Light Intensity Modulates the Functional Composition of Leaf Metabolite Groups and Phyllosphere Prokaryotic Community in Garden Lettuce (Lactuca sativa L.) Plants at the Vegetative Stage.

Light intensity primarily drives plant growth and morphogenesis, whereas the ecological impact of light intensity on the phyllosphere (leaf surface and endosphere) microbiome is poorly understood. In this study, garden lettuce (Lactuca sativa L.) plants were grown under low, medium, and high light intensities. High light intensity remarkably induced the leaf contents of soluble proteins and chlorophylls, whereas it reduced the contents of leaf nitrate. In comparison, medium light intensity exhibited the highest contents of soluble sugar, cellulose, and free amino acids. Meanwhile, light intensity resulted in significant changes in the composition of functional genes but not in the taxonomic compositions of the prokaryotic community (bacteria and archaea) in the phyllosphere. Notably, garden lettuce plants under high light intensity treatment harbored more sulfur-cycling mdh and carbon-cycling glyA genes than under low light intensity, both of which were among the 20 most abundant prokaryotic genes in the leaf phyllosphere. Furthermore, the correlations between prokaryotic functional genes and lettuce leaf metabolite groups were examined to disclose their interactions under varying light intensities. The relative abundance of the mdh gene was positively correlated with leaf total chlorophyll content but negatively correlated with leaf nitrate content. In comparison, the relative abundance of the glyA gene was positively correlated with leaf total chlorophyll and carotenoids. Overall, this study revealed that the functional composition of the phyllosphere prokaryotic community and leaf metabolite groups were tightly linked in response to changing light intensities. These findings provided novel insights into the interactions between plants and prokaryotic microbes in indoor farming systems, which will help optimize environmental management in indoor farms and harness beneficial plant-microbe relationships for crop production.

Genome-Wide Identification and Characterization of Effector Candidates with Conserved Motif in Falciphora oryzae.

Microbes employ effectors to disrupt immune responses and promote host colonization. Conserved motifs including RXLR, LFLAK-HVLVxxP (CRN), Y/F/WxC, CFEM, LysM, Chitin-bind, DPBB_1 (PNPi), and Cutinase have been discovered to play crucial roles in the functioning of effectors in filamentous fungi. Nevertheless, little is known about effectors with conserved motifs in endophytes. This research aims to discover the effector genes with conserved motifs in the genome of rice endophyte Falciphora oryzae. SignalP identified a total of 622 secreted proteins, out of which 227 were predicted as effector candidates by EffectorP. By utilizing HMM features, we discovered a total of 169 effector candidates with conserved motifs and three novel motifs. Effector candidates containing LysM, CFEM, DPBB_1, Cutinase, and Chitin_bind domains were conserved across species. In the transient expression assay, it was observed that one CFEM and one LysM activated cell death in tobacco leaves. Moreover, two CFEM and one Chitin_bind inhibited cell death induced by Bax protein. At various points during the infection, the genes' expression levels were increased. These results will help to identify functional effector proteins involving omics methods using new bioinformatics tools, thus providing a basis for the study of symbiosis mechanisms.

Host genetic determinants drive compartment-specific assembly of tea plant microbiomes.

Diverse host factors drive microbial variation in plant-associated environments, whereas their genetic mechanisms remain largely unexplored. To address this, we coupled the analyses of plant genetics and microbiomes in this study. Using 100 tea plant (Camellia sinensis) cultivars, the microbiomes of rhizosphere, root endosphere and phyllosphere showed clear compartment-specific assembly, whereas the subpopulation differentiation of tea cultivars exhibited small effects on microbial variation in each compartment. Through microbiome genome-wide association studies, we examined the interactions between tea genetic loci and microbial variation. Notably, genes related to the cell wall and carbon catabolism were heavily linked to root endosphere microbial composition, whereas genes related to the metabolism of metal ions and small organic molecules were overrepresented in association with rhizosphere microbial composition. Moreover, a set of tea genetic variants, including the cytoskeleton-related formin homology interacting protein 1 gene, were strongly associated with the ß-diversity of phyllosphere microbiomes, implying their interactions with the overall structure of microbial communities. Our results create a catalogue of tea genetic determinants interacting with microbiomes and reveal the compartment-specific microbiome assembly driven by host genetics.

Ethylene enhances root water transport and aquaporin expression in trembling aspen (Populus tremuloides) exposed to root hypoxia.

BACKGROUND: Root hypoxia has detrimental effects on physiological processes and growth in most plants. The effects of hypoxia can be partly alleviated by ethylene. However, the tolerance mechanisms contributing to the ethylene-mediated hypoxia tolerance in plants remain poorly understood. RESULTS: In this study, we examined the effects of root hypoxia and exogenous ethylene treatments on leaf gas exchange, root hydraulic conductance, and the expression levels of several aquaporins of the plasma membrane intrinsic protein group (PIP) in trembling aspen (Populus tremuloides) seedlings. Ethylene enhanced net photosynthetic rates, transpiration rates, and root hydraulic conductance in hypoxic plants. Of the two subgroups of PIPs (PIP1 and PIP2), the protein abundance of PIP2s and the transcript abundance of PIP2;4 and PIP2;5 were higher in ethylene-treated trembling aspen roots compared with non-treated roots under hypoxia. The increases in the expression levels of these aquaporins could potentially facilitate root water transport. The enhanced root water transport by ethylene was likely responsible for the increase in leaf gas exchange of the hypoxic plants. CONCLUSIONS: Exogenous ethylene enhanced root water transport and the expression levels of PIP2;4 and PIP2;5 in hypoxic roots of trembling aspen. The results suggest that ethylene facilitates the aquaporin-mediated water transport in plants exposed to root hypoxia.

A hyperspectral deep learning attention model for predicting lettuce chlorophyll content.

BACKGROUND: The phenotypic traits of leaves are the direct reflection of the agronomic traits in the growth process of leafy vegetables, which plays a vital role in the selection of high-quality leafy vegetable varieties. The current image-based phenotypic traits extraction research mainly focuses on the morphological and structural traits of plants or leaves, and there are few studies on the phenotypes of physiological traits of leaves. The current research has developed a deep learning model aimed at predicting the total chlorophyll of greenhouse lettuce directly from the full spectrum of hyperspectral images. RESULTS: A CNN-based one-dimensional deep learning model with spectral attention module was utilized for the estimate of the total chlorophyll of greenhouse lettuce from the full spectrum of hyperspectral images. Experimental results demonstrate that the deep neural network with spectral attention module outperformed the existing standard approaches, including partial least squares regression (PLSR) and random forest (RF), with an average R2 of 0.746 and an average RMSE of 2.018. CONCLUSIONS: This study unveils the capability of leveraging deep attention networks and hyperspectral imaging for estimating lettuce chlorophyll levels. This approach offers a convenient, non-destructive, and effective estimation method for the automatic monitoring and production management of leafy vegetables.

Biogeographic patterns and drivers of soil viromes.

Viruses are crucial in shaping soil microbial functions and ecosystems. However, studies on soil viromes have been limited in both spatial scale and biome coverage. Here we present a comprehensive synthesis of soil virome biogeographic patterns using the Global Soil Virome dataset (GSV) wherein we analysed 1,824 soil metagenomes worldwide, uncovering 80,750 partial genomes of DNA viruses, 96.7% of which are taxonomically unassigned. The biogeography of soil viral diversity and community structure varies across different biomes. Interestingly, the diversity of viruses does not align with microbial diversity and contrasts with it by showing low diversity in forest and shrubland soils. Soil texture and moisture conditions are further corroborated as key factors affecting diversity by our predicted soil viral diversity atlas, revealing higher diversity in humid and subhumid regions. In addition, the binomial degree distribution pattern suggests a random co-occurrence pattern of soil viruses. These findings are essential for elucidating soil viral ecology and for the comprehensive incorporation of viruses into soil ecosystem models.

Linking microbial biogeochemical cycling genes to the rhizosphere of pioneering plants in a glacier foreland.

Glacier retreat raises global concerns but brings about the moment to study soil and ecosystem development. In nutrient-limited glacier forelands, the adaptability of pioneering plant and microbial species is facilitated by their interactions, including rhizosphere effects, but the details of this adaptability are not yet understood. In the rhizosphere of five pioneering plants, we comprehensively deciphered the microbial taxonomic and functional compositions. Two nitrogen-fixing microbial genera, Bradyrhizobium and Mesorhizobium, were among the most abundant taxa in the rhizomicrobiome. Moreover, several rhizobial genera, including Rhizobium, Pararhizobium, Allohrizobium, and Sinorhizobium, head the list of major modules in microbial co-occurrence networks, highlighting the vital roles of nitrogen-cycling taxa in the rhizomicrobiome of pioneering plants. Microbial genes involved in nitrogen, sulfur, phosphorus, and methane cycles were simultaneously correlated with microbial community dissimilarity, and 12 functional pathways were detected with distinct relative abundances among soils. Zooming in on the nitrogen-cycling genes, nifW, narC, nasA, nasB, and nirA were mainly responsible for the significant differences between soils. Furthermore, soil pH and the carbon/nitrogen ratio were among the topsoil properties interacting with nitrogen and sulfur cycling gene dissimilarity. These results explicitly linked biogeochemical cycling genes to the rhizomicrobiome and soil properties, revealing the roles of these genes as microbial drivers in mediating rhizosphere soil-plant-microbiome interactions.

Perspective on the development of synthetic microbial community (SynCom) biosensors.

Synthetic microbial community (SynCom) biosensors are a promising technology for detecting and responding to environmental cues and target molecules. SynCom biosensors use engineered microorganisms to create a more complex and diverse sensing system, enabling them to respond to stimuli with enhanced sensitivity and accuracy. Here, we give a definition of SynCom biosensors, outline their construction workflow, and discuss current biosensing technology. We also highlight the challenges and future for developing and optimizing SynCom biosensors and the potential applications in agriculture and food management, biotherapeutic development, home sensing, urban and environmental monitoring, and the One Health foundation. We believe SynCom biosensors could be used in a real-time and remote-controlled manner to sense the chaos of constantly dynamic environments.

Estimation of rice seedling growth traits with an end-to-end multi-objective deep learning framework.

In recent years, rice seedling raising factories have gradually been promoted in China. The seedlings bred in the factory need to be selected manually and then transplanted to the field. Growth-related traits such as height and biomass are important indicators for quantifying the growth of rice seedlings. Nowadays, the development of image-based plant phenotyping has received increasing attention, however, there is still room for improvement in plant phenotyping methods to meet the demand for rapid, robust and low-cost extraction of phenotypic measurements from images in environmentally-controlled plant factories. In this study, a method based on convolutional neural networks (CNNs) and digital images was applied to estimate the growth of rice seedlings in a controlled environment. Specifically, an end-to-end framework consisting of hybrid CNNs took color images, scaling factor and image acquisition distance as input and directly predicted the shoot height (SH) and shoot fresh weight (SFW) after image segmentation. The results on the rice seedlings dataset collected by different optical sensors demonstrated that the proposed model outperformed compared random forest (RF) and regression CNN models (RCNN). The model achieved R2 values of 0.980 and 0.717, and normalized root mean square error (NRMSE) values of 2.64% and 17.23%, respectively. The hybrid CNNs method can learn the relationship between digital images and seedling growth traits, promising to provide a convenient and flexible estimation tool for the non-destructive monitoring of seedling growth in controlled environments.

Tackling Soil ARG-Carrying Pathogens with Global-Scale Metagenomics.

Antibiotic overuse and the subsequent environmental contamination of residual antibiotics poses a public health crisis via an acceleration in the spread of antibiotic resistance genes (ARGs) through horizontal gene transfer. Although the occurrence, distribution, and driving factors of ARGs in soils have been widely investigated, little is known about the antibiotic resistance of soilborne pathogens at a global scale. To explore this gap, contigs from 1643 globally sourced metagnomes are assembled, yielding 407 ARG-carrying pathogens (APs) with at least one ARG; APs are detected in 1443 samples (sample detection rate of 87.8%). The richness of APs is greater in agricultural soils (with a median of 20) than in non-agricultural ecosystems. Agricultural soils possess a high prevalence of clinical APs affiliated with Escherichia, Enterobacter, Streptococcus, and Enterococcus. The APs detected in agricultural soils tend to coexist with multidrug resistance genes and bacA. A global map of soil AP richness is generated, where anthropogenic and climatic factors explained AP hot spots in East Asia, South Asia, and the eastern United States. The results herein advance this understanding of the global distribution of soil APs and determine regions prioritized to control soilborne APs worldwide.

Stable expression of aquaporins and hypoxia-responsive genes in adventitious roots are linked to maintaining hydraulic conductance in tobacco (Nicotiana tabacum) exposed to root hypoxia.

Formation of adventitious roots in plants is a common response to hypoxia caused by flooding. In tobacco, after one week of root hypoxia treatment, plants produced twice as many adventitious roots as the aerated plants, but their maximum length was reduced. Hypoxia severely reduced net photosynthesis, transpiration rates, and photosynthetic light responses. Relative transcript abundance of the examined aquaporins in lateral roots was reduced by hypoxia, but in adventitious roots it remained unchanged. This apparent lack of an effect of root hypoxia on the aquaporin expression likely contributed to maintenance of high hydraulic conductance in adventitious roots. Lateral roots had lower porosity compared with adventitious roots and the expression of the ACS (1-aminocyclopropane-1-carboxylate synthase) gene was induced in hypoxic lateral roots, but not in adventitious roots, providing additional evidence that lateral roots were more affected by hypoxia compared with adventitious roots. ATP concentrations were markedly lower in both hypoxic lateral and adventitious roots compared with aerated roots, while the expression of fermentation-related genes, ADH1 (alcohol dehydrogenase 1) and PDC1 (pyruvate decarboxylase 1), was higher in lateral roots compared with adventitious roots. Since root porosity was greater in adventitious compared with lateral roots, the results suggest that the improved O2 delivery and stable root aquaporin expression in adventitious roots were likely the key factors helping flooded tobacco plants maintain high rates of root hydraulic conductance and, consequently, shoot gas exchange.

Plant water transport and aquaporins in oxygen-deprived environments.

Oxygen deprivation commonly affects plants exposed to flooding and soil compaction. The resulting root hypoxia has an immediate effect on plant water relations and upsets water balance. Hypoxia inhibits root water transport and triggers stomatal closure. The processes contributing to the inhibition of root hydraulic conductivity and conductance (hydraulic conductivity of the whole root system) are complex and involve changes in root morphology and the functions of aquaporins. Aquaporins (AQPs) comprise a group of membrane intrinsic proteins that are responsible for the transport of water, as well as some small neutral solutes and ions. They respond to a wide range of environmental stresses including O2 deprivation, but the underlying functional mechanisms are still elusive. The aquaporin-mediated water transport is affected by the acidification of the cytoplasm and depletion of ATP that is required for aquaporin phosphorylation and membrane functions. Cytoplasmic pH, phosphorylation, and intracellular Ca2+ concentration directly control AQP gating, all of which are related to O2 deprivation. This review addresses the structural determinants that are essential for pore conformational changes in AQPs, to highlight the underlying mechanisms triggered by O2 deprivation stress. Gene expression of AQPs is modified in hypoxic plants, which may constitute an important, yet little explored, mechanism of hypoxia tolerance. In addition to water transport, AQPs may contribute to hypoxia tolerance by transporting O2, H2O2, and lactic acid. Responses of plants to O2 deprivation, and especially those that contribute to maintenance of water transport, are highly complex and entail the signals originating in roots and shoots that lead to and follow the stomatal closure. These complex responses may involve ethylene, abscisic acid, and possibly other hormonal factors and signaling molecules in ways that remain to be elucidated.

Significance of oxygen transport through aquaporins.

Aquaporins are membrane integral proteins responsible for the transmembrane transport of water and other small neutral molecules. Despite their well-acknowledged importance in water transport, their significance in gas transport processes remains unclear. Growing evidence points to the involvement of plant aquaporins in CO2 delivery for photosynthesis. The role of these channel proteins in the transport of O2 and other gases may also be more important than previously envisioned. In this study, we examined O2 permeability of various human, plant, and fungal aquaporins by co-expressing heterologous aquaporin and myoglobin in yeast. Two of the most promising O2-transporters (Homo sapiens AQP1 and Nicotiana tabacum PIP1;3) were confirmed to facilitate O2 transport in the spectrophotometric assay using yeast protoplasts. The over-expression of NtPIP1;3 in yeasts significantly increased their O2 uptake rates in suspension culture. In N. tabacum roots subjected to hypoxic hydroponic conditions, the transcript levels of the O2-transporting aquaporin NtPIP1;3 significantly increased after the seven-day hypoxia treatment, which was accompanied by the increase of ATP levels in the apical root segments. Our results suggest that the functional significance of aquaporin-mediated O2 transport and the possibility of controlling the rate of transmembrane O2 transport should be further explored.

Dominance of an alien shrub Rhus typhina over a native shrub Vitex negundo var. heterophylla under variable water supply patterns.

Temporal heterogeneity of a resource supply can have a profound effect on the interactions between alien and native plant species and their potential invasiveness. Precipitation patterns may be variable and result in a higher heterogeneity of water supply with global climate change. In this study, an alien shrub species, Rhus typhina, introduced to China from North America and a native shrub species, Vitex negundo var. heterophylla, were grown in monoculture and mixed culture under different water supply regimes, with four levels of water supply frequencies but with a constant level of total supplied water. After 60 days of treatments, the alien species was found to be the superior competitor in the mixed culture and was unaffected by changes in the water supply pattern. The dominance of R. typhina was mainly owing to its greater biomass and effective modulation of leaf physiology. However, in the mixed culture, V. negundo var. heterophylla exhibited both leaf- and whole-plant-level acclimations, including higher leaf length to petiole length and root to shoot biomass ratios, and lower specific leaf weight and leaf length to leaf width ratio. Plant height of V. negundo var. heterophylla was comparable to that of R. typhina in the mixed culture, which is a strategy to escape shading. Although water treatments had little effect on most traits in both species, the possible influence of water regimes should not be neglected. Compared with high-frequency water supply treatments, more individuals of V. negundo var. heterophylla died in low-water-frequency treatments when in competition with R. typhina, which may lead to species turnover in the field. The authors recommended that caution should be exercised when introducing R. typhina to non-native areas in the context of global climate change.