In-depth multiomic characterization of the effects of obesity in high-fat diet-fed mice.

High-fat diet (HFD)-fed mice have been widely used in the clinical investigation of obesity. However, the long-term effect of HFD on gut microbiota and metabolites, plasma and liver metabolomics, colonic and liver transcriptomics remain largely unknown. In this study, 6-week-old C57BL/6J male mice fed with HFD for 14 weeks showed increased obesity-related indexes including alanine aminotransferase, aspartate aminotransferase, total cholesterol, total triglyceride, free fatty acids, lipopolysaccharides, IL-6, and TNFα. Furthermore, microbial diversity and richness were also significantly decreased. In the colon, genes involved in tryptophan metabolism, PPAR signaling pathway, cholesterol metabolism, and lipid localization and transport, were upregulated. While in the liver, MAPK signaling and unsaturated fatty acid biosynthesis were upregulated. Metabolomic analyses revealed decreased levels of glycerophospholipids and fatty acyl, but increased amino acids, coenzymes and vitamins, and organic acids in the colon, suggesting high absorption of oxidized lipids, while acyl-carnitine, lysophosphatidylcholine, lysophosphatidylethanolamine, and oxidized lipids were reduced in the liver, suggesting a more active lipid metabolism. Finally, correlation analyses revealed a positive correlation between gut microbiota and metabolites and the expression of genes associated with lipid localization, absorption, and transport in the colon, and nutrients and energy metabolism in the liver. Taken together, our results provide a comprehensive characterization of long-term HFD-induced obesity in mice.

A high-quality genome assembly highlights the evolutionary history of the great bustard (Otis tarda, Otidiformes).

Conservation genomics often relies on non-invasive methods to obtain DNA fragments which limit the power of multi-omic analyses for threatened species. Here, we report multi-omic analyses based on a well-preserved great bustard individual (Otis tarda, Otidiformes) that was found dead in the mountainous region in Gansu, China. We generate a near-complete genome assembly containing only 18 gaps scattering in 8 out of the 40 assembled chromosomes. We characterize the DNA methylation landscape which is correlated with GC content and gene expression. Our phylogenomic analysis suggests Otidiformes and Musophagiformes are sister groups that diverged from each other 46.3 million years ago. The genetic diversity of great bustard is found the lowest among the four available Otidiformes genomes, possibly due to population declines during past glacial periods. As one of the heaviest migratory birds, great bustard possesses several expanded gene families related to cardiac contraction, actin contraction, calcium ion signaling transduction, as well as positively selected genes enriched for metabolism. Finally, we identify an extremely young evolutionary stratum on the sex chromosome, a rare case among birds. Together, our study provides insights into the conservation genomics, adaption and chromosome evolution of the great bustard.

Metabolome and transcriptome signatures shed light on the anti-obesity effect of Polygonatum sibiricum.

Obesity has become one of the major threats to human health across the globe. The rhizomes of Polygonatum sibiricum have shown promising anti-obesity effect. However, the metabolic and genetic basis mediating this beneficial effect are not fully resolved. It is well known that older rhizomes of P. sibiricum exert stronger pharmacological effects. Here, we performed high-resolution metabolome profiling of P. sibiricum rhizomes at different growth stages, and identified that three candidate anti-obesity metabolites, namely phloretin, linoleic acid and α-linolenic acid, accumulated more in adult rhizomes. To elucidate the genetic basis controlling the accumulation of these metabolites, we performed transcriptome profiling of rhizomes from juvenile and adult P. sibiricum. Through third-generation long-read sequencing, we built a high-quality transcript pool of P. sibiricum, and resolved the genetic pathways involved in the biosynthesis and metabolism of phloretin, linoleic acid and α-linolenic acid. Comparative transcriptome analysis revealed altered expression of the genetic pathways in adult rhizomes, which likely lead to higher accumulation of these candidate metabolites. Overall, we identified several metabolic and genetic signatures related to the anti-obesity effect of P. sibiricum. The metabolic and transcriptional datasets generated in this work could also facilitate future research on other beneficial effects of this medicinal plant.

Light-induced stomatal opening in Arabidopsis is negatively regulated by chloroplast-originated OPDA signaling.

Stomatal movement is orchestrated by diverse signaling cascades and metabolic activities in guard cells. Light triggers the opening of the pores through the phototropin-mediated pathway, which leads to the activation of plasma membrane H+-ATPase and thereby facilitates potassium accumulation through Kin+ channels. However, it remains poorly understood how phototropin signaling is fine-tuned to prevent excessive stomatal opening and consequent water loss. Here, we show that the stomatal response to light is negatively regulated by 12-oxo-phytodienoic acid (OPDA), an oxylipin metabolite produced through enzymatic oxygenation of polyunsaturated fatty acids (PUFAs). We identify a set of phospholipase-encoding genes, phospholipase (PLIP)1/2/3, which are transactivated rapidly in guard cells upon illumination in a phototropin-dependent manner. These phospholipases release PUFAs from the chloroplast membrane, which is oxidized by guard-cell lipoxygenases and further metabolized to OPDA. The OPDA-deficient mutants had wider stomatal pores, whereas mutants containing elevated levels of OPDA showed the opposite effect on stomatal aperture. Transmembrane solute fluxes that drive stomatal aperture were enhanced in lox6-1 guard cells, indicating that OPDA signaling ultimately impacts on activities of proton pumps and Kin+ channels. Interestingly, the accelerated stomatal kinetics in lox6-1 leads to increased plant growth without cost in water or macronutrient use. Together, our results reveal a new role for chloroplast membrane oxylipin metabolism in stomatal regulation. Moreover, the accelerated stomatal opening kinetics in OPDA-deficient mutants benefits plant growth and water use efficiency.

Arsenic Accumulation and Physiological Response of Three Leafy Vegetable Varieties to As Stress.

Arsenic (As) in leafy vegetables may harm humans. Herein, we assessed As accumulation in leafy vegetables and the associated physiological resistance mechanisms using soil pot and hydroponic experiments. Garland chrysanthemum (Chrysanthemum coronarium L.), spinach (Spinacia oleracea L.), and lettuce (Lactuca sativa L.) were tested, and the soil As safety threshold values of the tested leafy vegetables were 91.7, 76.2, and 80.7 mg kg−1, respectively, i.e., higher than the soil environmental quality standard of China. According to growth indicators and oxidative stress markers (malondialdehyde, the ratio of reduced glutathione to oxidized glutathione, and soluble protein), the order of As tolerance was: GC > SP > LE. The high tolerance of GC was due to the low transport factor of As from the roots to the shoots; the high activity of superoxide dismutase, glutathione peroxidase, and catalase; and the high content of phytochelatin in the roots. Results of this work shed light on the use of As-contaminated soils and plant tolerance of As stress.

Calcium-dependent ABA signaling functions in stomatal immunity by regulating rapid SA responses in guard cells.

Stomatal immunity is mediated by ABA, an osmotic stress-responsive phytohormone that closes stomata via calcium-dependent and -independent signaling pathways. However, the functional involvement of ABA signal transducers in stomatal immunity remains poorly understood. Here, we demonstrate that stomatal immunity was compromised in mutants of the ABA signaling core. We also found that it is a subset of calcium-dependent protein kinases (CPK4/5/6), but not the calcium-independent kinase OST1, that relay the stomatal immune signaling. Surface-inoculated bacteria caused an endogenous ABA-dependent induction of local SA responses, whilst expression of the ABA biosynthetic genes and the ABA levels were not affected in leaf epidermis. Furthermore, flg22-elicited ROS burst was attenuated by mutations in CPK4 and CPK5, and pathogen-induced SA production in leaf epidermis was compromised in cpk4, cpk5, and cpk6 mutants. Our results suggest that CPKs function in stomatal immunity through fine-tuning apoplastic ROS levels as well as reinforcing the localized SA signal in guard cells. It is also envisioned that ABA mediates stomatal responses to biotic and abiotic stresses via two distinct but partially overlapping signaling modules.

The largest negative carbon isotope excursions in Neoproterozoic carbonates caused by recycled carbonatite volcanic ash.

The late Ediacaran Shuram Excursion (SE) records the most prominent negative δ13C excursions (δ13C = -12‰) during Earth's history. It has been hypothesized to have resulted from oxidation of dissolved organic matter, diagenetic or authigenic precipitates. However, the origin of the SE remains enigmatic; current models face challenges regarding the significant amount of atmospheric oxygen needed to balance such extensive oxidation and sustained inputs of light carbon with extremely negative C isotope compositions. Here, we show that the Doushantuo Formation at the Jiulongwan section in South China, a key stratum recording the SE event, contains mineralogical and geochemical signatures related to igneous processes. Both the occurrence of ankerite, feldspar, moissanite and euhedral quartz in the SE samples and the relatively consistent Ce anomalies of carbonate and O isotopes of quartz indicate a contribution from an igneous source. In particular, the SE samples have trace element and C isotope compositions similar to those of recycled carbonatites formed by decarbonation and melting of sedimentary carbonate rocks. These observations suggest that the deep cycle of ancient carbonate rocks, which were subjected to decarbonation during subduction, melting and eruption related to the breakup of the Rodinia supercontinent, contributed to the SE. This igneous model for the SE may provide a connection between the deep and shallow carbon cycles of the Earth.

The complete chloroplast genome of Hemerocallis citrina (Asphodelaceae), an ornamental and medicinal plant.

Hemerocallis citrina (Asphodelaceae) has been wildly cultivated as ornamental and medicinal plant. Here, we reported the first chloroplast genome sequence of H. citrina. The chloroplast genome size is 156,088 bp with GC content of 37.3%, including a large single-copy (LSC) of 84,843 bp, a small single-copy (SSC) of 18,507 bp, and a pair of 26,369 bp IR(inverted repeat) regions. A total of 133 genes were annotated including 87 protein-coding genes, 38 tRNA genes, and 8 rRNA genes. The phylogenetic analysis revealed that H. citrina belongs to the Hemerocallis genus in Asphodelaceae family.

The complete chloroplast genome of Acanthopanax brachypus (Araliaceae), a famous medicinal plant in China.

Acanthopanax brachypus (Araliaceae) is an important medicinal plant originated from China. Here, we reported the first chloroplast genome sequence of A. brachypus. The plastome of A. brachypu is 156,802 bp in length, including a large single-copy (LSC) of 86,742 bp, a small single-copy (SSC) of 18,184 bp, and a pair of inverted repeat regions (IRa and IRb) of 25,938 bp. It contains 113 unique genes consisting of 80 protein-coding genes, 29 tRNA genes, and 4 rRNA genes.

The complete chloroplast genome of Tagetes erecta (Asteroideae), a wildly cultivated ornamental plant.

Tagetes erecta (Asteraceae) has been wildly cultivated as ornamental and medicinal plant. Here, we reported the first chloroplast genome sequence of T. erecta. The chloroplast genome size is 152,065 bp with GC content of 37.4%, including a large single-copy (LSC) of 83,895 bp, a small single-copy (SSC) of 18,065 bp, and a pair of 25,048 bp IR (inverted repeat) regions. A total of 132 genes were annotated including 87 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The phylogenetic analysis revealed that T. erecta belongs to the subfamily Asteroideae.

Arabidopsis ADF5 promotes stomatal closure by regulating actin cytoskeleton remodeling in response to ABA and drought stress.

Stomatal movement plays an essential role in plant responses to drought stress, and the actin cytoskeleton and abscisic acid (ABA) are two important components of this process. Little is known about the mechanism underlying actin cytoskeleton remodeling and the dynamic changes occurring during stomatal movement in response to drought stress/ABA signaling. Actin-depolymerizing factors (ADFs) are conserved actin severing/depolymerizing proteins in eukaryotes, and in angiosperms ADFs have evolved actin-bundling activity. Here, we reveal that the transcriptional expression of neofunctionalized Arabidopsis ADF5 was induced by drought stress and ABA treatment. Furthermore, we demonstrated that ADF5 loss-of-function mutations increased water loss from detached leaves, reduced plant survival rates after drought stress, and delayed stomatal closure by regulating actin cytoskeleton remodeling via its F-actin-bundling activity. Biochemical assays revealed that an ABF/AREB transcription factor, DPBF3, could bind to the ADF5 promoter and activate its transcription via the ABA-responsive element core motif ACGT/C. Taken together, our findings indicate that ADF5 participates in drought stress by regulating stomatal closure, and may also serve as a potential downstream target of the drought stress/ABA signaling pathway via members of the ABF/AREB transcription factors family.

Actin filament reorganisation controlled by the SCAR/WAVE complex mediates stomatal response to darkness.

Stomata respond to darkness by closing to prevent excessive water loss during the night. Although the reorganisation of actin filaments during stomatal closure is documented, the underlying mechanisms responsible for dark-induced cytoskeletal arrangement remain largely unknown. We used genetic, physiological and cell biological approaches to show that reorganisation of the actin cytoskeleton is required for dark-induced stomatal closure. The opal5 mutant does not close in response to darkness but exhibits wild-type (WT) behaviour when exposed to abscisic acid (ABA) or CaCl2 . The mutation was mapped to At5g18410, encoding the PIR/SRA1/KLK subunit of the ArabidopsisSCAR/WAVE complex. Stomata of an independent allele of the PIR gene (Atpir-1) showed reduced sensitivity to darkness and F1 progenies of the cross between opal5 and Atpir-1 displayed distorted leaf trichomes, suggesting that the two mutants are allelic. Darkness induced changes in the extent of actin filament bundling in WT. These were abolished in opal5. Disruption of filamentous actin using latrunculin B or cytochalasin D restored wild-type stomatal sensitivity to darkness in opal5. Our findings suggest that the stomatal response to darkness is mediated by reorganisation of guard cell actin filaments, a process that is finely tuned by the conserved SCAR/WAVE-Arp2/3 actin regulatory module.

Stomata prioritize their responses to multiple biotic and abiotic signal inputs.

Stomata are microscopic pores in leaf epidermis that regulate gas exchange between plants and the environment. Being natural openings on the leaf surface, stomata also serve as ports for the invasion of foliar pathogenic bacteria. Each stomatal pore is enclosed by a pair of guard cells that are able to sense a wide spectrum of biotic and abiotic stresses and respond by precisely adjusting the pore width. However, it is not clear whether stomatal responses to simultaneously imposed biotic and abiotic signals are mutually dependent on each other. Here we show that a genetically engineered Escherichia coli strain DH5α could trigger stomatal closure in Vicia faba, an innate immune response that might depend on NADPH oxidase-mediated ROS burst. DH5α-induced stomatal closure could be abolished or disguised under certain environmental conditions like low [CO2], darkness, and drought, etc. Foliar spraying of high concentrations of ABA could reduce stomatal aperture in high humidity-treated faba bean plants. Consistently, the aggressive multiplication of DH5α bacteria in Vicia faba leaves under high humidity could be alleviated by exogenous application of ABA. Our data suggest that a successful colonization of bacteria on the leaf surface is correlated with stomatal aperture regulation by a specific set of environmental factors.