Digital descriptors sharpen classical descriptors, for improving genebank accession management: A case study on Arachis spp. and Phaseolus spp.

High-throughput phenotyping brings new opportunities for detailed genebank accessions characterization based on image-processing techniques and data analysis using machine learning algorithms. Our work proposes to improve the characterization processes of bean and peanut accessions in the CIAT genebank through the identification of phenomic descriptors comparable to classical descriptors including methodology integration into the genebank workflow. To cope with these goals morphometrics and colorimetry traits of 14 bean and 16 forage peanut accessions were determined and compared to the classical International Board for Plant Genetic Resources (IBPGR) descriptors. Descriptors discriminating most accessions were identified using a random forest algorithm. The most-valuable classification descriptors for peanuts were 100-seed weight and days to flowering, and for beans, days to flowering and primary seed color. The combination of phenomic and classical descriptors increased the accuracy of the classification of Phaseolus and Arachis accessions. Functional diversity indices are recommended to genebank curators to evaluate phenotypic variability to identify accessions with unique traits or identify accessions that represent the greatest phenotypic variation of the species (functional agrobiodiversity collections). The artificial intelligence algorithms are capable of characterizing accessions which reduces costs generated by additional phenotyping. Even though deep analysis of data requires new skills, associating genetic, morphological and ecogeographic diversity is giving us an opportunity to establish unique functional agrobiodiversity collections with new potential traits.

LysipheN: a gravimetric IoT device for near real-time high-frequency crop phenotyping: a case study on common beans.

Climate instability directly affects agro-environments. Water scarcity, high air temperature, and changes in soil biota are some factors caused by environmental changes. Verified and precise phenotypic traits are required for assessing the impact of various stress factors on crop performance while keeping phenotyping costs at a reasonable level. Experiments which use a lysimeter method to measure transpiration efficiency are often expensive and require complex infrastructures. This study presents the development and testing process of an automated, reliable, small, and low-cost prototype system using IoT with high-frequency potential in near-real time. Because of its waterproofness, our device-LysipheN-assesses each plant individually and can be deployed for experiments in different environmental conditions (farm, field, greenhouse, etc.). LysipheN integrates multiple sensors, automatic irrigation according to desired drought scenarios, and a remote, wireless connection to monitor each plant and device performance via a data platform. During testing, LysipheN proved to be sensitive enough to detect and measure plant transpiration, from early to ultimate plant developmental stages. Even though the results were generated on common beans, the LysipheN can be scaled up/adapted to other crops. This tool serves to screen transpiration, transpiration efficiency, and transpiration-related physiological traits. Because of its price, endurance, and waterproof design, LysipheN will be useful in screening populations in a realistic ecological and breeding context. It operates by phenotyping the most suitable parental lines, characterizing genebank accessions, and allowing breeders to make a target-specific selection using functional traits (related to the place where LysipheN units are located) in line with a realistic agronomic background.

Novel triterpenoid pyrones, phthalimides and phthalates are selectively cytotoxic in CCRF-CEM cancer cells - Synthesis, potency, and mitochondrial mechanism of action.

A series of triterpenoid pyrones was synthesized and subsequently modified to introduce phthalimide or phthalate moieties into the triterpenoid skeleton. These compounds underwent in vitro cytotoxicity screening, revealing that a subset of six compounds exhibited potent activity, with IC50 values in the low micromolar range. Further biological evaluations, including Annexin V and propidium iodide staining experiment revealed, that all compounds induce selective apoptosis in cancer cells. Measurements of mitochondrial potential, cell cycle analysis, and the expression of pro- and anti-apoptotic proteins confirmed, that apoptosis was mediated via the mitochondrial pathway. These findings were further supported by cell cycle modulation and DNA/RNA synthesis studies, which indicated a significant increase in cell accumulation in the G0/G1 phase and a marked reduction in S-phase cells, alongside a substantial inhibition of DNA synthesis. The activation of caspase-3 and the cleavage of PARP, coupled with a decrease in the expression of Bcl-2 and Bcl-XL proteins, underscored the induction of apoptosis through the mitochondrial pathway. Given their high activity and pronounced effect on mitochondria function, trifluoromethyl pyrones 1f and 2f, and dihydrophthalimide 2h have been selected for further development.

Linking photosynthesis and yield reveals a strategy to improve light use efficiency in a climbing bean breeding population.

Photosynthesis drives plant physiology, biomass accumulation, and yield. Photosynthetic efficiency, specifically the operating efficiency of PSII (Fq'/Fm'), is highly responsive to actual growth conditions, especially to fluctuating photosynthetic photon fluence rate (PPFR). Under field conditions, plants constantly balance energy uptake to optimize growth. The dynamic regulation complicates the quantification of cumulative photochemical energy uptake based on the intercepted solar energy, its transduction into biomass, and the identification of efficient breeding lines. Here, we show significant effects on biomass related to genetic variation in photosynthetic efficiency of 178 climbing bean (Phaseolus vulgaris L.) lines. Under fluctuating conditions, the Fq'/Fm' was monitored throughout the growing period using hand-held and automated chlorophyll fluorescence phenotyping. The seasonal response of Fq'/Fm' to PPFR (ResponseG:PPFR) achieved significant correlations with biomass and yield, ranging from 0.33 to 0.35 and from 0.22 to 0.31 in two glasshouse and three field trials, respectively. Phenomic yield prediction outperformed genomic predictions for new environments in four trials under different growing conditions. Investigating genetic control over photosynthesis, one single nucleotide polymorphism (Chr09_37766289_13052) on chromosome 9 was significantly associated with ResponseG:PPFR in proximity to a candidate gene controlling chloroplast thylakoid formation. In conclusion, photosynthetic screening facilitates and accelerates selection for high yield potential.

Influence of Increase in Phosphorus Supply on Agronomic, Phenological, and Physiological Performance of Two Common Bean Breeding Lines Grown in Acidic Soil under High Temperature Stress Conditions.

Many common bean (Phaseolus vulgaris L.) plants cultivated in areas of the world with acidic soils exhibit difficulties adapting to low phosphorus (P) availability, along with aluminum (Al) toxicity, causing yield loss. The objective of this study was to evaluate the influence of an increase in P supply level on the agronomic, phenological, and physiological performance of two common bean breeding lines grown in acidic soil, with low fertility and under high temperature conditions, in a screenhouse. A randomized complete block (RCB) design was used under a factorial arrangement (five levels of P × 2 genotypes) for a total of 10 treatments with four replications. The factors considered in the experiment were: (i) five P supply levels (kg ha-1): four levels of P0, P15, P30, and P45 through the application of rock phosphate (RP), and one P level supplied through the application of organic matter (PSOM) corresponding to 25 kg P ha-1 (P25); and (ii) two advanced bean lines (BFS 10 and SEF10). Both bean lines were grown under the combined stress conditions of high temperatures (day and night maximum temperatures of 42.5 °C/31.1 °C, respectively) and acidic soil. By increasing the supply of P, a significant effect was found, indicating an increase in the growth and development of different vegetative organs, as well as physiological efficiency in photosynthesis and photosynthate remobilization, which resulted in higher grain yield in both bean lines evaluated (BFS 10 and SEF10). The adaptive responses of the two bean lines were found to be related to phenological adjustments (days to flowering and physiological maturity; stomatal development), as well as to heat dissipation strategies in the form of heat (NPQ) or unregulated energy (qN) that contributed to greater agronomic performance. We found that, to some extent, increased P supply alleviated the negative effects of high temperature on the growth and development of the reproductive organs of bean lines. Both bean lines (BFS 10 and SEF 10) showed adaptive attributes suited to the combined stress conditions of high temperature and acidic soil, and these two lines can serve as useful parents in a bean breeding program to develop multiple stress tolerant cultivars.

Bioactive Compounds and Antioxidant Activity in Seeds of Bred Lines of Common Bean Developed from Interspecific Crosses.

Knowledge is limited about the level of bioactive compounds and antioxidant activity of seeds from bred lines of common beans developed from interspecific crosses using four different Phaseolus species (P. vulgaris L., P. coccineus L., P. acutifolius A. Gray. Gray., and P. dumosus). In this study, differences in the nutritional quality of seeds among 112 bean genotypes were evaluated by measuring the levels of phenolic compounds, pigments, antioxidant activity, and sugars. The bean genotypes were grown under high temperatures and acid soil conditions in the Amazon region of Colombia. Five typology groups of bean genotypes were identified based on the level of bioactive compounds and their functional capacity: (1) highly bioactive and functional (HBF); (2) moderately bioactive and functional (MBF); (3) moderate antioxidant content with pigment influence (MACP); (4) moderately antinutritional with limited antioxidant potential (MALAP); and (5) antinutritional, low bioactive, and functional (ALBF). We developed a nutritional quality index (NQI) with values ranging from 0 to 1 based on the nutritional and anti-nutritional balance of each genotype and the higher values of the NQI of a genotype indicating greater nutritional quality. We found three interspecific bred lines (SER 212, SER 213, and RRA 81), with NQI values higher than 0.8. These three lines belong to the typology group of HBF. The superior nutritional quality of these three interspecific bred lines is attributed to a greater level of bioactive compounds and antioxidant capacity. These three bred lines may serve as useful parents to develop nutritionally superior and stress-resilient beans from bean breeding programs. Further research is needed to explore the role of testa color in improving the nutritional quality of seeds of common bean genotypes grown under different climatic conditions.

High Temperature Tolerance in a Novel, High-Quality Phaseolus vulgaris Breeding Line Is Due to Maintenance of Pollen Viability and Successful Germination on the Stigma.

The common bean (Phaseolus vulgaris L.) is an important nutritional source globally but is sensitive to high temperatures and thus particularly vulnerable to climate change. Derived from a breeding program at CIAT (Colombia), a heat-tolerant breeding line, named heat-tolerant Andean-type 4 (HTA4), was developed by a series of crosses of parents with a small-bean tepary genotype (Phaseolus acutifolius L.) in their pedigree, which might be the donor of heat stress (HS) tolerance. Importantly, in HTA4, the large, commercially desirable Andean-type beans was restored. To assess underlying tolerance mechanisms, HTA4, together with a heat-sensitive Colombian variety (Calima), was exposed to HS (31 °C/24 °C HS vs. 26 °C/19 °C day/night) under controlled environment conditions. Vegetative growth and photosynthetic performance were not negatively impacted by HS in either genotype, although senescence was delayed in Calima. HS during the reproductive stage caused an increase in pod number in Calima but with few fully developed seeds and many pods aborted and/or abscised. In contrast, HTA4 maintained a similar filled pod number under HS and a higher seed weight per plant. Pollen showed high sterility in Calima, with many non-viable pollen grains (24.9% viability compared to 98.4% in control) with a thicker exine and fewer starch granules under HS. Calima pollen failed to adhere to the stigma and germinate under HS. In HTA4, pollen viability was significantly higher than in Calima (71.1% viability compared to 95.4% under control), and pollen successfully germinated and formed pollen tubes in the style under HS. It is concluded that HTA4 is heat tolerant and maintains a high level of reproductive output due to its ability to produce healthy pollen that is able to adhere to the stigma.

Interspecific common bean population derived from Phaseolus acutifolius using a bridging genotype demonstrate useful adaptation to heat tolerance.

Common bean (Phaseolus vulgaris L.) is an important legume crop worldwide and is a major nutrient source in the tropics. Common bean reproductive development is strongly affected by heat stress, particularly overnight temperatures above 20°C. The desert Tepary bean (Phaseolus acutifolius A. Gray) offers a promising source of adaptative genes due to its natural acclimation to arid conditions. Hybridization between both species is challenging, requiring in vitro embryo rescue and multiple backcrossing cycles to restore fertility. This labor-intensive process constrains developing mapping populations necessary for studying heat tolerance. Here we show the development of an interspecific mapping population using a novel technique based on a bridging genotype derived from P. vulgaris, P. Acutifolius and P. parvifolius named VAP1 and is compatible with both common and tepary bean. The population was based on two wild P. acutifolius accessions, repeatedly crossed with Mesoamerican elite common bush bean breeding lines. The population was genotyped through genotyping-by-sequencing and evaluated for heat tolerance by genome-wide association studies. We found that the population harbored 59.8% introgressions from wild tepary, but also genetic regions from Phaseolus parvifolius, a relative represented in some early bridging crosses. We found 27 significative quantitative trait loci, nine located inside tepary introgressed segments exhibiting allelic effects that reduced seed weight, and increased the number of empty pods, seeds per pod, stem production and yield under high temperature conditions. Our results demonstrate that the bridging genotype VAP1 can intercross common bean with tepary bean and positively influence the physiology of derived interspecific lines, which displayed useful variance for heat tolerance.

Using phenomics to identify and integrate traits of interest for better-performing common beans: A validation study on an interspecific hybrid and its Acutifolii parents.

Introduction: Evaluations of interspecific hybrids are limited, as classical genebank accession descriptors are semi-subjective, have qualitative traits and show complications when evaluating intermediate accessions. However, descriptors can be quantified using recognized phenomic traits. This digitalization can identify phenomic traits which correspond to the percentage of parental descriptors remaining expressed/visible/measurable in the particular interspecific hybrid. In this study, a line of P. vulgaris, P. acutifolius and P. parvifolius accessions and their crosses were sown in the mesh house according to CIAT seed regeneration procedures. Methodology: Three accessions and one derived breeding line originating from their interspecific crosses were characterized and classified by selected phenomic descriptors using multivariate and machine learning techniques. The phenomic proportions of the interspecific hybrid (line INB 47) with respect to its three parent accessions were determined using a random forest and a respective confusion matrix. Results: The seed and pod morphometric traits, physiological behavior and yield performance were evaluated. In the classification of the accession, the phenomic descriptors with highest prediction force were Fm', Fo', Fs', LTD, Chl, seed area, seed height, seed Major, seed MinFeret, seed Minor, pod AR, pod Feret, pod round, pod solidity, pod area, pod major, pod seed weight and pod weight. Physiological traits measured in the interspecific hybrid present 2.2% similarity with the P. acutifolius and 1% with the P. parvifolius accessions. In addition, in seed morphometric characteristics, the hybrid showed 4.5% similarity with the P. acutifolius accession. Conclusions: Here we were able to determine the phenomic proportions of individual parents in their interspecific hybrid accession. After some careful generalization the methodology can be used to: i) verify trait-of-interest transfer from P. acutifolius and P. parvifolius accessions into their hybrids; ii) confirm selected traits as "phenomic markers" which would allow conserving desired physiological traits of exotic parental accessions, without losing key seed characteristics from elite common bean accessions; and iii) propose a quantitative tool that helps genebank curators and breeders to make better-informed decisions based on quantitative analysis.

Impact of Web Blight on Photosynthetic Performance of an Elite Common Bean Line in the Western Amazon Region of Colombia.

Disease stress caused by plant pathogens impacts the functioning of the photosynthetic apparatus, and the symptoms caused by the degree of severity of the disease can generally be observed in different plant parts. The accurate assessment of plant symptoms can be used as a proxy indicator for managing disease incidence, estimating yield loss, and developing genotypes with disease resistance. The objective of this work was to determine the response of the photosynthetic apparatus to the increased disease severity caused by web blight Thanatephorus cucumeris (Frank) Donk on the common bean (Phaseolus vulgaris L.) leaves under acidic soil and the humid tropical conditions of the Colombian Amazon. Differences in chlorophyll fluorescence parameters, including Fv/Fm, Y(II), Y(NPQ), Y(NO), ETR, qP, and qN in leaves with different levels of severity of web blight in an elite line (BFS 10) of common bean were evaluated under field conditions. A significant effect of web blight on the photosynthetic apparatus was found. A reduction of up to 50% of energy use dedicated to the photosynthetic machinery was observed, even at the severity scale score of 2 (5% surface incidence). The results from this study indicate that the use of fluorescence imaging not only allows for the quantifying of the impact of web blight on photosynthetic performance, but also for detecting the incidence of disease earlier, before severe symptoms occur on the leaves.

Lupane derivatives containing various aryl substituents in the position 3 have selective cytostatic effect in leukemic cancer cells including resistant phenotypes.

In this work, a large set of betulinic acid derivatives modified with various aromatic substituents at the position C-3 were prepared via Suzuki-Myiaura cross-coupling. All compounds were tested for their in vitro cytotoxic activity in 8 cancer and 2 healthy cell lines. Derivatives 6h, 6i, and 6o had the lowest IC50 in the CCRF-CEM cell line (0.69-4.0 µM) and had high selectivity. In addition, 6h and 6i also showed significant activity in daunorubicin-resistant CEM and taxol-resistant K562 cell lines; therefore, they were selected for the evaluation of the mechanism of action. First, the effect of 6h, 6i, and 6o on cell death induction was studied. To our surprise, we have not detected almost any apoptotic cells, even following a long-time exposure of CCRF-CEM cells to the compounds. On the other hand, a dramatic cell number decrease was observed, proportional to the time of the compound's exposure. Based on this data it was concluded that the effect of compounds is cytostatic rather than cytotoxic, which was further confirmed by subsequent studies of the impact of 6h, 6i, and 6o on the cell cycle. Detailed cell cycle analysis revealed a block in the G1 phase accompanied by reduced expression of phosphorylated forms of the RB protein as well as cyclin A protein. Evaluation of the pharmacological properties of the most promising compounds revealed their high stability in the presence of phosphate buffer, human plasma, and microsomes and limited permeability determined using permeability through artificial membrane (PAMPA) and cell permeability assay: Caco-2 and MDCK-MDR1 cell lines. Compounds 6h, 6i, and 6o were selected for further drug development; their cytostatic effect may be advantageous in this process since we expect fewer non-specific interactions and toxicity than in highly cytotoxic compounds. In addition, the activity of 6h and 6i against resistant CEM-DNR and K562-TAX leukemic cell lines makes them promising as a possible future alternative to currently used therapies.

Triterpenoid pyrazines and pyridines - Synthesis, cytotoxicity, mechanism of action, preparation of prodrugs.

A set of fifteen triterpenoid pyrazines and pyridines was prepared from parent triterpenoid 3-oxoderivatives (betulonic acid, dihydrobetulonic acid, oleanonic acid, moronic acid, ursonic acid, heterobetulonic acid, and allobetulone). Cytotoxicity of all compounds was tested in eight cancer and two non-cancer cell lines. Evaluation of the structure-activity relationships revealed that the triterpenoid core determined whether the final molecule is active or not, while the heterocycle is able to increase the activity and modulate the specificity. Five compounds (1b, 1c, 2b, 2c, and 8) were found to be preferentially and highly cytotoxic (IC50 ≈ 1 µM) against leukemic cancer cell lines (CCRF-CEM, K562, CEM-DNR, or K562-TAX). Surprisingly, compounds 1c, 2b, and 2c are 10-fold more active in multidrug-resistant leukemia cells (CEM-DNR and K562-TAX) than in their non-resistant analogs (CCRF-CEM and K562). Pharmacological parameters were measured for the most promising candidates and two types of prodrugs were synthesized: 1) Sugar-containing conjugates, most of which had improved cell penetration and retained high cytotoxicity in the CCRF-CEM cell line, unfortunately, they lost the selectivity against resistant cells. 2) Medoxomil derivatives, among which compounds 26-28 gained activities of IC50 0.026-0.043 µM against K562 cells. Compounds 1b, 8, 21, 22, 23, and 24 were selected for the evaluation of the mechanism of action based on their highest cytotoxicity against CCRF-CEM cell line. Several experiments showed that the majority of them cause apoptosis via the mitochondrial pathway. Compounds 1b, 8, and 21 inhibit growth and disintegrate spheroid cultures of HCT116 and HeLa cells, which would be important for the treatment of solid tumors. In summary, compounds 1b, 1c, 2b, 2c, 24, and 26-28 are highly and selectively cytotoxic against cancer cell lines and were selected for future in vivo tests and further development of anticancer drugs.

Chlorophyll Fluorescence Imaging as a Tool for Evaluating Disease Resistance of Common Bean Lines in the Western Amazon Region of Colombia.

The evaluation of disease resistance is considered an important aspect of phenotyping for crop improvement. Identification of advanced lines of the common bean with disease resistance contributes to improved grain yields. This study aimed to determine the response of the photosynthetic apparatus to natural pathogen infection by using chlorophyll (Chla) fluorescence parameters and their relationship to the agronomic performance of 59 common bean lines and comparing the photosynthetic responses of naturally infected vs. healthy leaves. The study was conducted over two seasons under acid soil and high temperature conditions in the western Amazon region of Colombia. A disease susceptibility index (DSI) was developed and validated using chlorophyll a (Chla) fluorescence as a tool to identify Mesoamerican and Andean lines of common bean (Phaseolus vulgaris L.) that are resistant to pathogens. A negative effect on the functional status of the photosynthetic apparatus was found with the presence of pathogen infection, a situation that allowed the identification of four typologies based on the DSI values ((i) moderately resistant; (ii) moderately susceptible; (iii) susceptible; and (iv) highly susceptible). Moderately resistant lines, five of them from the Mesoamerican gene pool (ALB 350, SMC 200, BFS 10, SER 16, SMN 27) and one from the Andean gene pool (DAB 295), allocated a higher proportion of energy to photochemical processes, which increased the rate of electron transfer resulting in a lower sensitivity to disease stress. This photosynthetic response was associated with lower values of DSI, which translated into an increase in the accumulation of dry matter accumulation in different plant organs (leaves, stem, pods and roots). Thus, DSI values based on chlorophyll fluorescence response to pathogen infection could serve as a phenotyping tool for evaluating advanced common bean lines. Six common bean lines (ALB 350, BFS 10, DAB 295, SER 16, SMC 200 and SMN 27) were identified as less sensitive to disease stress under field conditions in the western Amazon region of Colombia, and these could serve as useful parents for improving the common bean for multiple stress resistance.

Water Use, Leaf Cooling and Carbon Assimilation Efficiency of Heat Resistant Common Beans Evaluated in Western Amazonia.

In our study, we analyzed 30years of climatological data revealing the bean production risks for Western Amazonia. Climatological profiling showed high daytime and nighttime temperatures combined with high relative humidity and low vapor pressure deficit. Our understanding of the target environment allows us to select trait combinations for reaching higher yields in Amazonian acid soils. Our research was conducted using 64 bean lines with different genetic backgrounds. In high temperatures, we identified three water use efficiency typologies in beans based on detailed data analysis on gasometric exchange. Profligate water spenders and not water conservative accessions showed leaf cooling, and effective photosynthate partitioning to seeds, and these attributes were found to be related to higher photosynthetic efficiency. Thus, water spenders and not savers were recognized as heat resistant in acid soil conditions in Western Amazonia. Genotypes such as BFS 10, SEN 52, SER 323, different SEFs (SEF 73, SEF 10, SEF 40, SEF 70), SCR 56, SMR 173, and SMN 99 presented less negative effects of heat stress on yield. These genotypes could be suitable as parental lines for improving dry seed production. The improved knowledge on water-use efficiency typologies can be used for bean crop improvement efforts as well as further studies aimed at a better understanding of the intrinsic mechanisms of heat resistance in legumes.

Adaptation of Interspecific Mesoamerican Common Bean Lines to Acid Soils and High Temperature in the Amazon Region of Colombia.

Knowledge of the physiological basis for improved genetic adaptation of common bean (Phaseolus vulgaris L.) lines to acid soils and high temperature conditions in the Amazon region of Colombia is limited. In this study, we evaluated the differences among 41 common bean lines in energy use, leaf cooling, photosynthate partitioning to pod formation and grain filling, and grain yield over two seasons under acid soil and high temperature stress in the Amazon region of Colombia. Common bean lines evaluated included medium and large seeded interspecific lines of Mesoamerican and Andean gene pools with different levels of adaptation to abiotic stress conditions and some lines are improved for iron and zinc (biofortified) concentration in seeds. We found three bean lines (GGR 147, SMG 21 and SMG 12) that were superior in their photosynthetic response, leaf cooling, photosynthate partitioning ability to pod formation and grain filling, resulting in grain yields exceeding 1900 kg ha-1 under acid soil and high temperature stress conditions. The superior photosynthetic performance was attributed to the efficient use of absorbed energy on the electron level in thylakoids, which is mainly oriented to a higher quantum yield of PSII (ΦII), lower energy dissipation in the form of heat (ΦNPQ), high linear electron flow (LEF) and high fraction of PSI centers in open state (PSIopen). We speculate that these photosynthetic and photosynthate partitioning responses of superior bean lines are part of the genetic adaptation to acidic soils and high temperature stress conditions. Among the evaluated bean lines, three lines (GGR 147, SMG 21 and SMG 12) combined the desirable attributes for genetic improvement of stress tolerance and biofortification. These lines can serve as parents to further improve traits (energy use efficiency and multiple stress resistance) that are important for bean production in the Amazon region.

Morphological and Molecular Characterization of Selected Chilean Runner Bean (Phaseolus coccineus L.) Genotypes Shows Moderate Agronomic and Genetic Variability.

The runner bean is the third most economically important Phaseolus species, which is cultivated on small-scale agriculture for the production of immature pods or to obtain dry seeds. However, despite the economic importance and agronomic potential of this species, the runner bean has been little studied from the genetic standpoint. Therefore, the main objective of this study was to characterize ten selected lines of runner bean obtained from Central (Santiago) and Southern (Valdivia and Villarica) Chile based on morphological and agronomic traits. In addition, the genetic variability of these lines was determined using 12 Inter-Simple Sequence Repeat (ISSR) markers to evaluate the potential of this germplasm for breeding and commercial purposes. As a result, the lines from Central Chile were characterized, and had a higher number of pods per plant compared to the Southern lines, although the size and weight of their seeds were lower. Moreover, a low level of genetic diversity (He = 0.251) was encountered in this population. Finally, this is one of the first studies that generate relevant and novel information on the morphological, agronomic and genetic characterization of the P. coccineus germplasm present in Chile.

Substituted dienes prepared from betulinic acid - Synthesis, cytotoxicity, mechanism of action, and pharmacological parameters.

A set of new substituted dienes were synthesized from betulinic acid by its oxidation to 30-oxobetulinic acid followed by the Wittig reaction. Cytotoxicity of all compounds was tested in vitro in eight cancer cell lines and two noncancer fibroblasts. Almost all dienes were more cytotoxic than betulinic acid. Compounds 4.22, 4.30, 4.33, 4.39 had IC50 below 5 µmol/L; 4.22 and 4.39 were selected for studies of the mechanism of action. Cell cycle analysis revealed an increase in the number of apoptotic cells at 5 × IC50 concentration, where activation of irreversible changes leading to cell death can be expected. Both 4.22 and 4.39 led to the accumulation of cells in the G0/G1 phase with partial inhibition of DNA/RNA synthesis at 1 × IC50 and almost complete inhibition at 5 × IC50. Interestingly, compound 4.39 at 5 × IC50 caused the accumulation of cells in the S phase. Higher concentrations of tested drugs probably inhibit more off-targets than lower concentrations. Mechanisms disrupting cellular metabolism can induce the accumulation of cells in the S phase. Both compounds 4.22 and 4.39 trigger selective apoptosis in cancer cells via intrinsic pathway, which we have demonstrated by changes in the expression of the crucial apoptosis-related protein. Pharmacological parameters of derivative 4.22 were superior to 4.39, therefore 4.22 was the finally selected candidate for the development of anticancer drug.

The Resistance of Oilseed Rape Microspore-Derived Embryos to Osmotic Stress Is Associated With the Accumulation of Energy Metabolism Proteins, Redox Homeostasis, Higher Abscisic Acid, and Cytokinin Contents.

The present study aims to investigate the response of rapeseed microspore-derived embryos (MDE) to osmotic stress at the proteome level. The PEG-induced osmotic stress was studied in the cotyledonary stage of MDE of two genotypes: Cadeli (D) and Viking (V), previously reported to exhibit contrasting leaf proteome responses under drought. Two-dimensional difference gel electrophoresis (2D-DIGE) revealed 156 representative protein spots that have been selected for MALDI-TOF/TOF analysis. Sixty-three proteins have been successfully identified and divided into eight functional groups. Data are available via ProteomeXchange with identifier PXD024552. Eight selected protein accumulation trends were compared with real-time quantitative PCR (RT-qPCR). Biomass accumulation in treated D was significantly higher (3-fold) than in V, which indicates D is resistant to osmotic stress. Cultivar D displayed resistance strategy by the accumulation of proteins in energy metabolism, redox homeostasis, protein destination, and signaling functional groups, high ABA, and active cytokinins (CKs) contents. In contrast, the V protein profile displayed high requirements of energy and nutrients with a significant number of stress-related proteins and cell structure changes accompanied by quick downregulation of active CKs, as well as salicylic and jasmonic acids. Genes that were suitable for gene-targeting showed significantly higher expression in treated samples and were identified as phospholipase D alpha, peroxiredoxin antioxidant, and lactoylglutathione lyase. The MDE proteome profile has been compared with the leaf proteome evaluated in our previous study. Different mechanisms to cope with osmotic stress were revealed between the genotypes studied. This proteomic study is the first step to validate MDE as a suitable model for follow-up research on the characterization of new crossings and can be used for preselection of resistant genotypes.

In pursuit of a better world: crop improvement and the CGIAR.

The CGIAR crop improvement (CI) programs, unlike commercial CI programs, which are mainly geared to profit though meeting farmers' needs, are charged with meeting multiple objectives with target populations that include both farmers and the community at large. We compiled the opinions from >30 experts in the private and public sector on key strategies, methodologies, and activities that could the help CGIAR meet the challenges of providing farmers with improved varieties while simultaneously meeting the goals of: (i) nutrition, health, and food security; (ii) poverty reduction, livelihoods, and jobs; (iii) gender equality, youth, and inclusion; (iv) climate adaptation and mitigation; and (v) environmental health and biodiversity. We review the crop improvement processes starting with crop choice, moving through to breeding objectives, production of potential new varieties, selection, and finally adoption by farmers. The importance of multidisciplinary teams working towards common objectives is stressed as a key factor to success. The role of the distinct disciplines, actors, and their interactions throughout the process from crop choice through to adoption by farmers is discussed and illustrated.

Biocatalysis in the Chemistry of Lupane Triterpenoids.

Pentacyclic triterpenes are important representatives of natural products that exhibit a wide variety of biological activities. These activities suggest that these compounds may represent potential medicines for the treatment of cancer and viral, bacterial, or protozoal infections. Naturally occurring triterpenes usually have several drawbacks, such as limited activity and insufficient solubility and bioavailability; therefore, they need to be modified to obtain compounds suitable for drug development. Modifications can be achieved either by methods of standard organic synthesis or with the use of biocatalysts, such as enzymes or enzyme systems within living organisms. In most cases, these modifications result in the preparation of esters, amides, saponins, or sugar conjugates. Notably, while standard organic synthesis has been heavily used and developed, the use of the latter methodology has been rather limited, but it appears that biocatalysis has recently sparked considerably wider interest within the scientific community. Among triterpenes, derivatives of lupane play important roles. This review therefore summarizes the natural occurrence and sources of lupane triterpenoids, their biosynthesis, and semisynthetic methods that may be used for the production of betulinic acid from abundant and inexpensive betulin. Most importantly, this article compares chemical transformations of lupane triterpenoids with analogous reactions performed by biocatalysts and highlights a large space for the future development of biocatalysis in this field. The results of this study may serve as a summary of the current state of research and demonstrate the potential of the method in future applications.