Heck, S., Parinthawong, N., Metraux, J.P. 2002EDS5, an essential component of salicylic acid-dependent signaling for disease resistance in Arabidopsis, is a member of the MATE transporter familyPlant Cell14275286CrossRefPubMedGoogle Scholar
  55. Overmyer, K., Tuominen, H., Kettunen, R., Betz, C., Langebartels, C., Sandermann, H., Kangasjarvi, J. 2000Ozone-sensitive Arabidopsis rcd1 mutant reveals opposite roles for ethylene and jasmonate signaling pathways in regulating superoxide-dependent cell deathPlant Cell1218491862CrossRefPubMedGoogle Scholar
  56. Pfannschmidt, T. 2003Chloroplast redox signals: how photosynthesis controls its own genesTrends Plant Sci.83341CrossRefPubMedGoogle Scholar
  57. Pursiheimo, S., Mulo, P., Rintamaki, E., Aro, E.M. 2001Coregulation of light-harvesting complex II phosphorylation and lhcb mRNA accumulation in winter ryePlant J.26317327CrossRefPubMedGoogle Scholar
  58. Pysh, L.D., Wysocka-Diller, J.W., Camilleri, C., Bouchez, D., Benfey, P.N. 1999The GRAS gene family in Arabidopsis: sequence characterization and basic expression analysis of the SCARECROW-LIKE genesPlant J.18111119CrossRefPubMedGoogle Scholar
  59. Rao, M.V., Lee, H., Creelman, R.A., Mullet, J.E., Davis, K.R. 2000Jasmonic acid signaling modulates ozone-induced hypersensitive cell deathPlant Cell1216331646CrossRefPubMedGoogle Scholar
  60. Richly, E., Dietzmann, A., Biehl, A., Kurth, J., Laloi, C., Apel, K., Salamini, F., Leister, D. 2003Covariations in the nuclear chloroplast transcriptome reveal a regulatory master-switchEMBO Rep.4491498CrossRefPubMedGoogle Scholar
  61. Rintamaki, E., Martinsuo, P., Pursiheimo, S., Aro, E.M. 2000Cooperative regulation of light-harvesting complex II phosphorylation via the plastoquinol and ferredoxin–thioredoxin system in chloroplastsProc. Natl. Acad. Sci. USA971164411649CrossRefPubMedGoogle Scholar
  62. Rochaix, J.D. 1996Post-transcriptional regulation of chloroplast gene expression in Chlamydomonas reinhardtiiPlant Mol. Biol.32327341CrossRefPubMedGoogle Scholar
  63. Runeckles, V.C., Chevonne, B.I. 1992Crop responses to ozoneLefohn, A.S. eds. Surface Level Ozone Exposures and Their Effects on VegetationLewis Publishers, Inc.Chelsea, MI189270Google Scholar
  64. Salvador, M.L., Klein, U. 1999The redox state regulates RNA degradation in the chloroplast of Chlamydomonas reinhardtiiPlant Physiol.12113671374CrossRefPubMedGoogle Scholar
  65. Sambrook, J., Fritsch, E.F., Maniatis, T. 1989Molecular Cloning: A Laboratory ManualCold Spring Harbor Laboratory PressCold Spring Harbor, NYGoogle Scholar
  66. Sandermann, H.,Jr. 2000Active oxygen species as mediators of plant immunity: three case studiesBiol. Chem.381649653CrossRefPubMedGoogle Scholar
  67. Sandermann, H.J., Ernst, E., Heller, W., Langebartels, C. 1998Ozone: an abiotic elicitor of plant defence reactionsTrends Plant Sci.34749CrossRefGoogle Scholar
  68. Sandermann, H. 1996Ozone and plant healthAnnu. Rev. Phytopathol.34347366CrossRefPubMedGoogle Scholar
  69. Scheel, D. 2002Oxidative burst and the role of reactive oxygen species in plant–pathogen interactionsInze, D.Montagu,  M. eds. Oxidative Stress in PlantsTaylor and FrancisNew York137153Google Scholar
  70. Schopfer, P., Liszkay, A., Bechtold, M., Frahry, G., Wagner, A. 2002Evidence that hydroxyl radicals mediate auxin-induced extension growthPlanta214821828CrossRefPubMedGoogle Scholar
  71. Shah, J., Klessig, D.F. 1996Identification of a salicylic acid-responsive element in the promoter of the tobacco pathogenesis-related beta-1,3-glucanase gene, PR-2dPlant J1010891101Google Scholar
  72. Shah, N.H., Fedoroff, N.V. 2004CLENCH: a program for calculating CLuster ENriCHment using the Gene OntologyBioinformatics2011961197CrossRefPubMedGoogle Scholar
  73. Shah, N.H., King, D.C., Shah, P.N., Fedoroff, N.V. 2003A tool-kit for cDNA microarray and promoter analysisBioinformatics1918461848CrossRefPubMedGoogle Scholar
  74. Sharma, Y.K., Davis, K.R. 1997The effects of ozone on antioxidant responses in plantsFree Radic. Biol. Med.23480488CrossRefPubMedGoogle Scholar
  75. Sharma, Y.K., Leon, J., Raskin, I., Davis, K.R. 1996Ozone-induced responses in Arabidopsis thaliana: the role of salicylic acid in the accumulation of defense-related transcripts and induced resistanceProc. Natl. Acad. Sci. USA9350995104CrossRefPubMedGoogle Scholar
  76. Sherameti, I., Sopory, S.K., Trebicka, A., Pfannschmidt, T., Oelmuller, R. 2002Photosynthetic electron transport determines nitrate reductase gene expression and activity in higher plantsJ. Biol. Chem.2774659446600CrossRefPubMedGoogle Scholar
  77. Shirasu, K., Dixon, R.A., Lamb, C. 1996Signal transduction in plant immunityCurr. Opin. Immunol.837CrossRefPubMedGoogle Scholar
  78. Stuart, G.W., Searle, P.F., Palmiter, R.D. 1985Identification of multiple metal regulatory elements in mouse metallothionein-i promoter by assaying synthetic sequencesNature317828831CrossRefPubMedGoogle Scholar
  79. Sullivan, J.A., Gray, J.C. 2002Multiple plastid signals regulate the expression of the pea plastocyanin gene in pea and transgenic tobacco plantsPlant J.32763774CrossRefPubMedGoogle Scholar
  80. Tamaoki, M., Nakajima, N., Kubo, A., Aono, M., Matsuyama, T., Saji, H. 2003Transcriptome analysis of O3-exposed Arabidopsis reveals that multiple signal pathways act mutually antagonistically to induce gene expressionPlant Mol. Biol.53443456CrossRefPubMedGoogle Scholar
  81. Tilly, J.J., Allen, D.W., Jack, T. 1998The CArG boxes in the promoter of the Arabidopsis floral organ identity gene APETALA3 mediate diverse regulatory effectsDevelopment12516471657PubMedGoogle Scholar
  82. Tuominen, H., Overmyer, K., Keinanen, M., Kollist, H., Kangasjarvi, J. 2004Mutual antagonism of ethylene and jasmonic acid regulates ozone-induced spreading cell death in ArabidopsisPlant J395969CrossRefPubMedGoogle Scholar
  83. Ulmasov, T., Hagen, G., Guilfoyle, T. 1994The ocs element in the soybean GH2/4 promoter is activated by both active and inactive auxin and salicylic acid analoguesPlant Mol. Biol.2610551064CrossRefPubMedGoogle Scholar
  84. Ulmasov, T., Hagen, G., Guilfoyle, T.J. 1997ARF1, a transcription factor that binds to auxin response elementsScience27618651868CrossRefPubMedGoogle Scholar
  85. Ulmasov, T., Liu, Z.B., Hagen, G., Guilfoyle, T.J. 1995Composite structure of auxin response elementsPlant Cell716111623CrossRefPubMedGoogle Scholar
  86. Vandenabeele, S., Kelen, K., Dat, J., Gadjev, I., Boonefaes, T., Morsa, S., Rottiers, P., Slooten, L., Van Montagu, M., Zabeau, M., Inze, D., Van Breusegem, F. 2003A comprehensive analysis of hydrogen peroxide-induced gene expression in tobaccoProceedings of the National Academy of Sciences of the United States of America1001611316118CrossRefPubMedGoogle Scholar
  87. Vandenabeele, S., Vanderauwera, S., Vuylsteke, M., Rombauts, S., Langebartels, C., Seidlitz, H.K., Zabeau, M., Van Montagu, M., Inze, D., Van Breusegem, F. 2004Catalase deficiency drastically affects gene expression induced by high light in Arabidopsis thalianaPlant J.394558CrossRefPubMedGoogle Scholar
  88. Vener, A.V., Ohad, I., Andersson, B. 1998Protein phosphorylation and redox sensing in chloroplast thylakoidsCurr. Opin. Plant Biol.1217223CrossRefPubMedGoogle Scholar
  89. Vener, A.V., Van Kan, P.J., Rich, P.R., Ohad, I.I., Andersson, B. 1997Plastoquinol at the quinol oxidation site of reduced cytochrome bf mediates signal transduction between light and protein phosphorylation: thylakoid protein kinase deactivation by a single-turnover flashProc. Natl. Acad. Sci. USA9415851590CrossRefPubMedGoogle Scholar
  90. Xu, W., Purugganan, M.M., Polisensky, D.H., Antosiewicz, D.M., Fry, S.C., Braam, J. 1995Arabidopsis TCH4, regulated by hormones and the environment, encodes a xyloglucan endotransglycosylasePlant Cell715551567CrossRefPubMedGoogle Scholar
  91. Zer, H., Ohad, I. 2003Light, redox state, thylakoid-protein phosphorylation and signaling gene expressionTrends Biochem. Sci.28467470CrossRefPubMedGoogle Scholar
  92. Zhou, J.M., Trifa, Y., Silva, H., Pontier, D., Lam, E., Shah, J., Klessig, D.F. 2000NPR1 differentially interacts with members of the TGA/OBF family of transcription factors that bind an element of the PR-1 gene required for induction by salicylic acidMol Plant Microbe Int.13191202Google Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Ramamurthy Mahalingam
    • 1
    Email author
  • Nigam Shah
    • 2
  • Alexandra Scrymgeour
    • 2
  • Nina Fedoroff
    • 2
  1. 1.Department of Biochemistry and Molecular Biology, 246 Noble Research CenterOklahoma State UniversityStillwaterUSA
  2. 2.Biology Department and Huck Institutes of the Life Sciences, Pennsylvania State UniversityUniversity ParkUSA

Personalised recommendations