Context: Fruit production systems face increasing pressure to enhance environmental sustainability while maintaining economic viability.Objective: This study aims to compare the environmental impacts of four orchard floor management systems for Fengtang plum production in Southwest China, identify key environmental hotspots, and integrate economic analysis to evaluate environmental-economic trade-offs, thereby identifying strategies that optimize both sustainability and profitability.Methods: This study applied life cycle assessment methodology to evaluate the environmental and economic performance of four orchard floor management systems for Fengtang plum (Prunus salicina) production in Southwest China: chemical herbicide application (CHI), polyethylene film mulching (PEM), cover cropping (CC), and natural weed coverage (NWC). Based on nine-year data from 198 orchards, the cradle-to-farm-gate analysis used 1 ton of Fengtang plum as the functional unit, grading fruit by diameter and evaluating economics via production costs, revenue, and benefit-cost ratios. Nine midpoint environmental impact categories were assessed using the CML-IA baseline method.Results and conclusions: Environmental impacts varied substantially among systems. For global warming potential (GWP), rankings were NWC > PEM > CC > CHI. CC achieved the highest economic returns (BCR = 2.0), with moderate environmental impacts and competitive profitability through increased production of Top-grade and First-grade fruit. Organic fertilizer overuse emerged as the major environmental hotspot, accounting for 72.3–78.0% of acidification potential and 65.2–71.4% of respiratory inorganics. Polyethylene film accounted for 61.5% of primary energy demand, underscoring the need for end-of-life management.Significance: The integrated findings offer actionable insights for sustainable intensification policies and farm-level decisions in mountainous agroecosystems.

Mulberry (Morus alba L.), a high-protein woody forage, is promising for alleviating animal husbandry’s protein feed shortage but suffers significant protein degradation during anaerobic fermentation and ruminal digestion. This study evaluated tannic acid (HT) and Acacia mangium tannin (CT) at 1–5 g/kg DM for effects on mulberry fermentation quality, bacterial community, and protein utilisation. Results showed HT dose-dependently reduced fermentation crude protein loss (max 2.72% vs. CK), promoted Lactiplantibacillus dominance, inhibited harmful microbes, and improved protein profiles (increased PB2、PB3, decreased NPN, enhanced methionine). CT exacerbated fermentation protein degradation and suppressed Lactiplantibacillus. Both tannins reduced ruminal protein degradation (by up to 17.94%) and mitigated methane production. Adding tannic acid is an effective strategy to enhance the nutritional value of ensiled mulberry.

The humid climate and frequent rainfall during the harvest season substantially hinder the utilization of triticale as feed. Although ensiling technology can effectively preserve nutrients, its fermentation quality depends on complex microbial interactions, the core mechanisms of which remain unclear. This study proposes and validates the hypothesis that “bacterial–fungal synergy” can enhance silage fermentation. By inoculating triticale silage with Aspergillus niger (AN), Lactiplantibacillus plantarum (LP) or their combination (ANLP) and performing multi-omics analyses, the mechanism underlying this synergistic effect was systematically elucidated in this study. Compared with the control treatment, triticale silage inoculated with ANLP presented significant decreases in the neutral detergent fiber (NDF), acid detergent fiber (ADF), and ammonia nitrogen (NH3-N) contents and significant increases in the water-soluble carbohydrate (WSC), crude protein (CP), and lactic acid (LA) contents (P < 0.05). More crucially, ANLP treatment specifically enriched Delftia, indicating a special functional role for this bacterium in triticale silage. Further metabolomic and correlation analyses revealed that the synergy between A. niger and L. plantarum not only promoted the proliferation of Delftia but also activated the phenylalanine, tyrosine, and tryptophan biosynthesis pathways. This activation drove the synthesis of phenolic acid compounds with antimicrobial and antioxidant activities, such as coumaric acid and indole derivatives. These bioactive metabolites effectively inhibited the growth of harmful microorganisms. In vitro digestibility trials confirmed that the ANLP-treated group achieved the highest dry matter and protein degradation rates, thereby validating the pathway from the microbial mechanism to end-use feed value. Overall, the synergistic effects of bacteria (L. plantarum) and fungi (A. niger) can improve the fermentation quality and nutritional content of triticale by promoting amino acid metabolism and increasing the production of bioactive substances, providing a new strategy for increasing its utilization as a feed resource for ruminants.

Coal gangue stockpiling leads to considerable soil heavy metal contamination and ecological degradation. Vetiveria zizanioides (L.) exhibits strong phytoremediation potential; however, the mechanisms underlying heavy metal immobilisation during soil function recovery remains unclear. In this study, a spatiotemporal substitution method was applied to investigate soil heavy metal distribution, microbial communities, and metabolite profiles during a nine-year cultivation period of V. zizanioides in coal gangue. Long-term cultivation effectively immobilised heavy metals (Cr, Cu, and Zn), reduced their migration and progressively restored soil pH while enhancing soil multifunctionality. Bacterial network vulnerability decreased, whereas network complexity and robustness increased, resulting in greater microbial diversity in the coal gangue-V. zizanioides system. Sulfurifustis, a dominant bacterial genus, strongly influenced microbial network structure and promoted heavy metal immobilisation. Keystone taxa (Proteobacteria, Actinobacteriota, and Chloroflexi) were essential in determining bacterial network structures. In addition, purine metabolism and its intermediate metabolite allantoin were significantly enhanced and were potentially associated with plant detoxification under heavy metal stress in soils. Overall, these findings demonstrate that V. zizanioides cultivation immobilises heavy metals, restores soil functionality, and maintains microecological balance through bacterial network assembly and metabolic reprogramming.

The experiment aimed to investigate the optimal processing technology for rice straw fermented with compound probiotics. Using molasses, distiller's grains, corn flour and wheat bran as exogenous additives, rice straw was fermented with compound probiotics consisting of Lactiplantibacillus plantarum, Enterococcus faecalis, Bacillus subtilis, Saccharomyces cerevisiae and Aspergillus niger. The viable count, pH value, lactic acid (LA) content, and crude protein (CP) content were used as evaluation indicators. Single-factor experiments and orthogonal experiments were conducted to optimize the fermentation substrate composition and fermentation conditions. The results showed that the optimal substrate composition was 46% rice straw, 3% molasses, 30% distiller's grains, 12% corn flour, and 9% wheat bran. The optimal fermentation conditions were initial pH value 6, inoculum size 20%, and fermentation time 10 days. Compared with the control, the concentrations of LA and CP of the optimized fermented rice straw were extremely increased (P<0.01), the concentrations of neutral detergent fiber (NDF) and acid detergent fiber (ADF) were significantly decreased. The study shows that this study provides a feasible fermentation process reference for the resource utilization of rice straw and a reference for the application of multi-strain synergistic fermentation in the conversion of agricultural waste.

Grassland receive multiple forms of reactive nitrogen (N) from fertilization and deposition, yet the form-specific responses of nitrifier guilds and their potential activity remain poorly resolved, particularly in fragile karst ecosystems. We conducted a greenhouse pot experiment using a karst grassland soil to compare different N forms (calcium nitrate, ammonium chloride, ammonium nitrate, urea, glycine and urea+glycine) applied at an equivalent N rate. We quantified soil properties, plant biomass, potential nitrification rate (PNR), and the abundance and community composition of ammonia-oxidizing archaea (AOA) and bacteria (AOB) to elucidate the mechanistic links between N form, ammonia oxidisers, and nitrification potential. All N additions significantly increased PNR (34.8%-121.2%) and AOB amoA gene abundance (19.3%-36.8%) relative to the control, with organic N forms (PNR: 79.1%-121.2%; AOB: 32.1%-36.8%), especially glycine, producing the strongest stimulation compared to inorganic forms (PNR: 34.8%-52.2%; AOB: 19.3%-24.5%). Organic N inputs also significantly elevated dissolved organic carbon (DOC) and root biomass and consistently shifted AOB communities from Nitrosospira Cluster 1 towards Cluster 9, whereas ammonium chloride induced strong acidification and reduced AOB diversity (-46.8%). AOA abundance and diversity responded more weakly to N form and were only weakly related to PNR. Multivariate analyses and random forest models further identified AOB gene abundance, AOB cluster composition and DOC as the main predictors of PNR. Together, these results show that N chemical form regulates nitrification potential in karst grassland soils by reshaping AOB guild structure through coupled changes in carbon availability and plant biomass allocation, highlighting the importance of form-aware N management and deposition assessment beyond total N inputs.

Mulberry (Morus alba L.), a high-protein woody forage, has considerable potential to alleviate protein feed shortages in animal husbandry. However, its utilization is constrained by substantial protein degradation during anaerobic fermentation and ruminal digestion. This study investigated the effects of tannic acid (HT) and Acacia mangium tannin (CT) at five graded inclusion levels (1, 2, 3, 4 and 5 g/kg dry matter, DM), together with a control treatment (CK), on the fermentation quality, bacterial community composition and protein utilization of mulberry silage after 60 days of ensiling, with four independent biological replicates per treatment. The results showed that HT dose-dependently reduced crude protein loss during fermentation, with a maximum reduction of 2.72% compared with the control. HT also promoted the dominance of Lactiplantibacillus, suppressed undesirable microorganisms, and optimized silage protein profiles by elevating PB2 and PB3 fractions and reducing non-protein nitrogen (NPN), which is poorly available to ruminants and leads to protein resource waste, increased urinary nitrogen excretion and associated environmental pollution risks. In contrast, CT promoted protein degradation during fermentation and suppressed the abundance of Lactiplantibacillus. Both tannin sources decreased ruminal protein degradation, with reductions of up to 17.94% (add 5 g/kg tannic acid). These findings reveal the regulatory mechanisms of tannins on microbiota and protein degradation in mulberry silage. Amid surging ruminant feed demand and severe environmental pressures from conventional grain feeds, this study lays a theoretical basis for promoting mulberry as sustainable ruminant forage with higher nutrient utilization. It also delivers a viable technique to mitigate silage nitrogen loss and lower nitrogen pollution from ruminant production.

Nitrogen (N) and phosphorus (P) are essential for ecosystem productivity and nutrient cycling, yet their balance is increasingly altered by human-driven N enrichment. However, how different N forms affect plant nutrient status and stoichiometry in karst grasslands remains unclear. We established a controlled pot experiment with two typical karst soils from southwestern China (yellow and limestone soils), applying four N forms (Ca(NO3)2, NH4Cl, NH4NO3, and urea) at two application rates. Biomass, N and P concentrations, and N:P mass ratio were measured in perennial ryegrass and white clover, as well as at the community level. N application significantly increased plant shoot biomass, with ammonium-based forms (urea and NH4Cl) producing the strongest effects and yellow soil exhibiting higher sensitivity. Perennial ryegrass biomass increased under all N treatments, while white clover biomass declined under high ammonium input. However, N addition generally reduced plant N and P concentrations in tissues due to dilution effects, which were most pronounced under NH4Cl treatments. Community-level N:P mass ratio remained relatively stable, primarily influenced by the proportion of white clover, indicating a buffering effect at the community scale. Ammonium-based N inputs produced the strongest biomass gains but also the largest declines in shoot N and P, consistent with growth dilution and reduced P availability under ammonium-induced acidification. Responses were consistently stronger in yellow soil than in limestone soil. At the community level, the N:P mass ratio was largely buffered by white clover, decreasing only under high NH₄Cl where clover abundance declined. These findings highlight that N form is a key, often overlooked driver of plant stoichiometry in karst grasslands and support N-form-aware fertilization with P co-management to sustain productivity while minimizing nutrient imbalance.

Nitrogen is crucial for plant growth and development. Karst areas face significant degrees of both nitrogen leaching and enrichment, yet paper mulberry (Broussonetia papyrifera) thrives in these areas. Here, the physiology, proteome, phosphoproteome, and ubiquitome of paper mulberry seedling leaves were investigated to understand the mechanisms of plant adaptation to nitrogen stress. We discovered that paper mulberry responds to nitrogen stress by regulating the expression of ammonium transporter protein (AMT1.1) and nitrate and peptide transporter proteins (NPF6.4, NPF8.1, and NPF8.3). Key pathways involved in the response of paper mulberry to different nitrogen levels include photosynthesis, carbon fixation, and nitrogen metabolism. The plant also enhances its antioxidant defences to reduce ROS stress, while protein phosphorylation serves as a signalling component. Lysine ubiquitination may play an essential role in the degradation of misfolded proteins in paper mulberry under low-nitrogen stress. Amino acid, starch, and sucrose metabolism provides the energy necessary for paper mulberry adaptation to high-nitrogen stress. This study provides insights into the possible biological adaptation mechanisms of paper mulberry under controlled nitrogen stress, which may inform future studies relevant to karst areas.

Mulberry (Morus alba L.) serves as a high-value woody protein feed resource to address the shortage of protein feed supplies, but inherent limitations, including excessive moisture, insufficient indigenous lactic acid bacteria, and intense protein degradation, constrain its silage preparation. To address these problems, this study developed pre-fermented juices using alternative raw materials, including grape pomace and red clover, which are rich in natural antioxidants, such as polyphenols and isoflavonoids. Four groups were set in the experiment: red clover pre-fermented juice (treatment R), grape pomace pre-fermented juice (treatment G), mulberry pre-fermented juice (treatment M), and blank control (CK). Combined with bacterial community and metabolomic analyses, the present research further investigated their effects on the fermentation quality of mulberry silage during a 60-day ensiling period. The results indicated that treatment R exhibited the optimal comprehensive effect, enriching Enterococcus, upregulating the accumulation of isoflavonoids (formononetin, glycitein, and biochanin A), reducing silage pH to 3.99, elevating lactic acid concentration, and restraining protein degradation by inhibiting protease activity. Treatment M facilitated the proliferation of Lactobacillus and the accumulation of beneficial metabolites such as 3,4-dihydroxybenzaldehyde, while treatment G exerted only a mild regulatory effect. Overall, all pre-fermented juice additions effectively improved fermentation performance and nutrient preservation of mulberry silage. This finding elucidates the mechanism of microbial-metabolite interactions during mulberry ensiling, offers a low-cost, environmentally friendly technical strategy for high-quality silage production, and facilitates the sustainable application of mulberry resources in ruminant feeding systems.

Phosphorus (P) limitation constrains productivity in subtropical red-soil croplands, yet conventional fertilisation often fails to restore biologically mediated P cycling. Here, we examined how twelve years of contrasting land management regulate soil C:N:P stoichiometry, phoD-harbouring bacterial communities, and alkaline phosphatase (ALP) activity across surface and subsoil layers in a sloping agroecosystem. Land management markedly altered soil stoichiometry and microbial P-acquisition potential. Although mineral fertilisation increased soil P pools, it reduced soil C:P ratios and suppressed phoD gene abundance and ALP activity under C-limited conditions. In contrast, cover-based systems restored soil C:P balance, restructured phoD functional communities, and enhanced ALP activity, supporting the recovery of ALP-mediated P-acquisition potential comparable to natural restoration. Random forest and structural equation modelling consistently identified soil C:P as the strongest predictor of phoD abundance, richness, community composition, and ALP activity, integrating management and depth effects. Moreover, white clover cover facilitated vertical coupling of stoichiometric signals, partially alleviating depth-related constraints on microbial P acquisition. These results indicate that, in this C-depleted subtropical red-soil sloping cropland, the bacterial phoD-ALP pathway was more strongly associated with soil C:P balance than with P availability alone. Our findings suggest that restoring soil C:P balance through cover-based management can enhance bacterial ALP-mediated P-acquisition potential and support more sustainable P management in subtropical sloping croplands.

This study aims to investigate the effects of three lactic acid bacteria (LAB) strains, Lactiplantibacillus plantarum, Lactiplantibacillus pentosus and Limosilactobacillus fermentum, isolated from traditional pickles in Guizhou, on the fermentation process and microbial community dynamics of ensiled whole-plant maize, soybean, and their mixtures. The results revealed that compared to the CK group, the lactic acid levels of Lactiplantibacillus plantarum and Lactiplantibacillus pentosus were significantly increased in the treatment groups (p < 0.05), resulting in a faster pH reduction, along with decreases in ammonia nitrogen (AN) and butyric acid (BA) content. In contrast, the Limosilactobacillus fermentum treatment (p < 0.05) promoted acetic acid (AA) production and inhibited the growth of harmful microbiota in soybean silage. Notably, inoculation with all LAB strains enhanced the aerobic stability of maize silage by promoting the proliferation of Lactiplantibacillus during the later stages of fermentation, thereby sustaining a low pH and mitigating the depletion of water-soluble carbohydrates (WSC). Furthermore, all treatments accelerated silage fermentation by enhancing the LAB population and competing with yeast and Escherichia coli for available nutrients in mixed silage. These findings indicate that three LAB strains, when used as microbial additives, demonstrated potential to improve silage quality in the Karst region.

Triticale (× Triticosecale wittmack) is a high-yielding and nutritionally balanced forage crop with considerable potential for silage production. This study examined the effects of growth stage and moisture content on the fermentation quality and bacterial community composition of triticale silage by harvesting at heading and dough stages and ensiling at 60

Background/Objectives: Using straw as feed is crucial for addressing resource scarcity and waste-based environmental pollution and alleviating the contradiction between forage production and food security. However, rice straw has disadvantages such as a high lignin, cellulose, and hemicellulose content, as well as a dense cell wall structure, which greatly limit its utilization as feed. Improving the forage quality of rice straw through genetic breeding is an important means of enhancing its utilization rate. Methods: To explore the molecular basis underlying the improved nutritional and cell wall compositional traits of the rice lines (oszfp30-1 and oszfp30-2) obtained in previous studies, we conducted phenotypic, physiological, transcriptomic, and metabolomic analyses on oszfp30-1, oszfp30-2 and the wild type (WT) after 50 days of pot cultivation. Results: oszfp30-1 and oszfp30-2 exhibited higher plant height, above-ground biomass, crude protein, and crude fat content, while their hemicellulose, cellulose, and lignin contents were significantly lower than those of the wild type. Transcriptomics was used to identify 737 common differentially expressed genes (DEGs), and metabolomics was used to identify 189 common differentially expressed metabolites (DEMs). KEGG analysis revealed that these DEGs and DEMs were significantly enriched in terpene biosynthesis, starch and sucrose metabolism, phenylpropanoid biosynthesis, and amino sugar and nucleotide sugar metabolism. Conclusions: Our research reveals that the improvement in forage-quality-related traits is closely related to key genes involved in gibberellin synthesis, including GA20ox, GA2ox, and GA3ox; key genes involved in cellulose synthesis, including OsSUS and UGPase; key genes involved in hemicellulose synthesis, including UXS and IRX10; and key genes involved in lignin synthesis, including CCR, CAD, CCoAOMT, and COMT. These identified DEGs and DEMs are associated with the OsZFP30 mutation and may be downstream targets of this transcription factor, providing a foundation for further research on the OsZFP30 regulatory network.

The global phasing-out of antibiotic growth promoters (AGPs) has catalyzed an urgent search for efficacious and sustainable alternatives to maintain livestock health and productivity. Houttuynia cordata Thunb. (HC), a traditional pharmacophagous herb has emerged, as a promising candidate due to its pleiotropic bioactive profile. This review outlines the molecular mechanisms and clinical efficacy of HC extracts (HCE) as multifunctional feed additives. We elucidate that HCE, rich in volatile oils, flavonoids, and polysaccharides, systemically modulates host physiology by activating the Nrf2/HO-1 antioxidant signaling axis while concurrently suppressing hyper-inflammation via the inhibition of TLR4/NF-κB and NLRP3 inflammasome pathways. Crucially, HCE reinforces epithelial integrity across various biological barriers (intestinal and mammary) through the upregulation of tight junction proteins. Across diverse production systems, HCE has significant efficacy under experimental conditions. In ruminants, it alleviates heat stress-induced redox imbalance and mitigates mastitis. In monogastric species, it enhances viral resistance (notably against PRRSV) and optimizes meat quality. In aquaculture, it fortifies non-specific mucosal immunity against viral challenges. Despite technical hurdles regarding component stability and standardized quality control, HCE represents a transformative, green nutritional strategy. Advancements in microencapsulation and nano-emulsification are poised to catalyze the industrial integration of HCE, positioning it as a cornerstone of sustainable animal agriculture in the post-antibiotic era.

The large-scale production and high putrefactive nature of wet distiller's grains (WDG) waste inevitably cause resource waste and environmental pollution. Here, anaerobic fermentation technology incorporating different WDG ratios and micro aeration technology were performed to assist in the clean recycling of WDG and reveal the related underlying microbiological mechanisms. To optimize the low nitrogen emissions production during anaerobic fermentation and micro aeration, the total mixed ration experimental were designed with three levels of WDG: 0, 15% and 30% on a DM basis (i.e., the CON, T15 and C30 treatments, respectively) were fermented for 60 days, and subsequent micro aeration experiment were conducted for 1, 3, 5, and 7 days. The results demonstrated that WDG treatment groups increased lactic acid and acetic acid during anaerobic fermentation by 23.78%-44.27% and 5.27%-7.14%, respectively. Additionally, high incorporation proportion of WDG (30% dry matter) effectively inhibited pH elevation rate (7.49% vs. 13.64%), lactic acid degradation and ammonia-N formation after 7 days of micro aeration. Microbiologically, high Lactobacillus abundance (80.23-91.57%), high Issatchenkia abundance (22.19-47.91%) and lower Weissella abundance (2.00-4.85%) were observed in T30-treated during micro aeration. Moreover, the WDG treatment groups stimulated the proliferation of Lactobacillus acetotolerans and Issatchenkia orientalis, thereby promoting acetic acid accumulation and enhancing the aerobic stability of TMR. Additionally, high incorporation proportion of WDG (30% dry matter) increased Lactobacillus and Issatchenkia abundance during micro aeration, which were the specific microbes that enhanced aerobic stability. Overall, wet distiller's grains incorporation at a proportion of 30% dry matter enhanced the anaerobic fermentation system, achieving the clean recycling and utilization of industrial waste, and the effect of wet distiller's grains on aerobic stability relies on the proportion used, which is linked to variations in the bacteria and fungi microbes.

Phosphorus is an essential element for plant growth, and its deficiency severely limits crop productivity. To explore genetic resources for improving phosphorus use efficiency, this study investigated the differential low-phosphorus tolerance mechanisms of two kudzu (Pueraria lobata) germplasms from Australia (tolerant) and Jiangsu, China (sensitive) using hydroponics, RNA-seq, and WGCNA. The results showed that the Australian germplasm exhibited superior low-phosphorus tolerance through root morphological plasticity, which was characterized by increased root length and tip number under low phosphorus (0.05 mmol L-1 KH2PO4); enhanced reactive oxygen species scavenging, with higher peroxidase and catalase activities under extremely low phosphorus (0.005 mmol L-1 KH2PO4), and extensive transcriptome reprogramming, including 8896 upregulated genes in response to phosphorus deficiency. In contrast, the Jiangsu germplasm showed limited adaptive responses, with reduced root hairs and biomass under stress. WGCNA partitioned 21,734 expressed genes into 20 co-expression modules, among which the turquoise and light green modules showed significant correlations with phosphorus treatments and phenotypic traits. Genes in the turquoise module were primarily enriched in oxidative phosphorylation and phenylpropanoid biosynthesis pathways, whereas the light green module was significantly enriched in ribosome-related pathways. Five hub genes, ABCG5, TALDO, VAMP7B, EEF1AS, and RPLP0, were identified as core components of these modules. Collectively, these findings establish the Australian kudzu as a valuable germplasm resource for improving phosphorus use efficiency in crops and provide key molecular targets for precision breeding.

Fine-scale spatial vegetation patterns are widespread and can significantly affect large-scale ecological processes. Quantifying these patterns may provide valuable insight into larger scale ecological processes, facilitating the development of effective management strategies. In this paper, to quantify fine-scale spatial vegetation patterns, we analyzed seventy plots from seven humid grasslands in southwest China, using images with pixel sizes of 0.04, 0.25, 1, and 4 cm and plant cover levels ranging from 30.8 to 99.3%. The results showed that with increasing pixel size, plant patch density and total edge density decreased, while patch size increased, and the shape of plant patches became more regular. When plant cover was below 50%, increasing cover caused the small stems, branches, and leaves of grasses to fragment non-plant patches into smaller units, leading to higher patch density and larger patches, thus enhancing spatial heterogeneity. However, when plant cover exceeded 50%, further increases in cover resulted in rapid patch expansion and reduced patch density, creating a more homogeneous landscape dominated by plant patches. These findings suggest that medium plant cover levels are associated with the highest fine-scale spatial heterogeneity, which may facilitate biodiversity, stability, and functioning, with implications for ecosystem management and conservation, and warranting further investigation.

Fermented distiller’s grains (FDG) are rich in nutrients and have high feed utilisation value, but research in broiler feeding is limited. In this experiment, 216 male yellow-feathered broilers (30-day-old, 584.0 ± 3.85 g) were randomly divided into three groups, with six replicates per group and 12 broilers per replicate. The diet treatments were the control group, the 5

The process of ensiling serves as a dual-purpose technique, functioning as a means of preserving biomass for biogas production while serving as a biological pretreatment method with the potential for little fermentation loss. In this study, we investigate the potential of Rosa roxburghii pomace (RP) and Lactobacillus acidophilus to improve the biomass preservation of biogas produced from alfalfa during anaerobic storage. Coinoculation of RP and Lactobacillus acidophilus resulted in better preservation of nutrients (biomass), with a significant increase of 47.38% within the lactic acid content and a notable decline of 40.34% in the ammonia nitrogen content relative to those in the control treatment. Moreover, coinoculation of RP and Lactobacillus acidophilus resulted in an elevated proportion of Lactobacillus, and the species Lactiplantibacillus plantarum dominated anaerobic fermentation. The synergistic effect of RP and Lactobacillus acidophilus continuously stimulated anaerobic fermentation, leading to 33.33% and 23.17% increases in methane production and acetate content, respectively, after 72 h of coinoculation with RP and Lactobacillus acidophilus compared to the control treatment. Overall, coinoculation of RP and Lactobacillus acidophilus offers an attractive opportunity to increase methane production from fruit waste while reducing costs and synergistically integrating with other pretreatment techniques to optimize the methane generation potential. IMPORTANCE:Considering the increasing global energy demand and urgent environmental issues, exploring prospective resources for bioenergy production is imperative. However, the biomass of legume perennials may serve as an inexpensive and stable source of clean energy for modern society due to its wide availability and broad range of sources. In addition, the combination of RP and Lactobacillus acidophilus application increased the abundance of Lactobacillus, inhibited the growth of Kosakonia, and promoted anaerobic fermentation, which had beneficial synergistic effects on biomass retention and biogas production in alfalfa samples. Coinoculation improvements with RP and Lactobacillus acidophilus observed here are expected to reduce costs associated with CH4 conversion bioprocesses and increase CH4 production.