Register      Login
Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

Use of multicolour fluorescence imaging for diagnosis of bacterial and fungal infection on zucchini by implementing machine learning

Mónica Pineda A * , María Luisa Pérez-Bueno A B * , Vanessa Paredes A and Matilde Barón A
+ Author Affiliations
- Author Affiliations

A Department of Biochemistry and Molecular and Cell Biology of Plants, Estación Experimental del Zaidín, Spanish Council of Scientific Research (CSIC), Profesor Albareda, 1, 18008, Granada, Spain.

B Corresponding author. Email: marisa.perez@eez.csic.es

This paper is part of the Plant Phenotyping special issue (http://www.publish.csiro.au/FP/issue/8242).

Functional Plant Biology 44(6) 563-572 https://doi.org/10.1071/FP16164
Submitted: 29 April 2016  Accepted: 19 February 2017   Published: 3 April 2017

Abstract

Zucchini (Cucurbita pepo L.) is a cucurbitaceous plant ranking high in economic importance among vegetable crops worldwide. Pathogen infections cause alterations in plants primary and secondary metabolism that lead to a significant decrease in crop quality and yield. Such changes can be monitored by remote and proximal sensing, providing spatial and temporal information about the infection process. Remote sensing can also provide specific signatures of disease that could be used in phenotyping and to detect a pest, forecast its evolution and predict crop yield. In this work, metabolic changes triggered by soft rot (caused by Dickeya dadantii) and powdery mildew (caused by Podosphaera fusca) on zucchini leaves have been studied by multicolour fluorescence imaging and by thermography. The fluorescence parameter F520/F680 showed statistically significant differences between infected (with D. dadantii or P. fusca) and mock-control leaves during the whole period of study. Artificial neural networks, logistic regression analyses and support vector machines trained with a set of features characterising the histograms of F520/F680 images could be used as classifiers, discriminating between healthy and infected leaves. These results show the applicability of multicolour fluorescence imaging on plant phenotyping.

Additional keywords: cucurbit, Dickeya dadantii, multicolour fluorescence imaging, Podosphaera fusca, precision agriculture, thermal imaging.


References

Arens N, Backhaus A, Döll S, Fischer S, Seiffert U, Mock H-P (2016) Non-invasive presymptomatic detection of Cercospora beticola infection and identification of early metabolic responses in sugar beet. Frontiers in Plant Science 7, 1377
Non-invasive presymptomatic detection of Cercospora beticola infection and identification of early metabolic responses in sugar beet.Crossref | GoogleScholarGoogle Scholar |

Ashourloo D, Aghighi H, Matkan AA, Mobasheri MR, Rad AM (2016) An investigation into machine learning regression techniques for the leaf rust disease detection using hyperspectral measurement. IEEE Journal of Selected Topics in Applied Earth Observations and Remote Sensing 9, 4344–4351.
An investigation into machine learning regression techniques for the leaf rust disease detection using hyperspectral measurement.Crossref | GoogleScholarGoogle Scholar |

Baranowski P, Jedryczka M, Mazurek W, Babula-Skowronska D, Siedliska A, Kaczmarek J (2015) Hyperspectral and thermal imaging of oilseed rape (Brassica napus) response to fungal species of the genus Alternaria. PLoS One 10, e0122913
Hyperspectral and thermal imaging of oilseed rape (Brassica napus) response to fungal species of the genus Alternaria.Crossref | GoogleScholarGoogle Scholar |

Barón M, Flexas J, Delucia EH (2012) Photosynthetic responses to biotic stress. In ‘Terrestrial photosynthesis in a changing environment: a molecular, physiological, and ecological approach. Vol. 1’. (Eds J Flexas, F Loreto, H Medrano) pp. 331–350. (Cambridge University Press: Cambridge, UK)

Barón M, Pineda M, Pérez-Bueno ML (2016) Picturing pathogen infection in plants. Zeitschrift fur Naturforschung C 71, 355–368.

Bauriegel E, Giebel A, Herppich WB (2011) Hyperspectral and chlorophyll fluorescence imaging to analyse the impact of Fusarium culmorum on the photosynthetic integrity of infected wheat ears. Sensors 11, 3765–3779.
Hyperspectral and chlorophyll fluorescence imaging to analyse the impact of Fusarium culmorum on the photosynthetic integrity of infected wheat ears.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXksF2jurY%3D&md5=017c2cbe8668e83309015601c8f9740dCAS |

Behmann J, Mahlein A-K, Rumpf T, Römer C, Plümer L (2015) A review of advanced machine learning methods for the detection of biotic stress in precision crop protection. Precision Agriculture 16, 239–260.
A review of advanced machine learning methods for the detection of biotic stress in precision crop protection.Crossref | GoogleScholarGoogle Scholar |

Bellón-Gómez D, Vela-Corcia D, Pérez-García A, Torés JA (2015) Sensitivity of Podosphaera xanthii populations to anti-powdery-mildew fungicides in Spain. Pest Management Science 71, 1407–1413.
Sensitivity of Podosphaera xanthii populations to anti-powdery-mildew fungicides in Spain.Crossref | GoogleScholarGoogle Scholar |

Bellow S, Latouche G, Brown SC, Poutaraud A, Cerovic ZG (2012) In vivo localization at the cellular level of stilbene fluorescence induced by Plasmopara viticola in grapevine leaves. Journal of Experimental Botany 63, 3697–3707.
In vivo localization at the cellular level of stilbene fluorescence induced by Plasmopara viticola in grapevine leaves.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XhtVShtL7F&md5=50c233888404c8f6581d90bed5a250acCAS |

Buschmann C, Lichtenthaler HK (1998) Principles and characteristics of multi-colour fluorescence imaging of plants. Journal of Plant Physiology 152, 297–314.
Principles and characteristics of multi-colour fluorescence imaging of plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXisVKqtbw%3D&md5=7bceec93db13abbc8b90fd07b152600eCAS |

Calderón R, Navas-Cortés JA, Zarco-Tejada PJ (2015) Early detection and quantification of Verticillium wilt in olive using hyperspectral and thermal imagery over large areas. Remote Sensing 7, 5584–5610.
Early detection and quantification of Verticillium wilt in olive using hyperspectral and thermal imagery over large areas.Crossref | GoogleScholarGoogle Scholar |

Cerovic ZG, Samson G, Morales F, Tremblay N, Moya I (1999) Ultraviolet-induced fluorescence for plant monitoring: present state and prospects. Agronomie 19, 543–578.
Ultraviolet-induced fluorescence for plant monitoring: present state and prospects.Crossref | GoogleScholarGoogle Scholar |

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 Physiology 45, 887–896.
Thermal and chlorophyll-fluorescence imaging distinguish plant-pathogen interactions at an early stage.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXmsFeqsbg%3D&md5=b7f8da8e1a60ad2454e623ca6c8ca5bfCAS |

Delalieux S, Auwerkerken A, Verstraeten W, Somers B, Valcke R, Lhermitte S, Keulemans J, Coppin P (2009) Hyperspectral reflectance and fluorescence imaging to detect scab induced stress in apple leaves. Remote Sensing 1, 858–874.
Hyperspectral reflectance and fluorescence imaging to detect scab induced stress in apple leaves.Crossref | GoogleScholarGoogle Scholar |

Fernández-Ortuño D, Pérez-García A, López-Ruiz F, Romero D, De Vicente A, Torés JA (2006) Occurrence and distribution of resistance to QoI fungicides in populations of Podosphaera fusca in south central Spain. European Journal of Plant Pathology 115, 215–222.
Occurrence and distribution of resistance to QoI fungicides in populations of Podosphaera fusca in south central Spain.Crossref | GoogleScholarGoogle Scholar |

Gitelson AA, Buschmann C, Lichtenthaler HK (1998) Leaf chlorophyll fluorescence corrected for re-absorption by means of absorption and reflectance measurements. Journal of Plant Physiology 152, 283–296.
Leaf chlorophyll fluorescence corrected for re-absorption by means of absorption and reflectance measurements.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXisVKqtb8%3D&md5=8ca114f343cf59dccaba2b50e80b3438CAS |

Graham AR (1983) Fungal autofluorescence with ultraviolet illumination. American Journal of Clinical Pathology 79, 231–234.
Fungal autofluorescence with ultraviolet illumination.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaL3s7itlyksg%3D%3D&md5=3493924fbdce0d2ef7fd8e929bfed880CAS |

Granum E, Pérez-Bueno ML, Calderón CE, Ramos C, de Vicente A, Cazorla FM, Barón M (2015) Metabolic responses of avocado plants to stress induced by Rosellinia necatrix analysed by fluorescence and thermal imaging. European Journal of Plant Pathology 142, 625–632.
Metabolic responses of avocado plants to stress induced by Rosellinia necatrix analysed by fluorescence and thermal imaging.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2MXkvFentrw%3D&md5=bdcd907a539444724ee070dd0238dc8dCAS |

Hahn F (2009) Actual pathogen detection: sensors and algorithms – a review. Algorithms 2, 301–338.
Actual pathogen detection: sensors and algorithms – a review.Crossref | GoogleScholarGoogle Scholar |

Heisel F, Sowinska M, Miehé JA, Lang M, Lichtenthaler HK (1996) Detection of nutrient deficiencies of maize by laser induced fluorescence imaging. Journal of Plant Physiology 148, 622–631.
Detection of nutrient deficiencies of maize by laser induced fluorescence imaging.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28XksFWrsLs%3D&md5=5bb54a46c134bc45a1ef51618b8fb4e3CAS |

Hosmer J, David W, Lemeshow S, Sturdivant RX (2013) ‘Applied logistic regression.’ (3rd edn) (Jon Wiley & Sons Inc.: Hoboken, NJ, USA)

Hou J, Li L, He J (2016) Detection of grapevine leafroll disease based on 11-index imagery and ant colony clustering algorithm. Precision Agriculture 17, 488–505.
Detection of grapevine leafroll disease based on 11-index imagery and ant colony clustering algorithm.Crossref | GoogleScholarGoogle Scholar |

Hückelhoven R, Panstruga R (2011) Cell biology of the plant–powdery mildew interaction. Current Opinion in Plant Biology 14, 738–746.
Cell biology of the plant–powdery mildew interaction.Crossref | GoogleScholarGoogle Scholar |

Martinelli F, Scalenghe R, Davino S, Panno S, Scuderi G, Ruisi P, Villa P, Stroppiana D, Boschetti M, Goulart LR, Davis CE, Dandekar AM (2015) Advanced methods of plant disease detection. A review. Agronomy for Sustainable Development 35, 1–25.
Advanced methods of plant disease detection. A review.Crossref | GoogleScholarGoogle Scholar |

McGrath MT (2001) Fungicide resistance in cucurbit powdery mildew: experiences and challenges. Plant Disease 85, 236–245.
Fungicide resistance in cucurbit powdery mildew: experiences and challenges.Crossref | GoogleScholarGoogle Scholar |

Palacio-Bielsa A, Rodríguez Mosquera M, Cambra Álvarez M, Berruete Rodríguez I, López-Solanilla E, Rodríguez-Palenzuela P (2010) Phenotypic diversity, host range and molecular phylogeny of Dickeya isolates from Spain. European Journal of Plant Pathology 127, 311–324.
Phenotypic diversity, host range and molecular phylogeny of Dickeya isolates from Spain.Crossref | GoogleScholarGoogle Scholar |

Pérez-Bueno ML, Pineda M, Díaz-Casado ME, Barón M (2015) Spatial and temporal dynamics of primary and secondary metabolism in Phaseolus vulgaris challenged by Pseudomonas syringae. Physiologia Plantarum 153, 161–174.
Spatial and temporal dynamics of primary and secondary metabolism in Phaseolus vulgaris challenged by Pseudomonas syringae.Crossref | GoogleScholarGoogle Scholar |

Pérez-Bueno ML, Granum E, Pineda M, Flors V, Rodríguez-Palenzuela P, López-Solanilla E, Barón M (2016) Temporal and spatial resolution of activated plant defense responses in leaves of Nicotiana benthamiana infected with Dickeya dadantii. Frontiers in Plant Science 6, 1209
Temporal and spatial resolution of activated plant defense responses in leaves of Nicotiana benthamiana infected with Dickeya dadantii.Crossref | GoogleScholarGoogle Scholar |

Pérez‐García A, Romero D, Fernández‐Ortuño D, López‐Ruíz F, De Vicente A, Torés JA (2009) The powdery mildew fungus Podosphaera fusca (synonym Podosphaera xanthii), a constant threat to cucurbits. Molecular Plant Pathology 10, 153–160.
The powdery mildew fungus Podosphaera fusca (synonym Podosphaera xanthii), a constant threat to cucurbits.Crossref | GoogleScholarGoogle Scholar |

Raza S-e-A, Prince G, Clarkson JP, Rajpoot NM (2015) Automatic detection of diseased tomato plants using thermal and stereo visible light images. PLoS One 10, e0123262
Automatic detection of diseased tomato plants using thermal and stereo visible light images.Crossref | GoogleScholarGoogle Scholar |

Reverchon S, Nasser W (2013) Dickeya ecology, environment sensing and regulation of virulence programme. Environmental Microbiology Reports 5, 622–636.

Romero D, Rivera ME, Cazorla FM, Codina JC, Fernandez-Ortuno D, Tores JA, Perez-Garcia A, de Vicente A (2008) Comparative histochemical analyses of oxidative burst and cell wall reinforcement in compatible and incompatible melon-powdery mildew (Podosphaera fusca) interactions. Journal of Plant Physiology 165, 1895–1905.
Comparative histochemical analyses of oxidative burst and cell wall reinforcement in compatible and incompatible melon-powdery mildew (Podosphaera fusca) interactions.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhtlCmsb0%3D&md5=3dfc9e0703cea79928201d6f6b0f196cCAS |

Sankaran S, Maja JM, Buchanon S, Ehsani R (2013) Huanglongbing (citrus greening) detection using visible, near infrared and thermal imaging techniques. Sensors 13, 2117–2130.
Huanglongbing (citrus greening) detection using visible, near infrared and thermal imaging techniques.Crossref | GoogleScholarGoogle Scholar |

Xie CQ, Shao YN, Li XL, He Y (2015) Detection of early blight and late blight diseases on tomato leaves using hyperspectral imaging. Scientific Reports 5, 16564
Detection of early blight and late blight diseases on tomato leaves using hyperspectral imaging.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2MXhvVOrur%2FI&md5=dc1768a6adecfe19f8c531a6489e2f00CAS |

Zeng W, Melotto M, He SY (2010) Plant stomata: a checkpoint of host immunity and pathogen virulence. Current Opinion in Biotechnology 21, 599–603.
Plant stomata: a checkpoint of host immunity and pathogen virulence.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXht1OmsrnI&md5=6dc10563b637663e52e695fefa997f7cCAS |

Zitter T, Hopkins D, Thom C (1996) ‘Compendium of cucurbit diseases.’ (APS Press: St Paul, MN, USA)