Multidimensional discriminant analysis of species, strains and culture age of closely related entomopathogenic fungi using reflectance spectroscopy.

The diversity of fungal strains is influenced by genetic and environmental factors, growth conditions and mycelium age, and the spectral features of fungal mycelia are associated with their biochemical, physiological, and structural traits. This study investigates whether intraspecific differences can be detected in two closely related entomopathogenic species, namely Cordyceps farinosa and Cordyceps fumosorosea, using ultraviolet A to shortwave infrared (UVA-SWIR) reflectance spectra. Phylogenetic analysis of all strains revealed a high degree of uniformity among the populations of both species. The characteristics resulting from variation in the species, as well as those resulting from the age of the cultures were determined. We cultured fungi on PDA medium and measured the reflectance of mycelia in the 350-2500 nm range after 10 and 17 days. We subjected the measurements to quadratic discriminant analysis (QDA) to identify the minimum number of bands containing meaningful information. We found that when the age of the fungal culture was known, species represented by a group of different strains could be distinguished with no more than 3-4 wavelengths, compared to 7-8 wavelengths when the age of the culture was unknown. At least 6-8 bands were required to distinguish cultures of a known species among different age groups. Distinguishing all strains within a species was more demanding: at least 10 bands were required for C. fumosorosea and 21 bands for C. farinosa. In conclusion, fungal differentiation using point reflectance spectroscopy gives reliable results when intraspecific and age variations are taken into account.

Reflectance spectroscopy and machine learning as a tool for the categorization of twin species based on the example of the Diachrysia genus.

In our work we used noninvasive point reflectance spectroscopy in the range from 400 to 2100 nm coupled with machine learning to study scales on the brown and golden iridescent areas on the dorsal side of the forewing of Diachrysia chrysitis and D. stenochrysis. We used our approach to distinguish between these species of moths. The basis for the study was a statistically significant collection of 95 specimens identified based on morphological feature and gathered during 23 years in Poland. The numerical part of an experiment included two independent discriminant analyses: stochastic and deterministic. The more sensitive stochastic approach achieved average compliance with the species identification made by entomologists at the level of 99-100%. It demonstrated high stability against the different configurations of training and validation sets, hence strong predictors of Diachrysia siblings distinctiveness. Both methods resulted in the same small set of relevant features, where minimal fully discriminating subsets of wavelengths were three for glass scales on the golden area and four for the brown. The differences between species in scales primarily concern their major components and ultrastructure. In melanin-absent glass scales, this is mainly chitin configuration, while in melanin-present brown scales, melanin reveals as an additional factor.

A random forest model for the classification of wheat and rye leaf rust symptoms based on pure spectra at leaf scale.

The pure spectra acquisition of plant disease symptoms is essential to improving the reliability of remote sensing methods in crop protection. The reflectance values read from the pure spectra can be used as valuable training data for development of algorithms designed for plant disease detection at leaf and canopy scale. The aim of this paper is to identify and distinguish spectrally the leaf rust symptoms caused by two closely related special forms (f. sp.) of Puccinia recondita f. sp. tritici on wheat and Puccinia recondita f. sp. recondita on rye at leaf scale. Spectral measurements were made with FieldSpec 3 spectrometer in the wavelength range of 350-2500 nm. The spectrometer was connected to a microscope by optical fiber. Raw spectra of uredinia, chlorotic discoloration, green leaves, senescent inoculated leaves and senescent uninoculated leaves of wheat and rye, all of which obtained for this study, were investigated with a view towards making an automized classification of plant species and their phases. The created Random Forest models were tested separately using pure spectra, and from these vegetation indices were derived as predictors. Three vegetation indices, namely CRI, PRI and GNDVI, appeared to be the most robust in terms of distinguishing uredinia from other symptoms on rye and wheat leaves. PRI, EVI, NDVI705, and GNDVI were the most suitable for distinguishing uredinia, chlorotic discoloration, and green leaf stages on rye. That tusk on wheat leaves can be recognized if seven indices (PRI, MSAWI, SAVI, NDVI, NDVI705, GNDVI and RVI) are used together. For the classification of all disease symptoms for both plant species, the most useful were wavelengths in the VIS range: 431-436, 696-703 and 646-686 nm. However, the ranges of SWIR wavelengths (1938, 1955) and NIR wavelengths (1099-1104) also have a high contribution to the discrimination accuracy of the model. In the classification of all disease symptoms, the most important vegetation indices were CRI, OSAVI, and GNDVI. Analysis of the results revealed the advantage of the model based on the selected spectral wavelengths (Hit Rate of 96.6%) in comparison with predictions based on vegetation indices alone (Hit Rate of 91.7%). Both approaches show the highly applicable character of utilizing high quality spectral products such as satellite images in reducing operational costs of crop protection.

An application of reflectance spectroscopy to differentiate of entomopathogenic fungi species.

Fourier Transform Infrared Spectroscopy (FTIR) methods are the most commonly used spectroscopic techniques for differentiation of fungi species, however reflectance spectroscopy as a non-invasive technique can also be used. The aim of the study was to develop a method to rapidly differentiate fungi by means of reflectance spectroscopy using visible-infrared spectrum. Spectral measurements were conducted on six entomopathogenic fungi: Beauveria bassiana, Isaria fumosorosea, I. farinosa, I. tenuipes, Lecanicillium lecanii, L. muscarium cultured on Petri-dishes. The FieldSpec3 ASD spectroradiometer. Recording reflected radiance in the range 350-2500 nm was used. Measurements were performed in two modes: contact and proximal and obtained spectra were transformed using two methods: Savitzky-Golay (SG) and baseline alignment (BA) smoothing and derivative. The success rate of 100% in differentiate between fungi species was achieved with spectra recorded in visible-near infrared range with contact and proximal measurement and after SG transformation. Two wavelengths (411 nm and 520 nm) were needed to differentiate fungi using SG and proximal measurement while seven wavelengths were necessary to get full separation with contact measurement. BA spectra transformation method gave separation accuracy of 84, and 90% with four to five wavelengths for contact and proximal measurements, respectively, however, BA do not require full spectrum of wavelengths to fungi discrimination. Proposed reflectance spectroscopy method could discriminate between fungi species very similar macroscopically e.g. L. lecanii and L. muscarium until recently recognized as one species.