Using Mathematical Optimization Models to Improve the Efficiency of Duckweeds (Wolffia arrhiza and Lemna minor) Micropropagation.

The method of plant micropropagation is widely used to obtain genetically homogeneous and infection-free plants for the needs of various industries and agriculture. Optimization of plant growth and development conditions plays a key role in economically successful micropropagation. Computer technologies have provided researchers with new approaches for modeling and a better understanding of the role of the factors involved in plant growth in vitro. To develop new models for optimizing growth conditions, we used plants with a high speed of vegetative in vitro reproduction, such as duckweed (Wolffia arrhiza and Lemna minor). Using the development of the optimal modeling of the biological processes, we have obtained the prescriptions for an individually balanced culture medium that enabled us to obtain 1.5-2.0 times more duckweed biomass with a 1.5 times higher protein concentration in the dry mass. Thus, we have demonstrated that the method of optimization modeling of the biological processes based on solving multinomial tasks from the series of quadratic equations can be used for the optimization of trophic needs of plants, specifically for micropropagation of duckweeds in vitro.

Removal of contaminants of emerging concern by Wolffia arrhiza and Lemna minor depending on the process conditions, pollutants concentration, and matrix type.

Research was carried out on the removal of a group of six contaminants of emerging concern: bisphenol A, N,N-diethyl-m-toluamide, diethylstilbestrol, triclosan, estrone and estradiol from the water matrix during contact with small floating macrophytes Wolffia arrhiza and Lemna minor. The optimal conditions for the process, such as pH, light exposure per day, and plant mass, were determined using the design of experiments chemometric approach based on central composite design. Experiments conducted under the designated optimal conditions showed that after 7 days, the removal efficiency equals 88-98% in the case of W. arrhiza and 87-97% in the case of L. minor, while after 14 days of the experiment, these values are 93-99.6% and 89-98%, respectively. The primary mechanism responsible for removing CECs is the plant uptake, with the mean uptake rate constant equal to 0.299 day-1 and 0.277 day-1 for W. arrhiza and L. minor, respectively. Experiments conducted using municipal wastewater as a sample matrix showed that the treatment efficiency remains high (the average values 84% and 75%; in the case of raw wastewater, 93% and 89%, and in the case of treated wastewater, for W. arrhiza and L. minor, respectively). Landfill leachate significantly reduces plants' ability to remove pollutants (the average removal efficiency equals 59% and 56%, for W. arrhiza and L. minor, respectively).

Chiral Herbicide 2,4-D Ethylhexyl Ester: Absolute Configuration, Stereoselective Herbicidal Activity, Crop Safety, and Metabolic Behavior on Maize and Flixweed.

This study represents the initial examination of the herbicidal efficacy, crop safety, and degradation patterns of 2,4-D ethylhexyl ester (2,4-D EHE) at the enantiomeric level. Baseline separation of 2,4-D EHE enantiomers was achieved using a superchiral R-AD column, with their absolute configurations determined through chemical reaction techniques. Evaluation of weed control efficacy against sensitive species such as sun spurge and flixweed demonstrated significantly higher inhibition rates for S-2,4-D EHE compared to R-2,4-D EHE. Conversely, no stereoselectivity was observed in the fresh-weight inhibition rates of both enantiomers on crops or nonsensitive weeds. A sensitive HPLC-MS/MS method was developed to simultaneously detect two enantiomers and the metabolite 2,4-D in plants. Investigation into degradation kinetics revealed no substantial difference in the half-lives of R- and S-2,4-D EHE in maize and flixweed. Notably, the metabolite 2,4-D exhibited prolonged persistence at elevated levels on flixweed, while it degraded rapidly on maize.

Duckweed: Beyond an Efficient Plant Model System.

Duckweed (Lemnaceae) rises as a crucial model system due to its unique characteristics and wide-ranging utility. The significance of physiological research and phytoremediation highlights the intricate potential of duckweed in the current era of plant biology. Special attention to duckweed has been brought due to its distinctive features of nutrient uptake, ion transport dynamics, detoxification, intricate signaling, and stress tolerance. In addition, duckweed can alleviate environmental pollutants and enhance sustainability by participating in bioremediation processes and wastewater treatment. Furthermore, insights into the genomic complexity of Lemnaceae species and the flourishing field of transgenic development highlight the opportunities for genetic manipulation and biotechnological innovations. Novel methods for the germplasm conservation of duckweed can be adopted to preserve genetic diversity for future research endeavors and breeding programs. This review centers around prospects in duckweed research promoting interdisciplinary collaborations and technological advancements to drive its full potential as a model organism.

Sulfamethoxazole removal and fuel-feedstock biomass production from wastewater in a phyto-Fenton process using duckweed culture.

The phyto-Fenton process, which generates hydroxyl radicals through Fenton and Fenton-like reactions using plant-derived hydrogen peroxide (H2O2) and ferrous iron (Fe (II)) can degrade organic pollutants. Duckweed, an aquatic plant, is promising for a co-beneficial phytoremediation process that combines wastewater treatment and biomass production for biofuel feedstock. However, the phyto-Fenton process using duckweed has not been extensively studied. Because sulfamethoxazole (SMX), a major antibiotic, is distributed widely and is an emerging contaminant, its effective removal from contaminated water is necessary. The present study investigated the possibility of the simultaneous efficient removal of SMX from polluted water and biomass production for fuel feedstock by the phyto-Fenton process using duckweed. This is the first attempt to demonstrate the co-benefits of SMX removal and biomass production using duckweed. Intracellular H2O2 was produced using four duckweeds, Lemna aequinoctialis, L. minor, Landolina punctata, and Spirodela polyrhiza, in the range of 16.7-24.6 µ mol g-1 fresh weight, and extracellular H2O2 was released into the water phase. Consequently, duckweed could be used as an H2O2 supply source for the phyto-Fenton process. Specifically, 0.5 g fresh duckweed almost completely eliminated 1 mg L-1 SMX after 5 d in 50 mL sterile modified Hoagland solution containing 10 mM Fe (II). Fe (II)-dependent elimination of SMX indicated the occurrence of phyto-Fenton reaction. The phyto-Fenton process using duckweed effectively removed SMX. S. polyrhiza duckweed similarly removed 1 mg L-1 SMX even in sewage effluent containing other organic contaminants. During this treatment, duckweed biomass was generated at 7.95 g dry weight m-2 d-1, which was converted into methane at 353 normal liters CH4 kg-1 volatile solids by anaerobic digestion. For the first time, this study clearly demonstrates the potential for simultaneous SMX removal and biomass production from SMX-contaminated wastewater using duckweed.

Preliminary studies of selected Lemna species on the oxygen production potential in relation to some ecological factors.

Dissolved oxygen is fundamental for chemical and biochemical processes occurring in natural waters and critical for the life of aquatic organisms. Many organisms are responsible for altering organic matter and oxygen transfers across ecosystem or habitat boundaries and, thus, engineering the oxygen balance of the system. Due to such Lemna features as small size, simple structure, vegetative reproduction and rapid growth, as well as frequent mass occurrence in the form of thick mats, they make them very effective in oxygenating water. The research was undertaken to assess the impact of various species of duckweed (L. minor and L. trisulca) on dissolved oxygen content and detritus production in water and the role of ecological factors (light, atmospheric pressure, conductivity, and temperature) in this process. For this purpose, experiments were carried out with combinations of L. minor and L. trisulca. On this basis, the content of oxygen dissolved in water was determined depending on the growth of duckweed. Linear regression models were developed to assess the dynamics of changes in oxygen content and, consequently, organic matter produced by the Lemna. The research showed that the presence of L. trisulca causes an increase in dissolved oxygen content in water. It was also shown that an increase in atmospheric pressure had a positive effect on the ability of duckweed to produce oxygen, regardless of its type. The negative correlation between conductivity and water oxygenation, obtained in conditions of limited light access, allows us to assume that higher water conductivity limits oxygen production by all combinations of duckweeds when the light supply is low. Based on the developed models, it was shown that the highest increase in organic matter would be observed in the case of mixed duckweed and the lowest in the presence of the L. minor species, regardless of light conditions. Moreover, it was shown that pleustophytes have different heat capacities, and L. trisulca has the highest ability to accumulate heat in water for the tested duckweed combinations. The provided knowledge may help determine the good habitat conditions of duckweed, indicating its role in purifying water reservoirs as an effect of producing organic matter and shaping oxygen conditions with the participation of various Lemna species.

Optimizing biochar, vermicompost, and duckweed amendments to mitigate arsenic uptake and accumulation in rice (Oryza sativa L.) cultivated on arsenic-contaminated soil.

The accumulation of arsenic (As) in rice (Oryza sativa L.) grain poses a significant health concern in Bangladesh. To address this, we investigated the efficacy of various organic amendments and phytoremediation techniques in reducing As buildup in O. sativa. We evaluated the impact of five doses of biochar (BC; BC0.1: 0.1%, BC0.28: 0.28%, BC0.55: 0.55%, BC0.82: 0.82% and BC1.0: 1.0%, w/w), vermicompost (VC; VC1.0: 1.0%, VC1.8: 1.8%, VC3.0: 3.0%, VC4.2: 4.2% and VC5.0: 5.0%, w/w), and floating duckweed (DW; DW100: 100, DW160: 160, DW250: 250, DW340: 340 and DW400: 400 g m- 2) on O. sativa cultivated in As-contaminated soil. Employing a three-factor five-level central composite design and response surface methodology (RSM), we optimized the application rates of BC-VC-DW. Our findings revealed that As contamination in the soil negatively impacted O. sativa growth. However, the addition of BC, VC, and DW significantly enhanced plant morphological parameters, SPAD value, and grain yield per pot. Notably, a combination of moderate BC-DW and high VC (BC0.55VC5DW250) increased grain yield by 44.4% compared to the control (BC0VC0DW0). As contamination increased root, straw, and grain As levels, and oxidative stress in O. sativa leaves. However, treatment BC0.82VC4.2DW340 significantly reduced grain As (G-As) by 56%, leaf hydrogen peroxide by 71%, and malondialdehyde by 50% compared to the control. Lower doses of BC-VC-DW (BC0.28VC1.8DW160) increased antioxidant enzyme activities, while moderate to high doses resulted in a decline in these activities. Bioconcentration and translocation factors below 1 indicated limited As uptake and translocation in plant tissues. Through RSM optimization, we determined that optimal doses of BC (0.76%), VC (4.62%), and DW (290.0 g m- 2) could maximize grain yield (32.96 g pot- 1, 44% higher than control) and minimize G-As content (0.189 mg kg- 1, 54% lower than control). These findings underscore effective strategies for enhancing yield and reducing As accumulation in grains from contaminated areas, thereby ensuring agricultural productivity, human health, and long-term sustainability. Overall, our study contributes to safer food production and improved public health in As-affected regions.

Community interactions among microbes give rise to host-microbiome mutualisms in an aquatic plant.

Microbiomes often benefit plants, conferring resistance to pathogens, improving stress tolerance, or promoting plant growth. As potential plant mutualists, however, microbiomes are not a single organism but a community of species with complex interactions among microbial taxa and between microbes and their shared host. The nature of ecological interactions among microbes in the microbiome can have important consequences for the net effects of microbiomes on hosts. Here, we compared the effects of individual microbial strains and 10-strain synthetic communities on microbial productivity and host growth using the common duckweed Lemna minor and a synthetic, simplified version of its native microbiome. Except for Pseudomonas protegens, which was a mutualist when tested alone, all of the single strains we tested were commensals on hosts, benefiting from plant presence but not increasing host growth relative to uninoculated controls. However, 10-strain synthetic microbial communities increased both microbial productivity and duckweed growth more than the average single-strain inoculation and uninoculated controls, meaning that host-microbiome mutualisms can emerge from community interactions among microbes on hosts. The effects of community inoculation were sub-additive, suggesting at least some competition among microbes in the duckweed microbiome. We also investigated the relationship between L. minor fitness and that of its microbes, providing some of the first empirical estimates of broad fitness alignment between plants and members of their microbiomes; hosts grew faster with more productive microbes or microbiomes. IMPORTANCE: There is currently substantial interest in engineering synthetic microbiomes for health or agricultural applications. One key question is how multi-strain microbial communities differ from single microbial strains in their productivity and effects on hosts. We tested 20 single bacterial strains and 2 distinct 10-strain synthetic communities on plant hosts and found that 10-strain communities led to faster host growth and greater microbial productivity than the average, but not the best, single strain. Furthermore, the microbial strains or communities that achieved the greatest cell densities were also the most beneficial to their hosts, showing that both specific single strains and multi-strain synthetic communities can engage in high-quality mutualisms with their hosts. Our results suggest that ~5% of single strains, as well as multi-strain synthetic communities comprised largely of commensal microbes, can benefit hosts and result in effective host-microbe mutualisms.

Morphological diversity of the velamen radicum in the genus Anthurium (Araceae).

Epiphytes develop anatomical features to improve efficiency of the uptake of water and nutrients, such as absorptive foliar scales or a velamen radicum. Despite substantial studies on the occurrence, morphology, development and phylogeny of the velamen, most of the available literature is focused on Orchidaceae, making current knowledge on velamen clearly biased. A recent publication firmly established that velamina are common in Anthurium species. Thus, this study provides further insights by describing velamen morphological characteristics of Anthurium species and classifying them into different velamen types. Furthermore, we investigate if the different velamen morphological traits are clade-specific and phylogenetically conserved within the genus. Using SEM, we performed a morphological study on 89 Anthurium species, describing six micromorphological traits of velamen and exodermis, following traits used to classify Orchidaceae velamen by Porembski & Barthlott (1988). We distinguished nine velamen types, including two that are unique to Anthurium and not similar to any type found in Orchidaceae. Comparing velamen morphology within the phylogenetic tree of Anthurium revealed clear phylogenetic signals. This study provides detailed morphological descriptions among 89 species of Anthurium from the Araceae, and substantially broadens our knowledge of this tissue. However, velamen function has been even less studied, with hardly anything known about functional significance of having secondary cell wall thickening and perforations on velamen cell walls. Therefore, a logical next step would be to connect these anatomical features to their functions.

In situ phytoextraction of Mn and NH4+-N from aqueous electrolytic manganese residue solution by Pistia stratiotes: Effects of Fe/Co presence and rhizospheric microbe synergistic involvement.

The electrolytic manganese industry produces a large amount of electrolytic manganese residue (EMR). Soluble Mn, NH4+-N, and other pollutants may be released from the open-air stacked EMR and transported to the environment along with rainfall or surface runoff. Aqueous EMR solution (AES) generally contains various elements required for plant growth, and phytoremediation can be applied to remove these pollutants from AES. Since the contents of Fe and Co vary greatly in AES depending on the ore sources as well as the pre-treatment processes, the presence of bioavailable Fe and Co at different levels may affect plant growth, the rhizosphere microbes, and pollutant removal. The present study investigated the in-situ removal of Mn(II) and NH4+-N from AES solution using free floating aquatic plant Pistia stratiotes, focusing especially on the effects of Fe/Co presence and rhizospheric microbe synergistic involvement on contaminant removal. The results showed that 69.08% of Mn and 94.99% of NH4+-N were removed by P. stratiotes in 24 d. Both the presence of Fe(II) and Co(II) facilitated the Mn(II) immobilization and increased Mn(II) removal by 19-31% due to the enhanced peroxidase activity and the increased Mn accumulating in roots The complete removal of Mn from AES was found in the presence of Fe(II) at 2 mg L-1 or Co(II) at 0.5 mg L-1 and more than 51% accumulated Mn in the roots was stored in the vacuole and cytoplasm. BioMnOx was found on the surface of the roots, revealing that rhizofiltration, rhizospheric plaque/biofilm formation, and Mn biogeochemical cycle exert synergic effects on Mn(II) immobilization. The findings of the present study demonstrate the feasibility of using P. stratiotes in the treatment of aqueous EMR solutions and the presence of an appropriate amount of bio-available Fe and Co can promote the removal of Mn(II) and NH4+-N.

Impacts of Fulvic Acid on the Toxicity of the Herbicide Atrazine to Lemna minor.

Fulvic acids (FA) are environmentally prevalent components of dissolved organic carbon. Little research has evaluated their potential influence on the bioavailability of herbicides to non-target aquatic plants. This study evaluated the potential impacts of FA on the bioavailability of atrazine (ATZ) to the aquatic plant Lemna minor. Plants were exposed to 0, 15, 30, 60, 125, and 750 µg/L ATZ in media containing three FA concentrations (0, 5, and 15 mg/L) in a factorial study under static conditions. Fronds were counted after 7- and 14-days exposure and intrinsic growth rates (IGR) and total frond yields were calculated for analysis. Atrazine NOAECs and LOAECs within each FA treatment series (0, 5, or 15 mg/L) were identified and EC50s were estimated. NOAEC/LOAECs for yield and IGR were 60/125 µg/L except for yield in the 0 mg/L-FA series (30/60) and IGR in the 5 mg/L-FA series (30/60). NOAEC/LOAECs were 30/60 µg/L for all treatments and both endpoints after 14 days exposure. EC50s ranged from 88.2 to 106.1 µg/L (frond production 7 DAT), 158.0-186.0 µg/L (IGR, 7 DAT), 74.7-86.3 µg/L (frond production, 14 DAT), and 144.1-151.3 µg/L (IGR, 14 DAT). FA concentrations did not influence the toxicity of ATZ.

Caloric restriction extends lifespan in a clonal plant.

When subjected to dietary caloric restriction (CR), individual animals often outlive well-fed conspecifics. Here, we address whether CR also extends lifespan in plants. Whereas caloric intake in animals comes from ingestion, in plants it derives from photosynthesis. Thus, factors that reduce photosynthesis, such as reduced light intensity, can induce CR. In two lab experiments investigating the aquatic macrophyte Lemna minor, we tracked hundreds of individuals longitudinally, with light intensity-and hence, CR-manipulated using neutral-density filters. In both experiments, CR dramatically increased lifespan through a process of temporal scaling. Moreover, the magnitude of lifespan extension accorded with the assumptions that (a) light intensity positively relates to photosynthesis following Michaelis-Menten kinetics, and (b) photosynthesis negatively relates to lifespan via a power law. Our results emphasize that CR-mediated lifespan extension applies to autotrophs as well as heterotrophs, and suggest that variation in light intensity has quantitatively predictable effects on plant aging trajectories.

Changes in Metabolism and Content of Chlorophyll in Common Duckweed (Lemna minor L.) Caused by Environmental Contamination with Fluorides.

The impact of fluorine on plants remains poorly understood. We examined duckweed growth in extracts of soil contaminated with fluorine leached from chicken manure. Additionally, fluorine levels were analyzed in fresh manure, outdoor-stored manure, and soil samples at varying distances from the manure pile. Fresh manure contained 37-48 mg F- × kg-1, while soil extracts contained 2.1 to 4.9 mg F- × kg-1. We evaluated the physiological effects of fluorine on duckweed cultured on soil extracts or in 50% Murashige-Skoog (MS) medium supplemented with fluorine concentrations matching those in soil samples (2.1 to 4.9 mg F- × L-1), as well as at 0, 4, and 210 mg × L-1. Duckweed exposed to fluorine displayed similar toxicity symptoms whether in soil extracts or supplemented medium. Fluoride at concentrations of 2.1 to 4.9 mg F- × L-1 reduced the intact chlorophyll content, binding the porphyrin ring at position 32 without affecting Mg2+. This reaction resulted in chlorophyll a absorption peak shifted towards shorter wavelengths and formation of a new band of the F--chlorophyll a complex at λ = 421 nm. Moreover, plants exposed to low concentrations of fluorine exhibited increased activities of aminolevulinic acid dehydratase and chlorophyllase, whereas the activities of both enzymes sharply declined when the fluoride concentration exceeded 4.9 mg × L-1. Consequently, fluorine damages chlorophyll a, disrupts the activity of chlorophyll-metabolizing enzymes, and diminishes the plant growth rate, even when the effects of these disruptions are too subtle to be discerned by the naked human eye.

Comparison of mercury (Hg) bioaccumulation with mono- and mixed Lemna minor and Spirodela polyrhiza cultures.

Mercury (Hg) is a prevalent and harmful contaminant that persists in the environment. For phytoremediation, it is important to discover which plants can bioaccumulate meaningful amounts of Hg while also tolerating its toxicity. Additionally, increasing biodiversity could create a more resilient and self-sustaining system for remediation. This study explores whether mixed populations of Lemna minor and Spirodela polyrhiza can better bioaccumulate and tolerate Hg than monocultures. Mono- and mixed cultures of L. minor and S. polyrhiza were grown in mesocosms of 0.5 µg/L or 100 µg/L Hg (HgCl2) spiked water for 96 h. Change in weight of duckweed was used to assess Hg tolerance. Diffusive gradients in thin-films (DGTs) were used as surrogate monitoring devices for bioavailable levels of Hg. For biomass growth, the mixed culture of the L. minor was greater than the monoculture at the high dose. The L. minor accumulated more Hg in the mixed culture at the low dose while the S. polyrhiza was higher in the mixed at the high dose. Hg speciation in water was modeled using Windermere Humic Aqueous Model 7 (WHAM7) to compare the bioavailable species indicated by the DGTs.  Potentially due to the controlled conditions, the WHAM7 output of bioavailable Hg was almost 1:1 to that estimated by the DGTs, indicating good predictive capability of geochemical modeling and passive sampler DGT on metal bioavailability. Overall, the mixed cultures statistically performed as well as or better than the monocultures when tolerating and bioaccumulating Hg. However, there needs to be further work to see if the significant differences translate into practical differences worth the extra resources to maintain multiple species.

Trophic interactions in microbiomes influence plant host population size and ecosystem function.

Plant microbiomes that comprise diverse microorganisms, including prokaryotes, eukaryotes and viruses, are the key determinants of plant population dynamics and ecosystem function. Despite their importance, little is known about how species interactions (especially trophic interactions) between microbes from different domains modify the importance of microbiomes for plant hosts and ecosystems. Using the common duckweed Lemna minor, we experimentally examined the effects of predation (by bacterivorous protists) and parasitism (by bacteriophages) within microbiomes on plant population size and ecosystem phosphorus removal. Our results revealed that the addition of predators increased plant population size and phosphorus removal, whereas the addition of parasites showed the opposite pattern. The structural equation modelling further pointed out that predation and parasitism affected plant population size and ecosystem function via distinct mechanisms that were both mediated by microbiomes. Our results highlight the importance of understanding microbial trophic interactions for predicting the outcomes and ecosystem impacts of plant-microbiome symbiosis.

Population genomics and epigenomics of Spirodela polyrhiza provide insights into the evolution of facultative asexuality.

Many plants are facultatively asexual, balancing short-term benefits with long-term costs of asexuality. During range expansion, natural selection likely influences the genetic controls of asexuality in these organisms. However, evidence of natural selection driving asexuality is limited, and the evolutionary consequences of asexuality on the genomic and epigenomic diversity remain controversial. We analyzed population genomes and epigenomes of Spirodela polyrhiza, (L.) Schleid., a facultatively asexual plant that flowers rarely, revealing remarkably low genomic diversity and DNA methylation levels. Within species, demographic history and the frequency of asexual reproduction jointly determined intra-specific variations of genomic diversity and DNA methylation levels. Genome-wide scans revealed that genes associated with stress adaptations, flowering and embryogenesis were under positive selection. These data are consistent with the hypothesize that natural selection can shape the evolution of asexuality during habitat expansions, which alters genomic and epigenomic diversity levels.

A novel multitrophic biofloc technology for duckweed and Megalobrama amblycephala integrated culture: Improving nutrient utilization and animal welfare.

Biofloc technology (BFT) is an eco-friendly aquaculture model that utilizes zero-exchange water. In this study, we investigated the integration of duckweed into BFT in an effort to enhance nitrogen, phosphorus, and carbon utilization and to improve animal welfare for cultivating Megalobrama amblycephala. The experiment spanned 75 days, comparing a group of M. amblycephala supplemented with duckweed (DM) to a control group (CG) with no supplementation, where duckweed consumption relied solely on the feeding behavior of the fish. The concentrations of nitrate, total nitrogen, and phosphorus accumulation were lower in the DM than in the CG from day 45 onwards, with differences of 16.19, 26.90, and 1.45 mg/L, respectively, at the end of the experiment. The DM showed simultaneous increases of 5.77, 11.20, and 5.07 % in the absolute utilization of nitrogen, phosphorus, and carbon, respectively. The abundance of TM7a (10.27 %), linked to nitrate absorption, became the dominant genus in the water of the DM. Additionally, the abundance of Cetobacterium, associated with carbohydrate digestion, was significantly higher in gut of the DM (23.83 %) than in the gut of CG (1.24 %, P < 0.05). Supplementing the diet of M. amblycephala with duckweed improved digestion and antioxidant enzyme activity. Transcriptome data showed that duckweed supplementation resulted in an increase in the expression of genes related to protein digestion and absorption and carbohydrate metabolism in M. amblycephala, and analysis of the significantly enriched pathways further supported improved antioxidant capacity. Based on the above results, we concluded that as M. amblycephala consumes more duckweed, the differences in nitrogen and phosphorus levels between the DM and CG would continue to increase, along with a simultaneous increase in fixed carbon. Thus, this study achieved the goal of recycling BFT resources and improving animal welfare by integrating duckweed.

Optimization of Hoagland solution macro-elements as a culture media, for increasing protein content of duckweeds (Lemna minor).

This study aimed to increase the protein content of duckweed, a promising alternative to animal proteins and a sustainable source of plant protein cultivated via soilless agriculture, by manipulating the culture medium conditions (Hoagland solution). The contribution percentages of KH2PO4 and Ca(NO3)2, pivotal macro-elements in Hoagland solution affecting duckweed protein content, were determined using Plackett-Burman factorial design as 33.06 % and 36.61 %, respectively. Additionally, optimization was conducted employing response surface methodology, incorporating pH alongside KH2PO4 and Ca(NO3)2. Under optimal conditions of 3.92 mM KH2PO4, 7.95 mM Ca(NO3)2, and 7.22 pH, the protein content of duckweed increased significantly, reaching 51.09 % from 39.81 %. The duckweed cultivated in modified Hoagland solution exhibited protein content of 41.74 %, while duckweed grown in commercial Hoagland solution displayed protein content of 33.01 %. This study showed protein content of duckweed could significantly increase according to the growth medium and showcasing its potential as a sustainable source of plant protein.

Spatio-Temporal Study of Galactolipid Biosynthesis in Duckweed Using Mass Spectrometry Imaging and in vivo Isotope Labeling.

Isotope labeling coupled with mass spectrometry imaging (MSI) presents a potent strategy for elucidating the dynamics of metabolism at cellular resolution, yet its application to plant systems is scarce. It has the potential to reveal the spatio-temporal dynamics of lipid biosynthesis during plant development. In this study, we explore its application to galactolipid biosynthesis of an aquatic plant, Lemna minor, with D2O labeling. Specifically, matrix-assisted laser desorption/ionization-MSI data of two major galactolipids in L. minor, monogalactosyldiacylglycerol and digalactosyldiacylglycerol, were studied after growing in 50% D2O media over a 15-day time period. When they were partially labeled after 5 d, three distinct binomial isotopologue distributions were observed corresponding to the labeling of partial structural moieties: galactose only, galactose and a fatty acyl chain and the entire molecule. The temporal change in the relative abundance of these distributions follows the expected linear pathway of galactolipid biosynthesis. Notably, their mass spectrometry images revealed the localization of each isotopologue group to the old parent frond, the intermediate tissues and the newly grown daughter fronds. Besides, two additional labeling experiments, (i) 13CO2 labeling and (ii) backward labeling of completely 50% D2O-labeled L. minor in H2O media, confirm the observations in forward labeling. Furthermore, these experiments unveiled hidden isotopologue distributions indicative of membrane lipid restructuring. This study suggests the potential of isotope labeling using MSI to provide spatio-temporal details in lipid biosynthesis in plant development.