Minimizing Escherichia coli O157:H7 contamination in indoor farming: effects of cultivar type and ultra-violet light quality.

BACKGROUND: Bacterial contamination of produce is a concern in indoor farming due to close plant spacing, recycling irrigation, warm temperatures, and high relative humidity during production. Cultivars that inherently resist contamination and photo-sanitization using ultraviolet (UV) radiation during the production phase can reduce bacterial contamination. However, there is limited information to support their use in indoor farming. RESULTS: Lettuce (Lactuca sativa) cultivars with varying plant architectures grown in a custom-built indoor farm exhibited differences in E. coli O157:H7 survival after inoculation. The survival of E. coli O157:H7 was lowest in the leaf cultivar (open architecture) and highest in the romaine and oakleaf cultivars (compact architecture). Of the different UV wavelengths that were tested (UV-A, UV-A + B, UV-A + C), UV A + C at an intensity of 54.5 µmol m-2 s-1 (with 3.5 µmol m-2 s-1 of UV-C), provided for 15 min every day, was found to be most efficacious in reducing the E. coli O157:H7 survival on romaine lettuce with no negative effects on plant growth and quality. CONCLUSION: Contamination of E. coli O157:H7 on lettuce plants can be reduced and the food safety levels in indoor farms can be increased by selecting cultivars with an open leaf architecture coupled with photo-sanitization using low and frequent exposure to UV A + C radiation. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Connecting the dots: Path model to identify key phenotypic traits for screening plants with tolerance to nitrogen deficiency.

Varieties that tolerate low nitrogen (N) application rates can reduce fertilizer costs, minimize nitrate leaching and runoff losses, and lower overall CO2 emissions associated with fertilizer manufacturing. The goal of our research is to show the usefulness of path models to identify key phenotypic traits for screening plants with a tolerance to low N application rates. We grew tolerant and sensitive cultivars of poinsettia (Euphorbia pulcherrima) using a water-soluble fertilizer (15-5-15 Cal Mag) in both optimal (electrical conductivity of 2.5 dS·m-1) and N-deficient (electrical conductivity of 0.75 dS·m-1) treatments and measured 24 different traits at the cellular, leaf, and whole-plant scales in both cultivars and treatments. The experiment was laid out as a split-plot design with N treatments as main plots and cultivars as sub-plots, with five replications. Path analysis was conducted to develop sequential relationships among these traits. Statistical comparisons between tolerant and sensitive cultivars in the N-deficient treatment indicated an increase in shoot biomass (19.9 vs 14.4 g), leaf area (2775 vs 1824 cm2), leaf dry weight (14.7 vs 10.0 g), lateral root dry weight (3.7 vs 2.4 g), light-saturated photosynthesis (14.5 vs 10.1 µmol∙m-2∙s-1), maximum electron transport rate (119 vs 89 µmol∙m-2∙s-1), chlorophyll content (28.1 vs 12.9 g∙100g-1), leaf N content (27.5 vs 19.9 mg∙g-1), and fine root N content (26.1 vs 20.9 mg∙g-1), and a decrease in anthocyanin content (0.07 vs 0.16 ΔOD∙g-1). The path model indicated that an increase in the lateral root growth and fine root N content can lead to an increase in the leaf N content, in the N-deficient treatment. There were three separate paths that connected higher leaf N content to increased shoot biomass. These paths were mediated by the levels of anthocyanin, chlorophylls, and light-saturated photosynthesis rate (or rubisco capacity). The light-saturated photosynthesis model suggested that the increased uptake of N by fine roots in the tolerant cultivar was likely supported by the photosynthates translocated from the shoot to the root. Leaf N content was associated with multiple plant responses in the N-deficient treatment, and can be a useful screening trait for developing new cultivars, especially in marker-assisted molecular breeding.

Fixed vs. variable light quality in vertical farming: Impacts on vegetative growth and nutritional quality of lettuce.

Lettuce (Lactuca sativa) is commonly produced in vertical farms. The levels of nutritionally important phytochemicals such as beta-carotene (precursor to vitamin A) are generally low in lettuce. In this study, we investigated the benefits of variable lighting strategy (i.e., varying the light quality during production) on maintaining plant growth and increasing the biosynthesis of beta-carotene and anthocyanin. We tested two variable lighting methods, using green and red romaine lettuce, namely (i) providing growth lighting (supports vegetative growth) initially (21 days) followed by a high percentage of blue light (supports biosynthesis of phytochemicals) at final stages (10 days) and (ii) providing a high percentage of blue light initially followed by growth lighting at final stages. Our results indicate that the variable lighting method with initial growth lighting and high percentage of blue at final stages can maintain vegetative growth and enhance phytochemicals such as beta-carotene in green romaine lettuce while both variable lighting methods were not effective in red romaine lettuce. In green romaine lettuce, we did not observe a significant reduction in shoot dry weight but there was an increase in beta-carotene (35.7%) in the variable compared to the fixed lighting method with growth lighting for the entire duration. The physiological bases for differences in vegetative growth and synthesis of beta-carotene and anthocyanin in the variable and fixed lighting methods are discussed.

Leveraging high-throughput hyperspectral imaging technology to detect cadmium stress in two leafy green crops and accelerate soil remediation efforts.

Cadmium (Cd) is a toxic metal that can accumulate in soils and negatively impact crop as well as human health. Amendments like biochar have potential to address these challenges by reducing Cd bioavailability in soil, though reliance on post-harvest wet chemical methods to quantify Cd uptake have slowed efforts to identify the most effective amendments. Hyperspectral imaging (HSI) is a novel technology that could overcome this limitation by quantifying symptoms of Cd stress while plants are still growing. The goals of this study were to: 1) determine whether HSI can detect Cd stress in two distinct leafy green crops, 2) quantify whether a locally sourced biochar derived from hardwoods can reduce Cd stress and uptake in these crops, and 3) identify vegetative indices (VIs) that best quantify changes in plant stress responses. Experiments were conducted in a tightly controlled automated phenotyping facility that allowed all environmental factors to be kept constant except Cd concentration (0, 5 10 and 15 mg kg-1). Symptoms of Cd stress were stronger in basil (Ocimum basilicum) than kale (Brassica oleracea), and were easier to detect using HSI. Several VIs detected Cd stress in basil, but only the anthocyanin reflectance index (ARI) detected all levels of Cd stress in both crop species. The biochar amendment did reduce Cd uptake, especially at low Cd concentrations in kale which took up more Cd than basil. Again, the ARI index was the most effective in quantifying changes in plant stress mediated by the biochar. These results indicate that the biochar evaluated in this study has potential to reduce Cd bioavailability in soil, and HSI could be further developed to identify rates that can best achieve this benefit. The technology also may be helping in elucidating mechanisms mediating how biochar can influence plant growth and stress responses.

Blue and Far-Red Light Affect Area and Number of Individual Leaves to Influence Vegetative Growth and Pigment Synthesis in Lettuce.

Published work indicates that high percentage of blue light can enhance pigment levels but decreases growth, while addition of far-red light to growth light can increase quantum efficiency and photosynthesis in leafy greens. Combining high-energy blue light with low-energy far-red light may increase both vegetative growth and pigment levels. However, the effect of high-energy blue and low-energy far-red light on the vegetative growth and pigments synthesis is unclear. This information can be potentially useful for enhancing the levels of pigments with nutritional value (e.g., beta-carotene and anthocyanins) in the produce grown in vertical farms. We grew romaine lettuce (cv. Amadeus) under similar light intensity (approximately 130 µmol⋅m-2⋅s-1) but different proportions of red: blue: far-red including 90:10: 0 ("High-R"), 50: 50: 0 ("High-B"), and 42: 42: 16 ("High-B+FR") for 31 days. Results indicated that canopy area and leaf photosynthetic rate of lettuce plants was reduced in the High-B, thereby reducing plant growth. We did not observe photosynthesis enhancement in the High-B+FR. Instead, plants clearly showed photomorphogenic effects. The phytochrome photostationary state (PSS) decreased with far-red addition, resulting in reduced leaf number per plant. This was likely to shift the allocation of resources toward elongation growth for shade avoidance. Further, we observed an increase in the area of individual leaves, canopy area, and shoot dry weight in the High-B+FR. However, these appear to be an indirect consequence of decreased leaf number per plant. Our results also indicate that changes in expansion growth at individual leaf scale largely regulated pigment concentration in plants. As individual leaf area became smaller (e.g., High-B) or larger (e.g., High-B+FR), the levels of pigments including chlorophylls and beta-carotene increased or decreased, respectively. Area of individual leaves also positively influenced canopy area (and likely light interception) and shoots dry weight (or vegetative growth). Our study provides additional insights into the effects of high-energy blue and low-energy far-red light on individual leaf number and leaf growth, which appear to control plant growth and pigment levels in lettuce.

Growth and photosynthetic responses of Chinese cabbage (Brassica rapa L. cv. Tokyo Bekana) to continuously elevated carbon dioxide in a simulated Space Station "Veggie" crop-production environment.

Among candidate leafy vegetable species initially considered for astronauts to pick and eat from the Veggie plant-growth unit on the International Space Station (ISS), Chinese cabbage (Brassica rapa L. cv. Tokyo Bekana) ranked high in ground-based screening studies. However, subsequent attempts to optimize growth within rigorous ISS-like growth environments on the ground were frustrated by development of leaf chlorosis, necrosis, and uneven growth. 'Tokyo Bekana' ('TB') grown on ISS during the VEG-03B and C flights developed similar stress symptoms. After lengthy troubleshooting efforts to identify causes of sub-par growth in highly controlled environments, the super-elevated CO2 concentrations that plants on ISS are exposed to continuously (average of 2,800 µmol/mol) emerged as a candidate environmental condition responsible for the observed plant-stress symptoms. Subsequent ground-based studies found continuous exposure to ISS levels of CO2 under Veggie environmental and cultural conditions to significantly inhibit growth of 'TB' compared to near-Earth-normal CO2 controls. The present study investigated growth and gas-exchange responses of 'TB' to sub-ISS but still elevated CO2 levels (900 or 1,350 µmol/mol) in combination with other potential stressors related to ISS/Veggie compared to 450 µmol/mol CO2 controls. Shoot dry mass of plants grown at 450 µmol•mol-1 CO2 for 28 days was 96% and 80% higher than that of plants grown at 900 µmol•mol-1 CO2 and 1,350 µmol•mol-1 CO2, respectively. Leaf number and leaf area of controls were significantly higher than those of plants grown at 1,350 µmol•mol-1 CO2. Photosynthetic rate measured using a leaf cuvette was significantly lower for plants grown at 900 µmol•mol-1 CO2 than for controls. The ratio of leaf internal CO2 concentration (Ci) to cuvette ambient CO2 concentration (Ca) was significantly lower for plants grown at 450 µmol•mol-1 CO2 than for plants grown at elevated CO2. Thus, continuously elevated CO2 in combination with a Veggie cultivation system decreased growth, leaf area, and photosynthetic efficiency of Chinese cabbage 'Tokyo Bekana'. The results of this study suggest that 'Tokyo Bekana' is very sensitive to continuously elevated CO2 in such a growth environment, and indicate the need for improved environmental control of CO2 and possibly root-zone factors for successful crop production in the ISS spaceflight environment. Differential sensitivity of other salad crops to an ISS/Veggie growth environment also is possible, so it is important to mimic controllable ISS-like environmental conditions as precisely as possible during ground-based screening.

Recycling Nutrient Solution Can Reduce Growth Due to Nutrient Deficiencies in Hydroponic Production.

It is common in hydroponics to supply nutrients to crops by maintaining electrical conductivity (EC) of the recycling solution at a target level. Levels of individual nutrients in the solution are generally not assessed as their regular measurement and adjustment can be both expensive and technically challenging. However, the approach of growing crops at a target EC can potentially result in nutrient imbalances in the solution and reduced growth. We quantified the effects of recycling on solution EC changes, tissue nutrient concentration, canopy growth rate, plant water status, and shoot and root weight of lettuce (Lactuca sativa) in a greenhouse. The tap water quality was moderately alkaline and similar to that commonly observed in many commercial greenhouses. In our research, recycling solution maintained at a target EC (1.8 dS⋅m-1) significantly reduced shoot fresh (22-36%) and dry weight compared to the control supplied regularly with freshly prepared solution at the target EC. Further, recycling significantly decreased N, P, K, and Fe and increased Na and Cu levels in the tissue, in addition to increasing solution EC between adjustments compared to the control. Using image analysis of groups of plants, we identified that the negative effects of recycling on canopy area started 2 weeks after transplanting. Based on these results, we hypothesized that certain unwanted compounds (e.g., bicarbonates) and slowly consumed elements (e.g., Ca, Mg) were added to the recycling solution through the alkaline tap water with time. Their accumulation "artificially" increased solution EC and "masked" the lower than optimal levels of major nutrients in the solution, leading to the reductions in the concentration of nutrients in the tissue and plant growth. Supporting this, the negative effects of recycling were not observed when the recycling solution was either discarded after 2 weeks of use or made using reverse osmosis water and continuously used. Our findings aid in proper management of recycling solution in hydroponic lettuce production.

Physiological responses related to increased grain yield under drought in the first biotechnology-derived drought-tolerant maize.

Maize (Zea mays ssp. mays L.) is highly susceptible to drought stress. This work focused on whole-plant physiological mechanisms by which a biotechnology-derived maize event expressing bacterial cold shock protein B (CspB), MON 87460, increased grain yield under drought. Plants of MON 87460 and a conventional control (hereafter 'control') were tested in the field under well-watered (WW) and water-limited (WL) treatments imposed during mid-vegetative to mid-reproductive stages during 2009-2011. Across years, average grain yield increased by 6% in MON 87460 compared with control under WL conditions. This was associated with higher soil water content at 0.5 m depth during the treatment phase, increased ear growth, decreased leaf area, leaf dry weight and sap flow rate during silking, increased kernel number and harvest index in MON 87460 than the control. No consistent differences were observed under WW conditions. This indicates that MON 87460 acclimated better under WL conditions than the control by lowering leaf growth which decreased water use during silking, thereby eliciting lower stress under WL conditions. These physiological responses in MON 87460 under WL conditions resulted in increased ear growth during silking, which subsequently increased the kernel number, harvest index and grain yield compared to the control.

The physiological basis for genetic variation in water use efficiency and carbon isotope composition in Arabidopsis thaliana.

Ecologists and physiologists have documented extensive variation in water use efficiency (WUE) in Arabidopsis thaliana, as well as association of WUE with climatic variation. Here, we demonstrate correlations of whole-plant transpiration efficiency and carbon isotope composition (δ(13)C) among life history classes of A. thaliana. We also use a whole-plant cuvette to examine patterns of co-variation in component traits of WUE and δ(13)C. We find that stomatal conductance (g s) explains more variation in WUE than does A. Overall, there was a strong genetic correlation between A and g s, consistent with selection acting on the ratio of these traits. At a more detailed level, genetic variation in A was due to underlying variation in both maximal rate of carboxylation (V cmax) and maximum electron transport rate (Jmax). We also found strong effects of leaf anatomy, where lines with lower WUE had higher leaf water content (LWC) and specific leaf area (SLA), suggesting a role for mesophyll conductance (g m) in variation of WUE. We hypothesize that this is due to an effect through g m, and test this hypothesis using the abi4 mutant. We show that mutants of ABI4 have higher SLA, LWC, and g m than wild-type, consistent with variation in leaf anatomy causing variation in g m and δ(13)C. These functional data also add further support to the central, integrative role of ABI4 in simultaneously altering ABA sensitivity, sugar signaling, and CO2 assimilation. Together our results highlight the need for a more holistic approach in functional studies, both for more accurate annotation of gene function and to understand co-limitations to plant growth and productivity.

Genetics of drought adaptation in Arabidopsis thaliana II. QTL analysis of a new mapping population, KAS-1 x TSU-1.

Despite compelling evidence that adaptation to local climate is common in plant populations, little is known about the evolutionary genetics of traits that contribute to climatic adaptation. A screen of natural accessions of Arabidopsis thaliana revealed Tsu-1 and Kas-1 to be opposite extremes for water-use efficiency and climate at collection sites for these accessions differs greatly. To provide a tool to understand the genetic basis of this putative adaptation, Kas-1 and Tsu-1 were reciprocally crossed to create a new mapping population. Analysis of F(3) families showed segregating variation in both delta(13)C and transpiration rate, and as expected these traits had a negative genetic correlation (r(g)=- 0.3). 346 RILs, 148 with Kas-1 cytoplasm and 198 with Tsu-1 cytoplasm, were advanced to the F(9) and genotyped using 48 microsatellites and 55 SNPs for a total of 103 markers. This mapping population was used for QTL analysis of delta(13)C using F(9) RIL means. Analysis of this reciprocal cross showed a large effect of cytoplasmic background, as well as two QTL for delta(13)C. The Kas-1 x Tsu-1 mapping population provides a powerful new resource for mapping QTL underlying natural variation and for dissecting the genetic basis of water-use efficiency differences.