Plastic pollution control in aquatic environments has mainly emphasized source reduction and end-of-pipe treatment, whereas the interception of rainfall-driven migration pathways remains less examined. Urban surface runoff is a critical pathway by which plastic debris is transported from streets and other impervious surfaces into receiving waters. Unlike most previous studies that focus primarily on wetlands, bioretention systems, or microplastics within a single pavement configuration, this study systematically evaluates two widely used permeable paving materials, permeable bricks and permeable asphalt, under simulated rainfall conditions representative of East Asian monsoon regions. Results indicate that both materials promoted rainfall infiltration and substantially reduced plastic transport under the tested laboratory conditions, intercepting over 90% of plastic pollutants by count and exceeding 99% by mass. Under the tested specimen configurations, permeable bricks showed a 10.98% higher particle-count interception efficiency than permeable asphalt across various rainfall intensities. Permeable bricks effectively intercepted particles larger than 0.25mm, whereas permeable asphalt allowed the passage of particles up to 2mm. Fibrous plastics exhibited the highest likelihood of passage through permeable pavements, followed by fragments, whereas foamy plastics rarely passed through. The majority of intercepted plastics accumulated on pavement surfaces and could be substantially removed by timely sweeping or vacuum cleaning in the short-term repeated-rainfall test. This study highlights permeable pavement as a practical first-barrier measure for interrupting runoff-driven plastic transport and provides evidence for material selection, pore-structure optimization, and maintenance-oriented stormwater management.

Urban stormwater runoff is a major pathway for anthropogenic pollutants into aquatic ecosystems. Phthalate esters (PAEs), widely produced endocrine-disrupting plasticizers, readily enter the environment; however, their transport via stormwater and contribution to nearshore marine pollution remain largely overlooked. The stormwater system of Macao discharges directly to the sea, representing a potentially significant PAE pathway, yet systematic investigations on PAEs pollution in its aquatic environment are lacking. This study conducted the first comprehensive investigation on the occurrence and distribution of 16 PAEs congeners in six stormwater drainage channels (covering residential, industrial, and ecological functional zones) and four seawater sampling sites in Macao using GC-MS, aiming to characterize PAEs pollution, identify sources, and estimate annual fluxes. Results showed widespread PAEs contamination: Σ16PAEs in stormwater ranged from 1.51 × 102-3.35 × 104 ng/L (0.152-33.5 μg/L), and 89.1-858 ng/L in seawater. High levels of dimethoxyethyl phthalate (DMEP) were detected, revealing unique local pollution characteristics. Among stormwater sites, Σ16PAEs loads were significantly higher in residential/industrial areas than ecological areas, with distinct seasonal variations. Linear correlations between some stormwater and seawater PAEs concentrations may imply a potential source-sink relationship. Principal component analysis indicated non-point sources primarily from residential and industrial discharges. Importantly, pumping stations were innovatively identified as key dry-season PAEs sources. The annual average Σ16PAEs input from the stormwater system of Macao to nearshore waters was 4.31 × 10-3-9.5 kg. As the first systematic report on aquatic PAEs pollution of Macao, these findings fill the regional research gap, support targeted dry/wet season control measures, and provide a reference for small coastal cities.

Earth's fragile high-altitude ecosystems are facing challenges from nutrient pollution, where data scarcity and complex topography create significant knowledge gaps. This study conducted the first large-scale survey and established a comprehensive inventory of human activities in the Qinghai Lake basin, the largest lake on the Qinghai-Tibet Plateau. We developed a hybrid framework that integrated detailed surveys, process-based modeling, and interpretable machine learning to quantify nitrogen and phosphorus fluxes despite data gaps. Notably, ungauged basins around the lake contribute substantially (34% of phosphorus and 21% of nitrogen) to the lake load, highlighting a serious underestimation of a blind spot in global monitoring networks. Contemporary anthropogenic emissions contributed only 41.12% of TN flux and 45.22% of TP flux. Further machine learning analysis indicates that non-direct human factors (runoff, climate, and geographical conditions) have a dominant influence on nutrient movement. These findings provide a replicable framework for data-poor regions and call for shifting management paradigms from anthropocentric to ecosystem-based approaches across high-altitude areas.

Biogeochemical niche (BN) is defined as the n-dimensional space occupied by the characteristic concentrations of elements in individuals of that species, which helps identify a species’ environmental response strategies based on its elemental composition. While most studies focus on BN partitioning among coexisting species, few studies explore how the same species adapts to varied environments through stoichiometric strategies. This study investigated BN differentiation of Stuckenia filiformis, a widespread submersed macrophyte in the Qinghai-Tibet Plateau, across nine Alpine wetlands. We quantified thirteen chemical elements in S. filiformis to characterize its BN. Our results showed significant BN differentiation of S. filiformis across different wetlands, indicated by large BN distances and low BN overlap. C, N, and P in S. filiformis exhibited stronger homeostatic control compared to trace elements. BN distances of S. filiformis between wetlands were significantly positively correlated with altitude, latitude, and environmental distance. The weakly plastic element Mn in S. filiformis significantly contributed to BN differentiation across wetlands, which was influenced by mean annual air temperature. Elemental composition variability within particular wetlands was dominated by differences in Na, K, P, and Zn, and was correlated with sediment Na content. Our study provides evidence of biogeochemical niche differentiation of a widespread submersed macrophyte species in Alpine wetlands, shedding light on individual adaptation strategies to changing environments, and can help understand the role of stoichiometric variation in influencing the biogeochemistry and ecosystem functioning of wetland ecosystems.

The phycosphere is an important ecological niche for bacteria and antibiotic resistance genes (ARGs). However, whether and how the interaction between microalgae and bacteria changed, and its further effect on the transmission of ARGs under pollutant stress remains enigmatic. Here, Auxenochlorella pyrenoidosa was co-cultured with bacteria screened from lake water to explore the algal–bacteria interaction and ARGs’ transmission in the presence of florfenicol (FF) and polylactic acid microplastics (PLA MPs). Our study demonstrated that the growth and metabolism of A. pyrenoidosa were promoted under FF treatment or co-treatment with PLA MPs, validated by phenotypic, transcriptome, and metabolome analyses. In contrast, the abundance of phycospheric bacteria was decreased as a result of niche competition. Nonetheless, the transmission of ARGs in the phycosphere was promoted due to the enrichment of antibiotic-resistant bacteria, especially Pseudomonas, rather than horizontal gene transfer. The algal-bacteria co-culture experiment further suggested that vitamin B6 secreted by Pseudomonas sp. likely contributes to underpinning A. pyrenoidosa’ survival under FF and PLA MPs stress. These findings underscore the dynamic interplay and co-evolution between algae and bacteria under pollutant exposure, and reveal a potential mechanism of vitamin B6-mediated mutualism. This study provides new insights into the assembly of phycospheric bacterial communities and the adaptive strategies of microalgae in contaminated aquatic environments.

Microbial conversion of CO and CO2 offers a promising route to C1 gas valorization, but acetogenic syngas fermentation remains constrained by bioenergetic bottlenecks. To address this, a biohybrid system integrating bovine serum albumin-stabilized gold nanoparticles with Clostridium autoethanogenum was constructed, increasing intracellular ATP levels, modulating redox balance, and promoting cell growth under illumination. Transcriptomic analysis of this system systematically identified key energy complexes and positive regulators associated with enhanced cellular energy metabolism. Based on these elements, a Push-Pull-Block strategy was implemented to generate the target strain CAB181, which increased OD600-normalized production of ethanol, acetate, and total acids and alcohols by 46.5%, 35.1%, and 34.2%, respectively. Multiomics analysis confirmed molecular-level metabolic reprogramming in CAB181. This work provides an effective strategy to improve the bioenergetic robustness and fermentation performance of C. autoethanogenum-based cell factories, and offers a reference framework for future engineering of other acetogens.

Silver carp (Hypophthalmichthys molitrix) stocking is a widely employed biomanipulation measure to tackle algal bloom in eutrophicated lakes, yet its role in nutrient cycling is still unclear. Through mesocosm experiments, we examined how fish density and environmental factors influenced water quality by comparing fish-mediated nutrient supply with sediment internal loading. Results suggested rapid nitrogen (N) and phosphorus (P) recycling within the water column, even under low ambient nutrient conditions. Fish-mediated nutrient fluxes exceeded sediment internal P loading by 9-fold (high fish density, HD) and 7.5-fold (low fish density, LD), and the HD treatment also enhanced N excretion by 1.7-fold compared with the LD treatment. The HD treatment demonstrated significantly improved phosphorus retention (33.6% vs. 22.8%) and feed efficiency (1.76 g/g vs. 3.06 g/g FCR) but 27% lower growth rates compared to the LD treatment. Elevated temperature intensified N limitation while alleviating P constraints, disrupting the nutrient–algae balance. Binary logistic regression and linear mixed model analyses confirmed that temperature–density interdependence critically regulated chlorophyll a (Chl.a) dynamics. HD stocking suppressed Chl.a, but warming water temperature attenuated this effect. We propose a conceptual framework for consumer-driven nutrient dynamics in carp-manipulated systems, demonstrating that summer stocking may prove ineffective due to thermal disruption of nutrient–algae coupling, while low densities risk stimulating blooms via incomplete grazing and nutrient supply. These findings highlight the necessity of high-density, seasonally-informed stocking strategies for effective lake restoration.

Cladophora blooms, exacerbated by climate change and littoral eutrophication, pose a significant ecological threat. Of particular concern is their potential to disrupt phytoplankton and bacterial assemblages, triggering a cascade of effects that may include shifts in nutrient cycling and the dissemination of resistomes. However, the mechanistic links between Cladophora’s life-stage-dependent dissolved organic matter (DOM) release, its role in restructuring epiphytic communities, and its promotion of resistome dissemination in natural, oligotrophic lakes remain poorly understood. To address this, this study integrates field and laboratory investigations of Cladophora qinghaiensis sp. nov.. The algal phycosphere functions as a dynamic “gene incubator”, driven by chemical shifts in algal‑derived DOM. During decay under low‑oxygen conditions, DOM composition transitions from tyrosine‑like proteins to recalcitrant fulvic‑acid‑like compounds, selectively enriching competitive, intrinsically resistant taxa such as Halomonas and Phacus. Microbes such as Acinetobacter drive nutrient cycling (e.g., nitrogen metabolism) and serve as hotspots for resistomes within the phycosphere. Contrary to the expectation that high cell density favors horizontal gene transfer (HGT), genomic analyses show that vertical gene transfer (VGT) dominates antibiotic resistance gene (ARG) proliferation in this niche, a pattern explained by strong DOM‑mediated host selection and subsequent propagation. In contrast, the resistome in the surrounding water is more diverse and primarily shaped by HGT via mobile genetic elements. These results establish a mechanistic link between life‑stage‑specific algal DOM components, selective epiphytic communities enrichment, and divergent pathways of resistome evolution, positioning the phycosphere as a key source of ARGs that amplifies ecological risk in nearshore environments.

The global challenge of food waste (FW) management and protein scarcity necessitate innovative valorization strategies. This study presents a two-stage FW bioconversion integrating microaeration/thermal pretreatment to enhance biodegradation into organic acids (OAs), including volatile fatty acids (VFAs) and lactic acid, followed by their utilization by Schizochytrium limacinum SR21 for single-cell protein (SCP) production. The optimized pretreatment conditions achieved an OAs yield of 37.0 g/L at an organic loading rate (OLR) of 20 gVS/L, producing 258.23 kg of total organic acids (wet weight) from one ton of FW (25.82% conversion). S. limacinum SR21 demonstrated efficient OAs consumption, with 90% OAs degradation within 48 h under optimal conditions. Genetic engineering via lactate oxidase overexpression further enhanced strain performance, resulting in 17.46 g/L biomass, 14.87% lipid content, and 6.53 g/L protein yield, which were 14.19% and 12% greater than those of the wild-type strains for biomass and protein, respectively, alongside a 107.1% increase in lipid content. The recombinant strain SdLDH achieved 46.72 kg SCP per ton of dry FW (18.18% conversion). Life cycle assessment (LCA) revealed 67.5-73.4% lower impacts compared with soybean protein and 27.2-59.0% compared with soybean meal, and 18.3% lower impacts than conventional FW treatment, including 90.3% lower global warming potential and lowering land-use impacts to similar to 1% of soybean-based production. This research establishes a sustainable circular economy pathway for FW-to-protein conversion, offering a low-carbon solution to address global protein deficits and waste management.

Rising water levels in Qinghai Lake, a key hydrological disturbance, have submerged extensive lakeshore grasslands, triggering ecological changes in the newly inundated zones. This study investigated how this hydrological process alters soil microbial community succession and co-occurrence networks through inputs of dissolved organic matter (DOM) and nitrogen-phosphorus (N-P) dynamics in the lakeshore zone of Qinghai Lake. The results demonstrated that grassland inundation caused by rising water levels enriched the soil with specific DOM components (fulvic and humic acid), driving significant changes in N and P concentrations. High-throughput sequencing results indicated that following grassland inundation, α-diversity declined, whereas β-diversity of soil microorganisms initially rose and then decreased as the duration of flooding prolonged. Furthermore, microbial community assembly shifted over time: in the initial inundation stages, higher proportions of readily bioavailable DOM components and N-P nutrients coincided with the predominance of stochastic processes. Over time, the proportion of recalcitrant DOM components increased, leading to a gradual dominance of deterministic processes. These microbial successional patterns were closely linked to DOM quality and N-P availability. Simultaneously, DOM and N-P content shaped more stable and complex co-occurrence network structures by influencing the formation of core microorganisms. Collectively, these findings highlight the central role of soil microorganisms in mediating ecosystem responses to hydrological disturbances. Our findings elucidate how hydrological disturbances in lakeshore ecosystems reshape soil microbial assembly-transitioning from stochastic responses to deterministic selection-and enhance the stability of co-occurrence networks through interactions between DOM, N, P, and microbial communities. This framework, linking hydrological processes to coupled soil biogeochemical and microbial ecological responses, establishes a vital scientific basis for predicting and managing the ecological consequences of climate-driven lake expansion. It also provides critical insights for developing adaptive management strategies aimed at preserving ecosystem resilience and biogeochemical functioning in lakeshore zones experiencing dynamic hydrological regimes.

Estuarine wetlands are essential regulators of water quality, yet purification performance in plateau regions is often constrained by low temperatures and high salinity. Here, we constructed a synergistic purification system integrating artificial substrates with native aquatic plants (Potamogeton crispus, Phragmites australis and Myriophyllum spicatum) in the Shaliu and Heima River estuarine wetlands of Qinghai Lake. Water quality monitoring, 16S rRNA sequencing and metagenomic profiling were combined to elucidate microbial community structure and nitrogen-phosphorus (N-P) removal mechanisms. The system exhibited stable purification efficiency under cold-saline conditions, achieving TN and TP removal of 70.43% and 92.27% at the Shaliu River site and 66.58% and 88.91% at the Heima River site, respectively. Artificial substrates significantly enhanced microbial diversity and selectively enriched functional taxa including Nitrospira, Sphingomonas, Nocardioides and Ferruginibacter, which were key drivers of nitrification, denitrification and organic matter degradation. Rhizosphere sediments of Phragmites and Myriophyllum showed increased pathways related to nitrogen metabolism, amino acid biosynthesis and carbon cycling, suggesting that plant rhizospheres primarily enhanced nutrient removal by supporting microbially mediated transformation and redox regulation. Overall, the substrate-plant-microbe synergistic system optimized microbial community assembly and functional gene expression, thereby reinforcing coupled N and P cycling. This study clarifies the microbial ecological mechanisms supporting nutrient removal in plateau estuarine wetlands and provides a scientific basis for designing sustainable purification strategies in cold and saline environments.

Soilborne diseases are important problems in modern agricultural production. Fusarium oxysporum f. sp. cucumerinum (FOC) is one of the predominant soilborne pathogens threatening cucumber cultivation, especially in Hainan, China. This study assessed FOC-resistant rootstocks using incidence rate, disease severity index (DSI), and area under the disease severity index curve (AUDRC), revealing “JinJiaZhen (Mc-4)” as resistant and “JinGangZhuan 1901 (Mc-18)” as susceptible. Comprehensive transcriptome and metabolome analyses were conducted to investigate the defense mechanisms of these rootstocks, revealing key pathways, such as the mitogen-activated protein kinase (MAPK) signaling pathway, starch and sucrose metabolism, and phenylpropanoid biosynthesis, which are crucial for plant disease resistance. Additionally, the study compared the resistance mechanisms of two other rootstocks, Mc-4 and Mc-18, against FOC infection through transcriptomic and metabolomic analyses. Mc-4 exhibited a higher number of differentially expressed genes (DEGs) related to phenylpropanoid biosynthesis compared to Mc-18. Untargeted metabolomics identified 4093 metabolites, with phenylpropanoid biosynthesis, isoquinoline alkaloid biosynthesis, and porphyrin metabolism as primary annotated pathways. On the sixth day post-inoculation, when the number of DEGs and differentially accumulated metabolites (DAMs) was highest, phenylpropanoid biosynthesis emerged as a key pathway in Mc-4, with 37 DEGs and 8 DAMs identified. Notably, Mc-4 showed upregulated expression of genes encoding enzymes involved in phenylpropanoid biosynthesis and increased accumulation of related metabolites, such as coniferyl-aldehyde, coniferyl alcohol, and coniferyl acetate. These findings highlight the differential defense mechanisms between resistant and sensitive rootstocks and provide insights into plant–pathogen interactions. This study’s results will contribute to the development of better and disease-free cucumber varieties, promoting sustainable agriculture.

Algal-bacterial interactions represent fundamental ecological processes in aquatic environments, crucially governing nutrient cycling and energy flow within food webs. Beyond their ecological roles, the algal phycosphere has recently been identified as a critical hotspot for the proliferation and enrichment of antibiotic resistance genes (ARGs). It’s reported that the total abundance of ARGs in the phycosphere of microalgae is up to 47-fold higher than in the surrounding water. However, a systematic understanding of how the phycosphere drives ARG dynamics in aquatic ecosystems remains limited. This review synthesizes current evidence to evaluate the mechanisms by which algae influence ARG proliferation within aquatic ecosystems. Findings indicate that in the phycosphere, algal-bacterial interactions shape ARG fate by modulating bacterial community composition. The symbiotic bacteria are specifically enriched in the phycosphere and play important roles in the proliferation of ARGs. Furthermore, exogenous factors (e.g., nutrients, antibiotics, microplastics, and warming) alter these interactions, thereby changing the phycospheric bacterial community and further affecting ARG evolution. Algal blooms typically enhance the dominance of key ARG hosts, promoting aquatic ARG proliferation. The review concludes by outlining research priorities essential for advancing mechanistic insights into algal-associated ARG dynamics.

Liangzi Lake, a typical shallow lake in the middle reaches of the Yangtze River, is important for water resource and biodiversity conservation. With the development of urbanization, anthropogenic activities have posed serious threats to the water quality and biodiversity of Liangzi Lake. To assess the aquatic ecosystem health of Liangzi Lake, the structure, the environmental response, and the interactions of plankton were investigated in 2022 and 2023. The results indicated that water temperature was a pivotal factor regulating plankton dynamics, with the assemblage patterns predominantly shaped by the phytoplankton species, which were Bacillariophyta in spring and Chlorophyta in summer. In terms of the phytoplankton, dissolved oxygen and the N:P ratio significantly affect cyanobacteria distribution. The high biomass and abundance of cyanobacteria in summer highlight the potential risk of harmful algal blooms. In contrast to the phytoplankton, the zooplankton exhibited enhanced resilience to changes in the surrounding environment. Rotifera was the dominant group in summer in terms of both abundance and biomass. Most core genera of plankton were jointly identified by eDNA metabarcoding and microscopical analysis, and eDNA metabarcoding had advantages in revealing a higher diversity. However, some taxa among rotifers such as Liliferotrocha were only identified using microscopical analysis. Therefore, a combination of both the methods is recommended to better understand the structuring mechanisms of plankton assemblages in lake ecosystems.

Filamentous algal blooms (or algal mats) are increasingly recognized as a growing threat to clear lakes worldwide, particularly in the context of climate change and lake eutrophication. Nevertheless, knowledge about filamentous algal mats and their environmental consequences is still limited. In this study, we investigated the structural characteristics and environmental impact of filamentous algae (Cladophora) mats in the brackish water of Qinghai Lake on the Tibetan Plateau. Our results classify the development of Cladophora blooms into three distinct stages: the attachment stage (May to July), the floating stage (August to September), and the decomposition stage (October to November), corresponding to attached, floating, and decaying mats, respectively. The attached mats consist of single layer, while the floating and decaying mats exhibit more complex structures, with two-layer and three-layer formations, respectively. Each layer displays distinct physiological states in the vertical direction, highlighting their structural diversity. The layered structure enables Cladophora mats to better adapt to environmental changes, ensuring long-term stability in the lake ecosystem through the synergistic effects of upper-layer protection, middle-layer growth, and bottom-layer decomposition. Notably, the water surrounding the decaying mats showed significantly elevated concentrations of nitrogen, phosphorus, and dissolved organic matter. Partial Least Squares Path Modeling analysis further revealed that Cladophora mats have a substantial influence on dissolved organic carbon and fluorescent dissolved organic matter, with path coefficients of 0.84 and 0.65, respectively. These findings significantly enhance our understanding of the dynamics of filamentous algal blooms and their environmental impacts, and are crucial for the conservation of lakes with high water quality.

Harmful cyanobacterial blooms pose significant threats to lake ecosystems, and the stocking of filter-feeding fish has often been used for their control. However, filter-feeding fish like silver carp excrete feces that not only retain viable cyanobacterial cells but also increase nutrient loading to the sediment. Furthermore, the quantity and frequency of fecal input vary depending on the biomass of algae and fish and the stocking strategy. In this study, a two-by-two factorial microcosm experiment was carried out to investigate the effects of silver carp feces on P release in shallow lakes. Results showed that fecal input quantity was the key determinant of P release. The peak flux reached 8.82 mg m−2 d−1 in high input treatments, compared to 1.01 mg m−2 d−1 in low input treatments. Phased-input exacerbated these effects compared to single-input. The dominant mechanisms of sediment P release varied with input levels. Microbial reduction was strongly associated with P release at low fecal input, while high-input scenarios showed concurrent hypoxia, an increase in sediment pH (from 7.28 to 7.46), and competition for adsorption sites by dissolved organic matter (DOM up to 38.57 mg L−1). These results indicate that stocking of filter-feeding fish for cyanobacterial bloom control substantially altered P flux dynamics, with high input treatments exhibiting fluxes from −6.02 to 8.82 mg m−2 d−1 compared to −0.007 to 0.33 mg m−2 d−1 in controls, depending on the patterns of fecal input. For the prevention and control of cyanobacterial blooms and to ensure the sustainability of lakes, the stocking of filter-feeding fish should be carried out before the outbreak of blooms to avoid the impact of large amounts of fish feces input on P release and water quality during the blooms.

Identifying alternative protein sources is crucial in view of the shortage of protein resources. A new strain Geotrichum candidum IBB69 was isolated for microbial protein production in this study. The protein yield, biomass, and specific protein production (the ratio of total intracellular and extracellular protein to biomass) of G. candidum IBB69 were 7.6 g/L, 18.42 g/L and 60.8

Assessing the response of phycospheric antibiotic resistance genes (ARGs) to pollutants such as antibiotics and microplastics (MPs) under the background of climate warming is crucial for understanding ARG fate in aquatic ecosystems. In this work, we conducted mesocosm experiments to evaluate how these stressors influence phytoplankton dominance, phycospheric bacteria, and ARG evolution. Results showed that in Microcystis-dominant ecosystem, a single stressor strengthened Microcystis dominance (up to 82.37 % of total abundance), but only warming significantly enhanced phycospheric bacterial metabolic activity and promoted phycospheric ARG proliferation (1.34-fold higher on day 16). The increased propagation of Microcystis symbiotic bacteria (e.g., Roseomonas and Methylobacterium) and microcystin-degraders (e.g., Pseudomonas and Sphingomonas) drove the spread of ARGs. Though the single treatment of antibiotics (0.85- and 0.53-fold for days 16 and 30) or MPs (0.72- and 0.20-fold for days 16 and 30) decreased the abundance of ARGs, co-treatment with warming reversed this suppression (e.g., 1.55- and 1.96-fold for WA and MW groups on day 16). The results underscore the necessity of considering the combined warming-pollutant effects in ARG ecological risk assessment in natural waters, particularly under phytoplankton succession scenarios. Such insights are vital for managing antimicrobial resistance in evolving aquatic environments under global change pressures.

Astaxanthin, a highly potent antioxidant pigment with growing applications in food, cosmetics, and pharmaceuticals, has gained significant attention. Schizochytrium were recognized as excellent commercially available strains for DHA production, featuring a functional mevalonate (MVA) pathway but low-level endogenous carotenoid synthesis. The wild-type Schizochytrium sp. strains are not ideal as astaxanthin production chassis prior to activation of the MVA pathway. The precursor supply of the MVA pathway was enhanced through electroporation-induced mutagenesis, leading to the development of a Schizochytrium sp. AST32 chassis, thereby achieving a significant elevation in astaxanthin content from 7.56 mu g/g to 55.17 mu g/g. Subsequently, through rational design, the astaxanthin biosynthetic pathway was further enhanced based on the mutant strain. Five key genes (idi, crtE, crtIBY, crtZ, crtW) were all expressed in Schizochytrium sp. AST32 led to a significant increase of astaxanthin content to 374.85 mu g/g. Furthermore, the astaxanthin synthesis module was targeted into cellular lipid body through innovative application of PLIN fusion protein, and a marked improvement in astaxanthin production, 400.38 mu g/g, were reached. Following fermentation optimization, a highest astaxanthin yielding, 29.53 mg/L (1913.07 mu g/g, 169.72-fold improvement over initial mutants) while maintaining DHA production at 3.83 g/L (45 % of total lipid content), was achieved. This study establishes a critical technical foundation for advancing the commercial scalability of astaxanthin biosynthesis in Schizochytrium sp..