Revealing mechanism of phenolic-amine reaction to form humus in compost based on high-resolution liquid chromatography mass spectrometry and spectroscopy.

Humus is the stable form of carbon storage in straw compost. The phenolic-amine reaction is a pathway for humus formation in straw compost. In this study, two reaction systems, GP group (pyrogallol and glycine) and GCP group (catechol, pyrogallol, and glycine), were constructed in a simulated composting environment and revealed the molecular binding mechanism of the phenolic-amine reaction through spectroscopy and mass spectrometry. The results showed that phenolic self-polymerization was faster than phenolic-amine reaction. Therefore, the aromatization degree of GP was 27.14 % higher than that of GCP. The phenolic-amine reaction first produced fulvic acid, and then formed humus units rich in active functional group structures (i.e., phenolic hydroxyl and carboxyl groups). These units further captured small molecule compounds to eventually form humic acid. This study would provide theoretical support for exploring the humus formation process and the promotion of the straw humification by adding phenol or amino acids to compost.

Photosynthetic carbon allocation in native and invasive salt marshes undergoing hydrological change: A mesocosm experiment.

Biogeochemical processes mediated by plants and soil in coastal marshes are vulnerable to environmental changes and biological invasion. In particular, tidal inundation and salinity stress will intensify under future rising sea level scenarios. In this study, the interactive effects of flooding regimes (non-waterlogging vs. waterlogging) and salinity (0, 5, 15, and 30 parts per thousand (ppt)) on photosynthetic carbon allocation in plant, rhizodeposition, and microbial communities in native (Phragmites australis) and invasive (Spartina alterniflora) marshes were investigated using mesocosm experiments and 13CO2 pulse-labeling techniques. The results showed that waterlogging and elevated salinity treatments decreased specific root allocation (SRA) of 13C, rhizodeposition allocation (RA) 13C, soil 13C content, grouped microbial PLFAs, and the fungal 13C proportion relative to total PLFAs-13C. The lowest SRA, RA, and fungal 13C proportion occurred under the combined waterlogging and high (30 ppt) salinity treatments. Relative to S. alterniflora, P. australis displayed greater sensitivity to hydrological changes, with a greater reduction in rhizodeposition, soil 13C content, and fungal PLFAs. S. alterniflora showed an earlier peak SRA but a lower root/shoot 13C ratio than P. australis. This suggests that S. alterniflora may transfer more photosynthetic carbon to the shoot and rhizosphere to facilitate invasion under stress. Waterlogging and high salinity treatments shifted C allocation towards bacteria over fungi for both plant species, with a higher allocation shift in S. alterniflora soil, revealing the species-specific microbial response to hydrological stresses. Potential shifts towards less efficient bacterial pathways might result in accelerated carbon loss. Over the study period, salinity was the primary driver for both species, explaining 33.2-50.8 % of 13C allocation in the plant-soil-microbe system. We propose that future carbon dynamics in coastal salt marshes under sea-level rise conditions depend on species-specific adaptive strategies and carbon allocation patterns of native and invasive plant-soil systems.

Novel homozygous ADK out-of-frame deletion causes adenosine kinase deficiency with rare phenotypes of sepsis, metabolites disruption and neutrophil dysfunction.

Adenosine kinase deficiency (OMIM #614300) is a type of inborn errors of metabolism with multiorgan symptoms primarily neurological disorders, hepatic impairment, global developmental delay, and mild dysmorphism. The genetic causes of adenosine kinase deficiency are homozygous or compound heterozygous loss-of-function variants of ADK. To date, fewer than 25 cases of adenosine kinase deficiency have been reported worldwide and none have been reported in China. In this research, trio whole-exome sequencing (Trio-WES) identified a novel homozygous ADK (NM_001123.4) out-of-frame deletion, c.518_519delCA (p.Thr173Serfs*15), in a Chinese patient with rare phenotypes of sepsis, metabolites disruption and neutrophil dysfunction. This variant was dysfunctional, with marked reduction of ADK level in both the patient's peripheral blood and cells transfected with the corresponding variant. Additionally, metabolomics detected by high-throughput mass spectrometry showed disturbances in the methionine (Met) and energy pathway. RNA sequencing (RNA-seq) of the patient's peripheral blood suggested a defective anti-inflammatory response characterized by impaired neutrophil activation, migration, and degranulation, which might be the primary cause for the sepsis. To our knowledge, we identified the first Chinese patient of adenosine kinase deficiency with a novel homozygous out-of-frame deletion in ADK causing multiorgan disorders, metabolites disruption, rare phenotypes of sepsis, and neutrophil dysfunction. Our findings broaden the genetic spectrum and pathogenic mechanisms of adenosine kinase deficiency.

The relationship between the microclimate and efflorescence of revealed mural paintings and the later protection strategy.

Throughout history, there were many fine mural paintings concealed within ancient buildings, hidden beneath layers of plaster, wall or other structure. In recent decades, research and practical efforts had primarily focused on nondestructive diagnosis of these hidden murals and the removal of their surface coverings. However, limited attention had been given to the consequences of overlay removal on mural preservation. This study aims to address this gap by examining the revealed mural paintings in the Prince Shi's Palace as a case study, employing an analysis of mural ontology, investigation into the preservation environment, and simulation experiments to comprehensively analyze the factors contributing to the rapid deterioration of these revealed murals. The findings indicated that certain carbonated lime particles adhered to the mural surface, and in a humid environment, these particles absorbed sulfur dioxide from the atmosphere and converted into calcium sulfate. The strong hygroscopic property of calcium sulfate enhanced the humidity on the surface of murals, thereby increasing their susceptibility to sulfur dioxide absorption. Over time, the murals suffered more sulphate and moisture damage. Additionally, historically inaccurate interventions such as non-breathable shading curtains installation and organic coatings reinforcement exacerbated this deterioration process. Therefore, safeguarding the integrity of these exposed concealed murals necessitated prioritizing the preservation of environmental humidity and acid gas levels.

Orchestrating seed storage protein and starch accumulation toward overcoming yield-quality trade-off in cereal crops.

Achieving high yield and good quality in crops is essential for human food security and health. However, there is usually disharmony between yield and quality. Seed storage protein (SSP) and starch, the predominant components in cereal grains, determine yield and quality, and their coupled synthesis causes a yield-quality trade-off. Therefore, dissection of the underlying regulatory mechanism facilitates simultaneous improvement of yield and quality. Here, we summarize current findings about the synergistic molecular machinery underpinning SSP and starch synthesis in the leading staple cereal crops, including maize, rice and wheat. We further evaluate the functional conservation and differentiation of key regulators and specify feasible research approaches to identify additional regulators and expand insights. We also present major strategies to leverage resultant information for simultaneous improvement of yield and quality by molecular breeding. Finally, future perspectives on major challenges are proposed.

Malonic acid shapes bacterial community dynamics in compost to promote carbon sequestration and humic substance synthesis.

The incorporation of malonic acid (MA) into compost as a regulator of the tricarboxylic acid (TCA) cycle has the potential to increase carbon sequestration. However, the influence of MA on the transformation of the microbial community during the composting process remains unclear. In this investigation, MA was introduced at different stages of chicken manure (CM) composting to characterize the bacterial community within the compost using high-throughput sequencing. We assess the extent of increased carbon sequestration by comparing the concentration of total organic carbon (TOC). At the same time, this study examines whether increased carbon sequestration contributes to humus formation, which was elucidated by evaluating the content and composition of humus. Our results show that the addition of MA significantly improved carbon sequestration within the compost, reducing the carbon loss rate (C loss (%)) from 64.70% to 52.94%, while increasing HS content and stability. High throughput sequencing and Random Forest (RF) analysis show that the introduction of MA leads to a reduction in the diversity of the bacterial communities, but enhanced the ability of bacterial communities to synthesize humus. Furthermore, the addition of MA favors the proliferation of Firmicutes. Also, the hub of operational taxonomic units (OTUs) within the community co-occurrence network shifts from Proteobacteria to Firmicutes. Remarkably, our study finds a significant decrease in negative correlations between bacteria, potentially mitigating substrate consumption due to negative interactions such as competition. This phenomenon contributes to the improved retention of TOC in the compost. This research provides new insights into the mechanisms by which MA regulates bacterial communities in compost, and provides a valuable theoretical basis for the adoption of this innovative composting strategy.

Insight into bacterial role attribution in dissolved organic matter humification during rice straw composting with microbial inoculation.

This study aims to investigate the effect of microbial role distribution in microbial carbon pumps on dissolved organic matter (DOM) humification during rice straw composting with microbial inoculation. Three composting groups were designed, named CK (control), B4 (with Bacillus subtilis, OR058594) and Z1 (with Aspergillus fumigatus, AF202956.1). As a result of inoculation, the composition of microbial communities was changed, so that the microorganisms that promoted DOM humification were concentrated in the responders in the microbial carbon pump. DOM was divided into three components in three composting treatments: C1, C2 and C3. After inoculation with Bacillus subtilis, the C2 component was significantly affected, while after inoculation with Aspergillus fumigatus, the C3 component was significantly affected. The results of physicochemical factors affecting the transformation of DOM fluorescence components indicated that C1, C2 and C3 were related to the abundance of the cellulose-degrading enzyme-encoding gene GH7 in CK and B4 composting. However, the C2 was susceptible to organic matter in Z1 composting. This study explored the distribution of microbial communities from a new perspective, which provided new information for analyzing DOM humification and treating agricultural straws to achieve clean conditions for environmental friendliness.

The TaSOC1-TaVRN1 module integrates photoperiod and vernalization signals to regulate wheat flowering.

Wheat needs different durations of vernalization, which accelerates flowering by exposure to cold temperature, to ensure reproductive development at the optimum time, as that is critical for adaptability and high yield. TaVRN1 is the central flowering regulator in the vernalization pathway and encodes a MADS-box transcription factor (TF) that usually works by forming hetero- or homo-dimers. We previously identified that TaVRN1 bound to an MADS-box TF TaSOC1 whose orthologues are flowering activators in other plants. The specific function of TaSOC1 and the biological implication of its interaction with TaVRN1 remained unknown. Here, we demonstrated that TaSOC1 was a flowering repressor in the vernalization and photoperiod pathways by overexpression and knockout assays. We confirmed the physical interaction between TaSOC1 and TaVRN1 in wheat protoplasts and in planta, and further validated their genetic interplay. A Flowering Promoting Factor 1-like gene TaFPF1-2B was identified as a common downstream target of TaSOC1 and TaVRN1 through transcriptome and chromatin immunoprecipitation analyses. TaSOC1 competed with TaVRT2, another MADS-box flowering regulator, to bind to TaVRN1; their coding genes synergistically control TaFPF1-2B expression and flowering initiation in response to photoperiod and low temperature. We identified major haplotypes of TaSOC1 and found that TaSOC1-Hap1 conferred earlier flowering than TaSOC1-Hap2 and had been subjected to positive selection in wheat breeding. We also revealed that wheat SOC1 family members were important domestication loci and expanded by tandem and segmental duplication events. These findings offer new insights into the regulatory mechanism underlying flowering control along with useful genetic resources for wheat improvement.

Arabidopsis G-protein ß subunit AGB1 represses abscisic acid signaling via attenuation of the MPK3-VIP1 phosphorylation cascade.

Heterotrimeric G proteins play key roles in cellular processes. Although phenotypic analyses of Arabidopsis Gß (AGB1) mutants have implicated G proteins in abscisic acid (ABA) signaling, the AGB1-mediated modules involved in ABA responses remain unclear. We found that a partial AGB1 protein was localized to the nucleus where it interacted with ABA-activated VirE2-interacting protein 1 (VIP1) and mitogen-activated protein kinase 3 (MPK3). AGB1 acts as an upstream negative regulator of VIP1 activity by initiating responses to ABA and drought stress, and VIP1 regulates the ABA signaling pathway in an MPK3-dependent manner in Arabidopsis. AGB1 outcompeted VIP1 for interaction with the C-terminus of MPK3, and prevented phosphorylation of VIP1 by MPK3. Importantly, ABA treatment reduced AGB1 expression in the wild type, but increased in vip1 and mpk3 mutants. VIP1 associates with ABA response elements present in the AGB1 promoter, forming a negative feedback regulatory loop. Thus, our study defines a new mechanism for fine-tuning ABA signaling through the interplay between AGB1 and MPK3-VIP1. Furthermore, it suggests a common G protein mechanism to receive and transduce signals from the external environment.

Rht12b, a widely used ancient allele of TaGA2oxA13, reduces plant height and enhances yield potential in wheat.

KEY MESSAGE: We identified a new wheat dwarfing allele Rht12b conferring reduced height and higher grain yield, pinpointed its causal variations, developed a breeding-applicable marker, and traced its origin and worldwide distribution. Plant height control is essential to optimize lodging resistance and yield gain in crops. RHT12 is a reduced height (Rht) locus that is identified in a mutationally induced dwarfing mutant and encodes a gibberellin 2-oxidase TaGA2oxA13. However, the artificial dwarfing allele is not used in wheat breeding due to excessive height reduction. Here, we confirmed a stable Rht locus, overlapping with RHT12, in a panel of wheat cultivars and its dwarfing allele reduced plant height by 5.4-8.2 cm, equivalent to Rht12b, a new allele of RHT12. We validated the effect of Rht12b on plant height in a bi-parent mapping population. Importantly, wheat cultivars carrying Rht12b had higher grain yield than those with the contrasting Rht12a allele. Rht12b conferred higher expression level of TaGA2oxA13. Transient activation assays defined SNP-390(C/A) in the promoter of TaGA2oxA13 as the causal variation. An efficient kompetitive allele-specific PCR marker was developed to diagnose Rht12b. Conjoint analysis showed that Rht12b plus the widely used Rht-D1b, Rht8 and Rht24b was the predominant Rht combination and conferred a moderate plant height in tested wheat cultivars. Evolutionary tracking uncovered that RHT12 locus arose from a tandem duplication event with Rht12b firstly appearing in wild emmer. The frequency of Rht12b was approximately 70% (700/1005) in a worldwide wheat panel and comparable to or higher than those of other widely used Rht genes, suggesting it had been subjected to positive selection. These findings not only identify a valuable Rht gene for wheat improvement but also develop a functionally diagnostic tool for marker-assisted breeding.

Effect of ferrous ions combined with zeolite on humification degree during food waste composting.

The research aims to clarify role of ferrous sulfate (FeSO4) combined with zeolite (Z) on humification degree based on investigation of concentration and structural stability of humic acid (HA) during food waste composting. Four treatments were set up, namely CK (control), Fe (5 %), Z (5 %) and Fe + Z (2.5 %+2.5 %). Results demonstrated that concentration and polymerization degree of HA were 53.4 % and 97.3 % higher in composting amended with Fe + Z than in the control, respectively. Meanwhile, formation of aromatic functional groups and recalcitrant fluorescent components (HAC3) was significantly promoted, indicating that Fe + Z treatment enhanced HA structure stability. The bacterial networks became tighter, and the proportion of core bacteria in dominant modules increased at Fe + Z treatment. Additionally, key factors affecting HAC3 and product quality were identified by structural equation models, which verified potential mechanism of humification enhancement. Overall, this study provided theoretical support for improving humification degree and product quality.

Biochemistry and Protein Interactions of the CYREN Microprotein.

Over the past decade, advances in genomics have identified thousands of additional protein-coding small open reading frames (smORFs) missed by traditional gene finding approaches. These smORFs encode peptides and small proteins, commonly termed micropeptides or microproteins. Several of these newly discovered microproteins have biological functions and operate through interactions with proteins and protein complexes within the cell. CYREN1 is a characterized microprotein that regulates double-strand break repair in mammalian cells through interaction with Ku70/80 heterodimer. Ku70/80 binds to and stabilizes double-strand breaks and recruits the machinery needed for nonhomologous end join repair. In this study, we examined the biochemical properties of CYREN1 to better understand and explain its cellular protein interactions. Our findings support that CYREN1 is an intrinsically disordered microprotein and this disordered structure allows it to enriches several proteins, including a newly discovered interaction with SF3B1 via a distinct short linear motif (SLiMs) on CYREN1. Since many microproteins are predicted to be disordered, CYREN1 is an exemplar of how microproteins interact with other proteins and reveals an unknown scaffolding function of this microprotein that may link NHEJ and splicing.

Improved carbon sequestration by utilization of ferrous ions during different organic wastes composting.

The humic acid (HA) possesses a more recalcitrant structure, making it crucial carbon components that improve carbon sequestration. Moreover, ferrous ions could improve microbial activity and enhance compost humification, and their oxidation into iron oxides could adsorb carbon components for sequestration. Based on the advantages of low cost and easy availability of ferrous sulfate (FeSO4), this study investigated the effect of FeSO4 on carbon sequestration during composting. Chicken manure (CM) and food waste (FW) composting were carried out in four treatments, namely control (CM, FW) and 5% (w/w) FeSO4 treated groups (CM+, FW+). Results indicated that FeSO4 increased HA content, improved organic carbon stability. Carbon loss for CM, CM+, FW and FW + treatments were 48.5%, 46.2%, 45.0%, and 40.3%, respectively. Meanwhile, FeSO4 enhanced the function of bacterial taxa involved in HA synthesis in CM + treatment, and improved the number of core bacteria significantly associated with formation of HA and iron oxide. SEM analysis verified that role of FeSO4 was significant in promoting HA synthesis during CM + composting, while it was remarkably in enhancing HA sequestration during FW + composting. This article provided fundamental theoretical backing for enhancing HA production and improving carbon sequestration during different materials composting.

Origin, distribution and spatial characteristics of dissolved organic matter in the Heilongjiang River.

The Heilongjiang River Basin is a vast area with significant DOM sources and composition differences. The mechanism of DOM degradation under spatial variation remains unclear. This research investigated the degradation characteristics of DOM in different watersheds of the Heilongjiang River. DOM levels were higher in midstream waters, while DOM degradation rates were higher in midstream and downstream waters. The parallel factor analysis (PARAFAC) results showed that the upstream amino acid fraction was significantly depleted, the midstream was dominated by the degradation of DOM of terrestrial origin, and the downstream humic acid fraction was decreased considerably. Gene sequencing results indicated that the upstream, middle, and downstream water bodies' microbial community composition and structure differed significantly. The network analysis results revealed microorganisms in upstream water bodies mainly utilized amino acid-like substances and small molecule humic acids. Microorganisms in the middle and lower reaches of the water column were characterized by the utilization of humic acid-like fractions. In this study, we further screened the key driving microorganisms (e.g., Flavobacterium and Lacibacter) responsible for the difference in the DOM utilization function of upstream-to-midstream and midstream-to-downstream microorganisms in the Heilongjiang River. These findings will help identify the cycling process of DOM under spatial variation and predict the succession pattern of microbial communities.

Exploring the nitrogen fixing strategy of bacterial communities in nitrogen cycling by adding calcium superphosphate at various periods during composting.

Chicken manure, as an organic solid waste with a high nitrogen content, generates large amounts of ammonia during composting, which leads to pollution of the surrounding environment, and causes a reduction in the quality of the compost product. Nitrogen is transformed through the nitrogen cycle and bacterial communities are the main contributors to the transformation of the nitrogen cycle. The microbial composition changes dramatically at different stages during composting. Therefore, calcium superphosphate (SSP) was added to compost as a nitrogen-fixing agent to elucidate the strategy and function of the bacterial community involved in the nitrogen cycle. The results showed that the addition of SSP at the initial, high temperature and cooling stages increased the inorganic nitrogen (NH4+-N, NO3--N) content by 51.99 %, 202.72 % and 173.37 % compared to CK, respectively. In addition, nitrogen cycle functional genes (gdh, nifH, pmoA-amoA, hao, nxrA, nirK, napA, nosZ, narG) abundance were determined by real-time qPCR. The nitrogen cycle genetic results showed that SSP addition at high temperature phase resulted in a 62.43 % down-regulation of ammonification genes, while nitrogen fixation and nitrification genes were enhanced. Random forests revealed a shift in the participation strategy of bacterial communities (e.g., Mycobacterium, Izemoplasmatales, Paracoccus, Ruminococcus) within the nitrogen cycle, leading to altered importance rankings despite involvement in different nitrogen cycle pathways. Moreover, Regression analysis and structural equation modelling revealed that SSP addition at high temperature stage stimulated the bacterial community engaged in nitrogen fixation and nitrification, resulting in increased nitrogen accumulation as NO3--N during composting. This paper offers the potential to yield novel scientific insights into the impact of microbially mediated nitrogen transformation processes and reduce gaseous pollution.

Fine mapping and characterization of a major QTL for plant height on chromosome 5A in wheat.

KEY MESSAGE: We precisely mapped QPH.caas-5AL for plant height in wheat, predicted candidate genes and confirmed genetic effects in a panel of wheat cultivars. Plant height is an important agronomic trait, and appropriately reduced height can improve yield potential and stability in wheat, usually combined with sufficient water and fertilizer. We previously detected a stable major-effect quantitative trait locus QPH.caas-5AL for plant height on chromosome 5A in a recombinant inbred line population of the cross 'Doumai × Shi 4185' using the wheat 90 K SNP assay. Here , QPH.caas-5AL was confirmed using new phenotypic data in additional environment and new-developed markers. We identified nine heterozygous recombinant plants for fine mapping of QPH.caas-5AL and developed 14 breeder-friendly kompetitive allele-specific PCR markers in the region of QPH.caas-5AL based on the genome re-sequencing data of parents. Phenotyping and genotyping analyses of secondary populations derived from the self-pollinated heterozygous recombinant plants delimited QPH.caas-5AL into an approximate 3.0 Mb physical region (521.0-524.0 Mb) according to the Chinese Spring reference genome. This region contains 45 annotated genes, and six of them were predicted as the candidates of QPH.caas-5AL based on genome and transcriptome sequencing analyses. We further validated that QPH.caas-5AL has significant effects on plant height but not yield component traits in a diverse panel of wheat cultivars; its dwarfing allele is frequently used in modern wheat cultivars. These findings lay a solid foundation for the map-based cloning of QPH.caas-5AL and also provide a breeding-applicable tool for its marker-assisted selection. Keymessage We precisely mapped QPH.caas-5AL for plant height in wheat, predicted candidate genes and confirmed genetic effects in a panel of wheat cultivars.

Genetic dissection of grain morphology and yield components in a wheat line with defective grain filling.

KEY MESSAGE: We identified stable QTL for grain morphology and yield component traits in a wheat defective grain filling line and validated genetic effects in a panel of cultivars using breeding-relevant markers. Grain filling capacity is essential for grain yield and appearance quality in cereal crops. Identification of genetic loci for grain filling is important for wheat improvement. However, there are few genetic studies on grain filling in wheat. Here, a defective grain filling (DGF) line wdgf1 characterized by shrunken grains was identified in a population derived from multi-round crosses involving nine parents and a recombinant inbreed line (RIL) population was generated from the cross between wdgf1 and a sister line with normal grains. We constructed a genetic map of the RIL population using the wheat 15K single nucleotide polymorphism chip and detected 25 stable quantitative trait loci (QTL) for grain morphology and yield components, including three for DGF, eleven for grain size, six for thousand grain weight, three for grain number per spike and two for spike number per m2. Among them, QDGF.caas-7A is co-located with QTGW.caas-7A and can explain 39.4-64.6% of the phenotypic variances, indicating that this QTL is a major locus controlling DGF. Sequencing and linkage mapping showed that TaSus2-2B and Rht-B1 were candidate genes for QTGW.caas-2B and the QTL cluster (QTGW.caas-4B, QGNS.caas-4B, and QSN.caas-4B), respectively. We developed kompetitive allele-specific PCR markers tightly linked to the stable QTL without corresponding to known yield-related genes, and validated their genetic effects in a diverse panel of wheat cultivars. These findings not only lay a solid foundation for genetic dissection underlying grain filling and yield formation, but also provide useful tools for marker-assisted breeding.

The dominant role of cooperation in fungal community drives the humification process of chicken manure composting under addition of regulatory factors.

This study aimed to explore the action mechanism of fungal community on the enhancement of humification during chicken manure composting by regulating the core pathway of carbon metabolism - the tricarboxylic acid cycle. Regulators adenosine triphosphate (ATP) and malonic acid were added at the beginning of composting. The analysis of changes in humification parameters showed that the humification degree and stability of compost products were improved by adding regulators. Compared with CK, the humification parameters of adding regulators group increased by 10.98% on average. Meanwhile, adding regulators not only increased key nodes, but also strengthened the positive correlation between fungi, and network relationship was closer. Moreover, core fungi associated with humification parameters were identified by constructing OTU networks, and the division and cooperation mechanism of fungi were confirmed. Ultimately, the functional role of the fungal community acting on humification was confirmed by statistical means, that was, the fungal community promoting humification was the main group of composting process. And the contribution was more obvious in ATP treatment. This study was helpful to gain insight into the mechanism of regulators addition to advance the humification process, and provided new ideas for the safe, efficient and harmless disposal of organic solid waste.

The role of microbiota during chicken manure and pig manure co-composting.

Co-composting is an excellent and effective technology for treating livestock manure in which microorganisms play a crucial function. Therefore, this study aimed at investigating the changes of microbial interactions during co-composting. Six different addition ratios of chicken and pig manure were used in composting experiment. The results showed that the co-composting system using 60% chicken manure and 40% pig manure significantly altered the microbial diversity and community structure. In addition, the complexity and tightness of its microbial community network structure reached the maximum, as did the strength of its cooperative and competitive microbial interactions. The higher microbial abundance and microbial interaction have the potential to promote the decomposition and transformation of compost components. Therefore, this study preliminarily revealed the changes of microbial community in co-composting, which provided a theoretical basis for optimizing microbial community interaction in composting systems by mixing different ratios of materials in practice.

Efficient proteome-wide identification of transcription factors targeting Glu-1: A case study for functional validation of TaB3-2A1 in wheat.

High-molecular-weight glutenin subunits (HMW-GS), a major component of seed storage proteins (SSP) in wheat, largely determine processing quality. HMW-GS encoded by GLU-1 loci are mainly controlled at the transcriptional level by interactions between cis-elements and transcription factors (TFs). We previously identified a conserved cis-regulatory module CCRM1-1 as the most essential cis-element for Glu-1 endosperm-specific high expression. However, the TFs targeting CCRM1-1 remained unknown. Here, we built the first DNA pull-down plus liquid chromatography-mass spectrometry platform in wheat and identified 31 TFs interacting with CCRM1-1. TaB3-2A1 as proof of concept was confirmed to bind to CCRM1-1 by yeast one hybrid and electrophoretic mobility shift assays. Transactivation experiments demonstrated that TaB3-2A1 repressed CCRM1-1-driven transcription activity. TaB3-2A1 overexpression significantly reduced HMW-GS and other SSP, but enhanced starch content. Transcriptome analyses confirmed that enhanced expression of TaB3-2A1 down-regulated SSP genes and up-regulated starch synthesis-related genes, such as TaAGPL3, TaAGPS2, TaGBSSI, TaSUS1 and TaSUS5, suggesting that it is an integrator modulating the balance of carbon and nitrogen metabolism. TaB3-2A1 also had significant effects on agronomic traits, including heading date, plant height and grain weight. We identified two major haplotypes of TaB3-2A1 and found that TaB3-2A1-Hap1 conferred lower seed protein content, but higher starch content, plant height and grain weight than TaB3-2A1-Hap2 and was subjected to positive selection in a panel of elite wheat cultivars. These findings provide a high-efficiency tool to detect TFs binding to targeted promoters, considerable gene resources for dissecting regulatory mechanisms underlying Glu-1 expression, and a useful gene for wheat improvement.