Characterization of conidial autofluorescence in powdery mildew.

ABSTRACT

Autofluorescence is produced by endogenous fluorophores, such as NAD(P)H, lipofuscin, melanin, and riboflavin, indicating the accumulation of substances and the state of energy metabolism in organisms. As an obligate parasite, powdery mildew is wildly spread by air and parasitic crops. However, most identification studies have been based on morphology and molecular biology which were far too time- and labor-consuming, thus lacking characteristic, simple, and effective means. Using microscopy under the blue and cyan channels, we elaborated visible conidial autofluorescence in three powdery mildew species, Erysiphe quercicola, E. cichoracearum, and Podosphaera hibiscicola, with a sharp increase during the conidia senescence in E. quercicola. Additionally, the main spectral excitation detected by fluorescence spectrometery was 375 nm for these species, with a common emission peak at approximately 458-463 nm, and an additional trend at 487 nm for P. hibiscicola. Because NAD(P)H has a similar spectral feature, we further investigated the relation between NAD(P)H and conidial autofluorescence by fluorescence spectra. We observed that the reduced coenzymes prominently contributed to conidial autofluorescence; however, the conidial autofluorescence in P. hibiscicola displayed a different trend that may be affected by the oxidized coenzyme -NAD. Finally, the normalized average spectra of these three powdery mildew species and standard samples showed that the spectral trend of each species was similar but that the features in detail were specific and distinct based on principal component analysis. In conclusion, we showed and characterized conidial autofluorescence in three powdery mildew species for the first time. The specific conidial autofluorescence in these species provides a new idea for the development of field spore capture and identification devices for the discrimination of powdery mildew at the species level.