Plant Cell, Tissue and Organ Culture

, Volume 91, Issue 2, pp 97–106 | Cite as

Chlorophyll fluorescence imaging for disease-resistance screening of sugar beet



Both biotic and abiotic stresses cause considerable crop yield losses worldwide (Chrispeels, Sadava Plants, genes, and crop biotechnology 2003; Oerke, Dehne Crop Prot 23:275–285 2004). To speed up screening assays in stress resistance breeding, non-contact techniques such as chlorophyll fluorescence imaging can be advantageously used in the quantification of stress-inflicted damage. In comparison with visual spectrum images, chlorophyll fluorescence imaging reveals cell death with higher contrast and at earlier time-points. This technique has the potential to automatically quantify stress-inflicted damage during screening applications. From a physiological viewpoint, screening stress-responses using attached plant leaves is the ideal approach. However, leaf growth and circadian movements interfere with time-lapse monitoring of leaves, making it necessary to fix the leaves to be studied. From this viewpoint, a method to visualise the evolution of chlorophyll fluorescence from excised leaf pieces kept in closed petri dishes offers clear advantages. In this study, the plant–fungus interaction sugar beet–Cercospora beticola was assessed both in attached leaf and excised leaf strip assays. The attached leaf assay proved to be superior in revealing early, pre-visual symptoms and to better discriminate between the lines with different susceptibility to Cercospora.


Cercospora beticola Sacc. Chlorophyll fluorescence imaging Plant disease resistance quantification Plant–pathogen interaction Sugar beet Thermography 





Chlorophyll fluorescence imaging


Days postinfection


Chlorophyll fluorescence image captured after low intensity excitation


Chlorophyll fluorescence image captured after high intensity excitation


Fluorescence imaging system



L.C. is a post-doctoral fellow of the Research Foundation—Flanders. D.H. is a post-doc with financial support provided through the European Community’s Human Potential Programme under contract HPRN-CT-2002–00254, STRESSIMAGING. The authors are grateful to Roland Valcke, Laboratory for Molecular and Physical Plant Physiology, Hasselt University, for advice on chlorophyll fluorescence imaging.


  1. Barbagallo RP, Oxborough K, Pallett KE, Baker NR (2003) Rapid, noninvasive screening for perturbations of metabolism and plant growth using chlorophyll fluorescence imaging. Plant Physiol 132:485–493PubMedCrossRefGoogle Scholar
  2. Barna B, Adam AL, Kiraly Z (1997) Increased levels of cytokinin induce tolerance to necrotic diseases and various oxidative stress-causing agents in plants. Phyton-Annales Rei Botanicae 37:25–29Google Scholar
  3. Berger S, Papadopoulos M, Schreiber U, Kaiser W, Roitsch T (2004) Complex regulation of gene expression, photosynthesis and sugar levels by pathogen infection in tomato. Physiol Plant 122:419–428CrossRefGoogle Scholar
  4. Buschmann C (1999) Thermal dissipation during photosynthetic induction and subsequent dark recovery as measured by photoacoustic signals. Photosynthetica 36:149–161CrossRefGoogle Scholar
  5. Chaerle L, Van Der Straeten D (2000) Imaging techniques and the early detection of plant stress. Trends Plant Sci 5:495–501PubMedCrossRefGoogle Scholar
  6. Chaerle L, Van Der Straeten D (2001) Seeing is believing: imaging techniques to monitor plant health. Biochim Biophys Acta–Gene Struct Expression 1519:153–166CrossRefGoogle Scholar
  7. Chaerle L, Hagenbeek D, De Bruyne E, Valcke R, Van Der Straeten D (2004) Thermal and chlorophyll-fluorescence imaging distinguish plant-pathogen interactions at an early stage. Plant Cell Physiol 45:887–896PubMedCrossRefGoogle Scholar
  8. Chaerle L, Saibo N, Van Der Straeten D (2005) Tuning the pores: towards engineering plants for improved water use efficiency. Trends Biotechnol 23:308–315PubMedCrossRefGoogle Scholar
  9. Chrispeels MJ, Sadava DE (2003) Plants, genes, and crop biotechnology. Jones and Bartlett, BostonGoogle Scholar
  10. Cooper C, Crowther T, Smith BM, Isaac S, Collin HA (2006) Assessment of the response of carrot somaclones to Pythium violae, causal agent of cavity spot. Plant Pathol 55:427–432CrossRefGoogle Scholar
  11. Dita MA, Rispail N, Prats E, Rubiales D, Singh KB (2006) Biotechnology approaches to overcome biotic and abiotic stress constraints in legumes. Euphytica 147:1–24CrossRefGoogle Scholar
  12. Gan SS, Amasino RM (1995) Inhibition of leaf senescence by autoregulated production of cytokinin. Science 270:1986–1988PubMedCrossRefGoogle Scholar
  13. Fernie AR, Tadmor Y, Zamir D (2006) Natural genetic variation for improving crop quality. Curr Opin Plant Biol 9:196–202PubMedCrossRefGoogle Scholar
  14. Fila G, Badeck FW, Meyer S, Cerovic Z, Ghashghaie J (2006) Relationships between leaf conductance to CO2 diffusion and photosynthesis in micropropagated grapevine plants, before and after ex vitro acclimatization. J Exp Bot 57:2687–2695PubMedCrossRefGoogle Scholar
  15. Fuller MP, Metwali EMR, Eed MH, Jellings AJ (2006) Evaluation of abiotic stress resistance in mutated populations of cauliflower (Brassica oleracea var. Botrytis). Plant Cell Tissue Organ Cult 86:239–248CrossRefGoogle Scholar
  16. Haisel D, Pospisilova J, Synkova H, Schnablova R, Batkova P (2006) Effects of abscisic acid or benzyladenine on pigment contents, chlorophyll fluorescence, and chloroplast ultrastructure during water stress and after rehydration. Photosynthetica 44:606–614CrossRefGoogle Scholar
  17. Horie T, Matsuura S, Takai T, Kuwasaki K, Ohsumi A, Shiraiwa T (2006) Genotypic difference in canopy diffusive conductance measured by a new remote-sensing method and its association with the difference in rice yield potential. Plant Cell Environ 29:653–660PubMedCrossRefGoogle Scholar
  18. Huang S, Vleeshouwers V, Visser RGF, Jacobsen E (2005) An accurate in vitro assay for high-throughput disease testing of Phytophthora infestans in potato. Plant Dis 89:1263–1267CrossRefGoogle Scholar
  19. Jafra S, Jalink H, van der Schoor R, van der Wolf JM (2006) Pectobacterium carotovorum subsp. carotovorum strains show diversity in production of and response to N-acyl homoserine lactones. J Phytopathol 154:729–739CrossRefGoogle Scholar
  20. Jauhar PP (2006) Modern biotechnology as an integral supplement to conventional plant breeding: the prospects and challenges. Crop Sci 46:1841–1859CrossRefGoogle Scholar
  21. Jones HG (2004) Application of thermal imaging and infrared sensing in plant physiology and ecophysiology. Adv Bot Res 41:107–163CrossRefGoogle Scholar
  22. Krause GH, Weis E (1991) Chlorophyll fluorescence and photosynthesis: the basics. Annu Rev Plant Physiol Plant Mol Biol 42:313–349CrossRefGoogle Scholar
  23. Lenk S, Chaerle L, Pfündel E, Langsdorf G, Hagenbeek D, Lichtenthaler H, Van Der Straeten D, Buschmann C (2007) Multi-colour fluorescence and reflectance imaging at the leaf level and its possible applications. J Exp Bot 58:807–814PubMedCrossRefGoogle Scholar
  24. Maxwell K, Johnson GN (2000) Chlorophyll fluorescence—a practical guide. J Exp Bot 51:659–668PubMedCrossRefGoogle Scholar
  25. Moshou D, Bravo C, Oberti R, West J, Bodria L, McCartney A, Ramon H (2005) Plant disease detection based on data fusion of hyper-spectral and multi-spectral fluorescence imaging using Kohonen maps. Real-Time Imaging 11:75–83CrossRefGoogle Scholar
  26. Nejad AR, Harbinson J, van Meeteren U (2006) Dynamics of spatial heterogeneity of stomatal closure in Tradescantia virginiana altered by growth at high relative air humidity. J Exp Bot 57:3669–3678CrossRefGoogle Scholar
  27. Nilsson HE (1995) Remote sensing and image analysis in plant pathology. Annu Rev Phytopathol 33:489–527CrossRefGoogle Scholar
  28. Oerke EC, Dehne HW (2004) Safeguarding production—losses in major crops and the role of crop protection. Crop Prot 23:275–285CrossRefGoogle Scholar
  29. Oerke EC, Steiner U, Dehne HW, Lindenthal M (2006) Thermal imaging of cucumber leaves affected by downy mildew and environmental conditions. J Exp Bot 57:2121–2132PubMedCrossRefGoogle Scholar
  30. Oxborough K (2004) Imaging of chlorophyll a fluorescence: theoretical and practical aspects of an emerging technique for the monitoring of photosynthetic performance. J Exp Bot 55:1195–1205PubMedCrossRefGoogle Scholar
  31. Pawelec A, Dubourg C, Briard M (2006) Evaluation of carrot resistance to alternaria leaf blight in controlled environments. Plant Pathol 55:68–72CrossRefGoogle Scholar
  32. Pontier D, Gan SS, Amasino RM, Roby D, Lam E (1999) Markers for hypersensitive response and senescence show distinct patterns of expression. Plant Mol Biol 39:1243–1255PubMedCrossRefGoogle Scholar
  33. Quilliam RS, Swarbrick PJ, Scholes JD, Rolfe SA (2006) Imaging photosynthesis in wounded leaves of Arabidopsis thaliana. J Exp Bot 57:55–69PubMedCrossRefGoogle Scholar
  34. Scharte J, Schon H, Weis E (2005) Photosynthesis and carbohydrate metabolism in tobacco leaves during an incompatible interaction with Phytophthora nicotianae. Plant Cell Environ 28:1421–1435CrossRefGoogle Scholar
  35. Soukupova J, Smatanova S, Nedbal L, Jegorov A (2003) Plant response to destruxins visualised by imaging of chlorophyll fluorescence. Physiol Plant 118:399–405CrossRefGoogle Scholar
  36. Thevenaz P, Ruttimann UE, Unser M (1998) A pyramid approach to subpixel registration based on intensity. IEEE Trans Image Process 7:27–41CrossRefPubMedGoogle Scholar
  37. Xie X, Wang Y, Williamson L, Holroyd GH, Tagliavia C, Murchie E, Theobald J, Knight MR, Davies WJ, Leyser HMO, Hetherington AM (2006) The identification of genes involved in the stomatal response to reduced atmospheric relative humidity. Curr Biol 16:882–887PubMedCrossRefGoogle Scholar
  38. Xu JR, Peng YL, Dickman MB, Sharon A (2006) The dawn of fungal pathogen genomics. Annu Rev Phytopathol 44:337–366PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  1. 1.Unit Plant Hormone Signalling and Bio-imagingGhent UniversityGentBelgium
  2. 2.Phytopathology DepartmentSESVanderHaveTienenBelgium

Personalised recommendations