MRDV / Institute for Rural Development / See the attached abstract below

<p dir="ltr">Urban agriculture has grown in popularity in recent decades, due to its ability to provide access to healthy fruits and vegetables in urban zones, as well as its importance in fostering knowledge of agriculture within communities. However, urban agriculture may struggle with unique challenges due to its proximity to urban and industrial activities, such as food safety risks due to toxic heavy metals and metalloids which may be present in urban soils in high concentrations. Heavy metals and metalloids (HM) like arsenic, cadmium, and lead are absorbed by plants from the soil, and may accumulate in the plants’ edible tissues, which are consumed by humans. Carrot (<i>Daucus carota</i> L.), in particular, hyperaccumulates these toxic heavy metals in its edible taproots, leading to food safety risks on urban farms.</p><p dir="ltr">One potential way to help address this challenge is to breed carrot varieties with low uptake of HM. In recent years, researchers have identified lines with high and low uptake in greenhouse trials and single location breeding nurseries. However, to be viable, these lines must consistently vary in HM across sites despite differences in environmental and management factors that can also greatly influence HM bioavailability and uptake. Moreover, screening for differences in HM uptake is time-consuming and expensive, and breeders need new tools to select among segregating breeding populations. By using on-farm participatory research as well as advanced phenotyping technologies, we investigate the viability of breeding carrots for HM uptake and the potential of new tools to advance these efforts in order to mitigate the risks on urban farms.</p><p dir="ltr">In the summers of 2021 and 2022, participatory on-farm trials were conducted to determine the HM risks on Indiana urban farms and to investigate the consistency of differences in HM uptake between carrot breeding lines taken from breeding trials (Chapter 2). Results of these trials indicated that while carrot genotype had an effect, there was still significant variability in carrot uptake of arsenic, cadmium and lead between farm sites and years. Results indicated significant differences between site-years, and carrot HM concentrations that correlated strongly with soil concentrations for that particular element. However, there were some site-years with low soil HM content and other soil factors expected to reduce uptake such as pH and phytoavailable zinc concentrations (such as site-year H), that had high carrot HM content. There were significant differences in carrot cadmium (Cd) and arsenic (As) content between carrot breeding lines. For instance, breeding line 3271 had a high As average concentration but low Cd average concentration, while breeding lines 6220 and 2327 had low As and high Cd concentrations. We identify the possibility of other mediating factors, such as uptake of antagonistic micronutrients, or microbe-assisted HM uptake and amelioration that need further attention.</p><p dir="ltr">In the fall of 2022, a study was conducted to investigate the possibility of using phenotyping technologies such as RGB and hyperspectral imaging to detect Cd stress in carrot and attempt to predict uptake (Chapter 3). RGB (red green blue) is a digital color model in which cameras can capture important visual cues compiled from information about each pixel. Hyperspectral imaging uses cameras to capture wavelengths beyond the visible spectrum, which can detect plant stress indicators like increased anthocyanin content for specific environmental stresses. Results of this trial were useful, with some time points and indices noting differences between carrot lines. For instance, RGB factors hue and fluorescence as well as hyperspectral reflectance plots and vegetative indices swirNDVI and ANTH were the most diagnostic. Breeding lines 6636 and 8503 showed the greatest separation between Cd treated and control carrots in imaging indices. However, further studies will be needed to optimize this approach for breeding programs.</p><p dir="ltr">This research demonstrates that growing carrots on most urban farms in Indiana is safe. The studies also provide further evidence that it will be possible to help lower food safety risks by selecting carrot varieties with low HM uptake, and phenotyping can help to advance these efforts. At the same time, new research to understand how soil factors such as microbiomes influence HM bioavailability and uptake on urban farms are also needed to further reduce potential risks. In the meantime, farmers should continue to test their soil for HM and take appropriate actions to reduce risks such as using raised beds and soil amendments that can bind metals like biochar. Consumers should also continue to wash and peel their carrots before consumption, as well as consume a balanced diet with a diverse set of vegetables and other crops.</p>

Tesis por compendio / [ES] El cambio climático ha aumentado la exposición de los cultivos a estreses como la sequía o la salinidad y, por tanto, tiene un impacto negativo en el rendimiento de las plantas y los cultivos en un mundo con una población en aumento en el que la seguridad alimentaria está amenazada. Existe un interés en desarrollar variedades y cultivares con mayor tolerancia a estreses abióticos. Con esta finalidad desarrollamos un abordaje, en cultivos con un alto valor económico y nutricional como brócoli y melón, para identificar rasgos fisiológicos y bioquímicos posiblemente limitantes de estrés abiótico y, por tanto, podrían ser dianas biotecnológicas para mejorar la tolerancia a estrés abiótico o que pueden ser útiles para predecir la tolerancia de variedades no caracterizadas previamente. En la presente tesis, empleamos fisiología y metabolómica para identificar como rasgos distintivos de tolerancia a estrés por sequía en brócoli, la fotosíntesis neta, la eficiencia en el uso del agua, la conductancia estomática, el ácido abscísico, metabolitos relacionados con el metabolismo del azufre y otras moléculas como la urea, el ácido quínico y el ácido glucónico lactona. Por otra parte, en melón encontramos el potencial hídrico y varios aminoácidos como serina, fenilalanina, glicina, isoleucina, asparagina y triptófano como rasgos distintivos entre cultivares tolerantes o sensibles bajo estrés por sequía. Al respecto del estrés salino, identificamos el ratio Na+/K+ como rasgo distintivo tanto para brócoli como para melón mientras que sólo en brócoli, la transpiración y la conductancia estomática como rasgos fisiológicos útiles y, en cuanto a rasgos bioquímicos, hormonas como el ácido abscísico, el ácido jasmónico y el ácido indol-3-acético así como metabolitos del ciclo de Krebs y el contenido total de los relacionados glutatión, cisteína y metionina y prolina, se mostraron como indicadores de respuestas a estrés salino en brócoli. Sin embargo, en melón, encontramos prolina, fenilalanina e histidina como rasgo distintivo de tolerancia a estrés salino. También utilizamos herramientas metabolómicas para identificar que el ácido γ-aminobutírico correlaciona con un mejor sabor del brócoli. Además, dada la importancia del metabolismo del azufre y la tolerancia al estrés, así como su relación con las propiedades nutricionales del brócoli, llevamos a cabo la caracterización de las enzimas serina O-acetiltransferasas de brócoli. La evidencia que encontramos sugiere que las dianas biotecnológicas más prometedoras para mejorar la tolerancia al estrés son los parálogos BoSAT1d y BoSAT3. / [CA] El canvi climàtic ha augmentat l'exposició dels cultius a estressos com la sequera o la salinitat i, per tant, té un impacte negatiu en el rendiment de les plantes i els cultius en un món amb una població en augment on la seguretat alimentària està amenaçada . Hi ha un interès a desenvolupar varietats i cultivars amb més tolerància a estressos abiòtics. A aquest efecte desenvolupem un abordatge, en cultius amb un alt valor econòmic i nutricional com ara bròquil i meló, per identificar trets fisiològics i bioquímics possiblement limitants d'estrès abiòtic i, per tant, podrien ser dianes biotecnològiques per millorar la tolerància a estrès abiòtic o que poden ser útils per predir la tolerància de varietats no prèviament caracteritzades. En aquesta tesi, utilitzem fisiologia i metabolòmica per identificar com a trets distintius de tolerància a estrès per sequera en bròquil, la fotosíntesi neta, l'eficiència en l'ús de l'aigua, la conductància estomàtica, l'àcid abscísic, metabòlits relacionats amb el metabolisme del sofre i altres molècules com la urea, l'àcid quínic i l'àcid glucònic lactona. D'altra banda, en meló trobem el potencial hídric i diversos aminoàcids com a serina, fenilalanina, glicina, isoleucina, asparagina i triptòfan com a trets distintius entre cultivars tolerants o sensibles sota estrès per sequera. Pel que fa a l'estrès salí, identifiquem la ràtio Na+/K+ com a tret distintiu tant per a bròquil com per a meló mentre que només en bròquil, la transpiració i la conductància estomàtica com a trets fisiològics útils i, quant a trets bioquímics, hormones com l'àcid abscísic , l'àcid jasmònic i l'àcid indol-3-acètic així com metabòlits del cicle de Krebs i el contingut total dels relacionats glutatió, cisteïna i metionina i prolina, es van mostrar com a indicadors de respostes a estrès salí en bròquil. Tot i això, en meló, trobem prolina, fenilalanina i histidina com a tret distintiu de tolerància a estrès salí. També utilitzem eines metabolòmiques per identificar que l'àcid γ-aminobutíric correlaciona amb un millor sabor del bròquil. A més, atesa la importància del metabolisme del sofre i la tolerància a l'estrès, així com la seva relació amb les propietats nutricionals del bròquil, duem a terme la caracterització dels enzims serina O-acetiltransferases de bròquil. La seva evidència suggereix que les dianes biotecnològiques més prometedores per millorar la tolerància a l'estrès són els paràlegs BoSAT1d i BoSAT3. / [EN] Climate change has increased the exposure of crops to stresses like drought and salinity and, thus, it has a negative impact on plant performance and crop yield in a world with an increasing population. This threatens food security. There is an interest in developing new varieties and cultivars of crops with enhanced tolerance to abiotic stresses. With this aim, we developed an approach in crops with high economic and nutritional values, such as broccoli and melon, to identify physiological and biochemical traits that could be limiting factors for abiotic stress and, thus are likely biotechnological targets for improving abiotic stress tolerance or may be useful for predicting tolerance of uncharacterized varieties. In the present thesis, we used physiology and metabolomics to identify as distinctive traits for drought stress tolerance in broccoli, net photosynthesis, water use efficiency, stomatal conductance, abscisic acid, metabolites related to sulfur metabolism and other molecules such as urea, quinic acid and gluconic acid lactone. On the other hand, in melon, we found water potential and several amino acids as serine, phenylalanine, glycine, isoleucine, asparagine and tryptophan to be differential traits among tolerant and sensitive cultivars under drought stress. Regarding salt stress tolerance, we identified Na+/K+ ratios as a distinctive trait for both broccoli and melon whereas in broccoli, but not melon, transpiration, and stomatal conductance were useful physiological traits. Concerning biochemical traits, hormones, such as abscisic acid, jasmonic acid and indole-3-acetic acid, metabolites of the Krebs cycle and total content of the related glutathione, cysteine and methionine and proline proved to be indicative of saline stress responses in broccoli. However, in melon, we found only proline, phenylalanine and histidine as a distinctive trait of salt stress tolerance. We also used metabolomic tools to identify that γ-Aminobutyric acid correlates with a better taste in broccoli. Given the importance of sulfur metabolism in stress tolerance, as well as its link with the nutritional properties of broccoli, we also carried out the characterization of the serine O-acetyltransferase enzymes in broccoli. Among the eight isoforms found in this species, our data suggest that the most promising biotechnological targets for enhancing stress tolerance are the BoSAT1d and the BoSAT3 paralogues. / I am a recipient of the FPU19/01977 grant from the Spanish Ministerio de Universidades. This work was funded by the RTC-2017-6468-2-AR project (APROXIMACIONES MOLECULARES PARA INCREMENTAR LA TOLERANCIA A SALINIDAD Y SEQUÍA DEL BRÓCOLI) awarded by the “Agencia estatal de Investigación” / Chevilly Tena, S. (2024). Molecular approaches to characterize and improve abiotic stress tolerance in broccoli and other crops [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/213681 / Compendio