A facultative anaerobic rhodobacter and application thereof

Abstract

The application discloses a facultative anaerobic rose genus bacterium and application thereof, and the facultative anaerobic rose genus bacterium is rose genus bacterium (Ruegerkobes sp. Rossellomorea arthrocnemi ) NJS-3, which is preserved in the China General Microbiological Culture Collection Center on September 28, 2025, and has a preservation number of CGMCC No. 36101, and a 16S rRNA gene sequence of the rose genus bacterium is shown as SEQ ID No. 1. The facultative anaerobic rose genus bacterium rose genus bacterium (Ruegerkobes sp. Rossellomorea arthrocnemi ) NJS-3 provided by the application has high benzene series (benzene, toluene, ethylbenzene, dimethylbenzene and the like) degradation capacity, can be used for bioremediation of benzene series contaminated soil and underground water, and has the advantages of high efficiency, greenness, no pollution and the like.

Description

Technical Field

[0001] This invention belongs to the field of microbiology and biotechnology, specifically relating to a facultative anaerobic Rosaceae bacterium (Rossellomorea arthrocnemi) and its applications. Background Technology

[0002] Benzene derivatives (BTEX) are a collective term for benzene and its derivatives, including benzene, toluene, ethylbenzene, and xylene. BTEX are carcinogenic, teratogenic, and mutagenic, and are highly volatile. Leakage is unavoidable during industrial applications and transportation, causing soil and groundwater pollution. This not only causes serious environmental problems but also poses a threat to human health.

[0003] Currently, groundwater pollution is mainly classified into three categories: physical remediation, chemical treatment, and bioremediation. Physical remediation technologies utilize the physical properties of pollutants to break them down, separate them, or control them. Major remediation techniques include thermal desorption, solidification stabilization, and electrokinetic remediation. However, physical remediation is costly and energy-intensive. Chemical treatment includes soil leaching and chemical oxidation, but while removing benzene compounds, it also generates ecotoxic byproducts or introduces leaching agents that cause secondary pollution. Unlike physical and chemical methods, bioremediation technologies, including phytoremediation and microbial remediation, are not only low-cost but also eliminate the risk of secondary pollution. They are considered the most promising remediation technologies. Microbial remediation primarily utilizes the metabolic activities of microorganisms to convert benzene compounds into non-toxic substances.

[0004] However, existing microbial remediation technologies mostly focus on the application of aerobic bacteria. In actual deep soil or groundwater pollution plumes, the environment is often anoxic or anaerobic, which greatly limits the activity of aerobic bacteria, resulting in low remediation efficiency. Therefore, screening and isolating microbial strains capable of efficiently degrading benzene compounds under facultative anaerobic or microaerobic conditions is of significant application value for solving the in-situ bioremediation of deep soil and groundwater. Summary of the Invention

[0005] In view of this, embodiments of the present invention provide a facultative anaerobic Rosaceae strain (Rossellomorea arthrocnemi) and its applications.

[0006] In a first aspect, embodiments of the present invention provide a facultative anaerobic Rosaceae strain, namely Rosellomorea arthrocnemi NJS-3, which was deposited at the China General Microbiological Culture Collection Center on September 28, 2025, with accession number CGMCC No. 36101. The gene sequence of the 16S rRNA of the Rosellomorea strain is shown in SEQ ID No. 1.

[0007] Secondly, embodiments of the present invention provide an application of facultative anaerobic Rosa bacteria in the degradation of benzene compounds.

[0008] Furthermore, the benzene series compounds are one or more of benzene, toluene, ethylbenzene, o-xylene, m-xylene, and p-xylene.

[0009] Furthermore, the application process specifically includes:

[0010] The facultative anaerobic Rosa spp. NJS-3 was activated in sterilized seed culture medium and cultured anaerobically in a constant temperature shaking incubator at 30℃-40℃.

[0011] Inoculate the seed culture solution into the liquid culture medium at an inoculation rate of 0.5%-3%, and place it in a constant temperature shaking incubator at 30℃-40℃ for anaerobic culture for more than 3 days;

[0012] Take soil or groundwater contaminated with benzene series compounds, add it to the liquid culture medium, and place the mixed mud-water solution in a constant temperature shaking incubator at 30℃-40℃ for more than 5 days.

[0013] Furthermore, the ratio of soil contaminated with benzene series compounds to liquid culture medium was 1:1 to 1:4 (w / v), and the content of benzene series compounds in the contaminated soil was 10 to 2000 mg / kg;

[0014] or,

[0015] The ratio of benzene-contaminated groundwater to liquid culture medium was 1:1 to 1:4 (v / v), and the benzene content in the contaminated groundwater was 10 to 2000 mg / L.

[0016] Furthermore, the seed culture medium consists of: 10 g / L peptone, 5 g / L yeast extract, 10 g / L NaCl, and pH 7.0.

[0017] Furthermore, the liquid culture medium has the following composition: NaNO3 5-18 g / L, KH2PO4 1-3 g / L, Na2HPO4·12H2O 1-3 g / L, KCl 0.5-2 g / L, NaCl 0.5-2 g / L, CaCl2 0.5-2 g / L, MgSO4 0.5-2 g / L, and pH 6.0-8.0.

[0018] Thirdly, embodiments of the present invention provide a microbial agent containing the aforementioned facultative anaerobic Rosa species.

[0019] Fourthly, embodiments of the present invention provide a method for preparing a bacterial agent, the method comprising: fermenting and culturing the above-mentioned facultative anaerobic Rosa species.

[0020] Fifthly, embodiments of the present invention provide a method for remediating benzene-contaminated soil and / or groundwater, characterized in that the method includes adding facultative anaerobic Rosa bacteria or bacterial agents to the benzene-contaminated soil and / or groundwater to be remediated.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0022] (1) The *Rosellomorea arthrocnemi* NJS-3 provided by this invention is a facultative anaerobic bacterium. Compared with traditional aerobic remediation strains, this strain can not only survive in aerobic environments, but also maintain high metabolic activity in anoxic or anaerobic microenvironments such as deep soil and groundwater. This characteristic effectively solves the technical problem of reduced remediation efficiency due to limited oxygen transport in in-situ bioremediation, and is particularly suitable for the remediation of deep environments contaminated by benzene series compounds.

[0023] (2) The facultative anaerobic Rosa spp. strain NJS-3 obtained by screening in this invention has extremely high tolerance to benzene compounds. Experimental data show that even in extreme environments with xylene concentrations as high as 2000 mg / L, this strain can still survive and maintain a degradation rate of 59.2%. This overcomes the shortcomings of conventional microorganisms, which are prone to toxic inhibition, inability to grow, or death when faced with high concentrations of organic pollutants, making it applicable to the remediation of areas with high concentrations of pollution sources.

[0024] (3) The facultative anaerobic Rosa spp. NJS-3 provided by this invention has high degradation efficiency and short remediation cycle: This strain degrades benzene compounds rapidly and removes them completely. In optimized mud systems and water remediation experiments, only 5 days of cultivation are required, and the degradation rate of benzene compounds in contaminated soil can reach up to 95.32%, and the degradation rate of benzene compounds in groundwater can reach up to 96.52%. The high efficiency of degradation significantly shortens the engineering remediation cycle.

[0025] (4) The facultative anaerobic Rosa spp. NJS-3 obtained by screening in this invention has a wide range of environmental adaptability and simple culture conditions: the strain can grow well in a wide pH range of 6.0-8.0, with the optimal pH being 6.6, which matches most natural soil and groundwater environments, without the need for large amounts of acid-base regulators. In addition, the strain can be grown using simple inorganic salt culture media, which is easy to prepare in the industrial fermentation process, reducing application costs.

[0026] (5) As a bioremediation technology, this invention utilizes the natural metabolism of microorganisms to mineralize benzene compounds, avoiding the high energy consumption problem of physical remediation (such as thermal desorption) and the risk of secondary pollution caused by chemical remediation (such as chemical oxidation and rinsing) that may introduce chemical agents. It has the advantages of high efficiency, greenness and environmental protection. Attached Figure Description

[0027] To more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0028] Figure 1 Microscopic morphology of Rosa spp. NJS-3 provided in an embodiment of the present invention. Detailed Implementation

[0029] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0030] It should be noted that, unless otherwise specified, the features in the following embodiments and implementation methods can be combined with each other.

[0031] Example 1: Isolation, screening and identification of Rosa spp. NJS-3

[0032] 1. Dilute and spread to obtain single colonies

[0033] Take 2g of soil contaminated with petroleum hydrocarbons, place it in a bottle containing 40mL of sterile water, shake to disperse, and seal. Take 1mL of the upper layer of liquid and spread it on a plate. Incubate at 30℃ for 12h, and finally dilute 10... -5 Find suitable single colonies for the study on plate. Inoculate the obtained single colonies into LB liquid medium for expansion culture. The composition of LB liquid medium (g / L) is: 10g peptone, 5g yeast extract, 10g NaCl, pH=7.0.

[0034] 2. Screening of benzene-degrading strains

[0035] The bacterial strains were screened using benzene compounds (xylene) as the sole carbon source. 3 mL of the enriched bacterial solution was added to 100 mL of liquid culture medium containing 200 mg / L xylene and anaerobically cultured at 30°C. Strains that survived under these culture conditions were selected. The liquid culture medium (g / L) consisted of: NaNO3 6, KH2PO4 1.5, Na2HPO4·12H2O 1.5, KCl 0.5, NaCl 0.5, CaCl2 0.5, MgSO4 0.5, pH=7.0.

[0036] Select bacterial strains that survive in liquid culture medium and measure the OD of the bacterial solution in the liquid culture medium. 600 The OD values ​​of the bacterial suspension in the liquid culture medium were measured at 24 h, 72 h, and 120 h. 600 The values ​​were 0.383, 0.588, and 0.623, respectively, indicating that the bacterium can utilize xylene for growth. Furthermore, the xylene content in the liquid culture medium was measured, and a decrease in xylene content was observed, indicating that the bacterium can degrade xylene.

[0037] 3. Taxonomic identification of strains

[0038] Microscopic morphology of Rosa spp. NJS-3 is shown in the figure. Figure 1 As shown in the image, the bacterial cells appear deep purple / blue-purple. Microbiological staining indicates that this strain is Gram-positive (G+), with typical rod-shaped cells. The cell ends appear blunt and rounded, not spherical or spiral, consistent with the morphological characteristics of the genus *Rosa* (a group of *Protosporum*). The cell arrangement is quite diverse, showing solitary (single bacilli), paired (appearing in pairs), and forming short or long chains (similar to *Streptomyces*). In some areas, the cells are densely clustered, exhibiting an irregular clump distribution.

[0039] The screened degrading strain was identified using molecular biology methods. Its 16S rRNA sequence, as shown in SEQ ID No. 1, was compared with the GenBank nucleic acid database. Sequence alignment confirmed it to be *Rosellomorea arthrocnemi*, and it was named *Rosellomorea arthrocnemi* NJS-3. It was deposited on September 28, 2025, at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 36101. The deposit address is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0040] Example 2: Selection of the optimal liquid culture medium for the growth of Rosa spp. NJS-3

[0041] After activating Rosa spp. NJS-3 in LB medium for 24 h, the strain was inoculated into a liquid medium with glucose as the carbon source (5000 mg / L) and a pH of 7.0. The inorganic salt content of the liquid medium is shown in Table 1. The strain was incubated in a 30°C shaking incubator for 2 days, and then the growth was measured. The medium conditions under which the strain grew best were considered the optimal medium.

[0042] Table 1: Growth of NJS-3 in different culture media

[0043]

[0044] Specifically, from the first to the fifth group of experimental data, as the concentration of inorganic salt components gradually increases, the growth rate (OD) of the strains also increases. 600 The concentration of inorganic salts showed a typical trend of first increasing and then decreasing, indicating that there is an optimal threshold for inorganic salt concentration; both excessively low and excessively high concentrations are detrimental to bacterial growth. The first to third groups represent the growth phase, where bacterial growth accelerated significantly with increasing nutrient salt concentration, indicating that inorganic salts are a limiting factor for growth within the low concentration range. The third group reached its maximum growth point. The fourth and fifth groups represent the inhibition phase, where growth decreased with further increases in concentration, indicating that excessively high inorganic salt concentrations inhibited bacterial growth.

[0045] The first group had the lowest inorganic salt content (e.g., only 5 g / L for NaNO3 and only 1 g / L for KH2PO4). The low OD600 value (1.9 ± 0.2) indicates that the essential nutrients such as nitrogen, phosphorus, and metal ions in the culture medium are insufficient, becoming limiting factors for the growth of the strain and preventing effective biomass accumulation.

[0046] Compared to the first group, the inorganic salt concentration in the second group was moderately increased, and nutrient limitations were overcome. The growth rate of the strain was significantly improved (OD). 600 The concentration of 2.5 ± 0.5 indicates that the nutrient supply within this concentration range has begun to meet the needs of the strain for rapid division.

[0047] The third group represents the optimal growth state. Under this formulation (NaNO3 9g / L, KH2PO4 2g / L, etc.), the strain achieved the highest biomass (OD). 600 The concentration reached 3.6 ± 0.3. This indicates that the carbon-nitrogen-phosphorus ratio (C:N:P) and metal ion concentration of this component are most suitable for the metabolic needs of Rosa spp. NJS-3. At this point, the nutrient supply is sufficient and has not yet reached the threshold for toxicity or high osmotic pressure, representing the optimal growth window for the strain.

[0048] The fourth group showed a significant decline in growth, despite having richer nutrients (NaNO3 reaching 10 g / L), but with lower growth (OD). 600 The concentration of inorganic salts was 2.3 ± 0.6, which was lower than that of the second and third groups. This suggests that the concentration of inorganic salts had exceeded the optimal range, which may have led to an increase in the osmotic pressure of the culture medium. This forced the strain to consume energy to maintain the osmotic balance inside and outside the cell, thereby reducing the energy allocation for growth and reproduction.

[0049] The fifth group exhibited a strong inhibitory effect. High concentrations of salt (such as NaNO3 up to 18 g / L) may have caused significant salt stress or ion toxicity to the cells, severely inhibiting enzyme activity or disrupting cell membrane permeability, leading to poor bacterial growth and reduced biomass (OD). 600 (1.7±0.3) even lower than the first group with poor nutrition.

[0050] In summary, as shown in Table 1 above, the inorganic salt liquid culture medium composition (g / L) for the third group of cultures is more suitable for the growth of the strains, namely: NaNO3 9 g / L, KH2PO4 2 g / L, Na2HPO4·12H2O 2 g / L, KCl 1 g / L, NaCl 1 g / L, CaCl2 1 g / L, MgSO4 1 g / L. This achieves the optimal combination of nutrient supply and osmotic pressure balance. This formula not only brings the highest biomass (OD200)... 60 =3.6), and its growth performance is stable, confirming it as the optimal inorganic salt liquid culture medium composition for Rosa spp. NJS-3.

[0051] Example 3: pH selection for the optimal growth of Rosa spp. NJS-3

[0052] After activating *Rosa* strain NJS-3 in LB medium for 24 h, it was inoculated into a liquid medium with glucose as the carbon source at a concentration of 5000 mg / L. The medium composition (g / L) was: NaNO3 9 g, KH2PO4 2 g, Na2HPO4·12H2O 2 g, KCl 1 g, NaCl 1 g, CaCl2 1 g, MgSO4 1 g, with different pH values ​​set. The medium was incubated at 30℃ with constant temperature and shaking for 2 days, and then the growth of the strain was measured. The pH of the medium with the best growth was considered the optimal medium.

[0053] Table 2: Growth of NJS-3 at different pH values

[0054]

[0055] As shown in Table 2 above, from group 1 (pH 6.0) to group 8 (pH 8.0), the growth rate (OD) of the strains increased with the gradual increase of the pH value of the culture medium. 600The values ​​(values) exhibit a significant unimodal trend, meaning they first rise rapidly to a peak, then fluctuate and decline. This indicates that environmental pH is a key factor affecting the metabolic activity of this strain, and excessively acidic or alkaline environments are detrimental to its biomass accumulation.

[0056] At pH 6.0, OD 600 The pH value was 2.7 ± 0.3. Although the strain could grow, the biomass was relatively low, indicating that the slightly acidic environment inhibited its growth. As the pH was finely adjusted to 6.2 and 6.4, the strain's growth rapidly increased, and the OD... 600 The values ​​reached 3.4±0.4 and 3.5±0.1, respectively. This rapid growth indicates that after slight relief of acid stress, the enzyme activity of the strain was rapidly activated and the metabolic rate was significantly accelerated.

[0057] When the pH reaches 6.6, the bacterial growth peaks, and the OD... 600 The pH value reached a maximum of 3.8 ± 0.1 with a very small standard deviation (± 0.1), demonstrating high stability in the growth state. This indicates that pH 6.6 is the optimal physiological pH for Rosa spp. NJS-3. Under these conditions, cell membrane permeability is optimal, nutrient absorption efficiency is highest, and the catalytic activity of intracellular enzyme systems is at its best, thus maximizing biomass accumulation.

[0058] As the pH value crosses the optimum point and enters the near-neutral range (pH 6.8 - 7.0), OD 600 The value dropped back to between 3.3 and 3.4. It is worth noting that at pH 7.0, although the average OD value (3.4) is still acceptable, the growth stability is not as good as in a slightly acidic environment (pH 6.6), and metabolic regulation begins to be slightly subjected to suboptimal pH stress.

[0059] When the environment becomes weakly alkaline (pH 7.5-8.0), the growth of the strain is significantly inhibited. At pH 8.0, OD... 600 The pH value dropped to a minimum of 2.5 ± 0.2, even lower than the level at pH 6.0. This indicates that the strain is sensitive to alkaline environments. Excessively high pH values ​​may lead to changes in cell membrane potential or denaturation and inactivation of key metabolic enzymes, thereby severely hindering the growth and reproduction of the strain.

[0060] In summary, *Rosa* fungus NJS-3 exhibits a distinct slightly acidophilic characteristic. As shown in Table 2 above, pH 6.6 was identified as its optimal growth pH, ​​under which the strain grows fastest and most stably. Preferably, the liquid culture medium comprises: NaNO3 6-10 g / L, KH2PO4 1.5-2.5 g / L, Na2HPO4·12H2O 1.5-2.5 g / L, KCl 0.5-1.5 g / L, NaCl 0.5-1.5 g / L, CaCl2 0.5-1.5 g / L, MgSO4 0.5-1.5 g / L, with a pH of 6.0-7.0. More preferably, the liquid culture medium comprises: NaNO3 9 g / L, KH2PO4 2 g / L, Na2HPO4·12H2O 2 g / L, KCl 1 g / L, NaCl 1 g / L, CaCl2 1 g / L, MgSO4 1 g / L, with a pH of 6.6. This characteristic suggests that maintaining a slightly acidic environment (around pH 6.6) will help to maximize the degradation efficiency of this strain in subsequent soil or groundwater remediation applications.

[0061] Example 4: Determination of the ability of Rosa spp. NJS-3 to degrade benzene compounds

[0062] After activating Rosa spp. NJS-3 in LB medium for 24 h, it was inoculated into a liquid medium with xylene as the sole carbon source (xylene concentration: 100–2000 mg / L). The medium was incubated at 30°C with shaking for 5 days. The residual xylene content in the medium was then analyzed by high-performance liquid chromatography (HPLC) to demonstrate the degradation effect. The composition of the liquid medium (g / L) was: NaNO3 9 g, KH2PO4 2 g, Na2HPO4·12H2O 2 g, KCl 1 g, NaCl 1 g, CaCl2 1 g, MgSO4 1 g, pH = 6.60.

[0063] Table 3: Xylene degradation ability of NJS-3

[0064]

[0065] As shown in Table 3 above, in the culture system with xylene as the sole carbon source (30℃, pH 6.60), the degradation rate of xylene by Rosa spp. NJS-3 exhibits a significant negative correlation with its initial concentration. That is, the degradation rate gradually decreases as the initial xylene concentration increases. However, the data change is not a simple linear decrease, but rather shows three distinct stages: high activity, significant inhibition, and a tolerance plateau.

[0066] Xylene exhibits a highly efficient degradation phase at concentrations of 100-200 mg / L. The *Rosa* strain NJS-3 demonstrates extremely high metabolic activity. At a xylene concentration of 100 mg / L, the degradation rate reaches 95.7 ± 0.3%; even when the concentration is doubled to 200 mg / L, the degradation rate remains at 93.1 ± 0.7%. This indicates that within this concentration range, xylene is primarily utilized by the strain as a nutrient carbon source, with negligible cytotoxic effects, allowing *Rosa* strain NJS-3 to almost completely remove contaminants.

[0067] When xylene concentration is between 500 and 1000 mg / L, an inhibitory transition phase occurs. As xylene concentration increases to 500 mg / L and above, the degradation rate begins to decline significantly. At 500 mg / L, the degradation rate drops to 86.3 ± 1.2%; at 700 mg / L, it further decreases to 73.6 ± 0.2%; and at 1000 mg / L, it drops to 63.6 ± 0.5%. This significant decline in degradation rate (from over 90% to around 60%) indicates that high concentrations of xylene begin to produce substrate inhibition. As an organic solvent, high concentrations of xylene may disrupt bacterial cell membrane structure or inhibit the activity of key metabolic enzymes, thus limiting the growth and metabolic efficiency of the strain.

[0068] At xylene concentrations of 1500-2000 mg / L, a high tolerance plateau was observed. Despite a significant increase in substrate concentration (doubling from 1000 mg / L to 2000 mg / L), the decline in degradation rate leveled off: at 1500 mg / L, the degradation rate was 60.2 ± 0.3%; at 2000 mg / L, the degradation rate remained at 59.2 ± 0.2%. At extremely high xylene concentrations (2000 mg / L), the degradation rate decreased by only 1 percentage point compared to 1500 mg / L. This demonstrates that *Rosa* fungus NJS-3 exhibits extremely strong tolerance to organic solvents. In environments where ordinary microorganisms might die due to solvent toxicity, NJS-3 not only survived but also maintained nearly 60% of its basal degradation capacity without metabolic collapse.

[0069] In summary, Rosaceae NJS-3 exhibits excellent removal efficiency (>93%) at low concentrations (<200 mg / L), making it suitable for deep remediation; while at high concentrations (>1500 mg / L), it demonstrates excellent tolerance and stability (~60%), making it suitable for the initial treatment of high-load pollution sources. This combination of high efficiency and tolerance validates its application potential in the remediation of complex benzene series pollution.

[0070] Example 5: Determination of inoculum size for degrading benzene series compounds by Rosa spp. NJS-3

[0071] After activating Rosa spp. NJS-3 in LB medium for 24 h, it was inoculated into a liquid medium with xylene as the sole carbon source at a concentration of 200 mg / L. Different inoculation amounts were set. The medium was incubated at 30℃ with shaking for 2 days, and then the residual xylene content in the medium was analyzed by high performance liquid chromatography (HPLC) to show the degradation effect. The composition of the liquid medium (g / L) was: NaNO3 9, KH2PO4 2, Na2HPO4·12H2O 2, KCl 1, NaCl 1, CaCl2 1, MgSO4 1, pH=6.60.

[0072] Table 4: Degradation of NJS-3 at different inoculum amounts

[0073]

[0074] As shown in Table 4 above, under the condition of a fixed xylene concentration of 200 mg / L, the xylene degradation rate after 48 hours showed a positive correlation trend of continuous increase with the increase of the initial inoculum amount (0.5%~3%). This means that the initial cell density directly affects the degradation efficiency of pollutants. Although theoretically increasing the inoculum amount may further improve the effect, considering the balance between the cost and efficiency of bacterial agent preparation in industrial applications, the xylene degradation rate at 48 hours is highest when the inoculum amount is 3%. Therefore, 3% is the optimal inoculum amount under the experimental conditions, which can obtain the best degradation output with reasonable input.

[0075] Example 6: Treatment of benzene-contaminated soil with Rosa spp. NJS-3

[0076] Rosa spp. NJS-3 was inoculated into sterilized LB medium and cultured anaerobically in a 30°C constant temperature shaking incubator (260 rpm) to form a seed culture solution. The seed culture solution was then inoculated into liquid medium at a volume ratio of 3%, with the following composition (g / L): NaNO3 9, KH2PO4 2, Na2HPO4·12H2O 2, KCl 1, NaCl 1, CaCl2 1, MgSO4 1, pH=6.60.

[0077] Take 10g of soil contaminated with benzene series compounds (hereinafter referred to as "contaminated soil") and add 10 mL, 20 mL, 30 mL, and 40 mL of liquid culture medium, respectively. The benzene series compound content in the contaminated soil is 253 mg / kg. The mixed mud-water solution is placed in a 30℃ constant temperature shaking incubator (260 rpm) for incubation. After 5 days, the residual benzene series compound content in the contaminated soil is analyzed.

[0078] Table 5: Degradation of NJS-3 under different soil and culture medium ratios

[0079]

[0080] The results in Table 5 above show that when the ratio of soil to liquid culture medium contaminated with benzene series compounds is 1:1 to 1:4 (w / v), the levels of benzene series compounds in the soil decrease. The ratio of soil to liquid culture medium can be selected according to the actual situation. From an engineering economic perspective, the 1:1 ratio has extremely high cost-effectiveness (removal rate ~94%). Although the degradation rate is about 1.5 percentage points lower than that of 1:4, the amount of culture medium used is reduced by 75%. In actual ex-situ soil remediation projects, it is recommended to prioritize a 1:1 or 1:2 ratio to reduce water treatment costs, while choosing 1:4 in scenarios pursuing the ultimate remediation goal.

[0081] Example 7: Treatment of benzene-contaminated groundwater by Rosa spp. NJS-3

[0082] Rosa spp. NJS-3 was inoculated into sterilized LB medium and cultured anaerobically in a 30°C constant temperature shaking incubator (260 rpm) to form a seed culture solution. The seed culture solution was then inoculated into liquid medium at a volume ratio of 3%, with the following composition (g / L): NaNO3 9, KH2PO4 2, Na2HPO4·12H2O 2, KCl 1, NaCl 1, CaCl2 1, MgSO4 1, pH=6.60.

[0083] Take 20 mL of water contaminated with benzene series compounds (toluene, xylene, benzene), and add 20 mL, 40 mL, 60 mL, and 80 mL of culture medium, respectively. The benzene series compound content in the contaminated water is 156 mg / L. Place the mixed aqueous solution in a 30℃ constant temperature shaking incubator (260 rpm) for incubation. After 5 days, analyze the residual benzene series compound content in the water.

[0084] Table 6: Degradation of NJS-3 under different groundwater and culture medium ratios

[0085]

[0086] The results in Table 6 above show that when the ratio of benzene-contaminated groundwater to liquid culture medium is 1:1-1:4 (v / v), the benzene levels in the groundwater are reduced. The ratio can be selected based on the specific circumstances. If the remediation target requires extremely low residual concentrations, a 1:4 ratio is recommended to achieve the highest removal rate of 96.52%. In conventional groundwater remediation projects, a 1:1 ratio can be used. Although its degradation rate (94.29%) is slightly lower than the 1:4 group (96.52%), the amount of culture medium used is reduced by 75% (from 4 times the volume to 1 times the volume). This means a significant reduction in the costs of microbial agent production, transportation, and injection, while achieving highly competitive remediation results.

[0087] In summary, the facultative anaerobic Rosaceae bacterium Rossellomorea arthrocnemi NJS-3 provided by this invention has high degradation efficiency for benzene series compounds and strong tolerance to benzene series compounds. It can be used for the bioremediation of benzene series contaminated soil and groundwater, and has the advantages of high efficiency, greenness and no pollution.

[0088] The above description is merely a preferred example of the present invention and is not intended to limit the scope of protection of the present invention. In addition to the above embodiments, the present invention may have other implementations. All technical solutions formed by equivalent substitutions or equivalent changes fall within the scope of protection claimed by the present invention.

Claims

1. A facultative anaerobic Rosa fungus, characterized in that, The facultative anaerobic Rosa species are Rosa species. Rossellomorea arthrocnemi NJS-3 was deposited at the China General Microbiological Culture Collection Center on September 28, 2025, with accession number CGMCC No. 36101. The gene sequence of the 16S rRNA of the Rosa species is shown in SEQ ID No.

1.

2. The application of the facultative anaerobic Rosa species as described in claim 1 in the degradation of benzene compounds.

3. The application according to claim 2, characterized in that, The benzene series compounds are one or more of benzene, toluene, ethylbenzene, o-xylene, m-xylene, and p-xylene.

4. The application according to claim 2 or 3, characterized in that, The application process specifically includes: The facultative anaerobic Rosa spp. NJS-3 was activated in sterilized seed culture medium and cultured anaerobically in a constant temperature shaking incubator at 30℃-40℃. Inoculate the seed culture solution into the liquid culture medium at an inoculation rate of 0.5%-3%, and place it in a constant temperature shaking incubator at 30℃-40℃ for anaerobic culture for more than 3 days; Take soil or groundwater contaminated with benzene series compounds, add it to the liquid culture medium, and place the mixed mud-water solution in a constant temperature shaking incubator at 30℃-40℃ for more than 5 days.

5. The application according to claim 4, characterized in that, The mass-to-volume ratio of soil contaminated with benzene series compounds to liquid culture medium was 1:1 to 1:4, and the content of benzene series compounds in the contaminated soil was 10 to 2000 mg / kg. or, The volume ratio of groundwater contaminated with benzene series compounds to liquid culture medium is 1:1 to 1:4, and the content of benzene series compounds in the contaminated groundwater is 10 to 2000 mg / L.

6. The application according to claim 4, characterized in that, The seed culture medium consists of: 10 g / L peptone, 5 g / L yeast extract, 10 g / L NaCl, and pH 7.

0.

7. The application according to claim 4, characterized in that, The liquid culture medium has the following composition: NaNO3 5-18 g / L, KH2PO4 1-3 g / L, Na2HPO4·12H2O 1-3 g / L, KCl 0.5-2 g / L, NaCl 0.5-2 g / L, CaCl2 0.5-2 g / L, MgSO4 0.5-2 g / L, and pH 6.0-8.

0.

8. A microbial agent, characterized in that, The bacterial agent contains the facultative anaerobic Rosa species as described in claim 1.

9. A method for preparing a microbial agent, characterized in that, The method includes: fermenting and culturing the facultative anaerobic Rosa species as described in claim 1.

10. A method for remediating benzene-contaminated soil and / or groundwater, characterized in that, The method includes adding the facultative anaerobic Rosa species of claim 1 or the microbial agent of claim 8 to the soil and / or groundwater contaminated with benzene series compounds to be remediated.

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