Optical detection of downy mildew in grapevine leaves: daily kinetics of autofluorescence upon infection

A 15-day survey of autofluorescence has been conducted upon infection by downy mildew [Plasmopara viticola (Berk. & M.A. Curtis) Berl. & de Toni] of leaves of a susceptible grapevine genotype. Different autofluorescence signals were followed from the cellular to the whole-leaf level by using four types of devices for fluorosensing: a macroscope, a spectrofluorimeter, a portable field optical sensor (the Multiplex 3), and a field fluorescence sensor prototype with 335nm excitation. It was shown for the first time, by the three different techniques and at three different scales, that the stilbene-dependent violet–blue autofluorescence (VBF) had a transitory behaviour, increasing to a maximum 6 days post-inoculation (DPI) and then decreasing to a constant lower level, nevertheless significantly higher than in the control leaf. This behaviour could be sensed from both sides of the leaf. On the abaxial side, VBF could discriminate the presence of infection from 1 DPI, and on the adaxial side from 3 DPI. There was a constant increase in blue-excited green fluorescence starting from 8 DPI, concomitant with a decrease in leaf chlorophyll content sensed by one reflectance and two fluorescence indices available on the Multiplex 3 sensor. These results show that a pre-symptomatic and symptomatic sensing of downy mildew is possible by autofluorescence-based sensors, and this is potentially applicable in the field.

[1]  P. Hugueney,et al.  Metabolism and roles of stilbenes in plants , 2009 .

[2]  M. Sbaghi,et al.  Degradation of stilbene‐type phytoalexins in relation to the pathogenicity of Botrytis cinerea to grapevines , 1996 .

[3]  Markus Riederer,et al.  Biology of the plant cuticle , 2006 .

[4]  M. Sbaghi,et al.  Phytoalexins from the Vitaceae: biosynthesis, phytoalexin gene expression in transgenic plants, antifungal activity, and metabolism. , 2002, Journal of agricultural and food chemistry.

[5]  C. Clément,et al.  Biosynthesis, metabolism, molecular engineering, and biological functions of stilbene phytoalexins in plants , 2010, BioFactors.

[6]  Z. Cerovic,et al.  Fast and local assessment of stilbene content in grapevine leaf by in vivo fluorometry. , 2007, Journal of agricultural and food chemistry.

[7]  Z. Cerovic,et al.  Quantitative study of fluorescence excitation and emission spectra of bean leaves. , 2006, Journal of photochemistry and photobiology. B, Biology.

[8]  C. Andary,et al.  Histochemical studies on the interaction between three species of grapevine, Vitis vinifera, V. rupestris and V. rotundifolia and the downy mildew fungus, Plasmopara viticola , 1995 .

[9]  Ismael Moya,et al.  Ultraviolet-induced fluorescence for plant monitoring: present state and prospects , 1999 .

[10]  Nicolas Tremblay,et al.  Sensing crop nitrogen status with fluorescence indicators. A review , 2011, Agronomy for Sustainable Development.

[11]  Karel Matouš,et al.  Pre-symptomatic detection of Plasmopara viticola infection in grapevine leaves using chlorophyll fluorescence imaging , 2009, European Journal of Plant Pathology.

[12]  R. Pezet,et al.  Evidence for oxidative detoxication of pterostilbene and resveratrol by a laccase-like stilbene oxidase produced by Botrytis cinerea , 1991 .

[13]  Ismael Moya,et al.  The use of chlorophyll fluorescence excitation spectra for the non‐destructive in situ assessment of UV‐absorbing compounds in leaves , 2002 .

[14]  W. Hillis,et al.  The chromatographic and spectral properties of stilbene derivatives. , 1968, Journal of chromatography.

[15]  E. Pfündel,et al.  Effects of natural intensities of visible and ultraviolet radiation on epidermal ultraviolet screening and photosynthesis in grape leaves. , 2001, Plant physiology.

[16]  J. Harborne,et al.  A survey of antifungal compounds from higher plants, 1982–1993 , 1994 .

[17]  Z. Cerovic,et al.  Optical Properties of Plant Surfaces , 2007 .

[18]  C. Andary,et al.  Histochemical responses of leaves of in vitro plantlets of Vitis spp. to infection with Plasmopara viticola , 1995 .

[19]  Reza Ehsani,et al.  Review: A review of advanced techniques for detecting plant diseases , 2010 .

[20]  Gwendal Latouche,et al.  Non-Destructive Optical Monitoring of Grape Maturation by Proximal Sensing , 2010, Sensors.

[21]  C. Büche,et al.  The Course of Colonization of Two Different Vitis Genotypes by Plasmopara viticola Indicates Compatible and Incompatible Host-Pathogen Interactions. , 2007, Phytopathology.

[22]  S. Orlandini,et al.  Optically-assessed preformed flavonoids and susceptibility of grapevine to Plasmopara viticola under different light regimes. , 2008, Functional plant biology : FPB.

[23]  D. Merdinoglu,et al.  Identification of effector genes from the phytopathogenic Oomycete Plasmopara viticola through the analysis of gene expression in germinated zoospores. , 2012, Fungal biology.

[24]  Gwendal Latouche,et al.  In vivo localization at the cellular level of stilbene fluorescence induced by Plasmopara viticola in grapevine leaves , 2012, Journal of Experimental Botany.