Salt stress inhibits plant growth, requiring salt-tolerant genes for the development of resilient plants. A key tolerance mechanism is potassium/sodium homeostasis, governed by Shaker K+ channels. Given that Shaker K+ channels from salt-sensitive species have been extensively studied while their counterparts in salt-tolerant plants remain largely unexplored, this study investigates the evolution and function of these channels in salt-tolerant bermudagrass to address this knowledge gap. Genomic analysis identified 25 Shaker K+ channel genes, an expanded family relative to other species. Phylogenetics placed them into five groups (I-V), with groups I, II, III, and V expanded via segmental duplication. Salt stress response screening revealed that only CdKAT1.1 was rapidly upregulated. Functional assays in yeast demonstrated that both CdKAT1.1 and its closest homolog CdKAT1.2 improve potassium uptake and salt tolerance, but the enhancement from CdKAT1.1 was significantly greater. This work elucidates the expansion and functional divergence of Shaker K+ channels in bermudagrass. CdKAT1.1 emerges as a superior regulator of potassium efficiency and salt tolerance, making it a prime candidate for molecular breeding to improve plant resilience in saline-alkaline soils.

Zoysia is a perennial grass in the Poaceae family and is recognized as an excellent warm-season turfgrass with good tolerance to salt, drought, and other abiotic stresses. However, the identification of zoysiagrass species is still difficult because of the lack of specific molecular markers. In this study, a total of 18,981 SSR loci were detected based on the reported genome sequence of Z. japonica 'Nagirizaki'. The SSR primers were designed with standard repeat units such as dinucleotides, trinucleotides, tetranucleotides, pentanucleotides and hexanucleotides; the minimum number of repeats was five and amplified fragments were in the size range of 150-280 bp. A total of 400 pairs of SSR primers were selected, and their polymorphism characteristics were detected using 6 Zoysia germplasm resources, 179 primer pairs produced polymorphic bands, yielding a polymorphism rate of 44.75%. Then, 39 pairs of SSR primers with abundant polymorphisms were screened to study genetic diversity. The results revealed that the polymorphism information content (PIC) of the amplified bands among the 36 tested materials ranged from 0.099 to 0.866, with an average of 0.656. On this basis, we found that 10 pairs of primers can construct DNA fingerprints for 36 zoysiagrass varieties and distinguish them from each other according to the size of the amplified bands. The SSR markers developed in this study are helpful for promoting the progress of related research on variety discrimination and marker-assisted breeding of zoysiagrass and have good application prospects.

Soil compaction stress on plants remains widespread despite the presence of various mitigation methods. To address this concern, we conducted a series of studies from 2021 to 2023. Preliminary tests assessed the effects of the combined application of yeast and glucose on the porosity of compacted soil and on turfgrass growth under conditions of soil compaction. In subsequent dose screening studies, two-factor tests were performed to identify the best combination of yeast and glucose doses that enhance soil porosity and promote turfgrass growth under compaction stress. Therewith, we evaluated the effects of the identified best combination of yeast and glucose on plant growth and on physicochemical properties of shallow soil. It was found that the best dose for enhancing turfgrass growth in compacted soil was 200 g of yeast and 200 g of glucose per square meter. This combination significantly improved turf quality by 40 %, reduced soil bulk density by 9.11 %, and enhanced soil nutrition within 28 days. Additionally, notable enhancements in plant growth were observed in coastal saline-alkali lands and alpine meadows. The findings suggest that this innovative microbial-based approach could substantially improve plant growth under soil compaction stress. Furthermore, it proposes that investigating composite microbial inoculants, incorporating cellulose-degrading microbes, could concurrently address crop straw accumulation and extensive soil compaction.

Centipedegrass [Eremochloa ophiuroides (Munro) Hack.] is a perennial warm-season (C4) grass, which is originated in China and characterized by good adaptation to acidic and infertile soils, insect resistance, low maintenance and high ornamental value. Based on the above advantages, centipedegrass can be widely used in landscaping works and ecological restoration. However, the establishment speed of centipedegrass lawn is limited by the rooting ability of the stolon nodes. Our previous study found that an E3 ubiquitin protein ligase EoSINAT5 may regulate the root development of centipedegrass. In this study, EoSINAT5 was found to be expressed highest in the leaves and nodes, and the nodes are the place where the adventitious roots taken. Overexpressing EoSINAT5 in centipedegrass led to longer roots as compared to wild-type plants. The transcription factor EoBBR directly bonded to the GAGA-rich elements of EoSINAT5 promoter and interacted with EoPROG1 to repress EoSINAT5 expression coordinately. Overexpressing EoBBR or EoPROG1 in rice all resulted in shorter roots and less root tips. This study constructed a novel genetic model EoPROG1/EoBBR-EoSINAT5 for regulating the centipedegrass’ root development, and also provided an important theoretical basis for molecular breeding of centipedegrass.

Most zoysiagrass varieties in China are commercially propagated through vegetative organs, such as stolons and underground stems. Native varieties that can be propagated by seed are scarce. The development of inflorescences in grasses represents a critical transition from vegetative to reproductive growth. The process of inflorescence development of Zoysia is closely related to the reproductive characteristics including the inflorescence density, the number of grains per spike, seed setting rate and the thousand-grain-weight and affects the final seed yield and quality. However, the inflorescence development of Zoysia has not yet been thoroughly characterized. In this study, we systematically observed the inflorescence development of zoysiagrass and divided it into eight landmark stages: vegetative formation period, growth cone elongation period, bract primordia formation period, spikelet primordia formation period, glume protuberance formation period, gynoecium and stamen formation period, anther separation and pistil stigma bifurcation period and flowering period. Unlike most grasses, the growth cone of zoysiagrass did not rapidly elongate before differentiating during the bract primordia formation stage; instead, it elongated and differentiated simultaneously. We identified a leaf stage during which the height of the new pulvinus of vertical stems is either parallel to or lower than that of the preceding pulvinus, marking the transitional phase from vegetative to reproductive growth. Growth cones formed in this leaf stage. No growth cone was found in the vertical stem of Zoysia without this phenotype after dissection. This phenotype can serve as an indicative marker for the transition from vegetative to reproductive growth in zoysiagrass. Implementing appropriate cultivation and management practices at this stage can enhance reproductive traits and ultimately improve seed yield and quality. This study provides a theoretical foundation for the establishment of high-yield cultivation technology systems for zoysiagrass and the breeding of new high-yield and high-quality seed varieties in China.

Centipedegrass [Eremochloa ophiuroides (Munro) Hack.] is the best warm-season grass that originated from China with high ornamental value, but with a large leaf angle and long internode, the density of centipedegrass is low, which limits the application of centipedegrass. To improve the density of centipedegrass, novel genes that are involved in plant architecture need to be identified and interpreted. In this study, we cloned and elucidated the function of EoPGP1 in centipedegrass. The results showed that EoPGP1 encodes a clade I P-Glycoprotein (PGP) protein consisting of two transmembrane domains (TMDs) and two nucleotide-binding domains (NBDs), arranged as TMD1-NBD1-Linker-TMD2-NBD2. The EoPGP1 gene is ubiquitously expressed in multiple tissues, and the EoPGP1 protein localizes to the plasma membrane. EoPGP1 could transport auxin in yeast cell. Under dim, red and blue light, EoPGP1 complemented the phenotypes induced by the loss-of-function of AtPGP1. Overexpression of EoPGP1 in rice increased leaf angle, free Indole-3-acetic acid (IAA) content and sensitivity to 1-N-naphthylphthalamic acid (NPA). In this study, we verified that EoPGP1 is involved in auxin transport and modifying plant architecture, providing a potential gene that is applicable for improving the density of centipedegrass.

Zoysiagrass (Zoysia matrella) is a warm-season turfgrass, but its slow growth rate and poor drought tolerance restrict its industrial use. In this research, zoysiagrass callus (undifferentiated parenchymal cell masses derived from dedifferentiated living cells with the capacity for whole-plant regeneration through redifferentiation) was irradiated with cold plasma at different dose. The results indicated that 483 W was the optimal dose for maximizing callus survival while maintaining mutagenic efficiency. Cold plasma induced diverse phenotypic variations, including alterations in leaf shape, stolon growth, and spike characteristics. After evaluations of turfgrass quality, growth performance, and sequence-related amplified polymorphism (SRAP) molecular marker analysis, four high-quality and fast-growing mutants was selected. Among them, two mutants also showed better drought tolerance. This study confirms that cold plasma-mediated callus irradiation is an effective approach for creating novel zoysiagrass germplasm. Cold plasma irradiation on zoysiagrass callus produced rapid-growth and droughttolerant mutants, offering a novel mutagenesis technology to advance turfgrass breeding and germplasm innovation.

Shaker K⁺ channels play a vital role in plant potassium homeostasis and stress adaptation. While existing research predominantly focuses on model plants like Arabidopsis and agricultural crops, investigations into Shaker K⁺ channels in grass species remain limited. Given the potential for distinct characteristics in grass Shaker K⁺ channel gene families, this study conducted a comprehensive genome-wide analysis of these channels in centipedegrass, a species widely used in landscaping. Seven Shaker K⁺ channel genes were identified, unevenly distributed across five chromosomes. Notably, centipedegrass harbors fewer members compared to other species, due to the absence of group III and IV subfamilies. Structural variations among these genes suggest functional diversity. Functional assays in potassium-deficient yeast revealed that only EoKAT1 and EoKAT2 displayed potassium uptake capabilities, with EoKAT1 exhibiting markedly higher absorption than EoKAT2. Surprisingly, EoAKT1 failed to demonstrate potassium absorption functionality, contrasting with its homologs in other species. Salt tolerance assays in yeast further showed that EoKAT1 significantly enhanced cellular salt tolerance, while EoKAT2 had no effect—a finding consistent with their differential potassium uptake capacities. EoKAT1 and EoKAT2 share a collinear relationship, and disparities in their potassium absorption, salt tolerance, and promoter elements highlight functional divergence, potentially enabling distinct environmental responses. Subcellular localization confirmed EoKAT1 is anchored to the plasma membrane. Collectively, this study provides insights into the unique characteristics of the Shaker K⁺ channel family in centipedegrass, differing from other species. EoKAT1 emerges as a promising candidate for improving plant potassium utilization and salt stress resilience. The centipedegrass Shaker K+ channel gene family displays distinctive features that might mirror the unique evolutionary attributes of grasses. EoKAT1 plays a role in improving potassium uptake and salt resistance.

As one of the most salt-tolerant grasses, characterizing salt-tolerance genes of Zoysia matrella [L.] Merr. not only broaden the theoretical information of salt tolerance, but also provide new salt-resistant genetic resources for crop breeding. The salt-inducible protein disulfide isomerase (ZmPDI) of Zoysia matrella [L.] Merr. was proved to enhance salt tolerance in homologous overexpression transgenic plants. In order to evaluate its potential application in crops, we conducted the salt tolerance evaluation in heterologous overexpression transgenic rice (OX-ZmPDI), Wild-type (WT) rice, and LOC_Os11g09280 (OsPDI, homologous gene of ZmPDI in rice) knock-out rice generated by CRISPR-Cas9 system (CR-OsPDI). Our findings revealed that OX-ZmPDI rice was higher and exhibited longer main root length, more proline (Pro) and malondialdehyde (MDA), and higher peroxidase (POD) activity than WT control after salt treatment, while CR-OsPDI resulted in contrary phenotypes. These results indicated that ZmPDI can significantly enhance the salt tolerance in rice, whereas loss-of-function of OsPDI reduces the salt tolerance. To further investigate these differences at the molecular level, we collected roots from OX-ZmPDI transgenic, CR-OsPDI transgenic, and wild-type (WT) plants at 0 and 24 h after salt treatment for RNA-seq and data-independent acquisition (DIA) proteome sequencing. Combined analysis of the transcriptome and proteome revealed that ZmPDI has the potential to enhance the salt tolerance of rice by modulating the expression of laccase-6, zingipain-2, WIP3, FKBP65, AKR4C10, GBSSII, Pho1, and TRXf1. Those results provided new information for the molecular regulation mechanism by which ZmPDI improves salt tolerance, and prove the potential of ZmPDI for application in crop breeding.

Perennial grasses seasonal nitrogen (N) cycle extends the residence and reuse time of N within the plant system, thereby enhancing N use efficiency. Currently, the mechanism of N metabolism has been extensively examined in model plants and annual grasses, and although perennial grasses exhibit similarities, they also possess distinct characteristics. Apart from assimilating and utilizing N throughout the growing season, perennial grasses also translocate N from aerial parts to perennial tissues, such as rhizomes, after autumn senescence. Subsequently, they remobilize the N from these perennial tissues to support new growth in the subsequent year, thereby ensuring their persistence. Previous studies indicate that the seasonal storage and remobilization of N in perennial grasses are not significantly associated with winter survival despite some amino acids and proteins associated with low temperature tolerance accumulating, but primarily with regrowth during the subsequent spring green-up stage. Further investigation can be conducted in perennial grasses to explore the correlation between stored N and dormant bud outgrowth in perennial tissues, such as rhizomes, during the spring green-up stage, building upon previous research on the relationship between N and axillary bud outgrowth in annual grasses. This exploration on seasonal N cycling in perennial grasses can offer valuable theoretical insights for new perennial grasses varieties with high N use efficiency through the application of gene editing and other advanced technologies.

As an important warm-season turfgrass species, bermudagrass (Cynodon dactylon L.) flourishes in warm areas around the world due to the existence of the C4 photosynthetic pathway. However, how C4 photosynthesis operates in bermudagrass leaves is still poorly understood. In this study, we performed single-cell RNA-sequencing on 5296 cells from bermudagrass leaf blades. Eight cell clusters corresponding to mesophyll, bundle sheath, epidermis and vascular bundle cells were successfully identified using known cell marker genes. Expression profiling indicated that genes encoding NADP-dependent malic enzymes (NADP-MEs) were highly expressed in bundle sheath cells, whereas NAD-ME genes were weakly expressed in all cell types, suggesting C4 photosynthesis of bermudagrass leaf blades might be NADP-ME type rather than NAD-ME type. The results also indicated that starch synthesis-related genes showed preferential expression in bundle sheath cells, whereas starch degradation-related genes were highly expressed in mesophyll cells, which agrees with the observed accumulation of starch-filled chloroplasts in bundle sheath cells. Gene co-expression analysis further revealed that different families of transcription factors were co-expressed with multiple C4 photosynthesis-related genes, suggesting a complex transcription regulatory network of C4 photosynthesis might exist in bermudagrass leaf blades. These findings collectively provided new insights into the cell-specific expression patterns and transcriptional regulation of photosynthetic genes in bermudagrass.

Potassium is crucial in plant metabolism processes, and sufficient potassium can improve plant tolerance to abiotic stress. We studied the effects of different KCl concentration treatments (0, 1, 5, 20 mM) on the biomass, photosynthetic characteristics, and ion content of Zoysia matrella under salt stress (NaCl 300 mM). The results showed that the plant dry weight, stomatal conductance, transpiration rate, photosynthesis rate, K+ content in plants, and K+/Na+ ratio in leaves of Zoysia matrella under NaCl stress were significantly lower than those under no NaCl conditions. The addition of K can promote an increase in plant dry weight and significantly improve the stomatal conductance, transpiration rate, and photosynthesis rate of plants. In addition, under salt stress, the addition of 20 mM KCl can significantly reduce the accumulation of Na+ in plants and promote the secretion of Na+ in leaves, thus improving the salt tolerance of Zoysia matrella.

The key to improving the efficiency by which acid-soil-growing plants utilize nitrogen is to improve their efficiency of utilizing ammonium. The carbon deficiency is recognized as the main cause of the failure to increase ammonium efficiency by simply strengthening the ammonium absorption capacity of the roots. Whether increasing carbon input by manipulating the stomatal aperture can enhance the utilization of ammonium in plants that receive high-intensity ammonium absorption in roots is our goal. We use a high ammonium treatment to mimic the excessive ammonium uptake condition and manipulate the stomata by spraying potassium or potassium uptake inhibitors through a foliar method. This was performed to investigate the effect of manipulating the stomatal aperture on the ammonium utilization capacity of centipedegrass, a typical acid soil-growing excellent turf grass, under the root excessive ammonium uptake condition. Once a high-ammonium root environment is encountered, the stomatal aperture rapidly decreases. Leaf potassium supply, rather than sodium ion supply, increases the stomatal aperture. Under hydroponic conditions, foliar application of potassium solution alleviates the inhibition of stomatal opening, restores photosynthetic capacity, and thus increases plant tolerance to high ammonium levels. When additional potassium absorption inhibitor Cs is added, the plant loses the relieving effect caused by foliar potassium spraying. Foliar spraying of potassium solution improves the high ammonium tolerance of centipedegrass by increasing the stomatal aperture. Co-reinforcing the leaf stomatal opening process and the root ammonium absorption capacity process is a feasible strategy to achieve the high efficiency of ammonium.

This study was designed to investigate the relationship between the caloric value and salt tolerance of two varieties of Miscanthus sacchariflorus (Amur silvergrass: M127 and M022). The salt tolerance capacity, photosynthetic characteristics, Na+ and K+ uptake by the roots and aboveground parts, and caloric value of different parts of the aboveground parts were obtained under hydroponic conditions. The results showed that M022 was more tolerant to salt stress than M127 and the former had a higher photosynthetic efficiency as well as a lower aboveground Na+ accumulation, K+ efflux, and larger K+/Na+ ratio. The calorific values of stems, spear leaves, aging leaves, and functional leaves of the two varieties showed a decreasing trend with increasing NaCl concentration. At 270 mM NaCl, the calorific values of the stems, aging leaves, functional leaves, and spear leaves was reduced by 18.10%, 46.73%, 26.11%, and 18.35% for M022 and 41.99%, 39.41%, 34.82%, and 45.09% for M127 compared to the controls, respectively. We observed that the aging leaves of M022 had a faster decline rate in calorific value than those of M127, indicating that the aging leaves of M022 preferentially isolated the harmful Na+ ion, reduced its accumulation in other parts, and increased the K+/Na+ ratio in the corresponding parts, thus inhibiting the decrease in calorific value. Following this result, it can be inferred that M022 inhibited the decline in calorific values during stress by efficiently compartmentalizing the distribution of Na+ and K+. Our results provide a theoretical basis and technical support for the efficient cultivation of salt-tolerant energy plants in saline–alkaline soil.

Tillering directly determines the seed production and propagation capacity of clonal plants. However, the molecular mechanisms involved in the tiller development of clonal plants are still not fully understood. In this study, we conducted a proteome comparison between the tiller buds and stem node of a multiple-tiller mutant mtn1 (more tillering number 1) and a wild type of centipedegrass. The results showed significant increases of 29.03% and 27.89% in the first and secondary tiller numbers, respectively, in the mtn1 mutant compared to the wild type. The photosynthetic rate increased by 31.44%, while the starch, soluble sugar, and sucrose contents in the tiller buds and stem node showed increases of 13.79%, 39.10%, 97.64%, 37.97%, 55.64%, and 7.68%, respectively, compared to the wild type. Two groups comprising 438 and 589 protein species, respectively, were differentially accumulated in the tiller buds and stem node in the mtn1 mutant. Consistent with the physiological characteristics, sucrose and starch metabolism as well as plant hormone signaling were found to be enriched with differentially abundant proteins (DAPs) in the mtn1 mutant. These results revealed that sugars and plant hormones may play important regulatory roles in the tiller development in centipedegrass. These results expanded our understanding of tiller development in clonal plants.

Carbon ion beam irradiation (CIBI) is a highly efficient mutagenesis for generating mutations that can be used to expand germplasm resources and create superior new germplasm. The study investigated the effects of different doses of CIBI (50 Gy, 100 Gy, 150 Gy, 200 Gy and 300 Gy) on seed germination and seedling survival, seedling morphological and physiological traits of an elite centipedegrass cultivar Ganbei. The results showed that irradiation greater than 50 Gy cause inhibition of seed germination, and the semi-lethal dose (LD50) is around 90 Gy for CIBI treated seeds of Ganbei. A carbon ion beam-mutagenized centipedegrass population was generated from Ganbei, with irradiation dosages from 50 Gy to 200 Gy. More than ten types of phenotypic variations and novel mutants with heritable tendencies mainly including putative mutants of stolon number, length and diameter, of internode length, of leaf length and width, of leaf chlorophyll content, of stolon growth rate, of aboveground tissue dry weight, of sward height were identified. While the total sugar content of the plants from irradiated seeds showed no obvious change in all treatments as compared to the control, the crude protein content displayed significant reduction at a high-dose treatment of 200 Gy. Genetic polymorphism was detected in mutagenized centipedegrass population using SSR-PCR analysis, suggesting that CIBI caused alteration of larger fragments of the DNA sequence. As a result, a preliminary batch of mutants was screened in this study. In summary, carbon ion beam mutagenesis is an effective way for developing centipedegrass germplasm with wider variation, and treating seeds with CIBI at a dosage of ~100 Gy could be effective in centipedegrass mutation breeding.

Centipedegrass (Eremochloa ophiuroides) is an important warm-season grass plant used as a turfgrass as well as pasture grass in tropical and subtropical regions, with wide application in land surface greening and soil conservation in South China and southern United States. In this study, the complete cp genome of E. ophiuroides was assembled using high-throughput Illumina sequencing technology. The circle pseudomolecule for E. ophiuroides cp genome is 139,107 bp in length, with a quadripartite structure consisting of a large single copyregion of 82,081 bp and a small single copy region of 12,566 bp separated by a pair of inverted repeat regions of 22,230 bp each. The overall A + T content of the whole genome is 61.60%, showing an asymmetric nucleotide composition. The genome encodes a total of 131 gene species, composed of 20 duplicated genes within the IR regions and 111 unique genes comprising 77 protein-coding genes, 30 transfer RNA genes, and 4 ribosome RNA genes. The complete cp genome sequence contains 51 long repeats and 197 simple sequence repeats, and a high degree of collinearity among E. ophiuroide and other Gramineae plants was disclosed. Phylogenetic analysis showed E. ophiuroides, together with the other two Eremochloa species, is closely related to Mnesithea helferi within the subtribe Rottboelliinae. These findings will be beneficial for the classification and identification of the Eremochloa taxa, phylogenetic resolution, novel gene discovery, and functional genomic studies for the genus Eremochloa.

The U-box protein family of ubiquitin ligases is important in the biological processes of plant growth, development, and biotic and abiotic stress responses. Plants in the genus Zoysia are recognized as excellent warm-season turfgrass species with drought, wear and salt tolerance. In this study, we conducted the genome-wide identification of plant U-box (PUB) genes in Zoysia japonica based on U-box domain searching. In total, 71 ZjPUB genes were identified, and a protein tree was constructed of AtPUBs, OsPUBs, and ZjPUBs, clustered into five groups. The gene structures, characteristics, cis-elements and protein interaction prediction network were analyzed. There were mainly ABRE, ERE, MYB and MYC cis-elements distributed in the promoter regions of ZjPUBs. ZjPUBs were predicted to interact with PDR1 and EXO70B1, related to the abscisic acid signaling pathway. To better understand the roles of ZjPUBs under salt stress, the expression levels of 18 ZjPUBs under salt stress were detected using transcriptome data and qRT-PCR analysis, revealing that 16 ZjPUBs were upregulated in the roots under salt treatment. This indicates that ZjPUBs might participate in the Z. japonica salt stress response. This research provides insight into the Z. japonica PUB gene family and may support the genetic improvement in the molecular breeding of salt-tolerant zoysiagrass varieties.

In the process of lawn production and planting, the application of different fertilizers has a significant impact on the growth of turfgrass and the soil environment. It is well known that organic fertilizer can promote the growth of turfgrasses, but its underlying mechanism still needs to be further analyzed. In our study, pot experiment was conducted, where three commonly used warm-season turfgrass species, including bermudagrass (Cynodon dactylon, 'Yangjiang', YJ), zoysiagrass (Zoysia japonica, 'Suzhi No.1′, S1) and paspalum grass (Paspalum vaginatum, 'Adalayd', AD), were planted for two growing seasons in PVC tubes with no fertilizer treatment (CK, sandy soil), chemical fertilizer treatment (CF, sandy soil with urea, Ca(H2PO4)2·H2O and K2SO4), low-nitrogen organic fertilizer treatment (LNOF, sandy soil with chicken manure composts) and high-nitrogen organic fertilizer treatment (HNOF, sandy soil with oil rapeseed cake composts). Each PVC tube contained 16 stolons of each turfgrass with the apical two nodes, and each treatment had six biological replications. Phenotypic and physiological data were analyzed using SPSS Statistics (Duncan's test) and revealed that HNOF treatment significantly enhanced turfgrass growth, maintained soil fertility, and improved nitrogen use efficiency (NUE) of plants. Conversely, the effects of LNOF on turfgrass and soil were similar to those of CF. The Principal Coordinate Analysis (PCoA) based on the Bray-Curtis distance metric showed that the bacterial community were mainly influenced by different fertilization treatments, while the fungal community were more associated with the different turfgrass species. The structural equation modeling (SEM) analysis revealed that different fertilization treatments primarily affect NUE by influencing the composition of the bacterial community, which has a significant impact on the total fresh weight of the turfgrasses. The Linear discriminant analysis effect size (LEfSe) and correlation analysis revealed that HNOF treatment induced the most significant difference operational taxonomic units (OTUs), among which, Bacillus OTUs were the largest number and all Bacillus OTUs were positive related with the NUE and fresh weight of the plants. Furthermore, the functions of "chitinolysis", "aerobic chemoheterotrophy", "aromatic compound degradation" and "chemoheterotrophy" showed a significant positive correlation with NUE, and some Bacillus strains have been reported to explicitly possess these functions. This research highlights the benefits of HNOF in promoting turfgrass growth and maintaining soil fertility, with Bacillus potentially playing a crucial role in those process. Our findings further suggest that the combination of stimulated Bacillus and HNOF (bio-organic fertilizers) may have a synergistic effect in promoting the growth of warm-season turfgrass, which needs to be investigated in the future studies.