Physiological Characterization Research Articles

Phenotypic characterization and gene mapping of hybrid necrosis in Triticum durum–Haynaldia villosa amphiploids

Key messagePhenotypical, physiological and genetic characterization was carried out on the hybrid necrosis gene from Haynaldia villosa, and the related gene Ne-V was mapped to chromosome arm 2VL.Introducing genetic variation from wild relatives into common wheat through wide crosses is a vital strategy for enriching genetic diversity and promoting wheat breeding. However, hybrid necrosis, a genetic autoimmunity syndrome, often occurs in the offspring of interspecific or intraspecific crosses, restricting both the selection of hybrid parents and the pyramiding of beneficial genes. To utilize the germplasms of Haynaldia villosa (2n = 2x = 14, VV), we conducted wide hybridization between durum wheat (2n = 4x = 28, AABB) and multiple H. villosa accessions to synthesize the amphiploids (2n = 6x = 42, AABBVV). This study revealed that 61.5% of amphiploids derived from the above crosses exhibited hybrid necrosis, with some amphiploids even dying before reaching maturity. However, the initiation time and severity of necrosis varied dramatically among the progenies, suggesting that there were multiple genetic loci or multiple alleles in the same genetic locus conferring to hybrid necrosis in H. villosa accessions. Genetic analysis was performed on the F2 and derived F2:3 populations, which were constructed between amphiploid STH59-1 with normal leaves and amphiploid STH59-2 with necrotic leaves. A semidominant hybrid necrosis-related gene, Ne-V, was mapped to an 11.8-cM genetic interval on the long arm of chromosome 2V, representing a novel genetic locus identified in Triticum-related species. In addition, the hybrid necrosis was correlated with enhanced H2O2 accumulation and cell death, and it was influenced by the temperature and light. Our findings provide a foundation for cloning the Ne-V gene and exploring its molecular mechanism.

New variants expand the neurological phenotype of COQ7 deficiency.

The protein encoded by COQ7 is required for CoQ10 synthesis in humans, hydroxylating 3-demethoxyubiquinol (DMQ10) in the second to last steps of the pathway. COQ7 mutations lead to a primary CoQ10 deficiency syndrome associated with a pleiotropic neurological disorder. This study shows the clinical, physiological, and molecular characterization of four new cases of CoQ10 primary deficiency caused by five mutations in COQ7, three of which have not yet been described, inducing mitochondrial dysfunction in all patients. However, the specific combination of the identified variants in each patient generated precise pathophysiological and molecular alterations in fibroblasts, which would explain the differential in vitro response to supplementation therapy. Our results suggest that COQ7 dysfunction could be caused by specific structural changes that affect the interaction with COQ9 required for the DMQ10 presentation to COQ7, the substrate access to the active site, and the maintenance of the active site structure. Remarkably, patients' fibroblasts share transcriptional remodeling, supporting a modification of energy metabolism towards glycolysis, which could be an adaptive mechanism against CoQ10 deficiency. However, transcriptional analysis of mitochondria-associated pathways showed distinct and dramatic differences between patient fibroblasts, which correlated with the extent of pathophysiological and neurological alterations observed in the probands. Overall, this study suggests that the combination of precise genetic diagnostics and the availability of new structural models of human proteins could help explain the origin of phenotypic pleiotropy observed in some genetic diseases and the different responses to available therapies.

Isolation and characterization of a native strain of the entomopathogenic fungus Beauveria bassiana for the control of the palm weevil Dynamis borassi (Coleoptera: Curculionidae) in the neotropics

This study aimed to isolate and characterize a native strain of Beauveria bassiana, coded as Bv065, showcasing its potential as a biological control agent targeting the palm weevil Dynamis borassi. Originating from a naturally infected D. borassi specimen collected in southwestern Colombia, the fungus underwent molecular identification and was identified as B. bassiana, exhibiting high sequence similarity with known reference strains. The physiological characterization revealed that Bv065 thrived within a temperature range of 25 to 30 °C and a pH range of 6 to 9. Moreover, the key carbon sources that allow optimal growth of the strain were identified through metabolic profiling, including sucrose, D-mannose, and γ-amino-butyric acid. These findings offer strategic insights for scalability and formulation methodologies. Additionally, enzymatic analyses unveiled robust protease activity within Bv065, crucial for catalysing insect cuticle degradation and facilitating host penetration, thus accentuating its entomopathogenic potential. Subsequent evaluations exposed Bv065’s pathogenicity against D. borassi, causing significant mortality within nine days of exposure, albeit exhibiting limited effectiveness against Rhynchophorus palmarum. This study underscores the importance of understanding optimal growth conditions and metabolic preferences of B. bassiana strains for developing effective biopesticides. The findings suggest Bv065 as a promising candidate for integrated pest management strategies in neotropical regions, particularly for controlling palm weevil infestations in coconut and peach palm cultivation. Future research avenues include refining mass production methodologies, formulating novel delivery systems, and conducting comprehensive field efficacy trials to unlock the full potential of Bv065 in fostering sustainable pest management practices. Overall, this study contributes to the growing body of knowledge on entomopathogenic fungi and their pivotal role in biological control, offering nuanced perspectives on eco-friendly alternatives to conventional insecticidal interventions.

Physiological and biochemical characterisation of split and healthy Daisy mandarin (Citrus reticulata Burm.) fruits

ABSTRACT Fruit splitting is a prevalent physiological disorder that has a significant impact on Daisy mandarin fruit production, posing challenges to farmers in terms of productivity and economic viability. Despite its impact, no prior research on fruit splitting in Daisy mandarin has been conducted. This study aimed to assess the effect of fruit splitting on quality, morphological characteristics, biochemical composition and nutrient content in Daisy mandarin. In 2022, a comprehensive analysis was conducted on both split and healthy Daisy mandarin fruits, budded on Rough lemon rootstock, to investigate their morphological characteristics, biochemical characteristics, mineral content and cell wall components. Additionally, the study examined the activity of cell wall-degrading enzymes, such as peroxidase (POD), cellulase (Cx) and polygalacturonase (PG). The Results indicated that split fruits exhibited larger fruit size and weight and increased water content in both the fruits and peel compared to healthy fruits. However, split fruits had thinner peel thickness and lower peel content. The peels of split fruits also contained higher level of total soluble sugars, amino acids, and proline content. Furthermore, the peels of split fruits displayed much higher activity of cell wall-degrading enzymes than those of healthy fruits. The nutrient analysis of the split fruits revealed a decrease in boron, calcium and potassium content in the peel compared to healthy fruits. The study provides crucial insights into the mechanisms underlying fruit splitting, identifying biochemical and physiological differences between split and healthy fruits. These findings offer valuable information for developing strategies to mitigate fruit splitting, enhancing fruit quality and reducing economic losses for citrus growers. Future research should explore the genetic and environmental factors contributing to fruit splitting and investigate potential interventions to improve nutrient management and fruit integrity in Daisy mandarin cultivation.

Isolation and characterization of novel acetogenic strains of the genera Terrisporobacter and Acetoanaerobium.

Due to their metabolic versatility in substrate utilization, acetogenic bacteria represent industrially significant production platforms for biotechnological applications such as syngas fermentation, microbial electrosynthesis or transformation of one-carbon substrates. However, acetogenic strains from the genera Terrisporobacter and Acetoanaerobium remained poorly investigated for biotechnological applications. We report the isolation and characterization of four acetogenic Terrisporobacter strains and one Acetoanaerobium strain. All Terrisporobacter isolates showed a characteristic growth pattern under a H2 + CO2 atmosphere. An initial heterotrophic growth phase was followed by a stationary growth phase, where continuous acetate production was indicative of H2-dependent acetogenesis. One of the novel Terrisporobacter isolates obtained from compost (strain COMT) additionally produced ethanol besides acetate in the stationary growth phase in H2-supplemented cultures. Genomic and physiological characterizations showed that strain COMT represented a novel Terrisporobacter species and the name Terrisporobacter vanillatitrophus is proposed (=DSM 116160T = CCOS 2104T). Phylogenomic analysis of the novel isolates and reference strains implied the reclassification of the T. petrolearius/T. hibernicus phylogenomic cluster to the species T. petrolearius and of the A. noterae/A. sticklandii phylogenomic cluster to the species A. sticklandii. Furthermore, we provide first insights into active prophages of acetogens from the genera Terrisporobacter and Acetoanaerobium.

Maribellus mangrovi sp. nov., an iron-reducing bacterium isolated from mangrove sediment.

A novel facultatively anaerobic and Gram-stain-negative bacterium, designated FJH33T, was isolated from mangrove sediment sampled in Zhangzhou, PR China. Cells of strain FJH33T were rod-shaped or slightly curved-shaped, with widths of 0.3-0.5 µm and lengths of 1.0-3.0 µm. Optimum growth of strain FJH33T occurred in the presence of 3 % NaCl (w/v), at 33 °C and at pH 7.0. Oxidase activity was negative, while catalase activity was positive. Its iron-reducing ability was determined. Based on 16S rRNA gene sequence similarity, strain FJH33T was most closely related to Maribellus luteus XSD2T (95.1 %), followed by Maribellus sediminis Y2-1-60T (95.0 %) and Maribellus maritimus 5E3T (94.9 %). Genome analysis of strains FJH33T and M. luteus XSD2T revealed low genome relatedness, with an average nucleotide identity value of 73.8% and a digital DNA-DNA hybridization value of 19.0%. Phylogenetic trees built from 16S rRNA genes and genome sequences showed that strain FJH33T represents a relatively independent phylogenetic lineage within the genus Maribellus. The major cellular fatty acids (≥10 %) were iso-C15 : 0 and C18 : 1 ω9c. The sole respiratory quinone was MK-7. The polar lipids consisted of phosphatidylethanolamine, diphosphatidylcholine, diphosphatidyglycerol and one unidentified lipid. The DNA G+C content was 41.4 mol%. Based on the integrated results of phylogenetic, physiological, biochemical and chemotaxonomic characterizations, we propose that strain FJH33T represents a novel species of the genus Maribellus, for which the name Maribellus mangrovi sp. nov. is proposed. The type strain is FJH33T (=KCTC 102210T=MCCC 1H01459T).

Genomic and functional insights of a mucin foraging Rhodopirellula halodulae sp. nov.

Nine novel strains were obtained from various algal and seagrass samples. The analysis of the 16S rRNA gene-based phylogenetic tree revealed monophyletic placement of all novel strains within the Rhodopirellula genus. The type strain was identified as JC737T, which shared 99.1 % 16S rRNA gene sequence identity with Rhodopirellula baltica SH1T, while strain JC740 was designated as an additional strain. The genome sizes of strains JC737T and JC740 were 6.6 and 6.7 Mb, respectively, and the G + C content was 56.2 %. The strains cladded distinctly in the phylogenomic tree, and the ANI and dDDH values of the strain JC737T were 75.8–76.1 % and 20.8–21.3 %, respectively, in comparison to other Rhodopirellula members. The strain demonstrated a versatile degradation capability, exhibiting a diverse array of complex polysaccharides, including mucin which had not been previously identified within the members of the phylum Planctomycetota. The phylogenomic, pan-genomic, morphological, physiological, and genomic characterization of the strain lead to the proposal to describe the strain as Rhodopirellula halodulae sp. nov.

Physiological and genomic analysis of Limnospira sp. AlgLouSa11 isolated from Tamanrasset, Algeria

Cyanobacteria designated as Spirulina, Arthrospira and Limnospira are worldwide well-known food and food supplement sources. Despite their long history of use for human consumption, many indigenous strains remain unstudied. Here we performed a genomic and physiological characterization of AlgLouSa11, an Algerian strain isolated from the Taguemart region in Tamanrasset. Its genome was sequenced and compared (via ANI and PaSit4) to 21 genomes of the Arthrospira and Limnospira genera, indicating that AlgLouSa11 belonged to the Limnospira genus, with Limnospira fusiformis KN01 being the closest relative (99.3 %). Growth was scored in three different culture media (Hiri, Zarrouk, and BG-11), with the highest cell density, 4.76 × 105 cells/ml after 11 days, obtained for Hiri medium. The optimal growth temperature was 34°C, with automatic agitation producing the highest yields (5.90 ± 0.25 g/l). The optimal growth pH was nine, with growth decreasing with increasing alkalinity. Finally, exploring the genome of Limnospira sp. AlgLouSa11 showed the presence of coding sequences involved in the biosynthesis of secondary metabolites such as cyanobactin, terpenes and lanthipeptides.

A neural m6A pathway regulates behavioral aggregation in migratory locusts.

RNA N6-methyladenosine (m6A), as the most abundant modification of messenger RNA, can modulate insect behaviors, but its specific roles in aggregation behaviors remain unexplored. Here, we conducted a comprehensive molecular and physiological characterization of the individual components of the methyltransferase and demethylase in the migratory locust Locusta migratoria. Our results demonstrated that METTL3, METTL14 and ALKBH5 were dominantly expressed in the brain and exhibited remarkable responses to crowding or isolation. The individual knockdown of methyltransferases (i.e., METTL3 and METTL14) promoted locust movement and conspecific attraction, whereas ALKBH5 knockdown induced a behavioral shift toward the solitary phase. Furthermore, global transcriptome profiles revealed that m6A modification could regulate the orchestration of gene expression to fine tune the behavioral aggregation of locusts. In summary, our in vivo characterization of the m6A functions in migratory locusts clearly demonstrated the crucial roles of the m6A pathway in effectively modulating aggregation behaviors.

In vitro culture of Campomanesia pubescens under different light qualities: a morphoanatomical and physiological characterization

Campomanesia pubescens (gabiroba) is a fruit tree that is native to the Cerrado and that has commercial potential and medicinal properties. It has recalcitrant seeds that do not tolerate desiccation and storage. Here, C. pubescens plants were grown in vitro under different light qualities. Light-emitting diodes (LEDs) were used at wavelengths for white (W) and blue in combination with red in the proportions BR (1:1), BR (1:3) and BR (3:1), at 100 ± 5 μmol m-2 s-1 over a 16-hour photoperiod. The leaf anatomy, chlorophyll a fluorescence, chloroplast pigments and malondialdehyde (MDA) content were evaluated. Studies on Cerrado plants grown in vitro focusing on light quality and its effects on plant development and growth are scarce. Records of anatomical and physiological C. pubescens characteristics when grown in vitro under different lighting qualities using LEDs are nonexistent. In this context, the aim of this study was to analyze the anatomical and physiological responses of C. pubescens seedlings from in vitro cultivation under different light qualities, to support future studies on the propagation and conservation of this species. This work is a pioneer for this species. When the plants were cultivated in combinations of blue/red LEDs 1:1 and 3:1, they yielded higher biomass, presenting higher epidermis and chlorenchyma values with greater stomatal density and functionality, better photosynthetic efficiency, higher Y(II) values, qP and higher total chlorophyll concentrations, and thus these lights did not cause oxidative damage compared to the white (control) LEDs. This study contributed to a better understanding of the anatomical and physiological changes in C. pubescens plants grown in vitro under different light qualities. C. pubescens plants exhibited varied anatomical and physiological characteristics depending on the spectrum of light used here. The combinations of BR (1:1) or BR (3:1) LEDs are promising for the propagation of the species, given their positive influence on most of the studied traits.

Physiological and transcriptome characterization provide new sights into cadmium tolerance and accumulation mechanisms in Tagetes patula L.

Tagetes patula L. (T. patula) is a Cd hyperaccumulator plant with high ornamental value and strong resistance. Understanding the molecular mechanisms of Cd tolerance in hyper-accumulating plants is important for improving phytoremediation efficiency. This investigation studied to how Cd affected T. patula's physiological characteristics and molecular alterations. With the increase of Cd concentration, chlorophyll content, and root biomass decreased; the activities of antioxidant enzymes such as GST, SOD and POD increased. Initially, Cd accumulated mainly in the root and then transferred upward to the leaves. The molecular mechanisms of T. patula after 150 mg/kg and 250 mg/kg treatments were analyzed by transcriptome. In total, 5429 and 6546 differentially expressed unigenes were identified. Phenylpropanoid biosynthesis and phenylalanine metabolism were identified as key pathways in reaction to Cd stress. In addition, many differential genes involved in the response to Cd stress were identified, including heavy metal transporter protein genes, genes and transcription factors related to Cd detoxification. These results indicate that the response of T. patula to Cd was multifaceted, laying the foundation for studies to improve its tolerance to toxic metals.

Genomics and physiology of Catenibacillus, human gut bacteria capable of polyphenol C-deglycosylation and flavonoid degradation.

The genus Catenibacillus (family Lachnospiraceae, phylum Bacillota) includes only one cultivated species so far, Catenibacillus scindens, isolated from human faeces and capable of deglycosylating dietary polyphenols and degrading flavonoid aglycones. Another human intestinal Catenibacillus strain not taxonomically resolved at that time was recently genome-sequenced. We analysed the genome of this novel isolate, designated Catenibacillus decagia, and showed its ability to deglycosylate C-coupled flavone and xanthone glucosides and O-coupled flavonoid glycosides. Most of the resulting aglycones were further degraded to the corresponding phenolic acids. Including the recently sequenced genome of C. scindens and ten faecal metagenome-assembled genomes assigned to the genus Catenibacillus, we performed a comparative genome analysis and searched for genes encoding potential C-glycosidases and other polyphenol-converting enzymes. According to genome data and physiological characterization, the core metabolism of Catenibacillus strains is based on a fermentative lifestyle with butyrate production and hydrogen evolution. Both C. scindens and C. decagia encode a flavonoid O-glycosidase, a flavone reductase, a flavanone/flavanonol-cleaving reductase and a phloretin hydrolase. Several gene clusters encode enzymes similar to those of the flavonoid C-deglycosylation system of Dorea strain PUE (DgpBC), while separately located genes encode putative polyphenol-glucoside oxidases (DgpA) required for C-deglycosylation. The diversity of dgpA and dgpBC gene clusters might explain the broad C-glycoside substrate spectrum of C. scindens and C. decagia. The other Catenibacillus genomes encode only a few potential flavonoid-converting enzymes. Our results indicate that several Catenibacillus species are well-equipped to deglycosylate and degrade dietary plant polyphenols and might inhabit a corresponding, specific niche in the gut.

SCAPV Creates the Possibility of Less Irrigation and Higher Productivity

In agrivoltaic (APV), photovoltaic (PV) panels are positioned above farmland to produce energy and food simultaneously. However, PV panels above farmland block most sunlight from reaching plants for photosynthesis. Plants require sunlight for photosynthesis. We proposed Spectrum-splitting and Concentrated APV (SCAPV) to address contradictions between photosynthesis and energy production simultaneously. This study examines the effect of SCAPV on the evapotranspiration and growth of peanuts and soybeans. Peanuts and soybeans were planted under SCAPV and open-air (CK) treatments, and a weather station was placed in each treatment. Results showed that evapotranspiration under SCAPV significantly decreased by 31% compared to CK. Thus, it improved physiological characterization, enhanced quality, and increased the yield of peanuts and soybeans. Peanuts' protein, fat, and linoleic acid increased by 5.54%, 0.28%, and 1.14% under SCAPV compared to CK. Fat, soluble sugar, linoleic acid, and alpha-linolenic acid of soybean were increased by 6.75%, 15.24%, 13.72%, and 15.14%, respectively, under SCAPV compared to CK. The average land equivalent ratio of SCAPV is 1.7. We trust that SCAPV could provide food and energy while reducing irritation on the same farmland.

Sustained bacterial N2O reduction at acidic pH

Nitrous oxide (N2O) is a climate-active gas with emissions predicted to increase due to agricultural intensification. Microbial reduction of N2O to dinitrogen (N2) is the major consumption process but microbial N2O reduction under acidic conditions is considered negligible, albeit strongly acidic soils harbor nosZ genes encoding N2O reductase. Here, we study a co-culture derived from acidic tropical forest soil that reduces N2O at pH 4.5. The co-culture exhibits bimodal growth with a Serratia sp. fermenting pyruvate followed by hydrogenotrophic N2O reduction by a Desulfosporosinus sp. Integrated omics and physiological characterization revealed interspecies nutritional interactions, with the pyruvate fermenting Serratia sp. supplying amino acids as essential growth factors to the N2O-reducing Desulfosporosinus sp. Thus, we demonstrate growth-linked N2O reduction between pH 4.5 and 6, highlighting microbial N2O reduction potential in acidic soils.

Transcription factor ZmEREB97 regulates nitrate uptake in maize (Zea mays) roots.

Maize (Zea mays L.) has very strong requirements for nitrogen. However, the molecular mechanisms underlying the regulations of nitrogen uptake and translocation in this species are not fully understood. Here, we report that an APETALA2/ETHYLENE RESPONSE FACTOR (AP2/ERF) transcription factor ZmEREB97 functions as an important regulator in the N-signaling network in maize. Predominantly expressed and accumulated in main root and lateral root primordia, ZmEREB97 rapidly responded to nitrate treatment. By overlapping the analyses of differentially expressed genes and conducting a DAP-seq assay, we identified 1446 potential target genes of ZmEREB97. Among these, 764 genes were co-regulated in two lines of zmereb97 mutants. Loss of function of ZmEREB97 substantially weakened plant growth under both hydroponic and soil conditions. Physiological characterization of zmereb97 mutant plants demonstrated that reduced biomass and grain yield were both associated with reduced nitrate influx, decreased nitrate content and less N accumulation. We further demonstrated that ZmEREB97 directly targets and regulates the expression of six ZmNRT genes by binding to the GCC box-related sequences in gene promoters. Collectively, these data suggest that ZmEREB97 is a major positive regulator of the nitrate response and that it plays an important role in optimizing nitrate uptake, offering a target for improvement of nitrogen use efficiency in crops.

Molecular and physiological characterization of AIP1, encoding the acetolactate synthase regulatory subunit in rice

Flooding deprives plants of oxygen and thereby causes severe stress by interfering with energy production, leading to growth retardation. Enzymes and metabolites may help protect plants from waterlogging and hypoxic environmental conditions. Acetolactate synthase (ALS) is a key enzyme in the biosynthesis of branched-chain amino acids (BCAAs), providing the building blocks for proteins and various secondary metabolites. Additionally, under energy-poor conditions, free BCAAs can be used as an alternative energy source by mitochondria through a catabolic enzyme chain reaction. In this study, we characterized ALS-INTERACTING PROTEIN 1 (OsAIP1), which encodes the regulatory subunit of ALS in rice (Oryza sativa). This gene was expressed in all parts of the rice plant, and its expression level was significantly higher in submerged and low-oxygen environments. Rice transformants overexpressing OsAIP1 showed a higher survival rate under hypoxic stress than did non-transgenic control plants under the same conditions. The OsAIP1-overexpressing plants accumulated increased levels of BCAAs, demonstrating that OsAIP1 is an important factor in the hypoxia resistance mechanism. These results suggest that ALS proteins are part of a defense mechanism that improves the tolerance of plants to low-oxygen environments.

Physiological characterization and bioactive compounds of promising accessions of cowpea (Vigna unguiculata L. Walp) in the Peruvian Amazon

Cowpea production in the Peruvian Amazon is increasing due to its ability to adapt to diverse environments, its contribution to soil conservation, and its versatility of uses. This crop is particularly valuable in human nutrition due to its high nutritional quality and nutraceutical properties of its bioactive compounds. The aim of the study was to evaluate the physiology of cowpea plants in accessions PER1005854, PER1005851, and PER12645, under the influence of rice husk mulch, screw tree litter, and oil palm fiber, as well as to determine the bioactive compounds present in the grains. This study aims to promote sustainable cowpea production, expose the nutritional aspects of the grains, and foster their consumption both locally and nationally. The experiment was conducted from November 2022 to March 2023, in Yurimaguas, Loreto, Peru. The application of organic mulches demonstrates a significantly positive impact on the growth of cowpea accessions during their vegetative phase. This effect is possibly attributed to increased water availability, as the mulch reduces soil evaporation. In particular, cowpea accessions PER12645 and PER1005851 exhibit desirable physiological characteristics for grain production, with short cycles of 71 days and higher yields of 1141 and 1125 kg/ha respectively. In contrast, accession PER1005854 is distinguished by its higher biomass production, reaching a value of 14497 kg/ha, a relevant trait for foliage production and its contribution to soil conservation. The bioactive compounds, such as proteins, lipids, and carbohydrates, present in the accessions PER1005854, PER1005851, and PER12645, are similar to those found in other legumes. However, the antioxidant activity of dark tegument cowpeas proved to be superior, particularly highlighted in accession PER12645 (black tegument) with 26.3 μmolTG/g, and in accession PER1005854 (dark red tegument) with 19.5 μmolTG/g. This characteristic is particularly important for consumption, as it is related to the ability to combat oxidative stress in the human body.

Attention LSTM U-Net model for Drosophila melanogaster heart tube segmentation in optical coherence microscopy images.

Optical coherence microscopy (OCM) imaging of the Drosophila melanogaster (fruit fly) heart tube has enabled the non-invasive characterization of fly heart physiology in vivo. OCM generates large volumes of data, making it necessary to automate image analysis. Deep-learning-based neural network models have been developed to improve the efficiency of fly heart image segmentation. However, image artifacts caused by sample motion or reflections reduce the accuracy of the analysis. To improve the precision and efficiency of image data analysis, we developed an Attention LSTM U-Net model (FlyNet3.0), which incorporates an attention learning mechanism to track the beating fly heart in OCM images. The new model has improved the intersection over union (IOU) compared to FlyNet2.0 + with reflection artifacts from 86% to 89% and with movement from 81% to 89%. We also extended the capabilities of OCM analysis through the introduction of an automated, in vivo heart wall thickness measurement method, which has been validated on a Drosophila model of cardiac hypertrophy. This work will enable the comprehensive, non-invasive characterization of fly heart physiology in a high-throughput manner.

Physiological and proteomic characterization revealed the response mechanisms underlying aluminium tolerance in lentil (Lens culinaris Medikus).

Aluminium (Al) toxicity causes major plant distress, affecting root growth, nutrient uptake and, ultimately, agricultural productivity. Lentil, which is a cheap source of vegetarian protein, is recognized to be sensitive to Al toxicity. Therefore, it is important to dissect the physiological and molecular mechanisms of Al tolerance in lentil. To understand the physiological system and proteome composition underlying Al tolerance, two genotypes [L-4602 (Al-tolerant) and BM-4 (Al-sensitive)] were studied at the seedling stage. L-4602 maintained a significantly higher root tolerance index and malate secretion with reduced Al accumulation than BM-4. Also, label-free proteomic analysis using ultra-performance liquid chromatography-tandem mass spectrometer exhibited significant regulation of Al-responsive proteins associated with antioxidants, signal transduction, calcium homeostasis, and regulation of glycolysis in L-4602 as compared to BM-4. Functional annotation suggested that transporter proteins (transmembrane protein, adenosine triphosphate-binding cassette transport-related protein and multi drug resistance protein), antioxidants associated proteins (nicotinamide adenine dinucleotide dependent oxidoreductase, oxidoreductase molybdopterin binding protein & peroxidases), kinases (calmodulin-domain kinase & protein kinase), and carbohydrate metabolism associated proteins (dihydrolipoamide acetyltransferase) were found to be abundant in tolerant genotype providing protection against Al toxicity. Overall, the root proteome uncovered in this study at seedling stage, along with the physiological parameters measured, allow a greater understanding of Al tolerance mechanism in lentil, thereby assisting in future crop improvement programmes.

Oerskovia flava sp. nov., a deoxynivalenol (DON)-degrading actinomycete isolated from the rhizosphere soil of long-term continuous cropping cucumber.

The deoxynivalenol (DON)-degrading bacterium JB1-3-2T was isolated from a rhizosphere soil sample of cucumber collected from a greenhouse located in Zhenjiang, Eastern China. The JB1-3-2T strain is a Gram-stain-positive, nonmotile and round actinomycete. Growth was observed at temperatures between 15 and 40 ℃ (optimum, 35 ℃), in the presence of 15% (w/v) NaCl (optimum, 3%), and at pH 3 and 11 (optimum, 7). The major cellular fatty acids identified were anteiso-C15:0, iso-C16:0 and anteiso-C17:0. Genome sequencing revealed a genome size of 4.11Mb and a DNA G + C content of 72.5mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the JB1-3-2T strain was most closely related to type strains of the Oerskovia species, with the highest sequence similarity to Oerskovia turbata NRRL B-8019T (98.2%), and shared 98.1% sequence identity with other valid type strains of this genus. Digital DNA‒DNA hybridization (dDDH) and average nucleotide identity (ANI) showed 21.8-22.2% and 77.2-77.3% relatedness, respectively, between JB1-3-2T and type strains of the genus Oerskovia. Based on genotypic, phylogenetic, chemotaxonomic, physiological and biochemical characterization, Oerskovia flava, a novel species in the genus Oerskovia, was proposed, and the type strain was JB1-3-2T (= CGMCC 1.18555T = JCM 35248T). Additionally, this novel strain has a DON degradation ability that other species in the genus Oerskovia do not possess, and glutathione-S-transferase was speculated to be the key enzyme for strain JB1-3-2T to degrade DON.