Low-Cost Imaging to Quantify Germination Rate and Seedling Vigor across Lettuce Cultivars.

The survival and growth of young plants hinge on various factors, such as seed quality and environmental conditions. Assessing seedling potential/vigor for a robust crop yield is crucial but often resource-intensive. This study explores cost-effective imaging techniques for rapid evaluation of seedling vigor, offering a practical solution to a common problem in agricultural research. In the first phase, nine lettuce (Lactuca sativa) cultivars were sown in trays and monitored using chlorophyll fluorescence imaging thrice weekly for two weeks. The second phase involved integrating embedded computers equipped with cameras for phenotyping. These systems captured and analyzed images four times daily, covering the entire growth cycle from seeding to harvest for four specific cultivars. All resulting data were promptly uploaded to the cloud, allowing for remote access and providing real-time information on plant performance. Results consistently showed the 'Muir' cultivar to have a larger canopy size and better germination, though 'Sparx' and 'Crispino' surpassed it in final dry weight. A non-linear model accurately predicted lettuce plant weight using seedling canopy size in the first study. The second study improved prediction accuracy with a sigmoidal growth curve from multiple harvests (R2 = 0.88, RMSE = 0.27, p < 0.001). Utilizing embedded computers in controlled environments offers efficient plant monitoring, provided there is a uniform canopy structure and minimal plant overlap.

Automated Imaging to Evaluate the Exogenous Gibberellin (Ga3) Impact on Seedlings from Salt-Stressed Lettuce Seeds.

Salinity stress is a common challenge in plant growth, impacting seed quality, germination, and general plant health. Sodium chloride (NaCl) ions disrupt membranes, causing ion leakage and reducing seed viability. Gibberellic acid (GA3) treatments have been found to promote germination and mitigate salinity stress on germination and plant growth. 'Bauer' and 'Muir' lettuce (Lactuca sativa) seeds were soaked in distilled water (control), 100 mM NaCl, 100 mM NaCl + 50 mg/L GA3, and 100 mM NaCl + 150 mg/L GA3 in Petri dishes and kept in a dark growth chamber at 25 °C for 24 h. After germination, seedlings were monitored using embedded cameras, capturing red, green, and blue (RGB) images from seeding to final harvest. Despite consistent germination rates, 'Bauer' seeds treated with NaCl showed reduced germination. Surprisingly, the 'Muir' cultivar's final dry weight differed across treatments, with the NaCl and high GA3 concentration combination yielding the poorest results (p < 0.05). This study highlights the efficacy of GA3 applications in improving germination rates. However, at elevated concentrations, it induced excessive hypocotyl elongation and pale seedlings, posing challenges for two-dimensional imaging. Nonetheless, a sigmoidal regression model using projected canopy size accurately predicted dry weight across growth stages and cultivars, emphasizing its reliability despite treatment variations (R2 = 0.96, RMSE = 0.11, p < 0.001).

The Use of Imaging to Quantify the Impact of Seed Aging on Lettuce Seed Germination and Seedling Vigor.

The decline in seed quality over time due to natural aging or mishandling requires assessing seed vigor for resilience in adverse conditions. Accelerated aging (AA) methods simulate seed deterioration by subjecting seeds to high temperatures and humidity. Saturated salt accelerated aging (SSAA) is an AA method adopted for small seeds like lettuce (Lactuca sativa). In this study, we subjected seeds of two lettuce cultivars ('Muir' and 'Bauer') to SSAA by sealing them in a box containing 40 g/100 mL of a sodium chloride (NaCl) solution in a dark growth chamber at 41 °C for 24, 48, and 72 h with a control. We monitored their vigor using embedded computer cameras, tracking the projected canopy size (PCS) daily from sowing to harvest. The cultivar 'Muir' exhibited consistent PCS values across the treatments, while 'Bauer' showed PCS variations, with notable declines after prolonged aging. The germination rates dropped significantly after 48 and 72 h of SSAA. A nonlinear regression model revealed a strong relationship between PCS and shoot dry weight across harvests and cultivars (R2 = 0.93, RMSE = 0.15, p < 0.001). The research found that the projected canopy size and shoot dry weight increased over time with significant differences in treatments for the cultivar 'Bauer' but not for 'Muir,' with the canopy size being a strong predictor of dry weight and no significant impact from the SSAA treatments. This study highlights cultivar-specific responses to aging and demonstrates the efficacy of our imaging tool in predicting lettuce dry weight despite treatment variations. Understanding how aging affects different lettuce varieties is crucial for seed management and crop sustainability.

CsiLAC4 modulates boron flow in Arabidopsis and Citrus via high-boron-dependent lignification of cell walls.

The mechanisms underlying plant tolerance to boron (B) excess are far from fully understood. Here we characterized the role of the miR397-CsiLAC4/CsiLAC17 (from Citrus sinensis) module in regulation of B flow. Live-cell imaging techniques were used in localization studies. A tobacco transient expression system tested modulations of CsiLAC4 and CsiLAC17 by miR397. Transgenic Arabidopsis were generated to analyze the biological functions of CsiLAC4 and CsiLAC17. CsiLAC4's role in xylem lignification was determined by mRNA hybridization and cytochemistry. In situ B distribution was analyzed by laser ablation inductively coupled plasma mass spectrometry. CsiLAC4 and CsiLAC17 are predominantly localized in the apoplast of tobacco epidermal cells. Overexpression of CsiLAC4 in Arabidopsis improves the plants' tolerance to boric acid excess by triggering high-B-dependent lignification of the vascular system's cell wall and reducing free B content in roots and shoots. In Citrus, CsiLAC4 is expressed explicitly in the xylem parenchyma and is modulated by B-responsive miR397. Upregulation of CsiLAC4 in Citrus results in lignification of the xylem cell walls, restricting B flow from xylem vessels to the phloem. CsiLAC4 contributes to plant tolerance to boric acid excess via high-B-dependent lignification of cell walls, which set up a 'physical barrier' preventing B flow.

The Energy Requirement for Supplemental Greenhouse Lighting Can Be Reduced by Considering 'Excess' Light from the Previous Day.

The sunlight greenhouse crops receive varies and is often insufficient for consistent year-round growth in greenhouses. Supplemental lighting is commonly applied in winter, but this practice has a significant energy cost, accounting for 10-30% of operating expenses and impacting greenhouse profitability. Greenhouse lights are traditionally adjusted based on sunlight intensity to meet crops' daily light requirements. However, if plants can withstand lower daily light integrals (DLI) after a sunny day without reducing the growth, there is potential to reduce the energy required for supplemental lighting and increase the profit. To determine whether excess light received one day can be 'carried over' to the next, we grew oakleaf lettuce (Lactuca sativa 'Green Salad Bowl' and 'Red Salad Bowl') under six lighting regimes inside a vertical farm. Plants in all treatments received an average DLI of 15 mol·m-2·d-1, but DLIs alternated from day-to-day (15/15, 17.5/12.5, 20/10, 22.5/7.5, 25/5, and 27.5/2.5 mol·m-2·d-1), resulting in DLI fluctuations from 0 to 25 mol·m-2·d-1. Plants had similar leaf area (~800 cm2/plant) and dry weight (~1.8 g/plant) when grown with DLI fluctuations from 0 to 15 mol·m-2·d-1, while higher DLI fluctuation reduced growth. To confirm this DLI "carrying-over" effect on plants grown under sunlight with supplemental light, we conducted a second study in a greenhouse with 'Green Salad Bowl' lettuce. In this study, plants were grown with five different DLI fluctuations (15/15, 16.75/13.25, 18.5/11.5, 20.25/9.75, and 22/8 mol·m-2·d-1), ranging from 0 to 14 mol·m-2·d-1, while maintaining an average DLI of 15 mol·m-2·d-1 in all the treatments. We observed similar leaf area (~750 cm2/plant) and dry weight (~1.8 g/plant) in lettuce plants grown with DLI fluctuations from 0 to 10.5 mol·m-2·d-1. Higher DLI fluctuations reduced growth. Hence, carrying excess light from a sunny to an overcast day is possible within limits. Our study concluded that the DLI requirement can be reduced by approximately 5.25 mol·m-2·d-1 on the day following a sunny day. By analyzing historical weather data from five US locations, we quantified the potential annual energy savings from incorporating this 'carrying-over DLI' concept. This approach resulted in annual energy savings of approximately 75-190 MWh/ha in greenhouse lettuce production. Such reductions in supplemental lighting energy will enhance the profitability and sustainability of the greenhouse industry.

Effect of Huanglongbing on the Volatile Organic Compound Profile of Fruit Juice and Peel Oil in 'Ray Ruby' Grapefruit.

Along with orange and mandarin, grapefruit production in Florida has declined sharply due to Huanglongbing (HLB), or citrus greening disease, caused by Candidatus Liberibacter asiaticus (CLas). HLB affects the volatile profiles of juice and peel oil in oranges, but there is limited information on grapefruit. In this research, 'Ray Ruby' grapefruit were harvested in 2020 and 2021 from healthy (HLB-) and HLB-affected (HLB+) trees. Peel oil was extracted by hydrodistillation, and the volatiles were analyzed by direct injection of the oil samples into gas chromatography-mass spectrometry (GC-MS). Volatiles in the juice were analyzed by headspace (HS)-solid-phase microextraction (SPME) coupled with GC-MS. HLB significantly altered the volatile profiles of peel oil and juice in 'Ray Ruby' grapefruit. Juice samples of HLB+ fruits had lower decanal, nonanal, and octanal, important citrus juice flavor compounds. HLB+ samples also showed reduced content of nonterpene compounds, other aliphatic and terpene aldehydes, and terpene ketones. Ethanol, acetaldehyde, ethyl acetate, and ethyl butanoate were increased in HLB+ juice samples, indicating an HLB-induced stress response. The most abundant compounds D-limonene and ß-caryophyllene, as well as other sesquiterpenes, were increased in HLB+ juice and peel oil samples. On the other hand, the oxidative/dehydrogenated terpenes were increased by HLB in peel oil but decreased in the juice sample. Nootkatone, the key grapefruit volatile was consistently reduced by HLB in both peel oil and juice samples. The impact of HLB on nootkatone deteriorated the quality of both juice and peel oil in grapefruits.

Grapefruit Root and Rhizosphere Responses to Varying Planting Densities, Fertilizer Concentrations and Application Methods.

Huanglongbing (HLB) disease has caused a severe decline in citrus production globally over the past decade. There is a need for improved nutrient regimens to better manage the productivity of HLB-affected trees, as current guidelines are based on healthy trees. The aim of this study was to evaluate the effects of different fertilizer application methods and rates with different planting densities on HLB-affected citrus root and soil health. Plant material consisted of 'Ray Ruby' (Citrus × paradisi) grapefruit trees grafted on 'Kuharske' citrange (Citrus × sinensis × Citrus trifoliata). The study consisted of 4 foliar fertilizer treatments, which included 0×, 1.5×, 3× and 6× the University of Florida Institute of Food and Agriculture (UF/IFAS) recommended guidelines for B, Mn and Zn. Additionally, 2 ground-applied fertilizer treatments were used, specifically controlled-release fertilizer (CRF1): 12-3-14 + B, Fe, Mn and Zn micronutrients at 1× UF/IFAS recommendation, and (CRF2): 12-3-14 + 2× Mg + 3× B, Fe, Mn and Zn micronutrients, with micronutrients applied as sulfur-coated products. The planting densities implemented were low (300 trees ha-1), medium (440 trees ha-1) and high (975 trees ha-1). The CRF fertilizer resulted in greater soil nutrient concentrations through all of the time sampling points, with significant differences in soil Zn and Mn. Grapefruit treated with ground-applied CRF2 and 3× foliar fertilizers resulted in the greatest bacterial alpha and beta diversity in the rhizosphere. Significantly greater abundances of Rhizobiales and Vicinamibacterales were found in the grapefruit rhizosphere of trees treated with 0× UF/IFAS foliar fertilizer compared to higher doses of foliar fertilizers.

Substrate pH Influences the Nutrient Absorption and Rhizosphere Microbiome of Huanglongbing-Affected Grapefruit Plants.

The substrate pH directly affects nutrient availability in the rhizosphere and nutrient uptake by plants. Macronutrients such as nitrogen, potassium, calcium, magnesium, and sulfur are highly available at pH 6.0-6.5, while micronutrients become less available at higher, alkaline pH (pH > 7.0). Recent research has indicated that low pHs can enhance nutrient uptake and improve sweet orange (Citrus sinensis) tree health. We designed a study to understand the influence of a wide range of substrate pH values on plant size and biomass, nutrient availability, leaf gas exchange, and rhizosphere microbiome of grapefruit (Citrus paradisi) affected by Huanglongbing (HLB). Two-year-old "Ray Ruby" grapefruit plants grafted on sour orange (Citrus aurantium) rootstock were cultivated indoors in 10-cm wide × 40-cm tall pots with peat:perlite commercial substrate (80:20 v/v). We tested two disease statuses [HLB-free or healthy (negative, HLB-) and HLB-affected (positive, HLB+)] and six substrate pH values (4, 5, 6, 7, 8, 9) in a 2 × 6 factorial arranged on a complete randomized design with four replications. The canopy volume of HLB+ plants was 20% lower than healthy plants, with pHs 7 and 9 resulting in 44% less canopy volume. The root and shoot ratio of dry weight was 25.8% lower in HLB+ than in healthy plants. Poor root growth and a decrease in fibrous roots were found, especially in pH 5 and 6 treatments in HLB+ plants (p < 0.0001). The disease status and the substrate pHs influenced the leaf nutrient concentration (p < 0.05). High substrate pH affects nutrient availability for root uptake, influencing the nutrient balance throughout the plant system. pH values did not affect plant photosynthesis, indicating that pH does not recover HLB+ plants to the photosynthetic levels of healthy plants-even though high pH positively influenced internal CO2. There were collectively over 200 rhizobacterial identified by the 16S rRNA gene sequencing in individual phylogenetic trees. Most rhizobacteria reads were identified in pH 9. Our results indicated no effect of substrate pHs on the plant disease status induced by enhanced nutrient uptake.