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Vegetable pea production hinges significantly on the duration of its growing season, primarily due to the fact that the period of optimal technological maturity of pea grains is short. Knowledge of the duration of the growing season for vegetable peas is of great importance from the aspect of production planning, especially on large-scale farms, in order to synchronize sowing and maturation timing with the capacities of mechanization for harvesting and processing. Interpretation of results based on weather conditions is of great importance as well, because environmental conditions closely determine pea yield and quality. In this paper, the length of the pea growing season was determined using the method of heat units for 12 different spring vegetable pea genotypes. The research included genotypes with varying growing season lengths, categorized as early, mid-early, mid-late, and late, spanning from 60 to 76 days. The examined pea genotypes accrued heat units ranging from 612.1 to 986.3, signifying varying degrees of heat accumulation. The methodology adopted for employing heat units in the planning of spring vegetable pea cultivation and maturation operates on the premise that heat accumulation during the growing season correlates linearly with growth and development intensity, as well as the duration of the growing season itself. Consequently, the summation of heat units emerges as a distinctive characteristic of each cultivar, facilitating informed decision-making in agricultural practices.

[ES] La arquitectura de la inflorescencia determina la posición y el número de flores (y frutos) en la planta. Esto afecta a la forma de la planta, lo que contribuye a la diversidad morfológica e influye en la producción de semillas. Por lo tanto, comprender cómo funciona la genética que está detrás del desarrollo de la inflorescencia es relevante no solo para saber más sobre la biología del desarrollo de las plantas, sino también para la agricultura, con el fin de diseñar nuevas estrategias para la mejora de plantas. La mayoría de las leguminosas tienen inflorescencias compuestas, en las que las flores no se forman en el tallo principal sino a partir de inflorescencias secundarias (I2) en los flancos de la inflorescencia primaria o principal (I1). Esto contrasta con las plantas con inflorescencias simples, como Arabidopsis, donde las flores se forman directamente a partir del I1. El guisante (Pisum sativum) pertenece a la familia de las Fabaceae y al clado galegoide de las leguminosas y tiene una inflorescencia compuesta. Es bien sabido que VEGETATIVE1/ FULc (VEG1) codifica un factor de transcripción que especifica la identidad del meristemo I2 en leguminosas, pero aún se desconoce cómo y a través de qué genes VEG1 controla el desarrollo del I2 y las vías genéticas en las que está involucrado. En este trabajo, nuestro objetivo fue identificar las dianas regulatorias de VEG1. Para ello, comparamos los transcriptomas provenientes de ápices de inflorescencia del control silvestre (wild-type) con los de mutantes de guisante con defectos en el desarrollo de la inflorescencia: proliferating inflorescence meristems (pim - con múltiples meristemos I2), veg1 y vegetative2 (veg2 - ninguno de los cuales produce ni meristemos I2 ni flores). Usando este enfoque, hemos aislado algunos genes que se expresan en el I2 e identificado algunas posibles dianas de VEG1, entre ellas algunos genes que parecen prometedores para ser usados a modo de herramientas genéticas para la mejora del, [CA] L'arquitectura de la inflorescència determina la posició i el nombre de flors (i fruits) en la planta. Això afecta a la forma de la planta, la qual cosa contribueix a la diversitat morfològica i influeix en la producció de llavors. Per tant, comprendre com funciona la genètica que està darrere del desenvolupament de la inflorescència és rellevant no sols per a saber més sobre la biologia del desenvolupament de les plantes, sinó també per a l'agricultura, amb la finalitat de dissenyar noves estratègies per a la millora de plantes. La majoria de les lleguminoses tenen inflorescències compostes, en les quals les flors no es formen en la tija principal sinó a partir d'inflorescències secundàries (I2) en els flancs de la inflorescència primària o principal (I1). Això contrasta amb les plantes amb inflorescències simples, com Arabidopsis, on les flors es formen directament a partir de l'I1. El pésol (Pisum sativum) pertany a la família de les Fabaceae i al clade galegoide de les lleguminoses i té una inflorescència composta. És ben sabut que VEGETATIVE1/ FULc (VEG1) codifica un factor de transcripció que especifica la identitat del meristemo I2 en lleguminoses, però encara es desconeix com i a través de quins gens VEG1 controla el desenvolupament de l'I2 i les vies genètiques en les quals està involucrat. En aquest treball, el nostre objectiu va ser identificar les dianes reguladores de VEG1. Per a això, comparem els transcriptomas provinents d'àpexs d'inflorescència del control silvestre (wild-type) amb els de mutants de pésol amb defectes en el desenvolupament de la inflorescència: proliferating inflorescence meristems (pim - amb múltiples meristemos I2), veg1 i vegetative2 (veg2 - cap dels quals produeix ni meristemos I2 ni flors). Usant aquest enfocament, hem aïllat alguns gens que s'expressen en l'I2 i identificat algunes possibles dianes de VEG1, entre elles alguns gens que semblen prometedors per a ser usats a manera d'eines genètiques per a la millora del ren, [EN] Inflorescence architecture determines position and number of flowers (and fruits) in the plant. This affects plant shape, contributing to morphological diversity, and also influences seed yield. Therefore, understanding the genetics behind inflorescence development is relevant not only to plant developmental biology but also to agriculture, to design new breeding strategies. Most legumes have compound inflorescences, in which the flowers do not form on the main stem but from secondary inflorescences (I2) at the flanks of the main primary inflorescence (I1). This is in contrast to plants with simple inflorescences, such as Arabidopsis, where the flowers directly form at the I1. Pea (Pisum sativum) belongs to the Fabaceae family and the galegoid clade of legumes and has a compound inflorescence. It is well known that VEGETATIVE1/ FULc (VEG1) encodes a transcription factor that specifies the identity of the I2 meristem in legumes, but it is still unknown how and through which genes VEG1 controls I2 development and the genetic pathways in which it is involved. In this work, we aimed to identify regulatory targets of VEG1. For that, we compared the transcriptomes of inflorescence apices from wild type and pea mutants with defects in inflorescence development: proliferating inflorescence meristems (pim - with multiple I2 meristems), veg1 and vegetative2 (veg2), none of which produce neither I2 meristems nor flowers). Using this approach, we have isolated I2-expressed meristem genes and identified some possible targets of VEG1, among them some genes that seem promising tools to improve yield in legumes. FLOWERING LOCUS T (FT) is a key regulator of the photoperiod inductive pathway that controls flowering time in Arabidopsis. In legumes, the FT clade has diversified into three subclades: FTa, FTb and FTc. Pea FTc is distant phylogenetically from the other FTs and has an unusual expression pattern, being expressed only at the inflorescence apex. In this work we have cha

[ES] La arquitectura de la inflorescencia determina la posición y el número de flores (y frutos) en la planta. Esto afecta a la forma de la planta, lo que contribuye a la diversidad morfológica e influye en la producción de semillas. Por lo tanto, comprender cómo funciona la genética que está detrás del desarrollo de la inflorescencia es relevante no solo para saber más sobre la biología del desarrollo de las plantas, sino también para la agricultura, con el fin de diseñar nuevas estrategias para la mejora de plantas. La mayoría de las leguminosas tienen inflorescencias compuestas, en las que las flores no se forman en el tallo principal sino a partir de inflorescencias secundarias (I2) en los flancos de la inflorescencia primaria o principal (I1). Esto contrasta con las plantas con inflorescencias simples, como Arabidopsis, donde las flores se forman directamente a partir del I1. El guisante (Pisum sativum) pertenece a la familia de las Fabaceae y al clado galegoide de las leguminosas y tiene una inflorescencia compuesta. Es bien sabido que VEGETATIVE1/ FULc (VEG1) codifica un factor de transcripción que especifica la identidad del meristemo I2 en leguminosas, pero aún se desconoce cómo y a través de qué genes VEG1 controla el desarrollo del I2 y las vías genéticas en las que está involucrado. En este trabajo, nuestro objetivo fue identificar las dianas regulatorias de VEG1. Para ello, comparamos los transcriptomas provenientes de ápices de inflorescencia del control silvestre (wild-type) con los de mutantes de guisante con defectos en el desarrollo de la inflorescencia: proliferating inflorescence meristems (pim - con múltiples meristemos I2), veg1 y vegetative2 (veg2 - ninguno de los cuales produce ni meristemos I2 ni flores). Usando este enfoque, hemos aislado algunos genes que se expresan en el I2 e identificado algunas posibles dianas de VEG1, entre ellas algunos genes que parecen prometedores para ser usados a modo de herramientas genéticas para la mejora del, [CA] L'arquitectura de la inflorescència determina la posició i el nombre de flors (i fruits) en la planta. Això afecta a la forma de la planta, la qual cosa contribueix a la diversitat morfològica i influeix en la producció de llavors. Per tant, comprendre com funciona la genètica que està darrere del desenvolupament de la inflorescència és rellevant no sols per a saber més sobre la biologia del desenvolupament de les plantes, sinó també per a l'agricultura, amb la finalitat de dissenyar noves estratègies per a la millora de plantes. La majoria de les lleguminoses tenen inflorescències compostes, en les quals les flors no es formen en la tija principal sinó a partir d'inflorescències secundàries (I2) en els flancs de la inflorescència primària o principal (I1). Això contrasta amb les plantes amb inflorescències simples, com Arabidopsis, on les flors es formen directament a partir de l'I1. El pésol (Pisum sativum) pertany a la família de les Fabaceae i al clade galegoide de les lleguminoses i té una inflorescència composta. És ben sabut que VEGETATIVE1/ FULc (VEG1) codifica un factor de transcripció que especifica la identitat del meristemo I2 en lleguminoses, però encara es desconeix com i a través de quins gens VEG1 controla el desenvolupament de l'I2 i les vies genètiques en les quals està involucrat. En aquest treball, el nostre objectiu va ser identificar les dianes reguladores de VEG1. Per a això, comparem els transcriptomas provinents d'àpexs d'inflorescència del control silvestre (wild-type) amb els de mutants de pésol amb defectes en el desenvolupament de la inflorescència: proliferating inflorescence meristems (pim - amb múltiples meristemos I2), veg1 i vegetative2 (veg2 - cap dels quals produeix ni meristemos I2 ni flors). Usant aquest enfocament, hem aïllat alguns gens que s'expressen en l'I2 i identificat algunes possibles dianes de VEG1, entre elles alguns gens que semblen prometedors per a ser usats a manera d'eines genètiques per a la millora del ren, [EN] Inflorescence architecture determines position and number of flowers (and fruits) in the plant. This affects plant shape, contributing to morphological diversity, and also influences seed yield. Therefore, understanding the genetics behind inflorescence development is relevant not only to plant developmental biology but also to agriculture, to design new breeding strategies. Most legumes have compound inflorescences, in which the flowers do not form on the main stem but from secondary inflorescences (I2) at the flanks of the main primary inflorescence (I1). This is in contrast to plants with simple inflorescences, such as Arabidopsis, where the flowers directly form at the I1. Pea (Pisum sativum) belongs to the Fabaceae family and the galegoid clade of legumes and has a compound inflorescence. It is well known that VEGETATIVE1/ FULc (VEG1) encodes a transcription factor that specifies the identity of the I2 meristem in legumes, but it is still unknown how and through which genes VEG1 controls I2 development and the genetic pathways in which it is involved. In this work, we aimed to identify regulatory targets of VEG1. For that, we compared the transcriptomes of inflorescence apices from wild type and pea mutants with defects in inflorescence development: proliferating inflorescence meristems (pim - with multiple I2 meristems), veg1 and vegetative2 (veg2), none of which produce neither I2 meristems nor flowers). Using this approach, we have isolated I2-expressed meristem genes and identified some possible targets of VEG1, among them some genes that seem promising tools to improve yield in legumes. FLOWERING LOCUS T (FT) is a key regulator of the photoperiod inductive pathway that controls flowering time in Arabidopsis. In legumes, the FT clade has diversified into three subclades: FTa, FTb and FTc. Pea FTc is distant phylogenetically from the other FTs and has an unusual expression pattern, being expressed only at the inflorescence apex. In this work we have cha

In this study, a total of 12 vegetable pea genotypes of different growing seasons were subjected to phenotypic characterization. The vegetable pea genotypes are a part of the collection maintained at the Institute of Field and Vegetable Crops Novi Sad. The plant material included 10 promising lines and two released cultivars, Tamiš and Dunav. The trial was carried out in 2022. It was set up at the Rimski Šančevi site, as a randomized block design in five replications. A total of 14 agronomic traits were analyzed. The obtained research results revealed divergence in the investigated plant material. The statistical significance of all sources of variation was determined by the LSD test. The height of the first fertile node was found to be the most variable feature, with a coefficient of variation of 40.54%. The tested genotypes were clustered into two groups and two subgroups within the second group. The correlation analysis of the examined quantitative traits revealed the presence of several statistically significant positive and negative correlations. Some of the most significant positive correlations were established between the grain weight per plant and the number of grains per plant and the yield of technologically mature grain, while the pod width and the number of fertile nodes per plant had the most negative correlations with the other tested traits., U toku ovog istraživanja izvršena je fenotipska karakterizacija 12 genotipova povrtarskog graška, različite dužine vegetacije, koji pripadaju kolekciji Instituta za ratarstvo i povrtarstvo u Novom Sadu. Biljni materijal obuhvatao je 10 perspektivnih selekcionih linija i dve priznate sorte tamiš i dunav. Ogled je izveden tokom 2022. godine, postavljen je po slučajnom blok sistemu, u pet ponavljanja na lokalitetu Rimski Šančevi. Analizirano je 14 agronomskih osobina. Dobijeni rezultati istraživanja ukazuju na postojanje divergentnosti u ispitivanom biljnom materijalu. Analizom varijanse ispitivanih agronomskih osobina utvrđena je statistička značajnost svih izvora varijacije. Kao najvarijabilnija osobina istakla se visina prvog rodnog nodusa sa koeficijentom varijacije od 40,54%. Ispitivani genotipovi su primenom klaster analize grupisani u dve grupe sa dve podgrupe u okviru druge grupe. Korelacionom analizom ispitivanih kvantitativnih svojstava utvrđeno je postojanje nekoliko statistički značajnih pozitivnih i negativnih međuzavisnosti. Neke od najznačajnijih pozitivnih korelacija utvrđene su između osobina: masa zrna po biljci i prinos tehnološki zrelog zrna, kao i broja zrna po biljci i prinosa tehnološki zrelog zrna, dok svojstva širina mahune i suma rodnih nodusa po biljci imaju najviše negativnih korelacija sa ostalim ispitivanim osobinama.

Characterization data of a Pisum sp. germplams colection grown at Córdoba, Spain under three crop seasons

In common with other plant species, the garden pea (Pisum sativum) produces the auxin indole-3-acetic acid (IAA) from tryptophan via a single intermediate, indole-3-pyruvic acid (IPyA). IPyA is converted to IAA by PsYUC1, also known as Crispoid (Crd). Here, we extend our understanding of the developmental processes affected by the Crd gene by examining the phenotypic effects of crd gene mutations on leaves, flowers, and roots. We show that in pea, Crd/PsYUC1 is important for the initiation and identity of leaflets and tendrils, stamens, and lateral roots. We also report on aspects of auxin deactivation in pea.

In common with other plant species, the garden pea (Pisum sativum) produces the auxin indole-3-acetic acid (IAA) from tryptophan via a single intermediate, indole-3-pyruvic acid (IPyA). IPyA is converted to IAA by PsYUC1, also known as Crispoid (Crd). Here, we extend our understanding of the developmental processes affected by the Crd gene by examining the phenotypic effects of crd gene mutations on leaves, flowers, and roots. We show that in pea, Crd/PsYUC1 is important for the initiation and identity of leaflets and tendrils, stamens, and lateral roots. We also report on aspects of auxin deactivation in pea.

The use of guided instructions are often debated in science education research, but nevertheless often used in lower secondary school. The aim of this licentiate thesis is to contribute to knowledge about science education in lower secondary school when pupils’ do lab work using guided instructions. Two pupils are observed when they, during an activity, are expected to interpret an instruction and handle different artifacts. In study 1, the aim is to exploratory investigate and illustrate and discuss how a pragmatic analysis of learning (a combination of practical epistemology analysis and pragmatic theory) may contribute when developing principles for guided lab work. The result showed that central principles for teaching is needed. Teachers need to participate during pupils’ lab work and focus on how pupils handle their learning environment and what pupils acknowledge as a learning environment. In study 2 the aim is to develop, empirically test and discuss a methodology (i.e. a combination of an analytical method, a strategy for analysis and pragmatic theory). The pragmatic inspired methodology is developed and tested to create knowledge about what kinds of different engagement may develop when pupils participate in guided lab work. In this study, which is based on earlier contributions to our understanding of engagement, interest and taste, two different intrinsic engagements were found; one is linked to the purpose of the activity and the other to enjoyment. The result showed that the first type of engagement is developed while the pupils solve a practical problem furthermore the pupils’ solution carries artistic qualities, (i.e. a personal investment beyond what is expected from the teacher regarding the laboratory work that they are involved in). Finally, I draw on studies 1 and 2 to conclude principles for guided lab work in lower secondary school. These principles may be useful when teachers want pupils to engage artistically in an activity.

Hormones are integral molecules in the human body, controlling most body systems and maintaining homeostasis in the body. They function as signal molecules for communication between cells, and are often secreted by one tissue with another as target. Protein hormones make up the majority of the hormones in the body, are mostly transported in the bloodstream, and bind to receptors on the cell surface since they can not diffuse through cell membranes. Hormones and their associated proteins are linked to many diseases, and could therefore have a lot of potential as biomarkers. With the use of the company Olink’s Proximity Extension Assay (PEA) technology the concentrations of human proteins in blood can be measured. These proteins can be combined into different sets, called panels, to measure biomarkers related to specific diseases or conditions. This project aimed to identify the protein hormones of the human body, and investigate their associated proteins and the pathways and potential diseases they were related to. Suggestions of new panels based on disease areas that were found to be particularly related to protein hormones were also to be put forward. Databases such as UniProt, Reactome, String, Signor, Olink Insight, and the Human Protein Atlas were used to investigate protein hormones and their associated proteins. The result of the research was compiled into three different panels: the overall Protein Hormone panel, the Wellness panel, and the Diabetes panel. These panels were based on the disease areas commonly found associated with protein hormones, and that were deemed to be of interest to Olink and to the research community. The research also resulted in a table detailing all found protein hormones with additional info on pathways and diseases, normal concentrations of the proteins in blood or plasma, and the associations between the hormones and other proteins. The financial viability and marketability of the suggested panels were also investigated in a bus

The general aim of this thesis was to study the prognostic clinical and biological markers of Diffuse Large B-cell Lymphoma (DLBCL). Paper I: Utilizing population-based data for patients with DLBCL in Sweden, the study aimed to establish whether event free survival at 24 months (EFS24) was a reproducible milestone. The disease-free survival for lymphoma patients was compared with that of age and sex matched Swedish general population. We demonstrated that overall survival was similar to age and sex matched general population only for younger patients (<60 years of age) achieving ES24. Patients older than that had worse prognosis. Death was mainly linked to cardiovascular disease and secondary malignancies. Paper II: Plasma samples collected via the bio bank U-CAN were analyzed using multiplex extension assay (PEA) utilizing preselected protein panels to examine the possibility of distinguishing lymphomas, leukemias and controls. The study confirmed that PEA technology could be used not only to effectively screen for large number of plasma protein biomarkers in low plasma sample volumes (1 µL), but even to discriminate between controls and different haematological malignancies. Paper III: Plasma protein pattern evolution in DLBCL patients was highlighted by PEA analysis of plasma proteins at different time points under treatment with Rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP). Significant distinctions in protein patterns at diagnosis compared to controls and striking differences in protein levels before and after treatment in patient who responded to treatment were evident. The three top proteins were TCL1A, CXCL13 and IL2RA. Paper IV: An interesting protein that emerged from the above studies was TCL1A. This plasma protein was analyzed in plasma samples by PEA. Validation by plasma enzyme immunosorbent assay (ELISA) was attempted. The cytoplasm and nucleus bound form of TCL1A were analyzed with the help of immunohistochemist

The use of guided instructions are often debated in science education research, but nevertheless often used in lower secondary school. The aim of this licentiate thesis is to contribute to knowledge about science education in lower secondary school when pupils’ do lab work using guided instructions. Two pupils are observed when they, during an activity, are expected to interpret an instruction and handle different artifacts. In study 1, the aim is to exploratory investigate and illustrate and discuss how a pragmatic analysis of learning (a combination of practical epistemology analysis and pragmatic theory) may contribute when developing principles for guided lab work. The result showed that central principles for teaching is needed. Teachers need to participate during pupils’ lab work and focus on how pupils handle their learning environment and what pupils acknowledge as a learning environment. In study 2 the aim is to develop, empirically test and discuss a methodology (i.e. a combination of an analytical method, a strategy for analysis and pragmatic theory). The pragmatic inspired methodology is developed and tested to create knowledge about what kinds of different engagement may develop when pupils participate in guided lab work. In this study, which is based on earlier contributions to our understanding of engagement, interest and taste, two different intrinsic engagements were found; one is linked to the purpose of the activity and the other to enjoyment. The result showed that the first type of engagement is developed while the pupils solve a practical problem furthermore the pupils’ solution carries artistic qualities, (i.e. a personal investment beyond what is expected from the teacher regarding the laboratory work that they are involved in). Finally, I draw on studies 1 and 2 to conclude principles for guided lab work in lower secondary school. These principles may be useful when teachers want pupils to engage artistically in an activity.

The general aim of this thesis was to study the prognostic clinical and biological markers of Diffuse Large B-cell Lymphoma (DLBCL). Paper I: Utilizing population-based data for patients with DLBCL in Sweden, the study aimed to establish whether event free survival at 24 months (EFS24) was a reproducible milestone. The disease-free survival for lymphoma patients was compared with that of age and sex matched Swedish general population. We demonstrated that overall survival was similar to age and sex matched general population only for younger patients (<60 years of age) achieving ES24. Patients older than that had worse prognosis. Death was mainly linked to cardiovascular disease and secondary malignancies. Paper II: Plasma samples collected via the bio bank U-CAN were analyzed using multiplex extension assay (PEA) utilizing preselected protein panels to examine the possibility of distinguishing lymphomas, leukemias and controls. The study confirmed that PEA technology could be used not only to effectively screen for large number of plasma protein biomarkers in low plasma sample volumes (1 µL), but even to discriminate between controls and different haematological malignancies. Paper III: Plasma protein pattern evolution in DLBCL patients was highlighted by PEA analysis of plasma proteins at different time points under treatment with Rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP). Significant distinctions in protein patterns at diagnosis compared to controls and striking differences in protein levels before and after treatment in patient who responded to treatment were evident. The three top proteins were TCL1A, CXCL13 and IL2RA. Paper IV: An interesting protein that emerged from the above studies was TCL1A. This plasma protein was analyzed in plasma samples by PEA. Validation by plasma enzyme immunosorbent assay (ELISA) was attempted. The cytoplasm and nucleus bound form of TCL1A were analyzed with the help of immunohistochemist

[ES] La intolerancia a la histamina es un trastorno relacionado con el déficit de enzima DAO a nivel intestinal. La suplementación oral o dietas restrictivas son las soluciones actuales. Se están estudiando nuevas estrategias, como la obtención de DAO de fuentes vegetales y su aplicación directa en alimentos para reducir el contenido de histamina. La investigación se enfoca en tecnologías como los pulsos eléctricos de alta intensidad (PEF), para mejorar el rendimiento de DAO durante la germinación. Esto podría aumentar la actividad de la misma, brindando una alternativa más atractiva y eficaz para tratar esta intolerancia. El objetivo de este trabajo fue obtener enzima DAO a partir de guisantes y garrofón durante su germinación tras el pretratamiento con tecnología PEF. Se observó que tanto la longitud del germen como el contenido proteico aumentaron con el tiempo de germinación en ambas semillas. La actividad enzimática de DAO alcanzó su máximo entre 3 y 6 días de germinación, disminuyendo después. El pH del agua de remojo influyó en la cantidad de DAO formada. Además, se evaluó el efecto de los pulsos eléctricos de alta intensidad (PEF) como pretratamiento antes de la germinación. Se encontraron resultados variables en la germinación, contenido proteico y actividad enzimática. Las semillas de guisante mostraron un mayor contenido proteico y actividad enzimática en comparación con las de garrofón. Sin embargo, comparando las muestras control con las tratadas con PEF, en estas últimas se obtuvo un contenido proteico y actividad enzimática menor en el caso de los guisantes, y no se encontraron diferencias significativas en el caso del garrofón., [EN] Histamine intolerance is a disorder related to a deficiency of the DAO enzyme at the intestinal level. Current solutions include oral supplementation or restrictive diets. New strategies are being investigated, such as obtaining DAO from plant sources and incorporating it into foods to reduce histamine levels. Research is focused on technologies like high-intensity pulsed electric fields (PEF) to enhance DAO yield during germination. This could increase DAO activity, offering a more appealing and effective alternative for treating this intolerance. The objective of this study was to obtain the DAO enzyme from peas and fava beans during their germination following pre-treatment with PEF technology. It was observed that both the sprout length and protein content increased with germination time in both seeds. DAO enzymatic activity reached its peak between 3 and 6 days of germination, and then decreased. The pH of the soaking water influenced the amount of formed DAO. Furthermore, the effect of high-intensity pulsed electric fields (PEF) as a pre-treatment before germination was evaluated. Variable results were obtained in germination, protein content, and enzymatic activity after PEF treatments. Pea seeds exhibited higher protein content and enzymatic activity compared to fava beans. However, when comparing the control samples with the PEF treated ones, the treated pea samples showed lower protein content and enzymatic activity, while no significant differences were found in the case of fava beans.

In common with other plant species, the garden pea (Pisum sativum) produces the auxin indole-3-acetic acid (IAA) from tryptophan via a single intermediate, indole-3-pyruvic acid (IPyA). IPyA is converted to IAA by PsYUC1, also known as Crispoid (Crd). Here, we extend our understanding of the developmental processes affected by the Crd gene by examining the phenotypic effects of crd gene mutations on leaves, flowers, and roots. We show that in pea, Crd/PsYUC1 is important for the initiation and identity of leaflets and tendrils, stamens, and lateral roots. We also report on aspects of auxin deactivation in pea.

Traditional farming practices, focussing on cultivating single crops have detrimental effects such as high input of agrochemicals, soil erosion, and the development of pesticide-resistant pests. Furthermore, the use and production of nitrogen fertilisers in such systems can lead to the release of CO, contributing to climate change. Hence, agroecosystems that combine robust crop yields with reduced resource inputs are needed and there is a rising interest in cereal-legume intercropping. However, challenges remain, including the difficulty of managing the variable proportions of each species in the harvested mixture and the limited understanding of how individual crops perform within intercropping systems. The aim of this study was to estimate how intercropping of hull-less barley and oat with pea contribute to better crop performance under climatic conditions of Serbia. The trials were conducted at Rimski Šančevi (Vojvodina, North province of Serbia) using a RCBD design with four replicates. Sowing densities were 30% (hull-ess barley or oats):70% (pea). On average, the yield of hull-less barley (Golijat) and oats (Dunav) in 2022 and 2023 was notably higher in pure stands compared to intercropping practices.

Widespread usage of intercropping systems has been limited because of a lack of knowledge about the key factors that affect the performance of intercrop components. We used general linear modelling to explain the effect of different cropping systems on the relationships among yield, thousand kernel weight (TKW), and crude protein of cereal crops under the same agro-ecological conditions and naturally occurring inocula of obligate pathogens. The results of our study showed that the yield variation under extreme fluctuations in climatic conditions could be lowered through intercropping cultivation. The disease indices of leaf rust and powdery mildew were highly dependent on the type of cultivation. The relationships among the levels of pathogenic infection and yield performances were not straightforward and were highly dependent on the yielding potentials of the cultivars. Our study indicated that changes in yield, TKW, and crude protein, as well as their relationships during intercropping cultivation, were cultivar specific and, therefore, not the same among all cereal crops exposed to the same agro-ecological conditions.

Biopriming (BP) represents a new seed treatment technique that integrates biological and physiological aspects, ensuring increased resistance of the seed andyoung plant. Today, in BP, many species of fungi are used intensively , including Trichoderma, but the effect of filamentous fungi has not been described yet. The aim of this study was to examine the role of seed BP with polysaccharides (PSH) isolated from submerged cultures of two different strains of fungal species Schizophyllum commune, originating from Serbia (SRB) and Italy (IT), in the fight against oxidative stress in seedlings under optimal conditions.

Biopriming (BP) of seeds ensures increased resistance of seeds and young plants to stress, by integrating beneficial microorganisms and their products. In addition, research on filamentous fungi revealed their enormous potential in biotechnological processes and demonstrated their ability to increase plant resistance to stressful conditions. Aim: The aim was to examine the influence of BPof pea seeds with polysaccharides (PSH) isolated from submerged cultures of Schizophyllum commune originating from Serbia (SRB) and Italy (IT), on the antioxidant activity of the plant under drought.

Salinity is one of the greatest challenges of successful agricultural production worldwide. However, seed priming might be efficient practice for enhancing seed germination and initial plant growth and development. This experiment was undertaken with the aim of assessing the impact of different priming methods on seed quality parameters and early growth of garden pea (Pisum sativum L.) cultivars under salinity stress. Pea seeds of three different cultivars were primed with water (hydropriming), KNO3 solution (0.5%), and PEG 6000 solution (-0.5 MPa) for 24h in the dark. Unprimed and primed seeds were germinated between filter papers saturated with 120 mM NaCl using the germination test. The results clearly showed that the examined parameters of primed pea seeds were substantially greater than the parameters of unprimed seeds under saline stress. Moreover, seed priming with KNO3 and PEG solutions were more effective in improving seed quality and initial growth in garden peas than hydropriming.

In common with other plant species, the garden pea (Pisum sativum) produces the auxin indole-3-acetic acid (IAA) from tryptophan via a single intermediate, indole-3-pyruvic acid (IPyA). IPyA is converted to IAA by PsYUC1, also known as Crispoid (Crd). Here, we extend our understanding of the developmental processes affected by the Crd gene by examining the phenotypic effects of crd gene mutations on leaves, flowers, and roots. We show that in pea, Crd/PsYUC1 is important for the initiation and identity of leaflets and tendrils, stamens, and lateral roots. We also report on aspects of auxin deactivation in pea.

A factorial experiment was conducted as randomized complete blocks in three replications at the Agriculture Faculty research farm, Takhar University to quantify the effect of planting density and planting arrangement on some morphological traits. Experimental treatments included 15, 30, 45, and 60 cm row spacing. The harvest was done by hand when all the pods were yellow, and seeds were hard. At the time of the final harvest, ten bushes were randomly selected from each plot and used to determine the performance components (elevation of the bush, the number of pods per plant, the number of seeds per pod, the weight of 100 seeds, and the biological yield). The results showed that the effect of density on performance was not significant. The maximum yield of seeds in the planting arrangement of 30 to 45 cm row distance has been obtained at the rate of 935.76, and 996.15 kg per hectare. The maximum height of the plant was observed at the distance between the rows of 30 cm. The density decreased with increasing the distance between the planting rows from 30 to 45 cm. The number of pods increased from 14.44 to 16.33 pods per plant, i.e., 11.57 percent.

Aldehydes are important flavor molecules to consider in plant-based products. Here, the flavor retention of a series of saturated aldehydes and mono-unsaturated aldehydes (2-alkenals) with different chain lengths (C4, C6, C8, and C10) in dispersions with protein isolates of pea, soy, fava bean, chickpea, and whey (as reference) was analyzed with APCI-TOF-MS. The headspace concentrations of alkenals were lower than aldehydes, meaning alkenals were retained more than saturated aldehydes. The retention was modeled by assuming hydrophobic interactions and covalent interactions. The ratio between the hydrophobic interaction parameter and the covalent parameter showed that covalent interactions are mainly important for butanal and butenal (C4). For the other aldehydes, hydrophobic interactions became increasingly important. Correlations were found between the chemical interaction parameters and the cysteine and methionine content of the different proteins. The obtained model parameters for each set of proteins and flavors allow the prediction of flavor retention when developing a flavored product with high protein content.

Partial shifts from animal-based to plant-based proteins in human diets could reduce environmental pressure from food systems and serve human health. Grain legumes can play an important role here. They are one of the few agricultural commodities for which Europe is not nearly self-sufficient. Here, we assessed area expansion and yield increases needed for European self-sufficiency of faba bean, pea and soybean. We show that such production could use substantially less cropland (4–8%) and reduce GHG emissions (7–22% current meat production) when substituting for animal-derived food proteins. We discuss changes required in food and agricultural systems to make grain legumes competitive with cereals for farmers and how their cultivation can help to increase sustainability of European cropping systems.

Hormones are integral molecules in the human body, controlling most body systems and maintaining homeostasis in the body. They function as signal molecules for communication between cells, and are often secreted by one tissue with another as target. Protein hormones make up the majority of the hormones in the body, are mostly transported in the bloodstream, and bind to receptors on the cell surface since they can not diffuse through cell membranes. Hormones and their associated proteins are linked to many diseases, and could therefore have a lot of potential as biomarkers. With the use of the company Olink’s Proximity Extension Assay (PEA) technology the concentrations of human proteins in blood can be measured. These proteins can be combined into different sets, called panels, to measure biomarkers related to specific diseases or conditions. This project aimed to identify the protein hormones of the human body, and investigate their associated proteins and the pathways and potential diseases they were related to. Suggestions of new panels based on disease areas that were found to be particularly related to protein hormones were also to be put forward. Databases such as UniProt, Reactome, String, Signor, Olink Insight, and the Human Protein Atlas were used to investigate protein hormones and their associated proteins. The result of the research was compiled into three different panels: the overall Protein Hormone panel, the Wellness panel, and the Diabetes panel. These panels were based on the disease areas commonly found associated with protein hormones, and that were deemed to be of interest to Olink and to the research community. The research also resulted in a table detailing all found protein hormones with additional info on pathways and diseases, normal concentrations of the proteins in blood or plasma, and the associations between the hormones and other proteins. The financial viability and marketability of the suggested panels were also investigated in a bus

The use of guided instructions are often debated in science education research, but nevertheless often used in lower secondary school. The aim of this licentiate thesis is to contribute to knowledge about science education in lower secondary school when pupils’ do lab work using guided instructions. Two pupils are observed when they, during an activity, are expected to interpret an instruction and handle different artifacts. In study 1, the aim is to exploratory investigate and illustrate and discuss how a pragmatic analysis of learning (a combination of practical epistemology analysis and pragmatic theory) may contribute when developing principles for guided lab work. The result showed that central principles for teaching is needed. Teachers need to participate during pupils’ lab work and focus on how pupils handle their learning environment and what pupils acknowledge as a learning environment. In study 2 the aim is to develop, empirically test and discuss a methodology (i.e. a combination of an analytical method, a strategy for analysis and pragmatic theory). The pragmatic inspired methodology is developed and tested to create knowledge about what kinds of different engagement may develop when pupils participate in guided lab work. In this study, which is based on earlier contributions to our understanding of engagement, interest and taste, two different intrinsic engagements were found; one is linked to the purpose of the activity and the other to enjoyment. The result showed that the first type of engagement is developed while the pupils solve a practical problem furthermore the pupils’ solution carries artistic qualities, (i.e. a personal investment beyond what is expected from the teacher regarding the laboratory work that they are involved in). Finally, I draw on studies 1 and 2 to conclude principles for guided lab work in lower secondary school. These principles may be useful when teachers want pupils to engage artistically in an activity.

The interaction between flavors and proteins results in a reduced headspace concentration of the flavor, affecting flavor perception. We analyzed the retention of a series of esters and ketones with different chain lengths (C4, C6, C8, and C10) by protein isolates of yellow pea, soy, fava bean, and chickpea, with whey as a reference. An increase in protein concentration led to a decrease in flavor compound in the headspace as measured with atmospheric pressure chemical ionization time-of-flight mass spectroscopy (APCI-TOF-MS). Flavor retention was described with a flavor-partitioning model. It was found that flavor retention could be well predicted with the octanol-water partitioning coefficient and by fitting the hydrophobic interaction parameter. Hydrophobic interactions were highest for chickpea, followed by pea, fava bean, whey, and soy. However, the obtained predictive model was less appropriate for methyl decanoate, possibly due to its solubility. The obtained models and fitted parameters are relevant when designing flavored products with high protein concentrations.

The general aim of this thesis was to study the prognostic clinical and biological markers of Diffuse Large B-cell Lymphoma (DLBCL). Paper I: Utilizing population-based data for patients with DLBCL in Sweden, the study aimed to establish whether event free survival at 24 months (EFS24) was a reproducible milestone. The disease-free survival for lymphoma patients was compared with that of age and sex matched Swedish general population. We demonstrated that overall survival was similar to age and sex matched general population only for younger patients (<60 years of age) achieving ES24. Patients older than that had worse prognosis. Death was mainly linked to cardiovascular disease and secondary malignancies. Paper II: Plasma samples collected via the bio bank U-CAN were analyzed using multiplex extension assay (PEA) utilizing preselected protein panels to examine the possibility of distinguishing lymphomas, leukemias and controls. The study confirmed that PEA technology could be used not only to effectively screen for large number of plasma protein biomarkers in low plasma sample volumes (1 µL), but even to discriminate between controls and different haematological malignancies. Paper III: Plasma protein pattern evolution in DLBCL patients was highlighted by PEA analysis of plasma proteins at different time points under treatment with Rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP). Significant distinctions in protein patterns at diagnosis compared to controls and striking differences in protein levels before and after treatment in patient who responded to treatment were evident. The three top proteins were TCL1A, CXCL13 and IL2RA. Paper IV: An interesting protein that emerged from the above studies was TCL1A. This plasma protein was analyzed in plasma samples by PEA. Validation by plasma enzyme immunosorbent assay (ELISA) was attempted. The cytoplasm and nucleus bound form of TCL1A were analyzed with the help of immunohistochemist

The general aim of this thesis was to study the prognostic clinical and biological markers of Diffuse Large B-cell Lymphoma (DLBCL). Paper I: Utilizing population-based data for patients with DLBCL in Sweden, the study aimed to establish whether event free survival at 24 months (EFS24) was a reproducible milestone. The disease-free survival for lymphoma patients was compared with that of age and sex matched Swedish general population. We demonstrated that overall survival was similar to age and sex matched general population only for younger patients (<60 years of age) achieving ES24. Patients older than that had worse prognosis. Death was mainly linked to cardiovascular disease and secondary malignancies. Paper II: Plasma samples collected via the bio bank U-CAN were analyzed using multiplex extension assay (PEA) utilizing preselected protein panels to examine the possibility of distinguishing lymphomas, leukemias and controls. The study confirmed that PEA technology could be used not only to effectively screen for large number of plasma protein biomarkers in low plasma sample volumes (1 µL), but even to discriminate between controls and different haematological malignancies. Paper III: Plasma protein pattern evolution in DLBCL patients was highlighted by PEA analysis of plasma proteins at different time points under treatment with Rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP). Significant distinctions in protein patterns at diagnosis compared to controls and striking differences in protein levels before and after treatment in patient who responded to treatment were evident. The three top proteins were TCL1A, CXCL13 and IL2RA. Paper IV: An interesting protein that emerged from the above studies was TCL1A. This plasma protein was analyzed in plasma samples by PEA. Validation by plasma enzyme immunosorbent assay (ELISA) was attempted. The cytoplasm and nucleus bound form of TCL1A were analyzed with the help of immunohistochemist

Plant-derived ingredients for food formulation have gained increasing interest in recent years as animal products pose a higher burden on the environment. Among plant proteins, those from pea (Pisum sativum L.) are of particular interest because of their low allergenicity, low cost, high availability, and good reputation among consumers. However, the technofunctionality of pea proteins is often inferior to animal-derived proteins limiting a more widespread use in food products. These technofunctional properties include - among others - foaming, gelling, and binding of other ingredients and it depends on the food product, which functionality food scientists must utilize and optimize. Cost effective approaches to improve the technofunctionality of pea proteins are therefore desirable and would allow the industry to further implement the use of sustainable ingredients in foods. In line with these overall goals, the aim of the first section of this thesis was to characterize a commercial pea protein isolate and to modulate the physicochemical and technofunctional properties through homogenization for foaming application. The main goal of the second section was to mix pea proteins with pectin to obtain a suitable binder with desired properties for the application in meat alternatives. The mixing approach was based on previous research data that had shown that interacting protein-polysaccharide systems display a synergistic behaviour in terms of their functional properties. First section: Foams are two phase systems consisting of gas bubbles that are stabilized by surface-active ingredients such as proteins in the discontinuous, aqueous phase. The physico-chemical properties of proteins such as their solubility determines foaming performance. In Chapter I, a commercial pea protein isolate was fractionated into a water-soluble and a water-insoluble fraction for characterization. Although the two fractions were similar in protein composition, they showed distinct difference, Pflanzliche Inhaltsstoffe für die Herstellung von Lebensmitteln haben in den letzten Jahren zunehmend an Bedeutung gewonnen, da tierische Produkte eine größere Belastung für die Umwelt darstellen. Unter den pflanzlichen Proteinen sind diejenigen aus Erbsen (Pisum sativum L.) wegen ihrer geringen Allergenität, ihrer niedrigen Kosten, ihrer hohen Verfügbarkeit und ihrer Wertschätzung seitens der Verbraucher von besonderem Interesse. Die Technofunktionalität von Erbsenproteinen ist jedoch häufig schlechter als die von tierischen Proteinen, was eine breitere Verwendung in Lebensmitteln einschränkt. Zu diesen technofunktionellen Eigenschaften gehören unter anderem Schäumen, Gelieren und Binden anderer Zutaten, und es hängt vom Produkt ab, welche Funktionen Lebensmittelwissenschaftler nutzen und optimieren müssen. Kostengünstige Ansätze zur Verbesserung der Technofunktionalität von Erbsenproteinen sind daher wünschenswert und würden es der Industrie ermöglichen, die Verwendung nachhaltiger Zutaten in Lebensmitteln weiter zu beschleunigen. Basierend auf diesen übergeordneten Absichten bestand das Ziel des ersten Abschnitts dieser Arbeit darin, ein kommerzielles Erbsenproteinisolat zu charakterisieren und die physikochemischen und technofunktionellen Eigenschaften durch Homogenisierung für die Anwendung als Schaumbildner zu modulieren. Das Hauptziel des zweiten Abschnitts bestand darin, Erbsenproteine mit Pektin zu mischen, um ein geeignetes Bindemittel mit den gewünschten Eigenschaften für die Anwendung in Fleischalternativen zu erhalten. Der Mischungsansatz basierte auf früheren Forschungsdaten, die zeigten, dass interagierende Protein-Polysaccharid-Systeme eine Synergie in Bezug auf ihre funktionellen Eigenschaften aufweisen. Erster Abschnitt: Schäume sind Zweiphasensysteme, die aus Gasblasen bestehen und durch oberflächenaktive Bestandteile wie Proteine in der diskontinuierlichen, wässrigen Phase stabilisiert werden. Die physikochemischen Eigenschaften der Proteine, wie

The presence of so-called anti-nutritional factors can reduce the bioavailability of nutrients following consumption of seeds which are otherwise an excellent source of proteins, carbohydrates and micronutrients. Among the proteins associated with negative effects on quality in pea (Pisum sativum L.) seeds are lectin, pea albumin 2 (PA2) and trypsin inhibitors (TI). Here we have investigated the impact of these proteins on protein digestibility and amino acid availability, using naturally occurring and derived mutant lines of pea lacking these proteins. The mutations were stacked to generate a triple mutant which was compared with a wild-type progenitor and a line lacking the major seed trypsin inhibitors alone. In vitro digestions following the INFOGEST protocol revealed significant differences in the degree of hydrolysis, protein profile and apparent amino acid availability among the pea variants. Proteins resistant to digestion were identified by MALDI-TOF mass spectrometry and amino acid profiles of digested samples determined. The results indicate that pea seeds lacking certain proteins can be used in the development of novel foods which have improved protein digestibility, and without negative impact on seed protein concentration or yield.

Drought is one of the greatest threats to agricultural production. Legumes are relatively susceptible to drought and have the unique ability to establish symbiosis with nitrogen-fixing bacteria. Deciphering how symbiotic interactions affect legume stress tolerance is a relevant question that remains unsolved, especially in the context of maintaining crop production under climate change. Our study addresses the comparative analysis of drought response and drought tolerance in symbiosis-dependent versus N-fed plants. Our results support the existence of attenuation in the damage promoted by drought in nodulated pea and vetch plants. This increase in drought tolerance correlates with changes in physiological parameters, such the hydric status of the plant, changes in biochemical parameters, such as the accumulation of osmolytes and antioxidant pigments, or the protection against lipid peroxidation. We have also found changes at the molecular level, identifying different genomic pathways through transcriptomic analysis. Although this characterization strengthens the evidence for a common strategy of coping with water deprivation in nodulated and N-fed plants, the above-mentioned changes suggest divergent and specific strategies of drought tolerance in nodulated plants as well as differences between pea and common vetch. In addition, our transcriptomic studies help unravel the molecular mechanisms through which rhizobial symbiosis contributes to drought tolerance in Vicia sativa and Pisum sativum. These results highlight the potential of using effective Rhizobium inoculants as a tool to cope with drought and enhance N-fixing ability under agriculturally relevant conditions.

[EN] Hybrid organic-inorganic metal halide perovskites have emerged as promising materials in the development of optoelectronic devices, namely due to their high optical absorption coefficient, tuneable bandgap and charge carrier mobility. However, more efforts are needed to improve their stability against oxygen, moisture and light and find new ways to modulate their optical properties. In this work, we study the photoluminescence properties of lead-bromide-based layered (2D) perovskites with different composition and dimensionality as bulk crystals and mechanically exfoliated flakes. We highlight the tuning of the photoluminescence emission of 2D metal halide perovskites by design and enhance their ambient stability by encapsulating the exfoliated flakes with h-BN flakes. Finally, we carry out the fabrication of heterostructures using as building blocks perovskite flakes showing how to further modulate the photoluminescence emission colour.

The use of guided instructions are often debated in science education research, but nevertheless often used in lower secondary school. The aim of this licentiate thesis is to contribute to knowledge about science education in lower secondary school when pupils’ do lab work using guided instructions. Two pupils are observed when they, during an activity, are expected to interpret an instruction and handle different artifacts. In study 1, the aim is to exploratory investigate and illustrate and discuss how a pragmatic analysis of learning (a combination of practical epistemology analysis and pragmatic theory) may contribute when developing principles for guided lab work. The result showed that central principles for teaching is needed. Teachers need to participate during pupils’ lab work and focus on how pupils handle their learning environment and what pupils acknowledge as a learning environment. In study 2 the aim is to develop, empirically test and discuss a methodology (i.e. a combination of an analytical method, a strategy for analysis and pragmatic theory). The pragmatic inspired methodology is developed and tested to create knowledge about what kinds of different engagement may develop when pupils participate in guided lab work. In this study, which is based on earlier contributions to our understanding of engagement, interest and taste, two different intrinsic engagements were found; one is linked to the purpose of the activity and the other to enjoyment. The result showed that the first type of engagement is developed while the pupils solve a practical problem furthermore the pupils’ solution carries artistic qualities, (i.e. a personal investment beyond what is expected from the teacher regarding the laboratory work that they are involved in). Finally, I draw on studies 1 and 2 to conclude principles for guided lab work in lower secondary school. These principles may be useful when teachers want pupils to engage artistically in an activity.

The use of guided instructions are often debated in science education research, but nevertheless often used in lower secondary school. The aim of this licentiate thesis is to contribute to knowledge about science education in lower secondary school when pupils’ do lab work using guided instructions. Two pupils are observed when they, during an activity, are expected to interpret an instruction and handle different artifacts. In study 1, the aim is to exploratory investigate and illustrate and discuss how a pragmatic analysis of learning (a combination of practical epistemology analysis and pragmatic theory) may contribute when developing principles for guided lab work. The result showed that central principles for teaching is needed. Teachers need to participate during pupils’ lab work and focus on how pupils handle their learning environment and what pupils acknowledge as a learning environment. In study 2 the aim is to develop, empirically test and discuss a methodology (i.e. a combination of an analytical method, a strategy for analysis and pragmatic theory). The pragmatic inspired methodology is developed and tested to create knowledge about what kinds of different engagement may develop when pupils participate in guided lab work. In this study, which is based on earlier contributions to our understanding of engagement, interest and taste, two different intrinsic engagements were found; one is linked to the purpose of the activity and the other to enjoyment. The result showed that the first type of engagement is developed while the pupils solve a practical problem furthermore the pupils’ solution carries artistic qualities, (i.e. a personal investment beyond what is expected from the teacher regarding the laboratory work that they are involved in). Finally, I draw on studies 1 and 2 to conclude principles for guided lab work in lower secondary school. These principles may be useful when teachers want pupils to engage artistically in an activity.

In Nariño, the climbing pea varieties, due to their high production capacity, are unable to stand upright despite the use of trellising systems, resulting in lodging and losses in yield and product quality. A viable option to mitigate the problem is the use of climbing genotypes with the afila gene that replaces leaves with tendrils, favoring the grip of the plants on the strings that serve as tutors. In search of a solution to this problem, the adaptability and phenotypic stability for green pod yield of 20 climbing pea lines with the afila gene and the normal-leaf controls Obonuco Andina and Sindamanoy were evaluated in five municipalities in the southern part of the department of Nariño. A randomized complete block design with four replications was used at each location. The data obtained were analyzed using the Eberhart and Russell and AMMI models. In the Eberhart and Russell model, the most favorable environments were Puerres and Gualmatán and the most unfavorable environment was Potosí. Lines GR15, GR10 and the Sindamanoy variety were selected as high yielding, adaptable and predictable. The analysis of variance for the AMMI model indicated a high environmental effect, resulting from contrasting environments and low diversity between the lines with the afila gene evaluated. The selection index (GSIi) obtained from the stability value (ASV) and the classification by yield, allowed the identification and selection in order of merit of the genotypes Obonuco Andina, GR15, GR14, GR28, GR10, GR2, GR23, Sindamanoy and GR3 as those with the highest stability and yield. The coincidence in selection between the Eberhart and Russell and AMMI models was 33,3%., En Nariño, las variedades de arveja voluble, debido a su alta capacidad productiva, no logran mantenerse erguidas a pesar del uso de sistemas de tutorado, lo que genera acame y pérdidas en el rendimiento y la calidad del producto. Una opción viable para mitigar el problema es el uso de genotipos volubles con el gen afila que reemplaza hojas por zarcillos, favoreciendo el agarre de las plantas sobre las cuerdas que sirven de tutores. En busca de una solución a este problema, se evaluó la adaptabilidad y estabilidad fenotípica para el rendimiento en vaina verde de 20 líneas de arveja voluble con el gen afila y los testigos de hoja normal Obonuco Andina y Sindamanoy en cinco municipios del sur del departamento de Nariño. En cada localidad se aplicó un diseño de bloques completos al azar con cuatro repeticiones. Los datos obtenidos se analizaron utilizando los modelos de Eberhart y Russell y AMMI. En el modelo de Eberhart y Russell, los ambientes más favorables fueron Puerres y Gualmatán y el ambiente más desfavorable fue Potosí. Las líneas GR15, GR10 y la variedad Sindamanoy fueron seleccionadas como de alto rendimiento, adaptables y predecibles. El análisis de varianza para el modelo AMMI indicó un alto efecto ambiental resultante de ambientes contrastantes y baja diversidad entre las líneas con el gen afila evaluadas. El índice de selección (GSIi) obtenido a partir del valor de estabilidad (ASV) y la clasificación por rendimiento, permitió la identificación y selección por orden de mérito de los genotipos Obonuco Andina, GR15, GR14, GR28, GR10, GR2, GR23, Sindamanoy y GR3 como los de mayor estabilidad y rendimiento. La coincidencia en la selección entre los modelos Eberhart y Russell y AMMI fue del 33,3%.

Der er en stigende interesse for plantebaserede fødevarer. Ærter er en god kandidat til dyrkning af plantebaserede proteinrige afgrøder i Danmark. Ydermere kan ærter være værdifulde for jordens frugtbarhed og et robust sædskifte i økologisk landbrug. Københavns Universitets projekt "PEAS & LOVE" arbejder med at screene mere end 200 accessioner fra Nordisk Genressourcecenter for deres potentiale i forædling af fremtidige ærtesorter til konsum af økologiske modne ærter. Målet med denne undersøgelse er at identificere de mest lovende ærter til avl og anbefale dem til yderligere forskning, baseret på undersøgelse af markedsinteresse, sensoriske kvaliteter, proteinindhold, udbytte og generisk diversitet. For at udforske markedsinteressen blev et spørgeskema besvaret af otte relevante virksomheder i Danmark, som resulterede i to hovedinteresser: (i) virksomheder, der anvender ærteekstrakter, prioriterede højt proteinindhold og neutral smag; (ii) de virksomheder, der brugte hele eller flækkede ærter, prioriterede smag højest. Derudover, for at forstå de sensoriske kvaliteter, anvendte undersøgelsen en sensorisk analyse af 24 accessioner fra PEAS & LOVE projektet. Sensorisk analyse resulterede i, at ærterne blev scoret på ni forskellige parametre og opdelt i fire udseendekategorier. Endelig gennemgik studiet tidligere undersøgelsers resultater på udbytte, proteinindhold samt generisk diversitet. Konklusionen ud fra den sensoriske analyse sammen med data om udbytte, protein og genetisk diversitet holdt op imod de to markedsinteresser, blev de mest lovende ærtetilslutninger til avl identificeret til yderligere forskning. Disse var: SKR108, SKR131, SKR134, SKR138, SKR154, SKR161 og SKR184. Derudover blev SKR67, SKR88 og SKR135 bestemt som de næstmest anbefalelsesværdige ærteaccessioner., There is a growing interest in plant-based foods. Peas are a good candidate for growing plant-based protein rich crops in Denmark. Furthermore, peas can be valuable for soil fertility and a robust crop rotation in organic agriculture. Copenhagen University’s project “PEAS & LOVE” works with screening 200+ accessions from the Nordic Genetic Resource Center for their potential in breeding for future pea varieties for human consumption of organic mature peas. The goal of this study is to identify the most promising peas for breeding and recommend them for further research, based on investigation of market interest, sensory qualities, protein content, yield, and generic diversity. In order to explore market interest, a questionnaire was answered by eight relevant companies in Denmark, that resulted in two main interests: (i) companies using on pea extractions prioritized high protein content and neutral taste; (ii) the companies that used whole or split peas prioritized flavour highest. Additionally, to understand the sensory qualities, the study employed a sensory analysis of 24 accessions from the Pea & Love project. Sensory analysis resulted in the peas being scored on nine different parameters and divided in to four appearance categories. Finally, the study reviewed previous studies’ results on yield, protein content as well as generic diversity. As a conclusion from the sensory analysis together with data on yield, protein and genetic diversity held up against the two market interests, the most promising pea accessions for breeding were identified for further research. These were: SKR108, SKR131, SKR134, SKR138, SKR154, SKR161 and SKR184. In addition, SKR67, SKR88 and SKR135 were found to be the next most recommendable pea accessions.

Der er en stigende interesse for plantebaserede fødevarer. Ærter er en god kandidat til dyrkning af plantebaserede proteinrige afgrøder i Danmark. Ydermere kan ærter være værdifulde for jordens frugtbarhed og et robust sædskifte i økologisk landbrug. Københavns Universitets projekt "PEAS & LOVE" arbejder med at screene mere end 200 accessioner fra Nordisk Genressourcecenter for deres potentiale i forædling af fremtidige ærtesorter til konsum af økologiske modne ærter. Målet med denne undersøgelse er at identificere de mest lovende ærter til avl og anbefale dem til yderligere forskning, baseret på undersøgelse af markedsinteresse, sensoriske kvaliteter, proteinindhold, udbytte og generisk diversitet. For at udforske markedsinteressen blev et spørgeskema besvaret af otte relevante virksomheder i Danmark, som resulterede i to hovedinteresser: (i) virksomheder, der anvender ærteekstrakter, prioriterede højt proteinindhold og neutral smag; (ii) de virksomheder, der brugte hele eller flækkede ærter, prioriterede smag højest. Derudover, for at forstå de sensoriske kvaliteter, anvendte undersøgelsen en sensorisk analyse af 24 accessioner fra PEAS & LOVE projektet. Sensorisk analyse resulterede i, at ærterne blev scoret på ni forskellige parametre og opdelt i fire udseendekategorier. Endelig gennemgik studiet tidligere undersøgelsers resultater på udbytte, proteinindhold samt generisk diversitet. Konklusionen ud fra den sensoriske analyse sammen med data om udbytte, protein og genetisk diversitet holdt op imod de to markedsinteresser, blev de mest lovende ærtetilslutninger til avl identificeret til yderligere forskning. Disse var: SKR108, SKR131, SKR134, SKR138, SKR154, SKR161 og SKR184. Derudover blev SKR67, SKR88 og SKR135 bestemt som de næstmest anbefalelsesværdige ærteaccessioner., There is a growing interest in plant-based foods. Peas are a good candidate for growing plant-based protein rich crops in Denmark. Furthermore, peas can be valuable for soil fertility and a robust crop rotation in organic agriculture. Copenhagen University’s project “PEAS & LOVE” works with screening 200+ accessions from the Nordic Genetic Resource Center for their potential in breeding for future pea varieties for human consumption of organic mature peas. The goal of this study is to identify the most promising peas for breeding and recommend them for further research, based on investigation of market interest, sensory qualities, protein content, yield, and generic diversity. In order to explore market interest, a questionnaire was answered by eight relevant companies in Denmark, that resulted in two main interests: (i) companies using on pea extractions prioritized high protein content and neutral taste; (ii) the companies that used whole or split peas prioritized flavour highest. Additionally, to understand the sensory qualities, the study employed a sensory analysis of 24 accessions from the Pea & Love project. Sensory analysis resulted in the peas being scored on nine different parameters and divided in to four appearance categories. Finally, the study reviewed previous studies’ results on yield, protein content as well as generic diversity. As a conclusion from the sensory analysis together with data on yield, protein and genetic diversity held up against the two market interests, the most promising pea accessions for breeding were identified for further research. These were: SKR108, SKR131, SKR134, SKR138, SKR154, SKR161 and SKR184. In addition, SKR67, SKR88 and SKR135 were found to be the next most recommendable pea accessions.

Drought stress poorly impacts many morphological and physio-biochemical processes in plants. Pea (Pisum sativum L.) plants are highly nutritious crops destined for human consumption; however, their productivity is threatened under drought stress. Thiamine (vitamin B1) is well-known essential micronutrient, acting as a cofactor in key metabolic processes. Therefore, this study was designed to examine the protective effect of foliar application of thiamine (0, 250, and 500 ppm) on two varieties of pea plants under drought stress. Here, we conducted the pot experiment at the Government College Women University, Faisalabad, to investigate the physio-biochemical and morphological traits of two pea varieties (sarsabz and metior) grown under drought stress and thiamine treatment. Drought stress was applied to plants after germination period of 1 month. Results showed that root fresh and dry weight, shoot fresh and dry weight, number of pods, leaf area, total soluble sugars, total phenolics, total protein contents, catalase, peroxidase, and mineral ions were reduced against drought stress. However, the application of thiamine (both 250 and 500 ppm) overcome the stress and also enhances these parameters, and significantly increases the antioxidant activities (catalase and peroxidase). Moreover, the performance of sarsabz was better under control and drought stress conditions than metior variety. In conclusion, the exogenous application of thiamine enabled the plants to withstand drought stress conditions by regulating several physiological and biochemical mechanisms. In agriculture, it is a great latent to alleviate the antagonistic impact of drought stress on crops through the foliar application of thiamine.

Grain and forage legumes are important sources of food and feed, key for sustainable agriculture given the environmental services they provide. However, their cultivation is hampered in the Mediterranean Basin and Near East by the widespread occurrence of the root parasitic weed crenate broomrape (Orobanche crenata). Other broomrape species such as O. minor, O. foetida, and Phelipanche aegyptica are also of local importance. As for other parasitic weeds, a number of management strategies have been proposed, but considering that temperate legumes in the area are low-input crops, these strategies are largely uneconomical or hard to achieve, leaving the use of resistant cultivars as the most desirable option. Breeding for broomrape resistance is not an easy task, but significant progress has been achieved by classical breeding and selection and will profit from recent developments in phenomics and genomics. Here, achievements and prospects in broomrape management and resistance breeding are presented and critically discussed.

Background: Rust is a damaging disease affecting vital crops, including pea, and identifying highly resistant genotypes remains a challenge. Accurate measurement of infection levels in large germplasm collections is crucial for finding new resistance sources. Current evaluation methods rely on visual estimation of disease severity and infection type under field or controlled conditions. While they identify some resistance sources, they are error-prone and time-consuming. An image analysis system proves useful, providing an easy-to-use and affordable way to quickly count and measure rust-induced pustules on pea samples. This study aimed to develop an automated image analysis pipeline for accurately calculating rust disease progression parameters under controlled conditions, ensuring reliable data collection. Results: A highly efficient and automatic image-based method for assessing rust disease in pea leaves was developed using R. The method’s optimization and validation involved testing different segmentation indices and image resolutions on 600 pea leaflets with rust symptoms. The approach allows automatic estimation of parameters like pustule number, pustule size, leaf area, and percentage of pustule coverage. It reconstructs time series data for each leaf and integrates daily estimates into disease progression parameters, including latency period and area under the disease progression curve. Significant variation in disease responses was observed between genotypes using both visual ratings and image-based analysis. Among assessed segmentation indices, the Normalized Green Red Difference Index (NGRDI) proved fastest, analysing 600 leaflets at 60% resolution in 62 s with parallel processing. Lin’s concordance correlation coefficient between image-based and visual pustule counting showed over 0.98 accuracy at full resolution. While lower resolution slightly reduced accuracy, differences were statistically insignificant for most disease progression parameters, signif

Legumes have been sought as alternative protein sources to ensure food security and environmental sustainability. Characterizing their protein content and quality, including in underutilized grain legumes, e.g., grass pea, gives value to the legumes' underexplored variability. To fill the gap of knowledge in legumes' protein quality, for the first time, five extensive collections of cool season grain legumes were cropped under the same environmental conditions and further analyzed. Multivariate analysis showed the existent intra- and inter-species variability. The legume species with the highest protein content, grass pea, Lathyrus sativus (LS), was not the one with the overall highest individual amino acids content and in vitro protein digestibility. With these last characteristics lentil, Lens culinaris (LC), was highlighted. The highest average values of arginine (Arg), glutamic acid (Glu), and threonine (Thr) were found in LS and Vicia faba (VF). Cicer arietinum (CA) stood out as the species with the highest values of Thr and methionine (Met). Regarding the in vitro protein digestibility (IVPD), LC, followed by Pisum sativum (PS) and LS, were the legume species with the highest values. Ultimately, this study bought to the fore legume species that are not commonly used in western diets but have high adaptability to the European agricultural systems.

The incorporation of plant protein ingredients in foods is a means to promote the transition to vegetable proteins. Pea and lupin meet sustainability demands and their protein ingredients display promising technological properties, yet sometimes poor functionalities. However, the involved mechanisms are still unclear, partly because comprehensive and systematic characterization of those ingredients in terms of composition and physicochemical properties is still lacking. In this work, commercial protein fractions of pea and lupin (one isolate and one concentrate for each) were thoroughly characterized. A high-pressure homogenization (HPH) treatment was applied to their aqueous suspensions (pH 7.0) to improve their dispersibility. Although isolates displayed a higher protein content (up to 72 g/100 g (d.m.) against 39 g/100 g (d.m.) for the concentrates, with respective specific N factors), their solubility (i.e., the proteins remaining in the supernatant after centrifugation) was lower than for the concentrates (15–49 wt% of the total proteins, against 65 wt%). Substantial amounts of endogenous lipids in the powders were measured after chloroform/methanol extraction (3.4–10.3 g/100 g (d.m.)), of which about half were phospholipids. For all ingredients, detailed microscopic investigations (including confocal fluorescence microscopy), and light scattering measurements showed that HPH was useful to break down large powder grains. Thus, it altered the colloidal structures present, released endogenous lipid assemblies and enhanced protein solubility. Those new insights into the non-protein composition of plant protein fractions and their behaviour in aqueous media are key for improving their functionalities and facilitating food products’ rational formulation.

An experiment was undertaken to elucidate the genetic relationship between different quantitative traits for commercial cultivation and to evaluate selection criteria in pea breeding programs in five inbred parents. Their 17 F4’s derivatives in pea (Pisum sativum L.) evaluated ten characters during the winter season (November to February) of 2017-18 at the research farm, BSMRAU, Gazipur, Bangladesh. Analysis of variance explored significant differences among the genotypes for all the characters. Phenotypic coefficients of variation (PCV) were close to genotypic coefficients of variation (GCV) for all the characters indicating less influence on the environment and potentiality of selection. A high heritability relationship with high genetic advance was observed for plant height, pod per plant, hundred seed weight, and seed yield per plot. Pod length showed a highly significant positive correlation with pod width and hundred seeds weight. Only days to first flowering showed a highly negative correlation with pod length and hundred seed weight. Path coefficient analysis revealed that plant height, pod per plant, and seeds per pod had a highly positive effect on yield per plant. Therefore, associating and selecting those traits, yield improvement must be possible in pea, and the days to maturity, plant height, pods per plant, pod length, and seed showed a considerable positive and highly significant correlation with plant height, pod per plant, seed per pod, and yield per plant at both genotypic and phenotypic levels indicating yield could be increased with the increase of days to maturity, plant height, pods per plant, pod length, and seed.

An experiment was undertaken to elucidate the genetic relationship between different quantitative traits for commercial cultivation and to evaluate selection criteria in pea breeding programs in five inbred parents. Their 17 F4’s derivatives in pea (Pisum sativum L.) evaluated ten characters during the winter season (November to February) of 2017-18 at the research farm, BSMRAU, Gazipur, Bangladesh. Analysis of variance explored significant differences among the genotypes for all the characters. Phenotypic coefficients of variation (PCV) were close to genotypic coefficients of variation (GCV) for all the characters indicating less influence on the environment and potentiality of selection. A high heritability relationship with high genetic advance was observed for plant height, pod per plant, hundred seed weight, and seed yield per plot. Pod length showed a highly significant positive correlation with pod width and hundred seeds weight. Only days to first flowering showed a highly negative correlation with pod length and hundred seed weight. Path coefficient analysis revealed that plant height, pod per plant, and seeds per pod had a highly positive effect on yield per plant. Therefore, associating and selecting those traits, yield improvement must be possible in pea, and the days to maturity, plant height, pods per plant, pod length, and seed showed a considerable positive and highly significant correlation with plant height, pod per plant, seed per pod, and yield per plant at both genotypic and phenotypic levels indicating yield could be increased with the increase of days to maturity, plant height, pods per plant, pod length, and seed.

The pea, pea or pea is a crop that has a protein level that ranges between 20 and 24 %, which makes it an important food for food security. Given the high content of phosphorus and protein, it is demanding in fertilization, but highly cultivated in several municipalities of the department of Cauca, the research aimed to evaluate the effect of organic fertilizer based on coca leaf in the production of Pisum sativum L. in two municipalities of the department, through foliar and edaphic fertilization, to compare with conventional fertilization. A randomized complete block design was used with three repetitions and six treatments (Control; 4 % CocALOFA I; 8 % CocALOFA I; 4 % CocALOFA II: 8 % CocALOFA II; 8 % CocALOFA II) and as response variables were the performance components. Statistically significant differences were detected for plant height, green bean production and yield in Popayán, while in Timbío there were statistical differences in plant height, grains per pod, weight of one hundred grains, green bean production and yield. The treatment with the best results was 4 % CocALOFA I reaching heights between 1,57 and 1,55 m, weight of one hundred grains of 65 g, production per plant of 108,3 and 100,59 g and yields of 3009,28 and 2794,17 kg/ha. The yield with the mineralized organic liquid fertilizer based on 4 % coca leaf was 34 and 28 % higher than the commercial biofertilizer in the two experimental centers., La arveja, alverja o guisante es un cultivo que tiene un nivel proteico que oscila entre 20 y 24 %, lo cual lo convierte en un alimento importante para la seguridad alimentaria. Dado el alto contenido de fósforo y proteína es exigente en fertilización, pero altamente cultivado en varios municipios del departamento del Cauca, la investigación tuvo por objeto evaluar el efecto del abono orgánico a base de hoja de coca en la producción de Pisum sativum L. en dos municipios del departamento, mediante fertilización foliar y edáfica, para comparar con la fertilización convencional. Se empleó un diseño de bloques completos al azar con tres repeticiones y seis tratamientos (Testigo; CocALOFA I al 4 %; CocALOFA I al 8 %; CocALOFA II al 4 %: CocALOFA II al 8 %; CocALOFA II al 8 %) y como variables de respuesta se tuvieron los componentes de rendimiento. Se detectaron diferencias estadísticamente significativas para altura de la planta, producción en grano verde y rendimiento en Popayán, mientras en Timbío se presentaron diferencias estadísticas en altura de la planta, granos por vaina, peso de cien granos, producción en grano verde y rendimiento. El tratamiento con mejores resultados fue CocALOFA I al 4 % alcanzando alturas entre 1,57 y 1,55 m, peso de cien granos de 65 g, producción por planta de 108,3 y 100,59 g y rendimientos de 3009,28 y 2794,17 kg/ha. El rendimiento con el abono líquido orgánico mineralizado a base de hoja de coca al 4 % superó en un 34 y 28 % al biofertilizante comercial en los dos centros experimentales.

High-quality affinity probes are critical for sensitive and specific protein detection, in particular for detection of protein biomarkers in the early phases of disease development. Proximity extension assays (PEAs) have been used for high-throughput multiplexed protein detection of up to a few thousand different proteins in one or a few microliters of plasma. Clonal affinity reagents can offer advantages over the commonly used polyclonal antibodies (pAbs) in terms of reproducibility and standardization of such assays. Here, we explore nanobodies (Nbs) as an alternative to pAbs as affinity reagents for PEA. We describe an efficient site-specific approach for preparing high-quality oligo-conjugated Nb probes via enzyme coupling using Sortase A (SrtA). The procedure allows convenient removal of unconjugated affinity reagents after conjugation. The purified high-grade Nb probes were used in PEA, and the reactions provided an efficient means to select optimal pairs of binding reagents from a group of affinity reagents. We demonstrate that Nb-based PEA (nano-PEA) for interleukin-6 (IL6) detection can augment assay performance, compared to the use of pAb probes. We identify and validate Nb combinations capable of binding in pairs without competition for IL6 antigen detection by PEA., We thank Dr. Junhong Yan for discussions about Nbs oligo conjugation. The Single Cell Proteomic Facility at SciLifeLab, Uppsala for supports to setup PEA assay. This work was supported by a grant (2012-5852) from the Swedish Research Council’s special call for future therapy, Swedish Research Council under grant agreement No. 2020-02258, a Torsten Söderbergs Stiftelse (M130/16) and the European Research Council under the European Union's Seventh Framework Programme (FP/2007-2013)/ ERC Grant Agreement No. 294409, ProteinSeq and from The Swedish Collegium for Advanced Studies (SCAS)., Alternative recombinant protein binders selection via proximity assays

High-quality affinity probes are critical for sensitive and specific protein detection, in particular for detection of protein biomarkers in the early phases of disease development. Proximity extension assays (PEAs) have been used for high-throughput multiplexed protein detection of up to a few thousand different proteins in one or a few microliters of plasma. Clonal affinity reagents can offer advantages over the commonly used polyclonal antibodies (pAbs) in terms of reproducibility and standardization of such assays. Here, we explore nanobodies (Nbs) as an alternative to pAbs as affinity reagents for PEA. We describe an efficient site-specific approach for preparing high-quality oligo-conjugated Nb probes via enzyme coupling using Sortase A (SrtA). The procedure allows convenient removal of unconjugated affinity reagents after conjugation. The purified high-grade Nb probes were used in PEA, and the reactions provided an efficient means to select optimal pairs of binding reagents from a group of affinity reagents. We demonstrate that Nb-based PEA (nano-PEA) for interleukin-6 (IL6) detection can augment assay performance, compared to the use of pAb probes. We identify and validate Nb combinations capable of binding in pairs without competition for IL6 antigen detection by PEA., We thank Dr. Junhong Yan for discussions about Nbs oligo conjugation. The Single Cell Proteomic Facility at SciLifeLab, Uppsala for supports to setup PEA assay. This work was supported by a grant (2012-5852) from the Swedish Research Council’s special call for future therapy, Swedish Research Council under grant agreement No. 2020-02258, a Torsten Söderbergs Stiftelse (M130/16) and the European Research Council under the European Union's Seventh Framework Programme (FP/2007-2013)/ ERC Grant Agreement No. 294409, ProteinSeq and from The Swedish Collegium for Advanced Studies (SCAS)., Alternative recombinant protein binders selection via proximity assays

High-quality affinity probes are critical for sensitive and specific protein detection, in particular for detection of protein biomarkers in the early phases of disease development. Proximity extension assays (PEAs) have been used for high-throughput multiplexed protein detection of up to a few thousand different proteins in one or a few microliters of plasma. Clonal affinity reagents can offer advantages over the commonly used polyclonal antibodies (pAbs) in terms of reproducibility and standardization of such assays. Here, we explore nanobodies (Nbs) as an alternative to pAbs as affinity reagents for PEA. We describe an efficient site-specific approach for preparing high-quality oligo-conjugated Nb probes via enzyme coupling using Sortase A (SrtA). The procedure allows convenient removal of unconjugated affinity reagents after conjugation. The purified high-grade Nb probes were used in PEA, and the reactions provided an efficient means to select optimal pairs of binding reagents from a group of affinity reagents. We demonstrate that Nb-based PEA (nano-PEA) for interleukin-6 (IL6) detection can augment assay performance, compared to the use of pAb probes. We identify and validate Nb combinations capable of binding in pairs without competition for IL6 antigen detection by PEA., We thank Dr. Junhong Yan for discussions about Nbs oligo conjugation. The Single Cell Proteomic Facility at SciLifeLab, Uppsala for supports to setup PEA assay. This work was supported by a grant (2012-5852) from the Swedish Research Council’s special call for future therapy, Swedish Research Council under grant agreement No. 2020-02258, a Torsten Söderbergs Stiftelse (M130/16) and the European Research Council under the European Union's Seventh Framework Programme (FP/2007-2013)/ ERC Grant Agreement No. 294409, ProteinSeq and from The Swedish Collegium for Advanced Studies (SCAS)., Alternative recombinant protein binders selection via proximity assays

High-quality affinity probes are critical for sensitive and specific protein detection, in particular for detection of protein biomarkers in the early phases of disease development. Proximity extension assays (PEAs) have been used for high-throughput multiplexed protein detection of up to a few thousand different proteins in one or a few microliters of plasma. Clonal affinity reagents can offer advantages over the commonly used polyclonal antibodies (pAbs) in terms of reproducibility and standardization of such assays. Here, we explore nanobodies (Nbs) as an alternative to pAbs as affinity reagents for PEA. We describe an efficient site-specific approach for preparing high-quality oligo-conjugated Nb probes via enzyme coupling using Sortase A (SrtA). The procedure allows convenient removal of unconjugated affinity reagents after conjugation. The purified high-grade Nb probes were used in PEA, and the reactions provided an efficient means to select optimal pairs of binding reagents from a group of affinity reagents. We demonstrate that Nb-based PEA (nano-PEA) for interleukin-6 (IL6) detection can augment assay performance, compared to the use of pAb probes. We identify and validate Nb combinations capable of binding in pairs without competition for IL6 antigen detection by PEA., We thank Dr. Junhong Yan for discussions about Nbs oligo conjugation. The Single Cell Proteomic Facility at SciLifeLab, Uppsala for supports to setup PEA assay. This work was supported by a grant (2012-5852) from the Swedish Research Council’s special call for future therapy, Swedish Research Council under grant agreement No. 2020-02258, a Torsten Söderbergs Stiftelse (M130/16) and the European Research Council under the European Union's Seventh Framework Programme (FP/2007-2013)/ ERC Grant Agreement No. 294409, ProteinSeq and from The Swedish Collegium for Advanced Studies (SCAS)., Alternative recombinant protein binders selection via proximity assays