Transcriptomic and metabolomic analyses reveal the differential accumulation of phenylpropanoids and terpenoids in hemp autotetraploid and its diploid progenitor.

BACKGROUND: Cannabis sativa, a dioecious plant that has been cultivated worldwide for thousands of years, is known for its secondary metabolites, especially cannabinoids, which possess several medicinal effects. In this study, we investigated the autopolyploidization effects on the biosynthesis and accumulation of these metabolites, transcriptomic and metabolomic analyses were performed to explore the gene expression and metabolic variations in industrial hemp autotetraploids and their diploid progenitors. RESULTS: Through these analyses, we obtained 1,663 differentially expressed metabolites and 1,103 differentially expressed genes. Integrative analysis revealed that phenylpropanoid and terpenoid biosynthesis were regulated by polyploidization. No substantial differences were found in the cannabidiol or tetrahydrocannabinol content between tetraploids and diploids. Following polyploidization, some transcription factors, including nine bHLH and eight MYB transcription factors, affected the metabolic biosynthesis as regulators. Additionally, several pivotal catalytic genes, such as flavonol synthase/flavanone 3-hydroxylase, related to the phenylpropanoid metabolic pathway, were identified as being modulated by polyploidization. CONCLUSIONS: This study enhances the overall understanding of the impact of autopolyploidization in C. sativa and the findings may encourage the application of polyploid breeding for increasing the content of important secondary metabolites in industrial hemp.

Multi-omics provides new insights into the domestication and improvement of dark jute (Corchorus olitorius).

Jute (Corchorus sp.) is the most important bast fiber crop worldwide; however, the mechanisms underlying domestication and improvement remain largely unknown. We performed multi-omics analysis by integrating de novo sequencing, resequencing, and transcriptomic and epigenetic sequencing to clarify the domestication and improvement of dark jute Corchorus olitorius. We demonstrated that dark jute underwent early domestication and a relatively moderate genetic bottleneck during improvement breeding. A genome-wide association study of 11 important agronomic traits identified abundant candidate loci. We characterized the selective sweeps in the two breeding stages of jute, prominently, soil salinity differences played an important role in environmental adaptation during domestication, and the strongly selected genes for improvement had an increased frequency of favorable haplotypes. Furthermore, we speculated that an encoding auxin/indole-3-acetic acid protein COS07g_00652 could enhance the flexibility and strength of the stem to improve fiber yield. Our study not only provides valuable genetic resources for future fiber breeding in jute, but also is of great significance for reviewing the genetic basis of early crop breeding.

Establishment of an Agrobacterium-mediated genetic transformation and CRISPR/Cas9-mediated targeted mutagenesis in Hemp (Cannabis Sativa L.).

Hemp (Cannabis sativa L.) is an annual and typically dioecious crop. Due to the therapeutic potential for human diseases, phytocannabinoids as a medical therapy is getting more attention recently. Several candidate genes involved in cannabinoid biosynthesis have been elucidated using omics analysis. However, the gene function was not fully validated due to few reports of stable transformation for Cannabis tissues. In this study, we firstly report the successful generation of gene-edited plants using an Agrobacterium-mediated transformation method in C. sativa. DMG278 achieved the highest shoot induction rate, which was selected as the model strain for transformation. By overexpressing the cannabis developmental regulator chimera in the embryo hypocotyls of immature grains, the shoot regeneration efficiency was substantially increased. We used CRISPR/Cas9 technology to edit the phytoene desaturase gene and finally generated four edited cannabis seedlings with albino phenotype. Moreover, we propagated the transgenic plants and validated the stable integration of T-DNA in cannabis genome.

Insights into the mechanism of multi-walled carbon nanotubes phytotoxicity in Arabidopsis through transcriptome and m6A methylome analysis.

With the increasing production and wide application of carbon nanotubes (CNTs), they are inevitably released into the natural environment and ecosystems, where plants are the main primary producers. Hence, it is imperative to understand the toxic effects of CNTs on plants. The molecular mechanisms underlying the toxic effects of CNTs on plants are still unclear. Therefore, in the present study, we investigated the effects of high concentrations of multi-walled CNTs (MWCNTs) on Arabidopsis. Root elongation and leaf development were severely inhibited after MWCNT exposure. Excess production of H2O2, O2-, and malondialdehyde was observed, indicating that MWCNTs induced oxidative stress. The antioxidant system was activated to counter MWCNTs-induced oxidative stress. Combinatorial transcriptome and m6A methylome analysis revealed that MWCNTs suppressed auxin signaling and photosynthesis. Reactive oxygen species metabolism, toxin metabolism, and plant responses to pathogens were enhanced to cope with the phytotoxicity of MWCNTs. Our results provide new insights into the molecular mechanisms of CNT phytotoxicity and plant defense responses to CNTs.

Differential Proteomic Analysis to Identify Proteins Associated with Apomeiosis in Boehmeria tricuspis (Hance) Makino Using an iTRAQ-Based Strategy.

Numerous candidate genes related to apomixis have been identified through transcriptomics; however, the molecular mechanism underlying apomixis remains unclear. Elucidation of the underlying mechanisms is essential to expand its application in crop breeding. Therefore, here, we employed the isobaric tags for a relative and absolute quantification labeling technology to investigate the protein expression in Boehmeria tricuspis generated through different reproductive modes at the functional megaspore stage. We identified 40 differential abundance proteins associated with apomeiosis, most of which were involved in "response to stress". Functional analysis suggested that lower levels of reactive oxygen species (ROS) play a role in inducing the development of apomeiosis. Proteins related to ROS regulation, cell wall modifications, and stability under heat stress play a crucial role in the development of diplosporic apomeiosis. Our results give evidence to the insight that stress can induce a switch from apomixis to sexuality by ROS content, and an increased composition of stress tolerance as well as secondary metabolites can buffer ROS effects. Precise coordination of these proteins involved in inter-related regulatory control mechanisms may act together in the transition from the sexual to apomixis development.

Gene coexpression network analysis and tissue-specific profiling of gene expression in jute (Corchorus capsularis L.).

BACKGROUND: Jute (Corchorus spp.), belonging to the Malvaceae family, is an important natural fiber crop, second only to cotton, and a multipurpose economic crop. Corchorus capsularis L. is one of the only two commercially cultivated species of jute. Gene expression is spatiotemporal and is influenced by many factors. Therefore, to understand the molecular mechanisms of tissue development, it is necessary to study tissue-specific gene expression and regulation. We used weighted gene coexpression network analysis, to predict the functional roles of gene coexpression modules and individual genes, including those underlying the development of different tissue types. Although several transcriptome studies have been conducted on C. capsularis, there have not yet been any systematic and comprehensive transcriptome analyses for this species. RESULTS: There was significant variation in gene expression between plant tissues. Comparative transcriptome analysis and weighted gene coexpression network analysis were performed for different C. capsularis tissues at different developmental stages. We identified numerous tissue-specific differentially expressed genes for each tissue, and 12 coexpression modules, comprising 126 to 4203 genes, associated with the development of various tissues. There was high consistency between the genes in modules related to tissues, and the candidate upregulated genes for each tissue. Further, a gene network including 21 genes directly regulated by transcription factor OMO55970.1 was discovered. Some of the genes, such as OMO55970.1, OMO51203.1, OMO50871.1, and OMO87663.1, directly involved in the development of stem bast tissue. CONCLUSION: We identified genes that were differentially expressed between tissues of the same developmental stage. Some genes were consistently up- or downregulated, depending on the developmental stage of each tissue. Further, we identified numerous coexpression modules and genes associated with the development of various tissues. These findings elucidate the molecular mechanisms underlying the development of each tissue, and will promote multipurpose molecular breeding in jute and other fiber crops.

Construction of a high-resolution genetic map and identification of quantitative trait loci for salt tolerance in jute (Corchous spp.).

BACKGROUND: Jute (Corchorus spp.) is the most important natural fiber crop after cotton in terms of cultivation area and production. Salt stress greatly restricts plant development and growth. A high-density genetic linkage map is the basis of quantitative trait locus (QTLs) mapping. Several high-density genetic maps and QTLs mapping related to salt tolerance have been developed through next-generation sequencing in many crop species. However, such studies are rare for jute. Only several low-density genetic maps have been constructed and no salt tolerance-related QTL has been mapped in jute to date. RESULTS: We developed a high-density genetic map with 4839 single nucleotide polymorphism markers spanning 1375.41 cM and an average distance of 0.28 cM between adjacent markers on seven linkage groups (LGs) using an F2 jute population, LGs ranged from LG2 with 299 markers spanning 113.66 cM to LG7 with 1542 markers spanning 350.18 cM. In addition, 99.57% of gaps between adjacent markers were less than 5 cM. Three obvious and 13 minor QTLs involved in salt tolerance were identified on four LGs explaining 0.58-19.61% of the phenotypic variance. The interval length of QTL mapping varied from 1.3 to 20.2 cM. The major QTL, qJST-1, was detected under two salt stress conditions that explained 11.81 and 19.61% of the phenotypic variation, respectively, and peaked at 19.3 cM on LG4. CONCLUSIONS: We developed the first high-density and the most complete genetic map of jute to date using a genotyping-by-sequencing approach. The first QTL mapping related to salt tolerance was also carried out in jute. These results should provide useful resources for marker-assisted selection and transgenic breeding for salt tolerance at the germination stage in jute.

Combined genome-wide association analysis and transcriptome sequencing to identify candidate genes for flax seed fatty acid metabolism.

Flax seeds have a high oil content and are rich in unsaturated fatty acids, which have advantageous effects in preventing chronic diseases, such as cardiovascular diseases. At present, flax seeds are mainly developed for oil. Therefore, it is of practical significance to identify the candidate genes of fatty acid metabolism in flax seeds for breeding flax seeds with high oil content. In the present study, a natural population of flax containing 224 samples planted in 3 different environments was studied. The genome-wide association analysis (GWAS) of seed fatty acid content was conducted based on specific length amplified fragment sequencing (SLAF-seq) data. Transcriptome sequencing (RNA-seq) of samples from 3 different periods (14 d, 21 d and 28 d after anthesis) during seed development of the low oil variety Shuangya 4 and the high oil variety NEW was performed. The candidate genes for seed fatty acid metabolism were identified by combined analysis of these 2 methods. GWAS detected 16 SNP loci significantly associated with seed fatty acid content, and RNA-seq analysis identified 11,802 differentially expressed genes between high and low oil samples. Pathway enrichment analysis revealed that some differentially expressed genes were classified into fatty acid-related pathways. After comparison of these differentially expressed genes with the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, 20 genes homologous to other species were obtained. After analysis, 10 candidate genes were screened by GWAS and RNA-seq screening. Of these 10 genes, qRT-PCR assays using flax seeds in 5 different developmental stages showed that the expression levels of 6 candidate genes were significantly correlated with 5 fatty acid contents in seeds of the high oil variety NEW. Through metabolic pathway analysis found that 6 genes were involved in important fatty acid metabolic pathways, and some of them also have upstream and downstream regulation relations. The present study combined GWAS and RNA-seq methods to identify candidate genes for fatty acid metabolism in flax seeds, which provided reference for screening of candidate genes with complex traits.

A Full-Length Reference Floral Transcriptome of Boehmeria tricuspis Provides Insights into Apomeiosis and Polyploidy.

Boehmeria tricuspis (Hance) Makino constitutes a hardy herbaceous or shrubby perennial native to East Asia that includes different ploidy levels and reproductive modes (diplosporous to sexual). Although several apomeiosis-associated genes have been described, the genetic control and molecular mechanisms underlying apomeiosis remain poorly understood. Moreover, the basis of the correlation between polyploidy and apomixis has not yet been clarified. We utilized long-read sequencing to produce a full-length reference floral transcriptome of B. tricuspis. Based on the generated database, gene expression of the female flowers of different ploidy levels and reproductive mode cytotypes was compared. Overall, 1,387 genes related to apomeiosis, 217 genes related to ploidy, and 9 genes associated with both apomixis and ploidy were identified. Gene Ontology analyses of this set of transcripts indicated reproductive genes, especially those related to "cell differentiation" and "cell cycle process," as significant factors regulating apomeiosis. Furthermore, our results suggested that different expressions of stress response genes might be important in the preparation for apomeiosis transition. In addition, our observations indicated that the expression of apomeiosis may not depend on polyploidy but rather on deregulation of the sexual pathway in B. tricuspis.

Enhanced Escherichia coli inactivation and oxytetracycline hydrochloride degradation by a Z-scheme silver iodide decorated bismuth vanadate nanocomposite under visible light irradiation.

Novel Z-scheme AgI/BiVO4 photocatalysts were fabricated by a chemical deposition-precipitation approach. The photocatalytic activities of the obtained catalysts were evaluated by disinfection of Escherichia coli (E. coli) and degradation of oxytetracycline hydrochloride (OTC-HCl) under visible-light irradiation. The BA3 (contained 9.09% of AgI) exhibited the highest photocatalytic activity and maintained good stability. It could completely inactivate 7.0×107 CFU/mL of E. coli in 50 min and degrade 80% of OTC-HCl in 60 min. The enhanced photocatalytic activity of AgI/BiVO4 composites could be ascribed to the lower recombination rate of electron-hole pairs. Meanwhile, radical trapping experiments revealed that the superoxide radical (O2-) and holes (h+) were the dominant reactive species in photo-disinfection process. Furthermore, the effects of bacterial initial concentration and inorganic anions were also investigated to optimize the photocatalyst for practical application. This study will give a new insight to construct the effective Z-scheme system for bacterial inactivation and organic pollutants degradation.

Graphene sponge decorated with copper nanoparticles as a novel bactericidal filter for inactivation of Escherichia coli.

Nanotechnology has great potential in water purification. However, the limitations such as aggregation and toxicity of nanomaterials have blocked their practical application. In this work, a novel copper nanoparticles-decorated graphene sponge (Cu-GS) was synthesized using a facile hydrothermal method. Cu-GS consisting of three-dimensional (3D) porous graphene network and well-dispersed Cu nanoparticles exhibited high antibacterial efficiency against Esherichia coli when used as a bactericidal filter. The morphological changes determined by scanning electron microscope and fluorescence images measured by flow cytometry confirmed the involvement of membrane damage induced by Cu-GS in their antibacterial process. The oxidative ability of Cu-GS and intercellular reactive oxygen species (ROS) were also determined to elucidate the possible antibacterial mechanism of Cu-GS. Moreover, the concentration of released copper ions from Cu-GS was far below the drinking water standard, and the copper ions also have an effect on the antibacterial activity of Cu-GS. Results suggested that Cu-GS as a novel bactericidal filter possessed a potential application of water disinfection.

Effect of multi-walled carbon nanotubes on phytotoxicity of sediments contaminated by phenanthrene and cadmium.

To implement effective control and abatement programs for contaminants accumulating in sediments, strategies are needed for evaluating the quality of amended sediments. In this study, phytotoxicity of the sediments contaminated by cadmium and phenanthrene was evaluated after in situ remediation with multi-walled carbon nanotubes (MWCNTs) as adsorbents. Adsorption experiments and measurement of aqueous concentrations of the contaminants in overlying water were used to investigate the remediation effectiveness from physical and chemical aspects. The results indicated that MWCNTs showed a much better adsorption performance towards phenanthrene and Cd(II) compared with the sediments. The in situ remediation with MWCNTs could distinctly decrease the aqueous concentrations of phenanthrene and Cd(II) released from the sediments, reducing environmental risk towards overlying water. Influences of MWCNTs dose, MWCNTs diameter, and contact time on phtotoxicity of the contaminated sediments were studied. No significant inhibition of the amended sediments on germination of the test species was observed in the experiments, while the root growth was more sensitive than biomass production to the changes of contaminant concentrations. The analysis of Pearson correlation coefficients between evaluation indicators and associated remediation parameters suggested that phytotoxicity of sediments might inaccurately indicate the changes of pollutant content, but it was significant in reflecting the ecotoxicity of sediments after remediation.

Preparation of melamine sponge decorated with silver nanoparticles-modified graphene for water disinfection.

This paper reports the fabrication of melamine sponge decorated with silver nanoparticles-modified graphene (G/AgNPs-MS) for water disinfection. The G/AgNPs-MS composites with the high porosity and elasticity were used in an antibacterial process in which the composite was first immersed in bacterial suspension, and subsequently squeezed via hand compression. G/AgNPs-MS exhibited more excellent bactericidal activity against Escherichia coli and Staphylococcus aureus bacteria compared with melamine sponge (MS), melamine sponge decorated with graphene (G-MS), and melamine sponge decorated with silver nanoparticles (AgNPs-MS). The superior antibacterial effect was possibly ascribed to the coordination of graphene oxide (GO) and silver nanoparticles (Ag NPs). Compared to AgNPs-MS, G/AgNPs-MS displayed better stability with fewer Ag+ release. G/AgNPs-MS composites were highly reusable with no significant differences in the loss of bacterial viability over 12 operational cycles. The possible antibacterial mechanism of G/AgNPs-MS was also investigated. It was found that the destruction of bacterial membrane by G/AgNPs-MS played an important role in the bactericidal activity. The generation of intercellular ROS and scavenging assays confirmed the involvement of Ag+ and ROS in the antibacterial process of G/AgNPs-MS. All the results demonstrated that the prepared G/AgNPs-MS composites, as innovative antibacterial materials, showed a great potential for water disinfection.

The disinfection performance and mechanisms of Ag/lysozyme nanoparticles supported with montmorillonite clay.

The fabrication of montmorillonite (Mt) decorated with lysozyme-modified silver nanoparticles (Ag/lyz-Mt) was reported. The lysozyme (lyz) was served as both reducing and capping reagent. Coupling the bactericidal activity of the lyz with AgNPs, along with the high porous structure and large specific surface area of the Mt, prevented aggregation of AgNPs and promoted nanomaterial-bacteria interactions, resulting in a greatly enhanced bactericidal capability against both Gram positive and Gram negative bacteria. This paper systematically elucidated the bactericidal mechanisms of Ag/lyz-Mt. Direct contact between the Ag/lyz-Mt surface and the bacterial cell was essential to the disinfection. Physical disruption of bacterial membrane was considered to be one of the bactericidal mechanisms of Ag/lyz-Mt. Results revealed that Ag(+) was involved in the bactericidal activity of Ag/lyz-Mt via tests conducted using Ag(+) scavengers. A positive ROS (reactive oxygen species) scavenging test indirectly confirmed the involvement of ROS (O2(-), H2O2, and OH) in the bactericidal mechanism. Furthermore, the concentrations of individual ROS were quantified. Results showed that Ag/lyz-Mt nanomaterial could be a promising bactericide for water disinfection.

Magnetic chitosan-graphene oxide composite for anti-microbial and dye removal applications.

Magnetic chitosan-graphene oxide (MCGO) nanocomposite was prepared as a multi-functional nanomaterial for the applications of antibacterial and dye removal. The nanocomposite was characterized by scanning electronic microscope (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD) and Fourier transform infrared spectrometer (FTIR). The antibacterial performance for MCGO against Escherichia coli was varied depending on the concentration of MCGO. SEM images of E. coli cells demonstrated that the antimicrobial performance of MCGO nanocomposite was possibly due to the damage of cell membrane. This work also explored MCGO's adsorption performance for methyl orange (MO). The experimental parameters including adsorbent mass, pH value, contact time and concentration of MO on the adsorption capacity were investigated. The maximum adsorption capacity of MCGO for MO was 398.08 mg/g. This study showed that the MCGO offered enormous potential applications for water treatment.

Inactivation performance and mechanism of Escherichia coli in aqueous system exposed to iron oxide loaded graphene nanocomposites.

The challenge to achieve efficient disinfection and microbial control without harmful disinfection byproducts calls for developing new technologies. Magnetic-graphene oxide (M-GO) with magnetic iron oxide nanoparticles well dispersed on graphene oxide (GO) nanosheets exerted excellent antibacterial activity against Escherichia coli. The antibacterial performance of M-GO was dependent on the concentration and the component mass ratio of M/GO. The synergetic antibacterial effect of M-GO was observed with M/GO mass ratio of 9.09. TEM images illustrated the interaction between E. coli cells and M-GO nanocomposites. M-GO nanomaterials were possible to deposit on or penetrate into cells leading to leakage of intercellular contents and loss of cell integrity. The inactivation mechanism of E. coli by M-GO was supposed to result from both the membrane stress and oxidation stress during the incubation period. M-GO with excellent antibacterial efficiency against E. coli and separation-convenient property from water could be potent bactericidal nanomaterials for water disinfection.

Highly sensitive strategy for Hg2+ detection in environmental water samples using long lifetime fluorescence quantum dots and gold nanoparticles.

The authors herein described a time-gated fluorescence resonance energy transfer (TGFRET) sensing strategy employing water-soluble long lifetime fluorescence quantum dots and gold nanoparticles to detect trace Hg(2+) ions in aqueous solution. The water-soluble long lifetime fluorescence quantum dots and gold nanoparticles were functionalized by two complementary ssDNA, except for four deliberately designed T-T mismatches. The quantum dot acted as the energy-transfer donor, and the gold nanoparticle acted as the energy-transfer acceptor. When Hg(2+) ions were present in the aqueous solution, DNA hybridization will occur because of the formation of T-Hg(2+)-T complexes. As a result, the quantum dots and gold nanoparticles are brought into close proximity, which made the energy transfer occur from quantum dots to gold nanoparticles, leading to the fluorescence intensity of quantum dots to decrease obviously. The decrement fluorescence intensity is proportional to the concentration of Hg(2+) ions. Under the optimum conditions, the sensing system exhibits the same liner range from 1 × 10(-9) to 1 × 10(-8) M for Hg(2+) ions, with the detection limits of 0.49 nM in buffer and 0.87 nM in tap water samples. This sensor was also used to detect Hg(2+) ions from samples of tap water, river water, and lake water spiked with Hg(2+) ions, and the results showed good agreement with the found values determined by an atomic fluorescence spectrometer. In comparison to some reported colorimetric and fluorescent sensors, the proposed method displays the advantage of higher sensitivity. The TGFRET sensor also exhibits excellent selectivity and can provide promising potential for Hg(2+) ion detection.