As a synergistic integration between spectroscopy and imaging technologies, spectral imaging modalities have been emerged to tackle quality evaluation dilemmas by proposing different designs with effective and practical applications in food and agriculture. With the advantage of acquiring spatio-spectral data across a wide range of the electromagnetic spectrum, the state-of-the-art multispectral imaging in tandem with different multivariate chemometric analysis scenarios has been successfully implemented not only for food quality and safety control purposes, but also in dealing with critical research challenges in seed science and technology. This paper will shed some light on the fundamental configuration of the systems and give a birds-eye view of all recent approaches in the acquisition, processing and reproduction of multispectral images for various applications in seed quality assessment and seed phenotyping issues. This review article continues from where earlier review papers stopped but it only focused on fully-operated multispectral imaging systems for quality assessment of different sorts of seeds. Thence, the review comprehensively highlights research attempts devoted to real implementations of only fully-operated multispectral imaging systems and does not consider those ones that just utilized some key wavelengths extracted from hyperspectral data analyses without building independent multispectral imaging systems. This makes this article the first attempt in briefing all published papers in multispectral imaging applications in seed phenotyping and quality monitoring by providing some examples and research results in characterizing physicochemical quality traits, predicting physiological parameters, detection of defect, pest infestation and seed health.

Background: In order to select for quantitative plant resistance to pathogens, high throughput approaches that can precisely quantify disease severity are needed. Automation and use of calibrated image analysis should provide more accurate, objective and faster analyses than visual assessments. In contrast to conventional visible imaging, chlorophyll fluorescence imaging is not sensitive to environmental light variations and provides single-channel images prone to a segmentation analysis by simple thresholding approaches. Among the various parameters used in chlorophyll fluorescence imaging, the maximum quantum yield of photosystem II photochemistry (F_v/F_m) is well adapted to phenotyping disease severity. F_v/F_m is an indicator of plant stress that displays a robust contrast between infected and healthy tissues. In the present paper, we aimed at the segmentation of F_v/F_m images to quantify disease severity. Results: Based on the F_v/F_m values of each pixel of the image, a thresholding approach was developed to delimit diseased areas. A first step consisted in setting up thresholds to reproduce visual observations by trained raters of symptoms caused by Xanthomonas fuscans subsp. fuscans (Xff) CFBP4834-R on Phaseolus vulgaris cv. Flavert. In order to develop a thresholding approach valuable on any cultivars or species, a second step was based on modeling pixel-wise F_v/F_m-distributions as mixtures of Gaussian distributions. Such a modeling may discriminate various stages of the symptom development but over-weights artifacts that can occur on mock-inoculated samples. Therefore, we developed a thresholding approach based on the probability of misclassification of a healthy pixel. Then, a clustering step is performed on the diseased areas to discriminate between various stages of alteration of plant tissues. Notably, the use of chlorophyll fluorescence imaging could detect pre-symptomatic area. The interest of this image analysis procedure for assessing the levels of quantitative resistance is illustrated with the quantitation of disease severity on five commercial varieties of bean inoculated with Xff CFBP4834-R. Conclusions: In this paper, we describe an image analysis procedure for quantifying the leaf area impacted by the pathogen. In a perspective of high throughput phenotyping, the procedure was automated with the software R downloadable at http://www.r-project.org/. The R script is available at http://lisa.univ-angers.fr/PHENOTIC/telechargements.html.

The traditional methods for evaluating seeds are usually performed through destructive sampling followed by physical, physiological, biochemical and molecular determinations. Whilst proven to be effective, these approaches can be criticized as being destructive, time consuming, labor intensive and requiring experienced seed analysts. Thus, the objective of this study was to investigate the potential of computer vision and multispectral imaging systems supported with multivariate analysis for high-throughput classification of cowpea (Vigna unguiculata) seeds. An automated computer-vision germination system was utilized for uninterrupted monitoring of seeds during imbibition and germination to identify different categories of all individual seeds. By using spectral signatures of single cowpea seeds extracted from multispectral images, different multivariate analysis models based on linear discriminant analysis (LDA) were developed for classifying the seeds into different categories according to ageing, viability, seedling condition and speed of germination. The results revealed that the LDA models had good accuracy in distinguishing ‘Aged’ and ‘Non-aged’ seeds with an overall correct classification (OCC) of 97.51, 96.76 and 97%, ‘Germinated’ and ‘Non-germinated’ seeds with OCC of 81.80, 79.05 and 81.0%, ‘Early germinated’, ‘Medium germinated’ and ‘Dead’ seeds with OCC of 77.21, 74.93 and 68.00% and among seeds that give ‘Normal’ and ‘Abnormal’ seedlings with OCC of 68.08, 64.34 and 62.00% in training, cross-validation and independent validation data sets, respectively. Image processing routines were also developed to exploit the full power of the multispectral imaging system in visualizing the difference among seed categories by applying the discriminant model in a pixel-wise manner. The results demonstrated the capability of the multispectral imaging system in the ultraviolet, visible and shortwave near infrared range to provide the required information necessary for the discrimination of individual cowpea seeds to different classes. Considering the short time of image acquisition and limited sample preparation, this stat-of-the art multispectral imaging method and chemometric analysis in classifying seeds could be a valuable tool for on-line classification protocols in cost-effective real-time sorting and grading processes as it provides not only morphological and physical features but also chemical information for the seeds being examined. Implementing image processing algorithms specific for seed quality assessment along with the declining cost and increasing power of computer hardware is very efficient to make the development of such computer-integrated systems more attractive in automatic inspection of seed quality.

We review a set of recent multiscale imaging techniques, producing high-resolution images of interest for plant sciences. These techniques are promising because they match the multiscale structure of plants. However, the use of such high-resolution images is challenging in the perspective of their application to high-throughput phenotyping on large populations of plants, because of the memory cost for their data storage and the computational cost for their processing to extract information. We discuss how this renews the interest for multiscale image processing tools such as wavelets, fractals and recent variants to analyse such high-resolution images.

In this article, we assess the interest of the recently introduced multiscale scattering transform for texture classification applied for the first time in plant science. Scattering transform is shown to outperform monoscale approaches (gray-level co-occurrence matrix, local binary patterns) but also multiscale approaches (wavelet decomposition) which do not include combinatory steps. The regime in which scatter transform also outperforms a standard CNN architecture in terms of data-set size is evaluated ( ${10}^4$ instances). An approach on how to optimally design the scatter transform based on energy contrast is provided. This is illustrated on the hard and open problem of weed detection in culture crops of high density from the top view in intensity images. An annotated synthetic data-set available under the form of a data challenge and a simulator are proposed for reproducible science. Scatter transform only trained on synthetic data shows an accuracy of $85\%$ when tested on real data.

Image analysis is increasingly used in plant phenotyping. Among the various imaging techniques that can be used in plant phenotyping, chlorophyll fluorescence imaging allows imaging of the impact of biotic or abiotic stresses on leaves. Numerous chlorophyll fluorescence parameters may be measured or calculated, but only a few can produce a contrast in a given condition. Therefore, automated procedures that help screening chlorophyll fluorescence image datasets are needed, especially in the perspective of high-throughput plant phenotyping. We developed an automatic procedure aiming at facilitating the identification of chlorophyll fluorescence parameters impacted on leaves by a stress. First, for each chlorophyll fluorescence parameter, the procedure provides an overview of the data by automatically creating contact sheets of images and/or histograms. Such contact sheets enable a fast comparison of the impact on leaves of various treatments, or of the contrast dynamics during the experiments. Second, based on the global intensity of each chlorophyll fluorescence parameter, the procedure automatically produces radial plots and box plots allowing the user to identify chlorophyll fluorescence parameters that discriminate between treatments. Moreover, basic statistical analysis is automatically generated. Third, for each chlorophyll fluorescence parameter the procedure automatically performs a clustering analysis based on the histograms. This analysis clusters images of plants according to their health status. We applied this procedure to monitor the impact of the inoculation of the root parasitic plant Phelipanche ramosa on Arabidopsis thaliana ecotypes Col-0 and Ler. Using this automatic procedure, we identified eight chlorophyll fluorescence parameters discriminating between the two ecotypes of A. thaliana, and five impacted by the infection of Arabidopsis thaliana by P. ramosa. More generally, this procedure may help to identify chlorophyll fluorescence parameters impacted by various types of stresses. We implemented this procedure at http://www.phenoplant.org freely accessible to users of the plant phenotyping community.