The delayed senescence in harvested blueberry by hydrogen-based irrigation is functionally linked to metabolic reprogramming and antioxidant machinery.

Molecular hydrogen is beneficial for fruits quality improvement. However, the mechanism involved, especially cellular metabolic responses, has not been well established. Here, the integrated widely targeted metabolomics analysis (UPLC-MS/MS) and biochemical evidence revealed that hydrogen-based irrigation could orchestrate, either directly or indirectly, an array of physiological responses in blueberry (Vaccinium spp.) during harvesting stage, especially for the delayed senescence in harvested stage (4 °C for 12 d). The hubs to these changes are wide-ranging metabolic reprogramming and antioxidant machinery. A total of 1208 distinct annotated metabolites were identified, and the characterization of differential accumulated metabolites (DAMs) revealed that the reprogramming, particularly, involves phenolic acids and flavonoids accumulation. These changes were positively matched with the transcriptional profiles of representative genes for their synthesis during the growth stage. Together, our findings open a new window for development of hydrogen-based agriculture that increases the shelf-life of fruits in a smart and sustainable manner.

Argon-stimulated nitric oxide production and its function in alfalfa cadmium tolerance.

Recent results showed that argon may have great potential in both medicines (especially) and agriculture. However, how argon positively influences crop physiology remains elusive. Here, we observed that the stimulation of nitric oxide (NO) production upon cadmium (Cd) stress in hydroponic alfalfa root tissues was strengthened by argon-rich water and/or a NO-releasing compound. The pharmacological results further indicated that above potential source of NO stimulation achieved by argon might be attributed to NO synthase (NOS) and nitrate reductase (NR). Under hydroponic and pot conditions, the improvement of Cd tolerance elicited by argon, confirmed by the alleviation in the plant growth inhibition, oxidative damage, and Cd accumulation, was sensitive to the scavenger of NO. These results suggested a crucial role of argon-induced NO synthesis in response to Cd stress. Subsequent evidence showed that the improved iron homeostasis and increased S-nitrosylation were also dependent on argon-stimulated NO. Above results were matched with the transcriptional profiles of representative target genes involved in heavy metal detoxification, antioxidant defence, and iron homeostasis. Taken together, our results clearly indicated that argon-stimulated NO production contributes to Cd tolerance by favoring important defense strategies against heavy metal exposure.

An unexpected discovery toward argon-rich water amelioration of cadmium toxicity in Medicago sativa L.

Argon has organ-protective effects on animals. However, whether or how argon influences plant responses remains elusive. In this study, we discovered that the growth inhibition of hydroponically cultured alfalfa seedlings under 100 µM CdCl2 condition was significantly ameliorated by 100 % saturated argon-rich water (ARW). Less Cd uptake and accumulation were also observed in both root and shoot parts, which could be explained by the modified root cell walls, including the increased cell wall thickness, lignin content, and demethylation degree of covalently bound and ion-bound pectin, as well as the down-regulated expression of natural-resistance-associated-macrophage protein1 (Nramp1) encoding a heavy metal ion transporter in root tissue. The hindered Cd translocation from root to shoot achieved by ARW addition was validated by the decreased expression of heavy metal ATPase 2/4 (HMA2/4) in roots and decreased Cd content in xylem saps. The reestablished glutathione (GSH) homeostasis and redox balance, two important indicators of plant defense against Cd poisoning, were also observed. Further greenhouse experiments demonstrated that the phenotypic and physiological performances of alfalfa plants cultured in Cd-contaminated soil were significantly improved by irrigating with ARW. Above results implied that ARW confers plants tolerance against cadmium toxicity by impairing Cd uptake and accumulation and restoring GSH and redox homeostasis. These findings might open a new window for understanding argon biology in higher plants.

Packaging with hydrogen gas modified atmosphere can extend chicken egg storage.

BACKGROUND: Although hydrogen gas (H2 ) has been widely used in industry and gradually used in medicine, it is less applied to agriculture, especially in modified atmosphere packing (MAP). RESULTS: During chicken egg storage, H2 MAP not only slowed down or delayed the reduction in antioxidant capacities, but also alleviated the deterioration of egg quality. The extent of micro-cracks in the eggshell was also negatively influenced by H2 , which might result in eggs being less vulnerable to microbes. The earlier results demonstrated that the H2 MAP-extended shelf life of chicken eggs might be caused by the responses of eggshell and re-establishment of redox homeostasis. According to the data collected from different provinces of China, cost-economics analysis further suggested that the increase in the extra cost caused by H2 was trivial compared with the original price of eggs. CONCLUSION: Together, H2 MAP can prolong the shelf life of chicken eggs with the prospect of wider application. © 2021 Society of Chemical Industry.

Rice GLUTATHIONE PEROXIDASE1-mediated oxidation of bZIP68 positively regulates ABA-independent osmotic stress signaling.

Osmotic stress caused by drought and high salinity is a significant environmental threat that limits plant growth and agricultural yield. Redox regulation plays an important role in plant stress responses, but the mechanisms by which plants perceive and transduce redox signals are still underexplored. Here, we report a critical function for the thiol peroxidase GPX1 in osmotic stress response in rice, where it serves as a redox sensor and transducer. GPX1 is quickly oxidized upon exposure to osmotic stress and forms an intramolecular disulfide bond, which is required for the activation of bZIP68, a VRE-like basic leucine zipper (bZIP) transcription factor involved in the ABA-independent osmotic stress response pathway. The disulfide exchange between GPX1 and bZIP68 induces homo-tetramerization of bZIP68 and thus positively regulates osmotic stress response by regulating osmotic-responsive gene expression. Furthermore, we discovered that the nuclear translocation of GPX1 is regulated by its acetylation under osmotic stress. Taken together, our findings not only uncover the redox regulation of the GPX1-bZIP68 module during osmotic stress but also highlight the coordination of protein acetylation and redox signaling in plant osmotic stress responses.

Hydrogen-rich water prepared by ammonia borane can enhance rapeseed (Brassica napus L.) seedlings tolerance against salinity, drought or cadmium.

Hydrogen agriculture is recently recognized as an emerging and promising approach for low-carbon society. Since shorter retention time for hydrogen gas (H2) in conventional electrolytically produced hydrogen-rich water (HRW) limits its application, seeking a more suitable method to produce and maintain H2 level in HRW for longer time remain a challenge for scientific community. To solve above problems, we compared and concluded that the H2 in HRW prepared by ammonia borane (NH3·BH3) could meet above requirement. The biological effects of HRW prepared by NH3·BH3 were further evaluated in seedlings of rapeseed, the most important crop for producing vegetable oil worldwide. Under our experimental conditions, 2 mg/L NH3·BH3-prepared HRW could confer 3-day-old hydroponic seedlings tolerance against 150 mM sodium chloride (NaCl), 20% polyethylene glycol (PEG; w/v), or 100 µM CdCl2 stress, and intensify endogenous nitric oxide (NO) accumulation under above stresses. The alleviation of seedlings growth stunt was confirmed by reducing cell death and reestablishing redox homeostasis. Reconstructing ion homeostasis, increasing proline content, and reducing Cd accumulation were accordingly observed. Above responses were sensitive to the removal of endogenous NO with its scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-1-oxyl-3-oxide (cPTIO; 100 µM), reflecting the requirement of NO functioning in the regulation of plant physiology achieved by NH3·BH3-prepared HRW. The application of 1 mM tungstate, an inhibitor of nitrate reductase (NR; an important NO synthetic enzyme), showed the similar blocking responses in the phenotype, suggesting that NR might be the major source of NO involved in above H2 actions. Together, these results revealed that HRW prepared by NH3·BH3 could enhance rapeseed seedlings tolerance against abiotic stress, thus opening a new window for the application of H2 in agricultural production.

Fine mapping of the BnaC04.BIL1 gene controlling plant height in Brassica napus L.

BACKGROUND: Plant height is an important architecture trait which is a fundamental yield-determining trait in crops. Variety with dwarf or semi-dwarf phenotype is a major objective in the breeding because dwarfing architecture can help to increase harvest index, increase planting density, enhance lodging resistance, and thus be suitable for mechanization harvest. Although some germplasm or genes associated with dwarfing plant type have been carried out. The molecular mechanisms underlying dwarfism in oilseed rape (Brassica napus L.) are poorly understood, restricting the progress of breeding dwarf varieties in this species. Here, we report a new dwarf mutant Bndwarf2 from our B. napus germplasm. We studied its inheritance and mapped the dwarf locus BnDWARF2. RESULTS: The inheritance analysis showed that the dwarfism phenotype was controlled by one semi-dominant gene, which was mapped in an interval of 787.88 kb on the C04 chromosome of B. napus by Illumina Brassica 60 K Bead Chip Array. To fine-map BnDWARF2, 318 simple sequence repeat (SSR) primers were designed to uniformly cover the mapping interval. Among them, 15 polymorphic primers that narrowed down the BnDWARF2 locus to 34.62 kb were detected using a F2:3 family population with 889 individuals. Protein sequence analysis showed that only BnaC04.BIL1 (BnaC04g41660D) had two amino acid residues substitutions (Thr187Ser and Gln399His) between ZS11 and Bndwarf2, which encoding a GLYCOGEN SYNTHASE KINASE 3 (GSK3-like). The quantitative real-time PCR (qRT-PCR) analysis showed that the BnaC04.BIL1 gene expressed in all tissues of oilseed rape. Subcellular localization experiment showed that BnaC04.BIL1 was localized in the nucleus in tobacco leaf cells. Genetic transformation experiments confirmed that the BnaC04.BIL1 is responsible for the plant dwarf phenotype in the Bndwarf2 mutants. Overexpression of BnaC04.BIL1 reduced plant height, but also resulted in compact plant architecture. CONCLUSIONS: A dominant dwarfing gene, BnaC04.BIL1, encodes an GSK3-like that negatively regulates plant height, was mapped and isolated. Our identification of a distinct gene locus may help to improve lodging resistance in oilseed rape.

Glutathione produced by γ-glutamyl cysteine synthetase acts downstream of hydrogen to positively influence lateral root branching.

Hydrogen gas (H2) mediation of lateral root (LR) branching was previously described. However, related signaling pathway is largely unexplored. In this study, we discovered that application with H2 using hydrogen-rich water, mimicking the responses of exogenous glutathione (GSH), not only enhanced GSH synthesis, but also induced tomato LR development. The changes in the transcripts of auxin signaling-related genes and cell cycle regulatory genes were matched with above phenotypes. The addition of H2 could trigger higher transcript levels of SlGSH1 and SlGSH2, encoding γ-glutamylcysteine synthetase (γ-ECS) and glutathione synthetase (GS), confirming the stimulation of GSH synthesis. These responses were greatly abolished when the inhibitor of γ-ECS was applied. The inhibition in lateral root primordium development, especially in emergence stage, was also observed. Genetic evidence revealed that the defects in GSH production and lateral rooting in Arabidopsis cad2-1, a γ-ECS defective mutant, were obviously abolished in the presence of GSH compared to those in the presence of H2. Further evidence revealed that mRNA levels of target genes elicited by H2 in wild-type, were differentially impaired in mutant plants. Together, above data clearly demonstrated that γ-ECS-dependent GSH production might be closely associated with H2 control of LR branching.

Methane control of cadmium tolerance in alfalfa roots requires hydrogen sulfide.

Hydrogen sulfide (H2S) is well known as a gaseous signal in response to heavy metal stress, while methane (CH4), the most prevalent greenhouse gas, confers cadmium (Cd) tolerance. In this report, the causal link between CH4 and H2S controlling Cd tolerance in alfalfa (Medicago sativa) plants was assessed. Our results observed that the administration of CH4 not only intensifies H2S metabolism, but also attenuates Cd-triggered growth inhibition in alfalfa seedlings, which were parallel to the alleviated roles in the redox imbalance and cell death in root tissues. Above results were not observed in roots after the removal of endogenous H2S, either in the presence of either hypotaurine (HT; a H2S scavenger) or DL-propargylglycine (PAG; a H2S biosynthesis inhibitor). Using in situ noninvasive microtest technology (NMT) and inductively coupled plasma mass spectroscopy (ICP-MS), subsequent results confirmed the participation of H2S in CH4-inhibited Cd influx and accumulation in roots, which could be explained by reestablishing glutathione (GSH) pool (reduced/oxidized GSH and homoglutathione) homeostasis and promoting antioxidant defence. Overall, our results clearly revealed that H2S operates downstream of CH4 enhancing tolerance against Cd stress, which are significant for both fundamental and applied plant biology.

Hydrogen sulfide-linked persulfidation of ABI4 controls ABA responses through the transactivation of MAPKKK18 in Arabidopsis.

Hydrogen sulfide (H2S) is a signaling molecule that regulates plant hormone and stress responses. The phytohormone abscisic acid (ABA) plays an important role in plant adaptation to unfavorable environmental conditions and induces the persulfidation of L-CYSTEINE DESULFHYDRASE1 (DES1) and the production of H2S in guard cells. However, it remains largely unclear how H2S and protein persulfidation participate in the relay of ABA signals. In this study, we discovered that ABSCISIC ACID INSENSITIVE 4 (ABI4) acts downstream of DES1 in the control of ABA responses in Arabidopsis. ABI4 undergoes persulfidation at Cys250 that is triggered in a time-dependent manner by ABA, and loss of DES1 function impairs this process. Cys250 and its persulfidation are essential for ABI4 function in the regulation of plant responses to ABA and the H2S donor NaHS during germination, seedling establishment, and stomatal closure, which are abolished in the ABI4Cys250Ala mutated variant. Introduction of the ABI4Cys250Ala variant into the abi4 des1 mutant did not rescue its hyposensitivity to ABA. Cys250 is critical for the binding of ABI4 to its cognate motif in the promoter of Mitogen-Activated Protein Kinase Kinase Kinase 18 (MAPKKK18), which propagates the MAPK signaling cascade induced by ABA. Furthermore, the DES1-mediated persulfidation of ABI4 enhances the transactivation activity of ABI4 toward MAPKKK18, and ABI4 can bind the DES1 promoter, forming a regulatory loop. Taken together, these findings advance our understanding of a post-translational regulatory mechanism and suggest that ABI4 functions as an integrator of ABA and MAPK signals through a process in which DES1-produced H2S persulfidates ABI4 at Cys250.

Structural Characterization and Immunostimulatory Activity of Heteropolysaccharides from Fuzhuan Brick Tea.

Fuzhuan brick tea (FBT), one of the unique dark teas, has various health-promoting functions. In the present study, one polysaccharide fraction, namely FBTPS-2-1, was extracted and purified from FBT, and its structure and potential immunostimulatory activity were investigated. The results showed that FBTPS-2-1,one of typical heteropolysaccharides, was mainly composed of Gal, Ara, and Glc with little molar content of Man, Rha, GalA, and GlcA in molar ratio of 46.59:22.13:13.57:8.20:6.02:2.12:1.38 and molecular weight of 748 kDa. The backbone of FBTPS-2-1 contained →4)-ß-d-Galp-(1→4)-ß-d-Galp-(1→, →4)-ß-d-Galp-(1→4)-α-d-Glcp-(1→, →4)-α-d-Glcp-(1→4)-α-d-Glcp-(1→, →4)-α-d-Glcp-(1→4)-ß-d-Galp-(1→, →3)-ß-d-Galp-(1→4)-ß-d-Galp-(1→, →3,6)-ß-d-Galp-(1→3)-ß-d-Galp-(1→ and →3,6)-ß-d-Galp-(1→3,6)-ß-d-Galp-(1→. The linkages of branches in FBTPS-2-1 were mainly composed of α-l-Araf-(1→3,6)-ß-d-Galp-(1→, →5)-α-l-Araf-(1→3,6)-ß-d-Galp-(1→, →6)-ß-d-Galp-(1→3,6)-ß-d-Galp-(1→, α-l-Araf-(1→3,5)-α-l-Araf-(1→, →3,5)-α-l-Araf-(1→5)-α-l-Araf-(1→, α-d-Galp-(1→3,5)-α-l-Araf-(1→ and →5)-α-l-Araf-(1→6)-ß-d-Galp-(1→. Furthermore, FBTPS-2-1 could increase the phagocytosis of macrophages and promote the secretion of NO and a variety of inflammatory cytokines, including TNF-α, IL-1ß, and IL-6, indicating noticeable immune enhancement activity. Thus, FBTPS-2-1 could serve as a potentially functional food to improve human health by modulating the host immunoreaction.

The Importance of Nitric Oxide as the Molecular Basis of the Hydrogen Gas Fumigation-Induced Alleviation of Cd Stress on Ganoderma lucidum.

Whether or not hydrogen gas (H2) can reduce cadmium (Cd) toxicity in Ganoderma lucidum has remained largely unknown. Here, we report that Cd-induced growth inhibition in G. lucidum was significantly alleviated by H2 fumigation or hydrogen-rich water (HRW), evaluated by lower oxidative damage and Cd accumulation. Moreover, the amelioration effects of H2 fumigation were better than of HRW in an optimum concentration of H2 under our experimental conditions. Further results showed that H2-alleviated growth inhibition in G. lucidum was accompanied by increased nitric oxide (NO) level and nitrate reductase (NR) activity under Cd stress. On the other hand, the mitigation effects were reversed after removing endogenous NO with its scavenger cPTIO or inhibiting H2-induced NR activity with sodium tungstate. The role of NO in H2-alleviated growth inhibition under Cd stress was proved to be achieved through a restoration of redox balance, an increase in cysteine and proline contents, and a reduction in Cd accumulation. In summary, these results clearly revealed that NR-dependent NO might be involved in the H2-alleviated Cd toxicity in G. lucidum through rebuilding redox homeostasis, increasing cysteine and proline levels, and reducing Cd accumulation. These findings may open a new window for H2 application in Cd-stressed economically important fungi.

Multi-Walled Carbon Nanotubes Can Promote Brassica napus L. and Arabidopsis thaliana L. Root Hair Development through Nitric Oxide and Ethylene Pathways.

Here, we report that multi-walled carbon nanotubes (MWCNTs) can promote plant root hair growth in the species analyzed in this study; however, low and excessive concentrations of MWCNTs had no significant effect or even an inhibiting influence. Further results show that MWCNTs can enter rapeseed root cells. Meanwhile, nitrate reductase (NR)-dependent nitric oxide (NO) and ethylene syntheses, as well as root hair formation, were significantly stimulated by MWCNTs. Transcription of root hair growth-related genes were also modulated. The above responses were sensitive to the removal of endogenous NO or ethylene with a scavenger of NO or NO/ethylene synthesis inhibitors. Pharmacological and molecular evidence suggested that ethylene might act downstream of NR-dependent NO in MWCNTs-induced root hair morphogenesis. Genetic evidence in Arabidopsis further revealed that MWCNTs-triggered root hair growth was abolished in ethylene-insensitive mutants ein2-5 and ein3-1, and NR mutant nia1/2, but not in noa1 mutant. Further data placed NO synthesis linearly before ethylene production in root hair development triggered by MWCNTs. The above findings thus provide some insights into the molecular mechanism underlying MWCNTs control of root hair morphogenesis.

Persulfidation-based Modification of Cysteine Desulfhydrase and the NADPH Oxidase RBOHD Controls Guard Cell Abscisic Acid Signaling.

Hydrogen sulfide (H2S) is a gaseous signaling molecule that regulates diverse cellular signaling pathways through persulfidation, which involves the post-translational modification of specific Cys residues to form persulfides. However, the mechanisms that underlie this important redox-based modification remain poorly understood in higher plants. We have, therefore, analyzed how protein persulfidation acts as a specific and reversible signaling mechanism during the abscisic acid (ABA) response in Arabidopsis (Arabidopsis thaliana). Here we show that ABA stimulates the persulfidation of l-CYSTEINE DESULFHYDRASE1, an important endogenous H2S enzyme, at Cys44 and Cys205 in a redox-dependent manner. Moreover, sustainable H2S accumulation drives persulfidation of the NADPH oxidase RESPIRATORY BURST OXIDASE HOMOLOG PROTEIN D (RBOHD) at Cys825 and Cys890, enhancing its ability to produce reactive oxygen species. Physiologically, s-persulfidation-induced RBOHD activity is relevant to ABA-induced stomatal closure. Together, these processes form a negative feedback loop that fine-tunes guard cell redox homeostasis and ABA signaling. These findings not only expand our current knowledge of H2S function in the context of guard cell ABA signaling, but also demonstrate the presence of a rapid signal integration mechanism involving specific and reversible redox-based post-translational modifications that occur in response to changing environmental conditions.

Transcriptome analysis reveals insight into molecular hydrogen-induced cadmium tolerance in alfalfa: the prominent role of sulfur and (homo)glutathione metabolism.

BACKGROUND: Hydrogen gas (H2) is hypothesised to play a role in plants that are coping with stresses by regulating signal transduction and gene expression. Although the beneficial role of H2 in plant tolerance to cadmium (Cd) has been investigated previously, the corresponding mechanism has not been elucidated. In this report, the transcriptomes of alfalfa seedling roots under Cd and/or hydrogen-rich water (HRW) treatment were first analysed. Then, the sulfur metabolism pathways were focused on and further investigated by pharmacological and genetic approaches. RESULTS: A total of 1968 differentially expressed genes (DEGs) in alfalfa seedling roots under Cd and/or HRW treatment were identified by RNA-Seq. The DEGs were classified into many clusters, including glutathione (GSH) metabolism, oxidative stress, and ATP-binding cassette (ABC) transporters. The results validated by RT-qPCR showed that the levels of relevant genes involved in sulfur metabolism were enhanced by HRW under Cd treatment, especially the genes involved in (homo)glutathione metabolism. Additional experiments carried out with a glutathione synthesis inhibitor and Arabidopsis thaliana cad2-1 mutant plants suggested the prominent role of glutathione in HRW-induced Cd tolerance. These results were in accordance with the effects of HRW on the contents of (homo)glutathione and (homo)phytochelatins and in alleviating oxidative stress under Cd stress. In addition, the HRW-induced alleviation of Cd toxicity might also be caused by a decrease in available Cd in seedling roots, achieved through ABC transporter-mediated secretion. CONCLUSIONS: Taken together, the results of our study indicate that H2 regulated the expression of genes relevant to sulfur and glutathione metabolism and enhanced glutathione metabolism which resulted in Cd tolerance by activating antioxidation and Cd chelation. These results may help to elucidate the mechanism governing H2-induced Cd tolerance in alfalfa.

The role of methane in plant physiology: a review.

Methane (CH4), one of the most important greenhouse gases, has conventionally been considered as a physiologic inert gas. However, this perspective has been challenged by the observation that CH4 has diverse biological functions in animals, such as anti-inflammatory, antioxidant, and anti-apoptosis. Meanwhile, it has now been identified as a possible candidate of gaseous signaling molecule in plants, although its biosynthetic and metabolic pathways as well as the mechanism(s) of CH4 signaling have not fully understood yet. This paper aims to review the available evidence for the biological roles of CH4 in regulating plant physiology. Although currently available reports do not fully support the notion of CH4 as a gasotransmitter, they do show that CH4 might be produced by an aerobic, non-microbial pathway from plants, and plays important roles in enhancing plant tolerance against abiotic stresses, such as salinity, drought, heavy metal exposure, and promoting root development, as well as delaying senescence and browning. Further results showed that CH4 could interact with reactive oxygen species (ROS), other gaseous signaling molecules [e.g., nitric oxide (NO), carbon monoxide (CO), and hydrogen sulfide (H2S)], and glutathione (GSH). These reports thus support the idea that plant-produced CH4 might be a component of a survival strategy of plants. Finally, the possibility of CH4 application in agriculture is preliminarily discussed.

Increasing tolerance to bispyribac-sodium is able to allow glutathione homeostasis to recover in indica rice compared with japonica rice.

The high activity and broad weed spectrum of BS has made it widely used in China. However, accidental crop injuries, particularly occurring in Jiangsu, Hunan, Hubei and Heilongjiang provinces in recent years, have resulted in limiting the application of BS in China. In this study, glutathione homeostasis was measured in the contrasting sensitivity of indica and japonica rice cultivar after bispyribac-sodium (BS) treatment. The results showed that japonica rice cultivar Nanjing 9108 was more sensitive to BS than indica rice Nanjing 11 and indica-hybrid cultivar Guangliangyou 6326. In response to the exposure of BS in all rice cultivars, especially Nanjing 9108, the perturbation of glutathione homeostasis occurred, including the decreased reduced glutathione (GSH) and increased oxidized glutathione (GSSG). These results were supported by increased activities of glutathione S-transferases (GSTs) in Nanjing 11 and Guangliangyou 6326. Further tests revealed that when Nanjing 11 was pretreated with the glutathione-depleting agents L-buthionine-sulfoximine (BSO) or diethylmaleate (DEM), the GSH levels, the activity of GSTs, and the gene expression levels of GR and GSTs decreased, finally increasing the phytotoxicity of BS. The aforementioned DEM inhibitory responses were further rescued by exogenously applied GSH. In contrast, the pretreatment of glutathione or N-acetyl-L-cysteine (NAC) not only increased the contents of GSH, the activities of GSTs, and the expression level of GR and GSTs gene, but also alleviated BS phytotoxicity in Nanjing 9108. In both cultivars, DEM increased phytotoxicity and GSH partially reversed this. This study suggests that increasing tolerance to BS was able to allow glutathione homeostasis to recover in indica rice cultivar compared with japonica rice cultivar.

L-Cysteine desulfhydrase-dependent hydrogen sulfide is required for methane-induced lateral root formation.

KEY MESSAGE: Methane-triggered lateral root formation is not only a universal event, but also dependent on L-cysteine desulfhydrase-dependent hydrogen sulfide signaling. Whether or how methane (CH4) triggers lateral root (LR) formation has not been elucidated. In this report, CH4 induction of lateral rooting and the role of hydrogen sulfide (H2S) were dissected in tomato and Arabidopsis by using physiological, anatomical, molecular, and genetic approaches. First, we discovered that CH4 induction of lateral rooting is a universal event. Exogenously applied CH4 not only triggered tomato lateral rooting, but also increased activities of L-cysteine desulfhydrase (DES; a major synthetic enzyme of H2S) and induced endogenous H2S production, and contrasting responses were observed in the presence of hypotaurine (HT; a scavenger of H2S) or DL-propargylglycine (PAG; an inhibitor of DES) alone. CH4-triggered lateral rooting were sensitive to the inhibition of endogenous H2S with HT or PAG. The changes in the transcripts of representative cell cycle regulatory genes, miRNA and its target genes were matched with above phenotypes. In the presence of CH4, Arabidopsis mutant Atdes1 exhibited defects in lateral rooting, compared with the wild-type. Molecular evidence showed that the transcriptional profiles of representative target genes modulated by CH4 in wild-type plants were impaired in Atdes1 mutant. Overall, our data demonstrate the main branch of the DES-dependent H2S signaling cascade in CH4-triggered LR formation.

Methane Control of Adventitious Rooting Requires γ-Glutamyl Cysteine Synthetase-Mediated Glutathione Homeostasis.

Although the key role of methane (CH4) in the induction of cucumber adventitious rooting has been observed previously, the target molecules downstream of the CH4 action are yet to be fully elucidated. Here, we reported that exogenous glutathione (GSH) induced cucumber adventitious root formation; while l-buthionine-sulfoximine (BSO) treatment inhibited it. BSO is a known inhibitor of γ-glutamyl cysteine synthetase (γ-ECS), an enzyme involved in GSH biosynthesis. Further investigations showed that endogenous GSH content was rapidly increased by CH4 application, which was correlated with the increased CsGSH1 transcript and γ-ECS activity. Mimicking the responses of GSH, CH4 could upregulate cell cycle regulatory genes (CsCDC6, CsCDPK1, CsCDPK5 and CsDNAJ-1) and auxin-response genes (CsAux22D-like and CsAux22B-like). Meanwhile, adventitious rooting-related CsmiR160 and CsmiR167 were increased or decreased, respectively, and contrasting tendencies were observed in the changes of their target genes, that included CsARF17 and CsARF8. The responses above were impaired by the removal of endogenous GSH with BSO, indicating that CH4-triggered adventitious rooting was GSH-dependent. Genetic evidence revealed that in the presence of CH4, Arabidopsis mutants cad2 (a γ-ECS-defective mutant) exhibited, not only the decreased GSH content in vivo, but also the defects in adventitious root formation, both of which were rescued by GSH administration other than CH4. Together, it can be concluded that γ-ECS-dependent GSH homeostasis might be required for CH4-induced adventitious root formation.

Atomic-engineered gold@silvergold alloy nanoflowers for in vivo inhibition of bacteria.

The problems of multidrug-resistant bacteria and environmental pollution associated with the abuse of antibiotics call for effective antibiotic alternatives. Here, gold@silvergold alloy nanoflowers (Au@AgAu ANFs) with distinct atomic structures are first fabricated and then demonstrated for in vivo inhibition of bacteria. The Au@AgAu ANFs display high antibacterial activity against the model Gram-negative bacterium Escherichia coli, with a minimum inhibitory concentration value of 4.8 µg mL-1, which is 3.1 times lower than that of silver nanoparticles. The alloy structure with a rough surface enables Au@AgAu ANFs to firmly adhere to the bacterial surface and damage the cell membrane, resulting in long-term (48 h) and highly stable (30 days) antibacterial activity. Meanwhile, the Au@AgAu ANFs show remarkable biocompatibility with human cells even at a high concentration of 40 µg mL-1. Application of Au@AgAu ANFs in the treatment of bacterial infections in the mouse intestine significantly reduces the reproduction of bacteria compared to an untreated mouse, giving results similar to those of the current antibiotic treatment, with no cytotoxicity. Our study opens up a new avenue for the rational design of safe and highly efficient antibacterial materials.