Mineral protection mediates soil carbon temperature sensitivity of nine old-growth temperate forests across the latitude transect.

Temperature sensitivity (Q10) of soil microbial respiration serves as a crucial indicator for assessing the response of soil organic carbon (SOC) to global warming. However, the biogeographic variation in Q10 remains inconsistent. In this study, we examined Q10 and its potential drivers in nine old-growth mixed broad-leaved Korean pine (Pinus koraiensis Sieb. et Zucc.) forests (the climax community of Asian temperate mixed forest) under a wide range of climatic conditions. We found that stand characteristics (arbuscular mycorrhizal tree basal area to ectomycorrhizal tree basal area ratio and root to shoot ratio) contributed to soil C sequestration by facilitating the accumulation of soil recalcitrant C components. Contrary to the C quality-temperature hypothesis, Q10 was not correlated with C quality (soil C to nitrogen ratio and recalcitrant C to labile C ratio). Soil mineral protection parameters (Fe/Al oxides) had negative effect on Q10 because they inhibited microbial activities by decreasing substrate accessibility. Additionally, soils with high microbial biomass C and microbial biomass C to soil organic C ratio had high Q10. Overall, understanding the complex relationships among Q10, mineral protection, and microbial attributes on a spatial scale is essential for accurately predicting soil C cycling in forest ecosystems.

Genome-wide association analysis reveals genes controlling an antagonistic effect of biotic and osmotic stress on Arabidopsis thaliana growth.

While the response of Arabidopsis thaliana to drought, herbivory or fungal infection has been well-examined, the consequences of exposure to a series of such (a)biotic stresses are not well studied. This work reports on the genetic mechanisms underlying the Arabidopsis response to single osmotic stress, and to combinatorial stress, either fungal infection using Botrytis cinerea or herbivory using Pieris rapae caterpillars followed by an osmotic stress treatment. Several small-effect genetic loci associated with rosette dry weight (DW), rosette water content (WC), and the projected rosette leaf area in response to combinatorial stress were mapped using univariate and multi-environment genome-wide association approaches. A single-nucleotide polymorphism (SNP) associated with DROUGHT-INDUCED 19 (DI19) was identified by both approaches, supporting its potential involvement in the response to combinatorial stress. Several SNPs were found to be in linkage disequilibrium with known stress-responsive genes such as PEROXIDASE 34 (PRX34), BASIC LEUCINE ZIPPER 25 (bZIP25), RESISTANCE METHYLATED GENE 1 (RMG1) and WHITE RUST RESISTANCE 4 (WRR4). An antagonistic effect between biotic and osmotic stress was found for prx34 and arf4 mutants, which suggests PRX34 and ARF4 play an important role in the response to the combinatorial stress.

Effect of minute amounts of arsenic on the sulfamethoxazole removal and microbial community structure via the SBR system.

In this study, the effect of arsenic on the sulfamethoxazole (SMX) removal efficiency and microbial community structure was investigated over 60 days using the SBR process. The results showed that the presence of arsenic had no significant impact on the system performance, the removal efficiencies of two reactors, R1 (the control test) and R2 (with the addition of arsenic), were 13.36 ± 5.71 and 14.20 ± 5.27%, which were attributed to the adsorption of SMX by fulvic acid-like substances and tryptophan-like proteins of extracellular polymeric substances. Compared to the seed sludge, the species number indicated that R2 possessed the richer diversity, while R1 possessed the lower diversity on day 60, which might be relative to the transferring of antibiotic resistance genes (ARGs) in sludge bacterial communities; the minute amounts of arsenic could make the relative levels of Sul1 and Sul2 genes which encode ARGs of sulfonamides in R2 (2.07 and 2.47%) be higher than that in R1 (1.65 and 1.27%), which made the bacterial community of the R2 system more adaptable to SMX stress. Therefore, the minute amounts of arsenic weakened the effect of SMX on the system and enhanced the stability of the microbial community structure.

Efficient partial nitrification with hybrid nitrifying granular sludge based on a simultaneous fill/draw SBR mode.

In this study, a simultaneous fill/draw SBR was applied to investigate the feasibility of partial nitrification process with inoculation of matured aerobic granular sludge. The system operated stably over 120 days with the relatively high ammonium removal efficiency (≥ 98.83%) and nitrite accumulation rate (≥ 89.60%). Moreover, a hybrid flocs/granules system was formed stably after long-term operation. The nitrite-oxidizing bacteria (NOB) was suppressed effectively because of the combined effect of simultaneous fill/draw mode and intermittent aeration conditions. Furthermore, batch tests were separately tested with isolated granules (> 200 µm) and flocs (< 200 µm), showing that the specific ammonia oxidation rate of granules and flocs were 15.94 ± 2.85 and 66.77 ± 0.83 mg N/(g MLSS·h), respectively. Correspondingly, the abundance of Nitrosomonas as a typical AOB in granules (6.24%) and flocs (11.94%) was obtained via the microbial diversity analysis, while NOB was almost hardly detected in granules and flocs.

The chromosome-level wintersweet (Chimonanthus praecox) genome provides insights into floral scent biosynthesis and flowering in winter.

BACKGROUND: Wintersweet (Chimonanthus praecox), an important ornamental plant, has evolved unique fragrant aroma and winter-flowering properties, which are critical for its successful sexual reproduction. However, the molecular mechanisms underlying these traits are largely unknown in this species. In addition, wintersweet is also a typical representative species of the magnoliids, where the phylogenetic position of which relative to eudicots and monocots has not been conclusively resolved. RESULTS: Here, we present a chromosome-level wintersweet genome assembly with a total size of 695.36 Mb and a draft genome assembly of Calycanthus chinensis. Phylogenetic analyses of 17 representative angiosperm genomes suggest that Magnoliids and eudicots are sister to monocots. Whole-genome duplication signatures reveal two major duplication events in the evolutionary history of the wintersweet genome, with an ancient one shared by Laurales, and a more recent one shared by the Calycantaceae. Whole-genome duplication and tandem duplication events have significant impacts on copy numbers of genes related to terpene and benzenoid/phenylpropanoid (the main floral scent volatiles) biosynthesis, which may contribute to the characteristic aroma formation. An integrative analysis combining cytology with genomic and transcriptomic data reveals biological characteristics of wintersweet, such as floral transition in spring, floral organ specification, low temperature-mediated floral bud break, early blooming in winter, and strong cold tolerance. CONCLUSIONS: These findings provide insights into the evolutionary history of wintersweet and the relationships among the Magnoliids, monocots, and eudicots; the molecular basis underlying floral scent biosynthesis; and winter flowering, and highlight the utility of multi-omics data in deciphering important ornamental traits in wintersweet.

Effects of reaction conditions on products and elements distribution via hydrothermal liquefaction of duckweed for wastewater treatment.

Wastewater treatment by duckweed is a naturally sustainable technology. However, its development is limited due to the lack of a follow-up treatment of duckweed. The duckweed was proposed for the treatment of rural domestic wastewater and agricultural wastewater, and it was further processed to produce bio-oil via hydrothermal liquefaction at various temperatures (250 °C-370 °C) and residence times (15-60 min). The highest bio-oil yield of 35.6 wt% was obtained at 370 °C, 45 min. The higher heating value of bio-oil was 40.85 MJ/kg, and the H/C ratio (1.72-1.98) was similar to that of petroleum (1.84). The gas chromatography-mass spectrometry analysis results revealed that the bio-oil mainly consisted of N-heterocycles, cyclic ketones, esters, amides, long-chain hydrocarbons, phenols, and aromatic intermediates. Valuable compounds (3-pyridinol, 2-pyrrolidinone, and its analogues) of high concentration were identified in the water-soluble organic matter. Compared with other materials, this study produced higher-quality bio-oil and water-soluble organic matter.

Expression of the subgroup IIIf bHLH transcription factor CpbHLH1 from Chimonanthus praecox (L.) in transgenic model plants inhibits anthocyanin accumulation.

KEY MESSAGE: Overexpression of CpbHLH1 in Arabidopsis and tobacco resulted in a dramatic decrease in anthocyanin accumulation by repressing the expression of late biosynthesis genes in the flavonoid biosynthesis pathway. Many basic helix-loop-helix (bHLH) transcription factors (TFs) of subgroup IIIf have been characterized as anthocyanin-associated activators in higher plants, but information regarding bHLH TFs that inhibit anthocyanin accumulation remains scarce. In this study, the subgroup IIIf bHLH TF CpbHLH1 from Chimonanthus praecox (L.) was identified as a negative regulator of anthocyanin accumulation. Our results showed that overexpression of CpbHLH1 in model plant species, Arabidopsis and tobacco, resulted in a dramatic decrease in anthocyanin content, whereas the content of proanthocyanidin was little affected. Quantitative RT-PCR (qRT-PCR) assays of the structural genes in the flavonoid biosynthesis pathway revealed that CpbHLH1 inhibits anthocyanin accumulation mainly through repressing the expression of late biosynthesis genes (LBGs). Interactions between CpbHLH1 protein and AtPAP1/NtAN2 protein were detected via yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays. This is the first bHLH repressor of anthocyanin biosynthesis identified in dicotyledons. These results can help us better understand the anthocyanin regulatory network in plants and may provide insights into the diverse functions of bHLH proteins.

In-Situ imaging detection of cell membrane and intracellular cholesterol via cascade reactions.

Herein, an effective membrane-to-intracellular cholesterol detection strategy was designed based on cascade reactions. A biochip array was firstly fabricated by consecutively immobilizing luminol modified gold nanoparticles (Au@luminol), soybean peroxidase (SBP) and cholesterol oxidase (ChoX) on the cellulose acetate (CA) membrane functionalized home-made micropore array. When cholesterol existed, it was oxidized by ChoX generating H2O2, which further triggered the CL reaction under the SBP catalysis, the CL signals were collected by a charge-coupled device (CCD). The proposed strategy exhibited a wide linear range from 0.12 µM to 1000 µM and relatively low detection limit (LOD) of 0.08 µM. Furthermore，it could be used to in-situ detect membrane cholesterol and intracelluar esterified cholesterol in HepG2 cells. After activated HepG2 cells were added to the modified biochip, membrane cholesterol was detected directly. Intracelluar esterified cholesterol was detected through the introduction of triton X-100 and cholesteryl esterase (ChoE). Additionally, the cholesterol content in cells was changed after stimulated by drugs, such as apolipoprotein A-I (ApoA-I), pitavastatin or probucol. The correlation of the CL signal with the amount of cholesterol confirmed that our strategy was feasible to simultaneously detect membrane and intracellular cholesterol at different cellular states. The proposed strategy exhibited excellent sensitivity, selectivity, stability, and reproducibility in a simple, cheap way, which opened a new door for studying clinic treatment of the cholesterol-related diseases.

Transcriptomic and proteomic approaches to explore the differences in monoterpene and benzenoid biosynthesis between scented and unscented genotypes of wintersweet.

Wintersweet (Chimonanthus praecox L.) is an important ornamental plant in China with a pleasant floral scent. To explore the potential mechanisms underlying differences in the fragrances among genotypes of this plant, we analyzed floral volatile organic compounds (VOCs) from two different genotypes: SW001, which has little to no fragrance, and the scented genotype H29. The major VOCs in H29 were linalool, trans-ß-ocimene, benzyl acetate, methyl salicylate, benzyl alcohol (BAlc) and methyl benzoate. The most important aroma-active compound in H29, linalool, was emitted at a low concentration in SW001, which had markedly higher levels of trans-ß-ocimene than H29. Next, to investigate scent biosynthesis, we analyzed the transcriptome and proteome of fully open flowers of the two genotypes. A total of 14 443 differentially expressed unigenes and 196 differentially expressed proteins were identified. Further analyses indicated that 56 differentially expressed genes involved in the terpenoid and benzenoid biosynthesis pathways might play critical roles in regulating floral fragrance difference. Disequilibrium expression of four terpene synthase genes resulted in diverse emission of linalool and trans-ß-ocimene in both genotypes. In addition, the expressions of two CpMYC2 transcription factors were both upregulated in H29, implying that they may regulate linalool production. Notably, 16 of 20 genes in the benzenoid biosynthesis pathway were downregulated, corresponding to the relatively low level of benzenoid production in SW001. The lack of benzyl acetate might indicate that SW001 may lack substrate BAlc or functional acetyl-CoA:benzylalcohol acetyltransferase.

Simultaneous detection of three biomarkers related to acute myocardial infarction based on immunosensing biochip.

An immunosensing biochip for simultaneous detection of three biomarkers related to acute myocardial infarction (AMI) was developed based on anionic soybean peroxidase (SBP) functionalized nanoprobe and chemiluminescent imaging. The nanoprobes (Ab2-SiO2-SBP) were fabricated by co-immobilization of SBP and one of the detection polyclonal antibodies, anti-cardiac troponin I antigen (anti-cTnI), anti-creatine kinase-MB (anti-CK-MB) and anti-myoglobin (anti-Myo), on the silica nanoparticle surface. The detection sensitivity was enhanced since the large surface area of silica carriers increased the loading of SBP for per sandwiched immunoreaction. The immunosensing biochip designed as 3 × 8 wells array was constructed by simultaneously immobilizing three capture monoclonal antibodies on the same one microtiter well with 2 × 3 active spots. In the presence of target protein, the nanoprobes will be attached onto the spots with high specificity through the sandwiched immunoreactions, which triggered the chemiluminescence (CL) signals on each sensing site of the microtiter plates and allowed to CL imaging of three biomarkers in one well at the same time. Therefore, the proposed biochip was a promising convenient strategy for simultaneous detection of cTnI, CK-MB and Myo, which showed potential application for multianalyte determination in clinical diagnostics.

Comprehensive analysis of wintersweet flower reveals key structural genes involved in flavonoid biosynthetic pathway.

Wintersweet (Chimonanthus praecox (L.)), with an over-one-thousand-years long history in cultivation, is still a popular ornamental woody plant in China. The tepals of wintersweet flower are waxy in nature and the overall color of the flower is yellow, while the inner tepals range from yellow to red, which makes it an ideal plant to study floral color formation in ornamental shrubs. In our current work, HPLC analysis revealed that the principal pigments in tepals were the metabolite of flavonoids. All the tepals were containing quercetin, kaempferol 3­O­rutinoside and rutin while cyanidin­3­O­glucoside and cyanidin­3­O­rutinoside were only found in the in the red tepals. Moreover, we found the rutin as the principal component of all the pigments revealed. As well as in this study, a reference transcriptome library constructed from two varieties H29 and H64 flower. Further, 30 proteins of flavonoid biosynthesis pathway were identified in H29 flower using proteome analysis. Based on these dataset, the flavonoid biosynthesis pathway was also speculated. After quantitative analysis of gene expression, we found that ANS act as an on-off switch for the accumulation of red pigments and had positive correlations with various steps genes of the flavonoid pathway. This expression profiling demonstrates that no gene products compete for common substrates to redirect the metabolic flux in wintersweet. It is also demonstrated that high expression of F3'H would provide sufficient content of the precursor, dihydroquercetin, for both flavonol and anthocyanin biosynthesis. The results help us to deepen and enrich the gene resource of color formation in wintersweet flower, and provide specific breeding strategies for increasing diversity of flower color.

B and E MADS-box genes determine the perianth formation in Cymbidium goeringii Rchb.f.

Cymbidium goeringii Rchb.f. is an important ornamental plant with a striking well-differentiated lip. Its complex floral architecture presents an exciting opportunity to examine perianth development. In flowering plants, class A, B and E floral homeotic genes play key roles in the specification of perianth identity. In this study, we used a cDNA library of wild-type C. goeringii flower buds for transcriptome sequencing. Eighteen candidate class A, B and E genes (including AP1/FUL-, AP2-, DEF-, GLO-, SEP- and AGL6-like genes) were identified. Quantitative real time polymerase chain reaction (qRT-PCR) results showed that CgDEF1, CgSEP2 and CgAGL6-1 were strongly detected only in the sepals and petals and were significantly downregulated in the lips. CgDEF3, CgDEF4 and CgAGL6-3 were highly expressed in the lips and lip-like petals but were only minimally detected in the sepals. Yeast two-hybrid analysis indicated that CgDEF1 and CgGLO formed a heterodimer. CgAGL6-1/CgSEP2 and CgDEF1 formed higher-order protein complexes with the assistance of the CgGLO protein, and both CgAGL6-1 and CgSEP2 formed a heterodimer. CgDEF3/CgDEF4 could interact independently with CgGLO and CgAGL6-3, respectively, while CgDEF3 and CgDEF4 also formed heterodimers with the assistance of the CgGLO. Based on a comprehensive analysis relating these gene expression patterns to protein interaction profiles, the mechanism of sepal/petal/lip determination was studied in C. goeringii. Furthermore, a hypothesis explaining the sepal/petal/lip determination of C. goeringii is proposed. The lip-quartet (CgDEF3/CgDEF4/CgAGL6-3/CgGLO) promoted lip formation, whereas the sepal/petal-quartet (CgDEF1/CgAGL6-1/CgSEP2/CgGLO) promoted sepal/petal formation. These results enrich the current knowledge regarding the mechanism and pathways of perianth formation in orchids.

Molecular cloning and expression of Chimonanthus praecox farnesyl pyrophosphate synthase gene and its possible involvement in the biosynthesis of floral volatile sesquiterpenoids.

Farnesyl pyrophosphate (FPP) synthase catalyzes the biosynthesis of FPP, which is the precursors of sesquiterpenoids such as floral scent volatiles, from isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP). cDNA encoding wintersweet (Chimonanthus praecox L.) FPP synthase was isolated by the RT-PCR and RACE methods. The deduced amino acid sequence showed a high identity to plant FPP synthases. Expression of the gene in Escherichia coli yielded FPPS activity that catalyzed the synthesis of FPP as a main product. Tissue-specific and developmental analyses of the mRNA levels of CpFPPS and volatile sesquiterpenoids levels in C. praecox flowers revealed that the FPPS may play a regulatory role in floral volatile sesquiterpenoids of wintersweet.