An innovative way to treat cash crop wastes: The fermentation characteristics and functional microbial community using different substrates to produce Agricultural Jiaosu.

With the increase of global demand for cash crops, a large of cash crop waste was produced and caused severe environmental issues. To produce Agricultural Jiaosu (AJ) using these wastes is a sustainable waste disposal method. However, the fermentation mechanism, metabolites, and microbial characteristics of AJ fermented with different substrates remain unclear. In this study, the effects of different substrates (fruit and vegetable waste and Chinese herbal medicine waste) on the fermentation characteristics of AJ, including metabolites and microbial community properties, were investigated. The results revealed that AJ fermentation was a process of converting organic matter into organic acids and other metabolites, mainly including hydrolysis, acidogenesis, and maturation stages. At the genus level, Lactobacillus, Acetobacter, Hydrogenibacillus, Halomonas, and Prevotella_1 were the dominant bacteria in the fermentation system. The bacterial diversity of composite substrate AJ was higher than that of single substrate AJ. The organic acids and secondary metabolites concentration and the composition of key microorganisms depended on the substrate type. Furthermore, AJ's potential functional genes were mainly concentrated in cofactors and vitamin, carbohydrate, and amino acid metabolism. The findings of this study indicated that AJ is an innovative eco-friendly technology that can convert cash crop wastes into sustainable eco-products, and that its characteristics depend on the substrate type. Therefore, the substrate used to produce AJ should be carefully selected according to the application field.

Characterization of nitrogen and water status in oat leaves using optical sensing approach.

BACKGROUND: Optical sensing is a potential tool to estimate plant N status, but soil water deficits may interefere with forming a clear relationship. A greenhouse study was conducted with oat plants treated with three water regimes and four N levels to determine whether optical sensing could be used to estimate leaf N and relative water content (RWC). RESULTS: Leaf N was strongly correlated with reflectance at 550 nm and at around 705 nm, and N treatments caused a red-edge peak shift to lower wavelength. The ratio of the first derivative reflectance at 741-696 nm (FDRE) was identified to be a good estimator of leaf N at jointing (R(2) = 0.90) and heading (R(2) = 0.86) stages across water treatments. Leaf N also had a stronger association with the red-edge position (REP) at both stages (R(2) = 0.83 and 0.78), or with the ratio R4 (R760/R550) at jointing (R(2) = 0.88), than with chlorophyll meter (SPAD) readings. Under water stress, the predictive accuracy of leaf N increased with these reflectance indices, but decreased using SPAD readings. CONCLUSION: The results indicate that specific reflectance indices of FDRE, REP and R4 may be used for a rapid and non-destructive estimation of oat plant N status over a range of water regimes.

Discovery of indolylacryloyl-derived oxacins as novel potential broad-spectrum antibacterial candidates.

The emergence of serious bacterial resistance towards clinical oxacins poses a considerable threat to global public health, necessitating the development of novel structural antibacterial agents. Seven types of novel indolylacryloyl-derived oxacins (IDOs) were designed and synthesized for the first time from commercial 3,4-difluoroaniline via an eight-step procedure. The synthesized compounds were characterized by modern spectroscopic techniques. All target molecules were evaluated for antimicrobial activities. Most of the prepared IDOs showed a broad antibacterial spectrum and strong activities against the tested strains, especially ethoxycarbonyl IDO 10d (0.25-0.5 µg/mL) and hydroxyethyl IDO 10e (0.25-1 µg/mL) exhibited much superior antibacterial efficacies to reference drug norfloxacin. These highly active IDOs also displayed low hemolysis, cytotoxicity and resistance, as well as rapid bactericidal capacity. Further investigations indicated that ethoxycarbonyl IDO 10d and hydroxyethyl IDO 10e could effectively reduce the exopolysaccharide content and eradicate the formed biofilm, which might delay the development of drug resistance. Preliminary exploration of the antibacterial mechanism revealed that active IDOs could not only destroy membrane integrity, resulting in changes in membrane permeability, but also promote the accumulation of reactive oxygen species, leading to the production of malondialdehyde and decreased bacterial metabolism. Moreover, they exhibited the capability to bind with DNA and DNA gyrase, forming supramolecular complexes through various noncovalent interactions, thereby inhibiting DNA replication and causing bacterial death. All the above results suggested that the newly developed indolylacryloyl-derived oxacins should hold great promise as potential multitargeting broad-spectrum antibacterial candidates to overcome drug resistance.

Cyanomethylquinolones as a New Class of Potential Multitargeting Broad-Spectrum Antibacterial Agents.

This work identified a class of cyanomethylquinolones (CQs) and their carboxyl analogues as potential multitargeting antibacterial candidates. Most of the prepared compounds showed high antibacterial activities against most of the tested bacteria, exhibiting lower MIC values (0.125-2 µg/mL) than those of clinical norfloxacin, ciprofloxacin, and clinafloxacin. The low hemolysis, drug resistance, and cytotoxicity, as well as good predictive pharmacokinetics of active CQs and carboxyl analogues revealed their development potential. Furthermore, they could eradicate the established biofilm, facilitating bacterial exposure to these antibacterial candidates. These active compounds could induce bacterial death through multitargeting effects, including intercalating into DNA, up-regulating reactive oxygen species, damaging membranes directly, and impeding metabolism. Moreover, the highly active cyclopropyl CQ 15 exhibited more effective in vivo anti-MRSA potency than ciprofloxacin. These findings highlight the potential of CQs and their carboxyl analogues as multitargeting broad-spectrum antibacterial candidates for treating intractable bacterial infections.

Biogas Residues Improved Microbial Diversity and Disease Suppression Function under Extent Indigenous Soil Microbial Biomass.

Indigenous soil microbial biomass (ISMB) plays a key role in maintaining essential functions and biodiversity of soil health. One of the critical unknowns is how the indigenous microorganisms respond to different fertilizers which is directly related to agricultural production. Therefore, we used Mi-Seq sequencing and network analyses to compare the response of ISMB to biogas residue and chemical fertilizers. The results showed that crop production was profoundly influenced by levels of ISMB present and is further dependent on the strategy of fertilizer application. Higher ISMB primarily manifests through retention of richer microbial abundance, a balanced community structure, and tightened co-occurrence within a certain proportion of Nitrospirae, Rhizophlyctidaceae, and Gemmatimonadetes. Compared to chemical fertilizer, biogas residue resulted in higher production with more strongly linked nodes such as Actinobacteria, Chloroflexi and Gemmatimonadetes. Under the same level of ISMB, the microbial diversity was richer and co-occurrence was tighter when biogas residues were applied compared with chemical fertilizer. In addition, the higher level of ISMB with biogas residue applied had a lower abundance of potential fungal pathogens in both bulk and rhizosphere soil compared with chemical fertilizer. This study provides critical data to understand the influence of ISMB and biogas residue on soil ecological system.

Agricultural Jiaosu: An Eco-Friendly and Cost-Effective Control Strategy for Suppressing Fusarium Root Rot Disease in Astragalus membranaceus.

Root rot caused by the pathogenic fungi of the Fusarium genus poses a great threat to the yield and quality of medicinal plants. The application of Agricultural Jiaosu (AJ), which contains beneficial microbes and metabolites, represents a promising disease control strategy. However, the action-effect of AJ on Fusarium root rot disease remains unclear. In the present study, we evaluated the characteristics and antifungal activity of AJ fermented using waste leaves and stems of medicinal plants, and elucidated the mechanisms of AJ action by quantitative real-time PCR and redundancy analysis. The effects of AJ and antagonistic microbes isolated from it on disease suppression were further validated through a pot experiment. Our results indicate that the AJ was rich in beneficial microorganisms (Bacillus, Pseudomonas, and Lactobacillus), organic acids (acetic, formic, and butyric acids) and volatile organic compounds (alcohols and esters). It could effectively inhibit Fusarium oxysporum and the half-maximal inhibitory concentration (IC50) was 13.64%. The antifungal contribution rate of the microbial components of AJ reached 46.48%. Notably, the redundancy analysis revealed that the Bacillus and Pseudomonas genera occupied the main niche during the whole inhibition process. Moreover, the abundance of the Bacillus, Pseudomonas, and Lactobacillus genera were positively correlated with the pH-value, lactic, formic and butyric acids. The results showed that the combined effects of beneficial microbes and organic acid metabolites increased the efficacy of the AJ antifungal activity. The isolation and identification of AJ's antagonistic microbes detected 47 isolates that exhibited antagonistic activities against F. oxysporum in vitro. In particular, Bacillus subtilis and Bacillus velezensis presented the strongest antifungal activity. In the pot experiment, the application of AJ and these two Bacillus species significantly reduced the disease incidence of Fusarium root rot and promoted the growth of Astragalus. The present study provides a cost-effective method to control of Fusarium root rot disease, and establishes a whole-plant recycling pattern to promote the sustainable development of medicinal plant cultivation.

Synthesis and Antitumor Activity of Diosgenin Hydroxamic Acid and Quaternary Phosphonium Salt Derivatives.

Diosgenin, a component separated from Dioscorea plants, is an important starting material for steroid hormone drugs and semisynthetic steroids. In the work, two series of diosgenin derivatives were designed, synthesized, and evaluated for their cellular anticancer activities. Most of the target compounds exhibited good inhibitory activities against four cell lines, Aspc-1 (human colon adenocarcinoma cells), H358 (human nonsmall cell lung cancer cells), HCT116 (human colorectal adenocarcinoma cells), and SW620 (human metastatic pancreatic cancer cells). Among them, the representative compound 2.2f exhibited 7.9-341.7-fold antiproliferative activities against the above-mentioned four cell lines compared with the lead compound diosgenin.

Effects of water-saving superabsorbent polymer on antioxidant enzyme activities and lipid peroxidation in oat (Avena sativa L.) under drought stress.

BACKGROUND: Drought stress significantly limits oat (Avena sativa L.) growth and productivity. Thus an efficient management of soil moisture and study of metabolic changes in response to drought are important for improved production of oat. The objective was to gain a better understanding of drought tolerance mechanisms and improve soil water management strategies using water-saving superabsorbent polymer (SAP) at 60 kg ha(-1) under three irrigation levels (adequate, moderate and deficit) using a new type of hydraulic pressure-controlled auto irrigator. RESULTS: The results showed that the relative water content and leaf water potential (ψ(1) ) were much higher in oats treated with SAP. Although the SAP had little effect on plant biomass accumulation under adequate and moderate irrigation, it significantly increased the biomass by 52.7% under deficit irrigation. Plants treated with SAP under deficit irrigation showed a significant decrease in superoxide dismutase, catalase, peroxidase, ascorbate peroxidase and glutathione reductase activities in leaves compared with control plants. CONCLUSION: Our results suggested that drought stress leads to production of oxygen radicals, which results in increased lipid peroxidation and oxidative stress in the plant, and the application of SAP could conserve soil water, making same available to plants for increased biomass accumulation and reduced oxidative stress especially under severe water stress.

Effects of water-saving superabsorbent polymer on antioxidant enzyme activities and lipid peroxidation in corn (Zea mays L.) under drought stress.

BACKGROUND: Drought is the most important abiotic stress factor limiting corn (Zea mays L.) growth and productivity. Therefore efficient management of soil moisture and study of metabolic changes in response to drought are important for improved production of corn. The objective of the present study was to gain a better understanding of drought tolerance mechanisms and improve soil water management strategies using a water-saving superabsorbent polymer (SAP) at 30 kg ha(-1) under three irrigation levels (adequate, moderate and deficit) using a new type of hydraulic pressure-controlled auto-irrigator. RESULTS: The results showed that relative water content and leaf water potential were much higher in corn treated with SAP. Although application of SAP reduced biomass accumulation by 11.1% under adequate irrigation, it increased the biomass markedly by 39.0% under moderate irrigation and 98.7% under deficit irrigation. Plants treated with SAP under deficit irrigation showed a significant decrease in superoxide dismutase, catalase, peroxidase, ascorbate peroxidase and glutathione reductase activities in leaves compared with control plants. CONCLUSION: The results of this study suggest that drought stress causes the production of oxygen radicals, leading to increased lipid peroxidation and oxidative stress in plants, and the application of a superabsorbent polymer could conserve soil water, making it available to plants for quenching oxidative stress and increasing biomass accumulation, especially under severe water stress.

A lysimeter study of nitrate leaching, optimum fertilisation rate and growth responses of corn (Zea mays L.) following soil amendment with water-saving super-absorbent polymer.

BACKGROUND: Nitrate leaching and the resulting groundwater contamination from intensive cereal production has become a major concern for long-term farmland efficiency and environmental sustainability in northern China. The aim of this study was to evaluate a water-saving super-absorbent polymer (SAP) for minimising NO(3)(-) leaching from soil and optimising corn growth and yield. Thirty-six undisturbed soil lysimeters were installed in a field lysimeter facility in drought-affected northern China to study the growth and yield characteristics of summer corn (Zea mays L.) as well as the amount of NO(3)-leaching losses under different fertiliser (standard, medium or 75% and low, or 50% of conventional fertilisation rate) and SAP (control, 0; level-1, 15 kg ha(-1) and level-2, 30 kg ha(-1)) treatments. RESULTS: Corn yield fell by 19.7% under medium and 37.7% under low fertilisation; the application of SAP increased yield significantly by 44.4% on level-1 and 80.3% on level-2. Similarly, plant height, leaf area, number of grains as well as protein, soluble sugar and starch contents in the grain also increased with SAP treatment. Application of SAP at 30 kg ha(-1) plus half of conventional fertilisation can reduce maximum (64.1%) nitrate leaching losses from soil. CONCLUSIONS: Application of SAP at 30 kg ha(-1) plus only half the amount of conventional fertiliser rate (150 kg urea, and 50 kg each of superphosphate and potassium sulfate) would be a more appropriate practice both for minimising nitrate leaching and sustainable corn production under the arid and semiarid conditions of northern China.

Effectiveness of a water-saving super-absorbent polymer in soil water conservation for corn (Zea mays L.) based on eco-physiological parameters.

BACKGROUND: The objective was to study soil water conservation and physiological growth of corn (Zea mays L.) using water-saving super-absorbent polymer (SAP) at 30 kg ha(-1). The effectiveness of SAP was studied under three irrigation levels (adequate, moderate and deficit) using a new type of negative hydraulic pressure controlled auto-irrigator in the years 2009 and 2010 in a greenhouse at Beijing, P.R. China. RESULTS: Eight weeks after sowing, plant height and leaf area increased significantly by 41.6 and 79.6% under deficit irrigation for SAP treatment. The SAP had little effect on shoot dry mass under adequate and moderate irrigation but increased it significantly by 133.5% under deficit irrigation. Similarly, the efficiency of water use also increased by 97.1%. Leaf water potential under adequate and moderate irrigation differs slightly for SAP application, whereas under deficit irrigation the values were exceeded significantly by 27.8%. The superior growth and water use efficiency of corn treated with SAP under deficit irrigation was ascribed to maintenance of higher relative water contents in leaves as well as intercellular carbon dioxide concentration, net photosynthesis and transpiration rate. CONCLUSIONS: Our results suggested that plant growth and different physiological activities are restricted by drought stress and the application of super-absorbent polymer could conserve soil water, making same available to plants for increased growth and biomass accumulation especially under severe water stress. Thus, application of SAP is a suitable soil management practice for the locations characterised by severe water stress.

The effects of C/N (10-25) on the relationship of substrates, metabolites, and microorganisms in "inhibited steady-state" of anaerobic digestion.

The carbon/nitrogen ratio (C/N) is a key parameter that affects the performance of anaerobic digestion (AD). Recent AD research has focused on optimizing the C/N of feedstock. The so-called "inhibited steady-state" refers to a special state of ammonia inhibition of AD that often occurs at low-C/N (below 25) when degradable nitrogen-rich substrates, such as livestock manure, are used as feedstock. However, the mechanism behind the "inhibited steady-state" is still unknown. In the current study, co-digestion and recirculation were used to create a C/N gradient in the influent to explore the relationship between substrates, metabolites, and microorganisms in the "inhibited steady-state." Data were collected at the macro, microbial, and genetic levels. Three CSTRs were successfully made run into the "inhibited steady-state" using influent C/Ns of 10-12. Digestion performance levels of R10-R12 were low and stable, transitioning from an aceticlastic methane-producing pathway to a hydrogenotrophic pathway as the C/N gradually decreased. As the abundance of the hydrogenophilic methanogens increased, the abundance of syntrophic acetate-oxidizing bacteria (SAOB) also increased. The succession between populations of Methanosaeta and Methanosarcina may be used as a microbiological indicator of ammonia inhibition. Under high-C/Ns, cooperation among bacteria was high, while under low-C/Ns, competition among bacteria was high. These results clarify the processes underlying the "inhibited steady-state," which is a condition often faced in actual large-scale biogas facilities that use degradable nitrogen-rich substrates. Moreover, practical guidelines for evaluating ammonia inhibition are provided, and strategies to alleviate ammonia suppression are developed.

A microbial ecosystem: agricultural Jiaosu achieves effective and lasting antifungal activity against Botrytis cinerea.

Synthetic fungicides are eco-unfriendly to agriculture and the environment. Agricultural Jiaosu (AJ), which originates from organic wastes, has the potential to be a substitute for synthetic fungicides. In this study, the characteristics of AJ and its antifungal activity against Botrytis cinerea were investigated for the first time. AJ was rich in lactic acid (4.46 g/L), acetic acid (1.52 g/L), Lactobacillus (72.45%) and Acetobacter (15.23%), which was a microbial ecosystem consisting of acid-based substances (AS) and beneficial microorganisms (BM). The results of the antifungal assays suggested that B. cinerea was effectively inhibited by AJ, with the half-maximal inhibitory concentration (IC50) of 9.24%. AJ showed the strongest and most-lasting inhibitory effect compared to cell-free supernatant and microbial solution of AJ, indicating that AS and BM and their synergistic effect contributed to the antifungal activity of AJ. Two-step inhibition' is an antifungal mode of AJ. Firstly, AS not only inhibited the pathogen directly but also provided a dominant niche for BM of AJ. Then, BM in AJ, especially Acetobacter, proliferated and metabolized acetic acid continuously. Thus, AJ achieved high-efficiency and long-acting inhibition. AJ is a promising biological agent considering its features of an eco-friendly, low-cost and easy-to-operate biological agent in rural areas.

Enhancing naked oat (Avena nuda L.) productivity with minimal indirect nitrogen loss and maximum nitrogen use efficiency through integrated use of different nitrogen sources.

Oat (Avena nuda L.) is a nutritious grain crop, rich in dietary fibers and phytochemicals. Application of efficient nitrogen (N) sources and dose is very important to obtain higher crop productivity and to achieve environmental sustainability. The exploitation of natural beneficial microbes and organic nitrogen in combination with chemical nitrogen would be effective to boost soil N for plant uptake. Hence, a field experiment was conducted during 2016 and 2017 with the aim to ameliorate the use of chemical N (CN) with organic nitrogen (ON) and microbial fertilizer (MBF) without compromising the productivity of oat. T1 = control, T2 = 100% CN, T3 = 100% CN+MBF, T4 = 75% CN+ 25% ON+MBF, T5 = 50% CN+ 50% ON+MBF, T6 = 100% ON+MBF, T7 = 100% ON were the treatments. 50% CN + 50% ON + MBF treatment proved to be an efficient combination regarding enhanced biomass and grain yield, nitrogen uptake and NUE as compared to rest of the treatments in both years. During the critical stages of the crop, when most of the applied CN was leached from the top 20 cm soil depth, a substantial N came from the PM mineralization through enhanced microbial activity by the addition of MBF. Lastly, the application of ON supplemented with MBF improved the rhizosphere soil properties, i.e. mineral N concentration, total N (TN), soil organic carbon (SOC), microbial biomass carbon (MBC), microbial biomass nitrogen (MBN), soil respiration rate and enzymatic activity. A balanced and source conscious application of CN, ON and MBF reduced N losses and added a substantial amount of N into the soil N pool. We concluded that organic N combined with chemical N and MBF proved to be effective in improving soil properties ensuring less N loss and increasing oat production in the semi-arid region.

Variations of the nirS-, nirK-, and nosZ-denitrifying bacterial communities in a northern Chinese soil as affected by different long-term irrigation regimes.

Denitrification causes nitrogen loss from agricultural soils and emission of nitrous oxide (N2O). Water addition leads to an increase in soil moisture which greatly influenced soil denitrification. However, it is unclear how irrigation management affected the denitrifying bacterial communities in agricultural systems. In the present study, we investigated the abundance, diversity, and composition of the nirS-, nirK-, and nosZ-denitrifying bacterial communities in the soil under different long-term irrigation regimes by using real-time PCR (qPCR) and Illumina MiSeq sequencing approaches. Results showed that the abundance of nosZ gene was 3.94-6.01 and 35.09-60.21 times more than that of nirS and nirK genes, and the abundance of nirS gene was 5.84-15.30 times higher than that of nirK gene, respectively, in different irrigation treatments. However, the Alpha diversity indices of the nirK-denitrifying bacterial community were higher than those of the nirS- and nosZ-denitrifying bacterial communities. Proteobacteria was the predominant phylum for all the denitrifying bacterial communities, and significant differences were observed in relative abundance of Alphaproteobacteria and Betaproteobacteria in predominant class between different irrigation treatments for the nirS- and nosZ-denitrifying bacterial communities, respectively. Irrigation significantly affected the abundance, Shannon and Invsimpson indices, and structure of the nirS- and nosZ-denitrifying bacterial communities, whereas it only minor influenced the structure of the nirK-denitrifying bacterial community. Furthermore, the shifts in abundance, diversity, and structure of the nirS- and nosZ-denitrifying bacterial communities correlated significantly with the soil property variations; however, no soil property was significantly correlated with the abundance and Alpha diversity index of the nirK-denitrifying bacterial community. Our results demonstrate that different long-term irrigation regimes greatly altered the abundance, diversity, and structure of the nirS- and nosZ- rather than the nirK-denitrifying bacterial communities.

Co-digestion of oat straw and cow manure during anaerobic digestion: Stimulative and inhibitory effects on fermentation.

Impacts of adding different amounts of cow manure (CM) on the anaerobic digestion (AD) of oat straw (OS) with total solids content (TS) values of 4%, 6%, 8% and 10% was assessed over 50 days using batch experiments. A modified Gompertz model was introduced to predict the methane yield and determine the kinetic parameters. The optimum addition was a 1:2 ratio of CM to the OS added, which resulted in a suitable C/N ratio of 27 and a higher degradation rate of lignocellulose. The best cumulative methane yield of 841.77 mL/g volatile solids added (VSadded) was 26.64% greater than that of digesting OS alone. In addition, the amount of CM added produced larger effects than that of changes in the TS. However, higher CM concentrations were found to be inhibitory. Clustering analysis could provide significant guidance for demonstrating project process and combining farming and animal husbandry.

Characterization of Selenium Accumulation, Localization and Speciation in Buckwheat-Implications for Biofortification.

Buckwheat is an important crop species in areas of selenium (Se) deficiency. To obtain better insight into their Se metabolic properties, common buckwheat (Fagopyrum esculentum) and tartary buckwheat (F. tataricum) were supplied with different concentrations of Se, supplied as selenate, selenite, or Astragalus bisulcatus plant extract (methyl-selenocysteine). Se was supplied at different developmental stages, with different durations, and in the presence or absence of potentially competing ions, sulfate, and phosphate. The plants were analyzed for growth, Se uptake, translocation, accumulation, as well as for Se localization and chemical speciation in the seed. Plants of both buckwheat species were supplied with 20 µM of either of the three forms of Se twice over their growth period. Both species accumulated 15-40 mg Se kg-1 DW in seeds, leaves and stems, from all three selenocompounds. X-ray microprobe analysis showed that the Se in seeds was localized in the embryo, in organic C-Se-C form(s) resembling selenomethionine, methyl-selenocysteine, and γ-glutamyl-methylselenocysteine standards. In short-term (2 and 24 h) Se uptake studies, both buckwheat species showed higher Se uptake rate and shoot Se accumulation when supplied with plant extract (methyl-selenocysteine), compared to selenite or selenate. In long-term (7 days) uptake studies, both species were resistant to selenite up to 50 µM. Tartary buckwheat was also resistant to selenate up to 75 µM Se, but >30 µM selenate inhibited common buckwheat growth. Selenium accumulation was similar in both species. When selenite was supplied, Se levels were 10-20-fold higher in root (up to 900 mg Se kg-1 DW) than shoot, but 4-fold higher in shoot (up to 1,200 mg Se kg-1 DW) than root for selenate-supplied plants. Additionally, sulfate and phosphate supply affected Se uptake, and conversely selenate enhanced S and P accumulation in both species. These findings have relevance for crop Se biofortification applications.

Comparison of ATP sulfurylase 2 from selenium hyperaccumulator Stanleya pinnata and non-accumulator Stanleya elata reveals differential intracellular localization and enzyme activity levels.

BACKGROUND: The plant Stanleya pinnata hyperaccumulates Se up to 0.5% of its dry weight in organic forms, whereas the closely related Stanleya elata does not hyperaccumulate Se. ATP sulfurylase (ATPS) can catalyze the formation of adenosine 5'-phosphoselenate (APSe) from ATP and selenate. We investigated the S. pinnata ATPS2 isoform (SpATPS2) to assess its possible role in Se hyperaccumulation. METHODS: ATPS expression and activity was compared in the two Stanleya species. The ATPS2 protein sequences were modeled. Sub-cellular locations were analyzed using GFP fusions. Enzyme activity of purified recombinant SpATPS2 was measured. RESULTS: ATPS2 transcript levels were six-fold higher in roots of S. pinnata relative to S. elata. Overall root ATPS enzyme activity was two-fold elevated in S. pinnata. Cloning and sequencing of SpATPS2 and S. elata ATPS2 (SeATPS2) showed the predicted SeATPS2 to be canonical, while SpATPS2, although very similar in its core structure, has unique features, including an interrupted plastid targeting signal due to a stop codon in the 5' region of the coding sequence. Indeed GFP fusions revealed that SpATPS2 had exclusive cytosolic localization, while SeATPS2 showed dual localization in plastids and cytosol. SpATPS2 activity was inhibited by both sulfate and selenate, indicating that the enzyme acts on both substrates. CONCLUSIONS: The ATPS2 from S. pinnata differs from non-accumulator ATPS2 in its elevated expression and sub-cellular localization. It likely acts on both selente and sulfate substrates. GENERAL SIGNIFICANCE: These observations shed new light on the role of ATPS2 in the evolution of Se hyperaccumulation in plants. This article is part of a Special Issue entitled Selenium research in biochemistry and biophysics - 200 year anniversary issue, edited by Dr. Elias Arnér and Dr. Regina Brigelius-Flohe.

[Soil microbial diversity of artificial peashrub plantation on North Loess Plateau of China].

Peashrub (Caragana korshinskii Kom) is a kind of excellent shrub used for dune-fixation in Loess Plateau of China. In order to explore relationship between peashrub and soil microorganisms, microbial communities diversity associated with rhizoplane, rhizosphere and bulk soil of peashrub in Loess Plateau of China were characterized based on a culture-independent approach. Three 16S rDNA gene libraries were constructed, respectively, and each different profile was used to define an operational taxonomic unit (OTU). The numbers of microorganisms decreased as root proximity decreased and a few OTUs became dominant. Phylogenetic analyses indicated that Proteobacteria was the predominant group in rhizoplane, which included many alpha-Proteobacteria, partially consisted of rhizobia, and gamma-Proteobacteria beneficial to plant growth. In bulk soil, the most frequent OTUs were closely related to Archaea, while Acidobacteria was the dominant group in rhizosphere of peashrub. The diversity index (H') was higher in rhizosphere than in rhizoplane and bulk soil, whereas microbial populations in rhizoplane and bulk soil had the greater dominance indices (D). It was shown that there was a significant change in microbial species composition along the root gradient, shifting from complex plant-associated bacterial community in the root habitats to a simple bacterial community in the bulk soil. These results showed that plant roots and soil conditions created a selective environment for microbial populations.

Methane Emissions and Microbial Communities as Influenced by Dual Cropping of Azolla along with Early Rice.

Azolla caroliniana Willd. is widely used as a green manure accompanying rice, but its ecological importance remains unclear, except for its ability to fix nitrogen in association with cyanobacteria. To investigate the impacts of Azolla cultivation on methane emissions and environmental variables in paddy fields, we performed this study on the plain of Dongting Lake, China, in 2014. The results showed that the dual cropping of Azolla significantly suppressed the methane emissions from paddies, likely due to the increase in redox potential in the root region and dissolved oxygen concentration at the soil-water interface. Furthermore, the floodwater pH decreased in association with Azolla cultivation, which is also a factor significantly correlated with the decrease in methane emissions. An increase in methanotrophic bacteria population (pmoA gene copies) and a reduction in methanogenic archaea (16S rRNA gene copies) were observed in association with Azolla growth. During rice cultivation period, dual cropping of Azolla also intensified increasing trend of 1/Simpson of methanogens and significantly decreased species richness (Chao 1) and species diversity (1/Simpson, 1/D) of methanotrophs. These results clearly demonstrate the suppression of CH4 emissions by culturing Azolla and show the environmental and microbial responses in paddy soil under Azolla cultivation.