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Novel foods to treat food allergy and gastrointestinal infection.

The gastrointestinal tract communicates directly with the external environment. Necessary nutrients must be absorbed and commensal bacteria tolerated, and foreign proteins, antigens, and pathogens must be simultaneously excluded or destroyed. Immaturity or disruption of the mucosal immune defenses increases vulnerability to food allergy, intolerance, and infectious disease. Diseases resulting from ingested foreign proteins and organisms are increasing and cause morbidity and mortality worldwide. There is no specific treatment for food allergy other than avoidance. Vaccination for infectious disease is limited by the cost and logistics of distribution and administration, particularly in developing countries. Novel strategies are being explored to modulate the gut mucosal immune system by altering protein expression in food. Crops are being developed to remove deleterious allergens to prevent immunogenic exposure while preserving nutritional quality. Local food plants that express protein fragments of pathogens might provide an effective means to stimulate gut mucosal immunity while increasing vaccine accessibility.

Autoimmunity↗

Growing root, tuber and nut crops hydroponically for CELSS.

Among the crops selected by the National Aeronautics and Space Administration for growth in controlled ecological life support systems are four that have subsurface edible parts -- potatoes, sweet potatoes, sugar beets and peanuts. These crops have been produced in open and closed (recirculating), solid media and liquid, hydroponic systems. Fluorescent , fluorescent plus incandescent and high pressure sodium plus metal halide lamps have proven to be effective light sources. Continuous light with 16 degrees C and 28/22 degrees C (day/night) temperatures have produced highest yields for potato and sweet potato, respectively. Dry weight yields of up to 4685, 2541, 1151 and 207 g m-2 for for potatoes, sweet potatoes, sugar beets and peanuts, respectively, have been produced in controlled environment hydroponic systems.

Arachis↗

Optimization of the postcolumn hydrolysis reaction on solid phases for the routine high-performance liquid chromatographic determination of N-methylcarbamate pesticides in food products.

Various solid-phase materials were evaluated as catalysts for the postcolumn hydrolysis of N-methylcarbamate pesticides in a high-performance liquid chromatographic (HPLC) method for food products. Inexpensive magnesium oxide has been used for the first time in HPLC as a very efficient catalyst and as a good alternative for, e.g., expensive anion-exchange resins. The reaction mechanism and kinetics were studied through measurement of the reaction band broadening, which depends on the stationary phase, mobile phase composition and flow-rate, and reaction temperature. Also, relationships between carbamate structure and reactivity were examined. The usefulness of the optimized postcolumn system was thoroughly tested by application to crop sample analysis during an extended period of time. The range of application of the solid-phase reaction was extended to 22 carbamate pesticides and ten of their metabolites. For crop sample analysis, limits of determination in the range 1-10 ppb were obtained.

Carbamates↗

Polyhydroxyalknoate synthesis in plants as a tool for biotechnology and basic studies of lipid metabolism.

Polyhydroxyalkanoates (PHAs) are polyesters of hydroxyacids naturally synthesized in bacteria as a carbon reserve. PHAs have properties of biodegradable thermoplastics and elastomers and their synthesis in crop plants is seen as an attractive system for the sustained production of large amounts of polymers at low cost. A variety of PHAs having different physical properties have now been synthesized in a number of transgenic plants, including Arabidopsis thaliana, rape and corn. This has been accomplished through the creation of novel metabolic pathways either in the cytoplasm, plastid or peroxisome of plant cells. Beyond its impact in biotechnology, PHA production in plants can also be used to study some fundamental aspects of plant metabolism. Synthesis of PHA can be used both as an indicator and a modulator of the carbon flux to pathways competing for common substrates, such as acetyl-coenzyme A in fatty acid biosynthesis or 3-hydroxyacyl-coenzyme A in fatty acid degradation. Synthesis of PHAs in plant peroxisome has been used to demonstrate changes in the flux of fatty acids to the beta-oxidation cycle in transgenic plants and mutants affected in lipid biosynthesis, as well as to study the pathway of degradation of unusual fatty acids.

Biotechnology↗

Structure-activity relationship (SAR) studies of benzoxazinones, their degradation products, and analogues. Phytotoxicity on problematic weeds Avena fatua L. and Lolium rigidum Gaud.

Avena fatua L. (wild oat) and Lolium rigidum Gaud. (rigid ryegrass) are highly problematic weeds affecting a wide variety of cereal crops worldwide. The fact that both of these weeds have developed resistance to several herbicide groups made them optimal candidates as target organisms for ongoing research about the potential application of allelochemicals and analogue compounds as natural herbicide models. Benzoxazinones, a family of natural allelochemicals present in corn, wheat, and rye, including 2,4-dihydroxy-(2H)-1,4-benzoxazin-3(4H)-one and 2,4-dihydroxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one, together with some degradation products, found in crop soils as well as in other systems, and some synthetic analogues of them were tested on wild oat and rigid ryegrass seeds; the results were statistically treated, and some structure-activity relationships, useful in further development of natural herbicide models, were elucidated. The most active compounds were the synthetic benzoxazinone 2-acetoxy-(2H)-1,4-benzoxazin-3(4H)-one and the degradation product 2-aminophenoxazin-3-one, with highly significant inhibition on the development of both weeds. The ecological role of these compounds is discussed by considering both degradability and phytotoxicity. The bioactivity of aminophenoxazines has been correlated by their aqueous solubility-lipophilicity predicted by means of computational methods.

Benzoxazines↗

Co-evolution and plant resistance to natural enemies.

Co-evolution between plants and their natural enemies is generally believed to have generated much of the Earth's biological diversity. A process analogous to co-evolution occurs in agricultural systems, in which natural enemies adapt to crop resistance introduced by breeding or genetic engineering. Because of this similarity, the investigation of resistance mechanisms in crops is helping to elucidate the workings of co-evolution in nature, while evolutionary principles, including those derived from investigation of co-evolution in nature, are being applied in the management of resistance in genetically engineered crops.

Animals↗

The Oryza bacterial artificial chromosome library resource: construction and analysis of 12 deep-coverage large-insert BAC libraries that represent the 10 genome types of the genus Oryza.

Rice (Oryza sativa L.) is the most important food crop in the world and a model system for plant biology. With the completion of a finished genome sequence we must now functionally characterize the rice genome by a variety of methods, including comparative genomic analysis between cereal species and within the genus Oryza. Oryza contains two cultivated and 22 wild species that represent 10 distinct genome types. The wild species contain an essentially untapped reservoir of agriculturally important genes that must be harnessed if we are to maintain a safe and secure food supply for the 21st century. As a first step to functionally characterize the rice genome from a comparative standpoint, we report the construction and analysis of a comprehensive set of 12 BAC libraries that represent the 10 genome types of Oryza. To estimate the number of clones required to generate 10 genome equivalent BAC libraries we determined the genome sizes of nine of the 12 species using flow cytometry. Each library represents a minimum of 10 genome equivalents, has an average insert size range between 123 and 161 kb, an average organellar content of 0.4%-4.1% and nonrecombinant content between 0% and 5%. Genome coverage was estimated mathematically and empirically by hybridization and extensive contig and BAC end sequence analysis. A preliminary analysis of BAC end sequences of clones from these libraries indicated that LTR retrotransposons are the predominant class of repeat elements in Oryza and a roughly linear relationship of these elements with genome size was observed.

Base Sequence↗

Coregionalized single- and multiresolution spatially varying growth curve modeling with application to weed growth.

Modeling of longitudinal data from agricultural experiments using growth curves helps understand conditions conducive or unconducive to crop growth. Recent advances in Geographical Information Systems (GIS) now allow geocoding of agricultural data that help understand spatial patterns. A particularly common problem is capturing spatial variation in growth patterns over the entire experimental domain. Statistical modeling in these settings can be challenging because agricultural designs are often spatially replicated, with arrays of subplots, and interest lies in capturing spatial variation at possibly different resolutions. In this article, we develop a framework for modeling spatially varying growth curves as Gaussian processes that capture associations at single and multiple resolutions. We provide Bayesian hierarchical models for this setting, where flexible parameterization enables spatial estimation and prediction of growth curves. We illustrate using data from weed growth experiments conducted in Waseca, Minnesota, that recorded growth of the weed Setaria spp. in a spatially replicated design.

Bayes Theorem↗

Influence of production, handling, and storage on phytonutrient content of foods.

The goals of agricultural production have traditionally been to try to accommodate needs for: 1) adequate and reliable yields to provide a sufficient food supply in a growing world; 2) food safety; 3) taste; 4) convenience; 5) profit; and 6) variety. Alternative strategies to enhance any of these outcomes are typically evaluated as to their probable effects on the key outcome: yield. However, with the burgeoning consumer interest in foods that optimize health, attention is shifting from concerns over quantity alone to concerns over the constituents of foods that may promote health, and thus to the agricultural practices that will protect, and perhaps enhance these constituents of the food supply. This shift in focus requires new thinking and new strategies across all segments of the food production system. This paper summarizes selected aspects of crop production that are pivotal to the nutrient value of foods for human consumption and suggests some strategies for establishing a new research and production paradigm that will embrace nutrient quality among the priorities of agricultural research.

Agriculture↗

Development of a highly efficient prime editing platform for cucurbits enables breeding of multi-disease-resistant cucumber.

The prime editing (PE) system is a precise genome editing technology that works efficiently in monocots; however, its application is limited by low editing efficiency in dicots, particularly Cucurbitaceae and Solanaceae plants. Here, we first significantly improved the transformation efficiency by introducing spectinomycin in cucurbits, then used the tomato elongation factor 1-alpha (SlEF1α) promoter to enhance PE protein expression, and incorporated the Csy4 ribonuclease to process pegRNAs, collectively addressing multiple constraints limiting PE efficiency in cucurbits. The optimized PE systems, particularly Csy4-PE6d, achieved an average desired editing frequency of 80.83% at targeted loci in cucumber via stable genetic transformation, with frequencies reaching up to 100% at certain sites. Moreover, Csy4-PE6d generated homozygous edits in 36.43% of transgenic lines and demonstrated robust editing activity in melon, pumpkin, and potato. Using the Csy4-PE6d tool, we generated heritable edited cucumber lines with dual resistance to bacterial angular leaf spot and downy mildew by targeting the CsSGR gene. Collectively, this optimized system substantially enhances PE efficiency in Cucurbit crops, providing an effective solution to common challenges such as low editing efficiency and limited heritability in these species.

Disease Resistance↗

Use of bromodeoxyuridine immunocapture to identify active bacteria associated with arbuscular mycorrhizal hyphae.

Arbuscular mycorrhizae are beneficial for crops grown under low-till management systems. Increasingly, it is becoming apparent that bacteria associated with mycorrhizae can enhance the beneficial relationship between mycorrhizae and plants. However, it has been difficult to study these relationships by conventional techniques. In this study actively growing bacteria were identified in soil from an undisturbed fallow field known to contain arbuscular mycorrhizae by using molecular tools to eliminate the need for cultivation. A thymidine analog, bromodeoxyuridine (BrdU), was added to the soil and incubated for 2 days. DNA was extracted, and the newly synthesized DNA was isolated by immunocapture of the BrdU-containing DNA. The active bacteria in the community were identified by 16S rRNA gene PCR amplification and DNA sequence analysis. Based on 16S rRNA gene sequence information, a selective medium was chosen to isolate the corresponding active bacteria. Bacillus cereus strain VA1, one of the bacteria identified by the BrdU method, was isolated from the soil and tagged with green fluorescent protein. By using confocal microscopy, this bacterium was shown to clearly attach to arbuscular mycorrhizal hyphae. This study was the first to use this combination of molecular and traditional approaches to isolate, identify, and visualize a specific bacterium that is active in fallow soil and associates with arbuscular mycorrhizal hyphae.

Antibodies, Monoclonal↗

An efficient method for the physical mapping of transgenes in barley using in situ hybridization.

The genetic transformation of crops by particle bombardment and Agrobacterium tumefaciens systems have the potential to complement conventional plant breeding programmes. However, before deployment, transgenic plants need to be characterized in detail, and physical mapping is an integral part of this process. Therefore, it is important to have a highly efficient method for transgene detection by fluorescence in situ hybridization (FISH). This study describes a new approach, which provides efficient control of probe length and labelling, both of which play an important role in in situ hybridization of transgenes. The approach is based on reducing the size of the plasmid prior to labelling by nick translation, rather than using the whole or linearized plasmid, or varying the amounts of DNaseI in the nick translation mixture. This provided much more efficient labelling of the probe, which yielded optimal hybridization. minimal fluorescent background, and accurate physical location of the transgene.

DNA Probes↗

Host genetic regulation of xylem-resident Pseudomonas enhances cucumber growth.

BACKGROUND: Although endophytic microorganisms play a critical role in plant growth and stress resilience, the genetic basis underlying host selection of beneficial microbiota-particularly within the xylem-remains poorly understood. Cucumber (Cucumis sativus), as a crop model with a well-developed system for studying vascular biology, offers a valuable system to investigate the host genetic determinants of xylem microbiome assembly. RESULTS: By conducting population-level microbiome profiling across 109 cucumber accessions, we identified a conserved xylem microbiota dominated by Proteobacteria. Within this community, 20 core amplicon sequence variants (ASVs) were consistently present in xylem sap. Genome-wide association mapping identified a host genetic locus, CsXPR1, which encodes a tetratricopeptide repeat protein that regulates the abundance of the dominant xylem-colonized Pseudomonas ASV_4. Colonization patterns of ASV_4 varied across host genotypes and were correlated with CsXPR1 expression levels, suggesting a precision genetic regulation of bacterial entry into vascular tissues. Pseudomonas fulva strain 220, with 97% 16S rRNA gene identity with ASV_4, could colonize in cucumber xylem by inoculation of either roots or leaves. Genome analysis and plate assays revealed the biosynthesis of indole-3-acetic acid (IAA), solubilization of phosphate, and a range of plant beneficial traits in strain 220. Inoculation with strain 220 significantly enhanced growth in cucumber, but only in CsXPR1 haplotype that exhibited high gene expression and higher recruitment capacity of the strain. These benefits included notable increases in plant height (38%), stem diameter (36%), leaf area (61%), fresh and dry weight (51% and 85%, respectively), and a 4.57-fold increase in 4-methyleneglutamine content within the xylem sap. CONCLUSION: Our findings reveal a complete "gene-to-function" pathway where the host gene CsXPR1 mediates a genotype-dependent growth promotion. It achieves this by regulating the xylem colonization of a beneficial bacterium, Pseudomonas fulva, which in turn enhances plant growth by enriching the xylem sap with the key metabolite 4-methyleneglutamine. Video Abstract.

Cucumis sativus↗

Work measurement for estimating food preparation time of a bioregenerative diet.

During space missions, such as the prospective Mars mission, crew labor time is a strictly limited resource. The diet for such a mission (based on crops grown in a bioregenerative life support system) will require astronauts to prepare their meals essentially from raw ingredients. Time spent on food processing and preparation is time lost for other purposes. Recipe design and diet planning for a space mission should therefore incorporate the time required to prepare the recipes as a critical factor. In this study, videotape analysis of an experienced chef was used to develop a database of recipe preparation time. The measurements were highly consistent among different measurement teams. Data analysis revealed a wide variation between the active times of different recipes, underscoring the need for optimization of diet planning. Potential uses of the database developed in this study are discussed and illustrated in this work.

Astronauts↗

Reversible male sterility: a novel system for the production of hybrid corn.

Hybrid corn seed is traditionally produced using either mechanical/hand detasseling or cytoplasmic male sterility, or a combination of both. In recent years, the development of transgenic systems to produce hybrid seed in several crops has attracted much attention. Here we describe a transgenic mechanism for production of hybrid corn, reversible male sterility (RMS), in which the action of the cytotoxic gene used to introduce male sterility is suppressed by the application of a chemical to the plant. Reversion of the sterility allows the RMS parent to be self-fertilized, a step which overcomes the need to remove fertile sib plants prior to making the hybrid cross. The key enabling technology in RMS is the use of a plant gene promoter which is specifically induced by chemical application. We have exemplified RMS in transgenic corn plants and believe that it provides specific benefits in the production of hybrid corn seed.

Hybridization, Genetic↗

Ozonation and alkaline-peroxide pretreatment of wheat straw for Cryptococcus curvatus fermentation.

Crop residues in an Advanced Life Support System (ALS) contain many valuable components that could be recovered and used. Wheat is 60% inedible, with approximately 90% of the total sugars in the residue cellulose and hemicellulose. To release these sugars requires pretreatment followed by enzymatic hydrolysis. Cryptococcus curvatus, an oleaginous yeast, uses the sugars in cellulose and hemicellulose for growth and production of storage triglycerides. In this investigation, alkaline-peroxide and ozonation pretreatment methods were compared for their efficiency to release glucose and xylose to be used in the cultivation of C. curvatus. Leaching the biomass with water at 65 degrees C for 4 h prior to pretreatment facilitated saccharification. Alkaline-peroxide and ozone pretreatment were almost 100% and 80% saccharification efficient, respectively. The sugars derived from the hydrolysis of alkaline-peroxide-treated wheat straw supported the growth of C. curvatus and the production of edible single-cell oil.

Biomass↗

Biotechnology-derived nutritious foods for developing countries: needs, opportunities, and barriers: discussion summary.

Improvements in diet diversification and quality can be facilitated by greater cooperation between the agricultural and the nutrition communities, according to an expert panel that met in early 2002. Encouraged to think innovatively, the panelists agreed that modern technology offers the potential to increase the amount and nutritional content of the food supply in developing countries, especially if the enhancements are made to the highest-yielding indigenous staple crops and if a total food-systems approach is taken. All types of interventions should be evaluated for their cost-effectiveness in preventing nutritional deficiencies in the developing world and for their sustainability.

Agriculture↗