Depending on the continent, region of the world, location, type of the crop, cultivar and the weather conditions in the particular season crop plants lose from a few percent to the total amount of yield. A high number of plant pathogens belong to the kingdom of Fungi; they can cause enormous problems with plant health and can contribute to the decrease of not only yield quantity but also its quality. One of the solutions highly promoted in One Health concept are decision support systems helping in early detection and identification of pathogens or indicating the conditions which may trigger their development at a given site and time. Ideally, these early warning systems would detect minute amounts of pathogens in the air and warn the farmers before the problem occurs. The lecture will contain the examples of successful use of spore samplers to monitor fluctuations of the spores of fungal plant pathogens worldwide, with the special focus on the System for Forecasting Disease Epidemics operated by the author. Conference participants will learn how such systems are organized, how they work, what are the results of spore trapping and how it contributes to crop protection and plant health. More and more often monitoring becomes automated and the spore detection relies on DNA-based diagnostics. This all leads to the novel term of molecular aeromycology. The unique combination of spore trapping and molecular detection of pathogen DNA became a very useful modern way to combat plant diseases in the manner which is the least destructive to environment.

Sustainability of world agriculture is critically dependent on the development of new generation crops that will be more resilient to drought and capable of coping with increasing concentration of salt in the soil, without yield penalties. The latter can be done by incorporating some specific traits found in halophytes. In this talk, I will focus on two major mechanisms conferring salinity tolerance in halophytes that can be targeted in crop breeding programs. The first one is effective Na⁺ sequestration in either internal (vacuoles) or external (epidermal bladder cells; EBC) structures. The second mechanism is plant’s ability to maintain efficient photosynthetic gas exchange via stomata under hyperosmotic stress conditions. Working along the first lines, we show that internal (vacuolar) Na⁺ sequestration in halophytes is achieved by the orchestrated regulation of at least four major traits: (1) higher expression levels and operation of tonoplast NHX Na⁺/H⁺ exchangers; (2) higher tonoplast H⁺-pump activity; (3) better cytosolic K⁺ retention originating from intrinsically higher plasma membrane H⁺ pumping ability; and (4) efficient control of SV and FV tonoplast channels to prevent futile Na⁺ cycling between the cytosol and vacuole. I then analyse mechanism underlying sequestration of toxic Na⁺ and Cl^- species in external structures such as epidermal bladder cells. I describe the essentiality of the tissue-specific expression of specific transporters and show experimental evidence for the functional expression and operation of HKT, SOS1, and HAK transporters in epidermis-stalk cell- EBC continuum. In the final part of my presentation I discuss peculiarities of stomata operation in halophytes. Once molecular identity of key genes and control modes of the above processes are understood, plant breeders would be able to devise breeding strategies to incorporate desirable features into traditional crops via molecular or traditional breeding tools thus expanding the sustainable limits for productive use of soil and water resources.

Plant genetic resources (PGR) play a major role for global food security. The most significant and widespread mean of conserving PGR is ex situ storage. Since the majority of genebank accessions globally are stored in the form of seed, seed longevity is of particular importance for crop germplasm preservation. Research was initiated for a range of crops stored in the German genebank over decades. Variation between crop species was detected. However, there is also intraspecific variation within genebank collections. It was concluded that the differences in germination after long term storage are genetically based. Therefore, genetic analyses of seed longevity were initiated. Genetic mapping was performed for barley, wheat, oilseed rape and tobacco.

In addition, mass spectrometry based untargeted metabolite profiling experiments were performed, in order to detect biochemical changes coinciding with loss in seed germination. The lipidomic composition of a wheat panel was investigated using high-resolution liquid chromatography-mass spectrometry (LC-MS). A high proportion of tentative oxidized lipids was detected, suggesting lipid oxidation as the causal trigger for membrane degradation.

The agri-food sector generates massive amounts of waste, generating serious environmental and social problems. In the frame of the Circular Economy concept, Biotechnology is envisioned as a reservoir of techniques that can be adapted and applied to transform agricultural waste in new products. In this context, plant-waste extracts, rich in bioactive compounds, are emerging as key players able to improve plant growth and crop productivity to sustainably address food security. To use this recourse, there is the need for safe, cheap, and sustainable technologies to recover these bioactive molecules, while avoiding the use of toxic organic solvents or expensive equipment. One such approach includes the use of beneficial bacteria, like Bacillus subtilis, naturally equipped with enzymes able to target complex plant polymers. The use of bacteria is combined with extraction methods based on isothermal pressurization cycles. The obtained products, rich in polyphenols like rutin, quercetin, ferulic acid, ecc., can be then reincorporated into the agronomic practices as biostimulants (defined as natural bioactive compounds of plant origin, acting as enhancers of plant growth and development), applied as foliar spray, soil irrigation, or as seed treatments or like plant protection products (PPP) towards harmful microorganisms as well. Additionally, such eco-friendly agricultural practices seek to mitigate environmental harm, ensuring long-term food production and economic stability. At a molecular level, the stimulating effects of such treatments on plant growth and stress resilience are mainly related to a substantial boost in the antioxidant defense. Advanced molecular techniques such as transcriptomics, metabolomics, and proteomics can provide deeper insights into the biochemical and molecular responses of plants treated with these bioactive compounds. Therefore, these biotechnological innovations not only aligns with the principles of the Circular Economy but also represent a sustainable strategy to enhance agricultural productivity while minimizing environmental impact.

Sage (Salvia officinalis L.) is considered a functional food that has many other uses in medicine, cosmetics, and the food industry, among others. It grows in the wild but is also widely cultivated for human consumption. The application of plant growth-promoting (PGP) microbes in sage production can be a sustainable eco-friendly production measure contrasted to traditional systems where chemical-based management practices are used. Moreover, the usage of different products of microbial origin is constantly increasing due to growing awareness of the potentially harmful effects of artificial substances in food. Pseudomonads are well-known bacteria with PGP traits. Phytohormone production, nitrogen fixation, phosphorus solubilization, and iron sequestration by siderophores are only a few mechanisms of growth promotion. The aim of this study was to evaluate the efficiency of Pseudomonas sp. isolates (denoted as P8, P37, and P53) on seed germination and the initial growth of sage in semi-controlled conditions for four weeks. Morphological, physiological, biochemical, and PGP characterization of bacteria isolates were performed prior to the study. All isolates demonstrated a good ability to produce siderophore and solubilize phosphorus, while only P53 produced hydrogen cyanide (HCN). Seed inoculation led to better germination and the initial growth of sage. Seedling development was stimulated, reaching a maximum of 57 mm in treatment P53 after 5 days. On average, a 21% increase was achieved with P37 and P53 in relation to the untreated control. Regarding root length and plant height, the inoculated plants were 2.4 to 3.4 and 1.2 to 1.5-fold larger in size, respectively. All isolates showed stimulatory potential, positively influencing the initial growth of sage. The best stimulatory potential was achieved with P37. Pseudomonas sp. isolates proved to have good bio-stimulatory potential and can be used in sage production.

Helichrysum italicum (Roth) G. Don is an important medicinal plant native to the Mediterranean region. Positive effects of the biological compounds on human health have been demonstrated by in vitro and in vivo studies. However, most studies so far have focussed mainly on the chemical composition of the essential oil, while the importance of the genotype has been neglected due to the lack of effective molecular markers. The latter is particularly important due to the complex taxonomic classification (the existence of subspecies and interspecies hybrids poses a challenge for the classification of species and subspecies) and cross-pollination, which contributes to high genetic and phenotypic diversity within populations. The aim of our study was to initiate genomic studies and develop SSR markers to identify genotypes. Initially, genomic DNA was sequenced using Ion Torrent technology, utilising a shotgun approach. The assembled contigs of the nuclear genome and chloroplast genome¹ were used to develop 24 highly informative nuclear microsatellite markers² and 16 chloroplast microsatellite markers³. Considering that H. italicum is in the early stages of domestication and cultivation in the agroecosystem is usually based on unverified plant material, the newly developed molecular markers represent a valuable tool for the identification of genotypes with different chemical compositions. Microsatellites are also useful in plant breeding for the control of parentage, for the authentication and traceability of plant material on the market, for the protection of local products and geographical origin, and for the exploration of genetic resources in botanical studies. Due to their transferability, they are also useful for research on related species. This paper presents the benefits and applicability of microsatellites and the results of recent genetic research on the immortelle.

This study aimed to evaluate the antioxidant properties of powdered lentil seeds after sprouting and elicitation. Green lentil seeds served as the research material. The seeds were treated with hydrogen peroxide as an elicitor and germinated for four days in a controlled climate chamber at 20°C and 80% relative humidity. The resulting sprouts were then subjected to either freeze-drying at 20, 40, and 60°C or air-drying at 40 and 60°C before being ground into powder. The following characteristics were assessed in the dried powders: color coordinates, total phenolics compound content (TPC), and antioxidant activity (AA) against DPPH and ABTS radicals. The findings indicated that both the elicitation process and the drying method significantly affected TPC and the antioxidant properties of the sprouts. Sprouts dried by convection showed lower TPC and antioxidant activity than those that were freeze-dried. Elicitation increased both TPC and AA across all drying temperatures. The optimal results were achieved with elicited lentil seeds that were freeze-dried at 20 and 40°C. However, elicitation did not significantly impact the color coordinates of the dried material. Convection-dried sprouts appeared lighter and less red than those obtained through freeze-drying.

Abstract: Asclepias syriaca, commonly called common milkweed native to North America is a perennial weed that has shown negative impacts on biodiversity and also as a competitive weed in cultivated fields. Common milkweed plants that naturally exhibited symptoms of wilting and necrosis were taken to isolate and identify the fungus. Based on DNA sequencing, this species was determined to be Colletotrichum sp. The experiment was conducted under laboratory conditions to determine the efficacy of treatment. Colletotrichum sp. was never isolated from the control plants, and none of them showed any symptoms. When the treatment was carried out at a concentration of 10⁷ spores/ml, common milkweed was able to show visible symptoms on its leaves. These findings suggest that fungi isolation from A. syriaca can be an effective biocontrol method in the future. While the results are encouraging, further studies are needed to optimize the pathogens application and ensure its environmental safety under field conditions. This research underscores the potential of fungal pathogens as eco-friendly alternatives to conventional weed control methods and contributes to knowledge on biological control strategies for invasive plant species.

Climatic variability and extreme weather events are increasingly impacting crop yield and value. Camelina sativa is a versatile, low-input Brassicaceae oilseed crop valued for its high-quality seeds and its adaptability to a wide range of climate conditions and cropping systems. Camelina's innate resilience to abiotic stress offers an opportunity to uncover mechanisms behind it and explore crop plasticity. This understanding is crucial for breeding climate-tolerant oilseeds in the context of climate change. Within the project UNTWIST (Grant Agreement No. 862524; www.untwist.eu), 54 accessions were collected from across Eurasia to showcase the genetic and plastic diversity of camelina in response to abiotic stress (drought and heat) and to assess performance in multi-location field trials. Genome sequencing of the 54 accessions and population structure analysis revealed that the study panel captured most of the diversity observed in the larger publicly available population. A combinatorial approach of field trials and trials in a controlled environment was used to investigate the response of camelina to drought and heat stress and examine the diversity of responses shown by individual camelina biotypes. Camelina demonstrated resistance to heat and drought, and the population illustrated a range of different adaptive responses. Morphology and phenology showed contrasting responses to each stress. (Un)targeted metabolomic profiling revealed a reorganisation of the metabolism of camelina, with substantial variation in line-specific responses to each of the applied stresses. The same trend was found in leaf fatty acid composition remodeled by stress. Metabolic markers that could underpin future breeding have been identified, and parameters have been developed from field trials at several sites that provide a means of assessing the agronomic performance of various germplasm. Camelina productivity in the multi-location field trials was linked to the environment at key developmental stages.

Erwinia amylovora, the etiological agent of fire blight disease, is an important threat to pome fruit production worldwide, particularly affecting apple (Malus domestica) and pear (Pyrus communis) trees and a wide number of other hosts within the Rosaceae family. When entering the cells, the bacteria disrupt the plant’s natural defense mechanisms and, once inside the xylem, can form biofilms, leading to tissue death due to blocked water and essential minerals transport. Unfortunately, there are no effective methods to halt the spread of this disease, which is aggravated by environmental changes that create favorable climatic conditions and the increased international trade of contaminated plant material. Despite its global prevalence and ability to infect a plethora of ornamental and edible hosts, studies indicate a high level of genomic homogeneity among E. amylovora strains worldwide. Despite previous genomic characterization efforts in Portugal, the lack of comprehensive epidemiological data and understanding of local environmental influences on pathogen evolution prevents effective management and outbreak prediction. In this study, results from phylogenetic inference and CRISPR CRR1 identification raise the hypothesis that there are at least two different invasion routes for E. amylovora in Portugal, or that different dissemination routes occurred since the first entry in the country. Phenotypic data are being collected on 82 Portuguese isolates, which already disclosed strains with growth rates ranging from 0.155 to 0.315 h-1 and different virulence levels. This metadata will be integrated with genomic analysis such as the identification of single-nucleotide polymorphisms (SNPs); functional annotation of candidate genes like ams, rls, and hrp; and exploration of relevant plasmid presence to elucidate the phenotypic variability among isolates. The findings will address gaps in understanding E. amylovora’s genome regulation to shed light on the evolutionary mechanisms driving fire blight’s adaptation and success, ultimately contributing to outbreak mitigation and improving disease management.

This work received financial support from PT national funds (FCT/MECI, Fundação para a Ciência e Tecnologia and Ministério da Educação, Ciência e Inovação) through the projects UID/50006 -Laboratório Associado para a Química Verde - Tecnologias e Processos Limpos and 2023.01413.BD.