<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>

Hyper-Raman-Streuung folgt anderen Symmetrieauswahlregeln als Raman-Streuung und profitiert als nicht-linearer Zweiphotonenprozess noch mehr von verstärkten elektromagnetischen Feldern an der Oberfläche plasmonischer Nanostrukturen. Damit könnte die oberflächenverstärkte Hyper-Raman-Streuung (SEHRS) praktische Bedeutung in der Spektroskopie erlangen. Durch die Kombination von SEHRS und oberflächenverstärkter Raman-Streuung (SERS) können komplementäre Strukturinformationen erhalten werden. Diese eignen sich aufgrund der Lokalisierung der Verstärkung auf die unmittelbare Umgebung der Nanostrukturen besonders für die Charakterisierung der Wechselwirkung zwischen Molekülen und Metalloberflächen. Ziel dieser Arbeit war es, ein tieferes Verständnis des SEHRS-Effekts zu erlangen und dessen Anwendbarkeit für analytische Fragestellungen einzuschätzen. Dazu wurden SEHRS-Experimente mit Anregung bei 1064 nm und SERS-Experimente mit Anregung bei derselben Wellenlänge sowie mit Anregung bei 532 nm - für eine Detektion von SEHRS und SERS im gleichen Spektralbereich - durchgeführt. Als Beispiel für nicht-resonante Anregung wurden die vom pH-Wert abhängigen SEHRS- und SERS-Spektren von para-Mercaptobenzoesäure untersucht. Mit diesen Spektren wurde die Wechselwirkung verschiedener Silbernanostrukturen mit den Molekülen charakterisiert. Anhand von beta-Carotin wurden Einflüsse von Resonanzverstärkung im SEHRS-Experiment durch die gleichzeitige Anregung eines molekularen elektronischen Übergangs untersucht. Dabei wurde durch eine Thiolfunktionalisierung des Carotins eine intensivere Wechselwirkung mit der Silberoberfläche erzielt, sodass nicht nur resonante SEHRS- und SERS-Spektren, sondern auch nicht-resonante SERS-Spektren von Carotin erhalten werden konnten. Die Anwendbarkeit von SEHRS für hyperspektrale Kartierung in Verbindung mit Mikrospektroskopie wurde durch die Untersuchung von Verteilungen verschiedener Farbstoffe auf strukturierten plasmonischen Oberflächen demonstriert. / Hyper-Raman scattering follows different symmetry selection rules than Raman scattering and, as a non-linear two-photon process, profits even more than Raman scattering from enhanced electromagnetic fields at the surface of plasmonic nanostructures. Surface-enhanced hyper-Raman scattering (SEHRS) could thus gain practical importance for spectroscopy. The combination of SEHRS and surface-enhanced Raman scattering (SERS) offers complementary structural information. Specifically, due to the localization of the enhancement to the close proximity of the nanostructures, this information can be utilized for the characterization of the interaction between molecules and metal surfaces. The aim of this work was to increase the understanding of the SEHRS effect and to assess its applicability to answer analytical questions. For that purpose, SEHRS experiments with excitation at 1064 nm and SERS experiments with excitation at the same wavelength, as well as with excitation at 532 nm - to detect SEHRS and SERS in the same spectral region - were conducted. As an example for non-resonant excitation, pH-dependent SEHRS and SERS spectra of para-mercaptobenzoic acid were examined. Based on these spectra, the interaction of different silver nanostructures with the molecules was characterized. beta-Carotene was used to study the influence of resonance enhancement by the excitation of a molecular electronic transition during SEHRS experiments. By the thiol-functionalization of carotene, a more intense interaction with the silver surface was achieved, which enables to obtain not only resonant SEHRS and SERS but also non-resonant SERS spectra of carotene. Hyperspectral SEHRS imaging in combination with microspectroscopy was demonstrated by analyzing the distribution of different dyes on structured plasmonic surfaces.

Optics are a powerful probe of chemical structure that can often be linked to theoretical predictions, providing robustness as a measurement tool. Not only do optical interactions like second harmonic generation (SHG), single and two-photon excited fluorescence (TPEF), and infrared absorption provide chemical specificity at the molecular and macromolecular scale, but the ability to image enables mapping heterogeneous behavior across complex systems such as biological tissue. This thesis will discuss nonlinear and linear optics, leveraging theoretical predictions to provide frameworks for interpreting analytical measurement. In turn, the causal mechanistic understanding provided by these frameworks will enable structurally specific quantitative tools with a special emphasis on application in biological imaging. The thesis will begin with an introduction to 2nd order nonlinear optics and the polarization analysis thereof, covering both the Jones framework for polarization analysis and the design of experiment. Novel experimental architectures aimed at reducing 1/f noise in polarization analysis will be discussed, leveraging both rapid modulation in time through electro-optic modulators (Chapter 2), as well as fixed-optic spatial modulation approaches (Chapter 3). In addition, challenges in polarization-dependent imaging within turbid systems will be addressed with the discussion of a theoretical framework to model SHG occurring from unpolarized light (Chapter 4). The application of this framework to thick tissue imaging for analysis of collagen local structure can provide a method for characterizing changes in tissue morphology associated with some common cancers (Chapter 5). In addition to discussion of nonlinear optical phenomena, a novel mechanism for electric dipole allowed fluorescence-detected circular dichroism will be introduced (Chapter 6). Tackling challenges associated with label-free chemically specific imaging, the construction of a novel infrared hyperspectral microscope for chemical classification in complex mixtures will be presented (Chapter 7). The thesis will conclude with a discussion of the inherent disadvantages in taking the traditional paradigm of modeling and measuring chemistry separately and provide the multi-agent consensus equilibrium (MACE) framework as an alternative to the classic meet-in-the-middle approach (Chapter 8). Spanning topics from pure theoretical descriptions of light-matter interaction to full experimental work, this thesis aims to unify these two fronts. <br>

Tesis por compendio / [ES] El conocimiento del estado nutricional de los cultivos permite corregir o ajustar cualquier exceso o deficiencia nutricional en los mismos, a lo largo de su ciclo vegetativo, asegurando un alto rendimiento en la producción y una óptima calidad del fruto. Tradicionalmente, para la realización del diagnóstico nutricional se ha utilizado el análisis de la ionómica de diferentes órganos de la planta, especialmente las hojas, por su facilidad de muestreo y por ser el órgano fotosintético de excelencia en las plantas. Por ello es necesario implementar estrategias sostenibles que nos permitan ajustar la dosis de fertilización según las necesidades del cultivo con el mínimo riesgo de contaminación. El objetivo de esta tesis doctoral es desarrollar métodos y modelos que permitan el diagnóstico nutricional en cultivos mediante métodos ópticos no destructivos, como la espectroscopia y la imagen hiperespectral en el rango Vis-NIR, en combinación con técnicas de quimiometría. De este modo, el primer bloque centrado en el cultivo de caqui cv. 'Rojo Brillante', comprende los estudios publicados en dos artículos científicos. En el primero de estos artículos se estudió el potencial de la espectroscopia Vis-NIR (430-1040 nm), con el objetivo de predecir macros y micronutrientes utilizando modelos de regresión PLS. Los resultados mostraron que es posible predecir de forma precisa macronutrientes como, fósforo (P), calcio (Ca) y magnesio (Mg), con un coeficiente de determinación en la predicción (R2P) de 0,78 a 0,63. En los micronutrientes, el boro (B) y el manganeso (Mn) fueron los que obtuvieron mejores coeficientes de predicción, con R2P de 0,79 y 0,69, respectivamente. En el segundo artículo se ha evaluado, para la estimación de la concentración de nutrientes, el uso de imágenes hiperespectrales en el rango entre 500 y 980 nm. Los resultados mostraron la predicción de los macronutrientes como N, P, potasio (K), Ca y Mg con R2P de 0,80 a 0,62 y, para los micronutrientes, solo en el B se obtuvo un valor aceptable para la estimación (R2p = 0,69). Además, utilizando el método de reducción de variables de influencia en la proyección (VIP) se obtuvo una predicción fiable para los nutrientes de N (R2P = 0,76) y B (R2P = 0,61). En el segundo bloque, se ha estudiado otro cultivo emblemático en la Comunidad Valenciana por su importancia social y económica, como son los cítricos. En este caso, se desarrollaron herramientas de estimación del cv. 'clementina de Nules', descritas en otros dos artículos científicos. En el tercer artículo se ha estudiado la capacidad de la espectroscopía para determinar la concentración de nutrientes en las hojas de los cítricos en un ciclo vegetativo completo. Los resultados mostraron una predicción con un R2P de 0,70 a 0,65 para el P, K Ca y B. Utilizando el coeficiente de regresión de ponderado (BW) se determinó un subconjunto de bandas importantes para determinar la concentración de P, K y B. Los resultados mostraron que las bandas de mayor relevancia para estos nutrientes se situaron en la región del visible (430-750 nm), asociada a la absorción de pigmentos fotosintéticos. Finalmente, en el cuarto artículo se ha estudiado el potencial de la imagen hiperespectral para discriminar entre hojas jóvenes y hojas de ciclos vegetativos anteriores, lo que mejoraría el diagnóstico dado que las tablas de referencia en este cultivo están realizadas en hojas de la brotación de primavera. Partiendo de esa hipótesis, se obtuvo que es posible realizar la discriminación entre ambos tipos de hojas. Posteriormente se realizó la predicción de concentración de nutrientes de hojas jóvenes, utilizando 49 bandas espectrales, obteniendo mejores resultados para los nutrientes P, K, Ca, hierro (Fe) y Mn con R2P de 0,69 a 0,60. Además, se realizó la predicción de estos nutrientes minimizando el número de bandas a diez, con el BW y se obtuvo un R2P de 0,67 a 0,57. / [CA] El coneixement de l'estat nutricional dels cultius permet corregir o ajustar qualsevol excés o deficiència nutricional en estos, al llarg del seu cicle vegetatiu, assegurant un alt rendiment en la producció i una òptima qualitat del fruit. Tradicionalment, per a la realització del diagnòstic nutricional s'han utilitzat l'anàlisi de la ionómica de diferents òrgans de la planta, especialment les fulles, per la seua facilitat de mostreig i per ser l'òrgan fotosintètic d'excellència en les plantes. Per això és necessari implementar estratègies sostenibles que ens permeten ajustar la dosi de fertilització segons les necessitats del cultiu amb el mínim risc de contaminació. L'objectiu d'esta tesi doctoral és desenvolupar mètodes i models que permeten la predicció del diagnòstic nutricional en cultius mitjançant mètodes òptics no destructius, com l'espectroscòpia Vis-NIR, en combinació amb tècniques quimio mètriques. D'esta manera, el primer capítol de publicacions es centra en el cultiu del caqui cv. Rojo Brillante, comprés per dos articles (I i II). En el primer d'estos articles es va estudiar el potencial de l'espectroscòpia Vis-NIR (430-1040 nm), amb l'objectiu de predir macros i micronutrients utilitzant models de regressió PLS. En este estudi es van aplicar tractaments diferencials per als nutrients de N (0 %, 33 %, 50 % i 100 %) i per a K2O (0 %, 50 % i 100 %) de la demanda del cultiu. Els resultats van mostrar que, sí que és possible predir de manera precisa macronutrients com, fòsfor (P), calci (Ca)i magnesi (Mg), amb un coeficient de determinació en la predicció (R2P) de 0,78 a 0,63. En els micronutrients, com el bor (B) i el manganés (Mn) van ser els que van obtindre millors coeficients de predicció, amb R2P de 0,79 i 0,69, respectivament. En el segon article s'ha avaluat, per a l'estimació de la concentració de nutrients, l'ús d'imatges hiperespectrales en un rang (500-980 nm). Els resultats van mostrar la predicció dels macronutrients com a nitrogen (N), P, potassi (K), Ca i Mg amb R2P de 0,80 a 0,62 i, per als micronutrients, només en el B es va obtindre un valor acceptable per a l'estimació (R2p 0,69). A més, utilitzant el mètode de reducció de variables d'influència en la projecció (VIP) es va obtindre una predicció fiable per als nutrients de N (R2P 0,76) i B (R2P 0,61). En el segon capítol, s'ha estudiat un altre cultiu emblemàtic a la Comunitat Valenciana d'importància econòmica, com són els cítrics. En este cas, es van desenvolupar ferramentes d'estimació del cv. 'clementina de Nules' compreses en dos articles (III i IV). De tal manera, que en el tercer article s'ha estudiat la capacitat de les tècniques espectromètriques per a determinar la concentració de nutrients en un cicle vegetatiu complet. Els resultats van mostrar una predicció amb un R2P de 0,70 a 0,65 per al P, K, Ca i B. Utilitzant el coeficient de regressió de pes (BW) es va determinar un subconjunt de bandes més influents per als nutrients P, K i B. Els resultats van mostrar que les bandes de major importància, per a estos nutrients, es situen a la regió del Vis (430-750 nm), el qual està associada a l'absorció de pigments fotosintètics. Finalment, en el quart article s'ha estudiat el potencial de les HSI per a discriminar fulles joves de fulles de cicles vegetatius anteriors, la qual cosa milloraria el diagnòstic atés que les taules de referència en este cultiu estan realitzades en fulles de la brotada de primavera. Posteriorment es va realitzar la predicció de concentració de nutrients de fulles joves, utilitzant 49 bandes espectrals, obtenint millors resultats per als nutrients P, K, Ca, ferro (Fe) i Mn amb R2P de 0,69 a 0,60. A més, es va realitzar la predicció d'estos nutrients minimitzant el nombre de bandes a deu, amb el BW i es va obtindre un R2P de 0,67 a 0,57. / [EN] Knowledge of the nutritional status of crops allows for correcting or adjusting any nutritional excess or deficiency throughout their vegetative cycle, ensuring high yields in production and optimal fruit quality. Traditionally, the analysis of the ionomics of different plant organs has been used for nutritional diagnosis, especially the leaves, due to their ease of sampling and being the photosynthetic organ par excellence in plants. These analyses are carried out by expensive conventional laboratory methods that are destructive, polluting, time-consuming and costly. Therefore, it is necessary to implement sustainable strategies that allow the fertilisation dose to be adjusted according to the crop's needs with the minimum risk of contamination. This doctoral thesis aims to develop methods and models for nutritional diagnosis prediction in crops using non-destructive optical methods, such as Vis-NIR spectroscopy, combined with chemometric techniques. Thus, the first chapter of the publications focuses on cultivating persimmon cv. 'Rojo Brillante', comprising two articles (I and II). In the first of these articles, the potential of Vis-NIR spectroscopy (430-1040 nm) was studied to predict macronutrients and micronutrients using PLS regression models. This study applied differential treatments for N nutrients (0 %, 33 %, 50 % and 100 %) and K2O (0 %, 50 % and 100 %) of crop demand. The results showed that it is possible to accurately predict macronutrients such as phosphorus (P), calcium (Ca) and magnesium (Mg), with a coefficient of determination in the prediction (R2P) of 0.78 to 0.63. Boron (B) and manganese (Mn) obtained the best micronutrient prediction coefficients, with R2P of 0.79 and 0.69, respectively. The second article evaluated hyperspectral imaging (HSI) in the range (500-980 nm) for nutrient concentration estimation. The results showed the prediction of macronutrients such as nitrogen (N), P, potassium (K), Ca and Mg with R2P from 0.80 to 0.62 and, for micronutrients, only in B, an acceptable value for the estimation was obtained (R2p 0.69). In addition, using the projection influence variable reduction (VIP) method, a reliable prediction was obtained for N (R2P 0.76) and B (R2P 0.61) nutrients. In the second chapter, another emblematic crop of economic importance in the Valencian Community, citrus, was studied. Estimation tools were developed for citrus cv. 'Clementina de Nules' and the results were published in two articles (III and IV). Thus, in the third article, the capacity of spectrometric techniques to determine the concentration of nutrients in a complete vegetative cycle was studied. The results showed prediction with an R2P of 0.70 to 0.65 for P, K Ca and B. Using the weight regression coefficient (BW), a subset of more influential bands was determined for P, K and B nutrients. The results showed that the bands of greatest importance for these nutrients are located in the Vis region (430-750 nm), which is associated with photosynthetic pigment uptake. Finally, in the fourth article, the potential of HSI to discriminate young leaves from leaves of previous vegetative cycles has been studied, which would improve the diagnosis given that the reference tables in this crop are made on leaves of spring sprouting. Subsequently, the prediction of nutrient concentration of young leaves was carried out using 49 spectral bands, obtaining better results for the nutrients P, K, Ca, iron (Fe) and Mn with R2P from 0.69 to 0.60. In addition, these nutrients were predicted by minimizing the number of bands to ten, with the BW and an R2P of 0.67 to 0.57. ¿ / Maylin Acosta thanks IFARHU-SENACYT for the Professional Excellence Scholarships, contract No. 270-2021-020. Sandra Munera thanks the Juan de la Cierva-Formación contract (FJC2021-047786-I) co-funded by MCIN/AEI/10.13039/501100011033 and EU NextGenerationEU/PRTR. This work is co-funded by MICIN-AEI through project TED2021-130117B-C31, GVA-IVIA through projects 52203 and 52204, and the European Regional Development Fund (ERDF) of the Generalitat Valenciana 2021–2027. / Acosta Tello, MO. (2024). Predicción de macro y micronutrientes en hojas de cítricos y caqui utilizando métodos ópticos no destructivos [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/207010 / Compendio