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    • 专利摘要:
    The invention is related to the field of soil remediation technologies, and in particular to a process for soil remediation by using graphene and a fermentation broth from rhodolorula mucilagl'nosa OP1 1, comprising steps of mixing graphene With soil to be 5 treated at a mixing ratio of 0.01 to 1 % by mass to cultivate a plant, and watering the soil near roots of the plant with a fermentation broth from rhodolorula mucilagmosa OP1 1. The process enables active heavy metal content in the soil to be effectively reduced and the pH level of coastal soil to be increased, and thus enables the soil to be improved. Further, the process also enables the content of heavy metal accumulated in the plant to 10 be reduced.
    公开号:NL2027967A
    申请号:NL2027967
    申请日:2021-04-14
    公开日:2021-06-07
    发明作者:Wang Fangli；Li Shaojing
    申请人:Univ Qingdao Agricultural；
    IPC主号:
    专利说明:

    PROCESS FOR SOIL REMEDIATION BY USING GRAPHENE AND FERMENTATION BROTH FROM RHODOTORULA MUCILAGINOSA OP11 Technical Field The present invention is related to the field of soil remediation technologies, and in particular to a process for soil remediation by using graphene and a fermentation broth from rhodotorula mucilaginosa OP11. Background IO Coastal wetlands are one of the major areas where heavy metals are able to more easily accumulate, and the coastal ecosystem is becoming increasingly worse for various reactions. Phytoremediation, as a very promising technology for removal of heavy metals from tidal flat, develops rapidly. Certain plants exhibiting special effects on the rhizospheric microbiome across the coastal soil can absorb and accumulate heavy metals from the contaminated coastal soil, reducing the heavy metal contamination and thus the hazards of heavy metals to tidal flat ecosystem. However, with respect to the coastal soil remediation through the conventional phytoremediation technologies, activities of the plants employed may be inhibited and even lost due to the presence of certain metals or other materials in the soil, or absorption of heavy metals by the plants in different areas of the target soil may vary greatly due to the different heavy metal concentrations in the areas. Therefore, the current phytoremediation technologies cannot achieve a reduction in the heavy metal contents in the soil.
    Summary In view of the above problems, an objective of the invention is to provide a process for soil remediation by using graphene and a fermentation broth from rhodotorula mucilaginosa OP11, which can enable the pH level of and the heavy metal contents in coastal soil to be reduced and increased, respectively, and can facilitate the absorption of heavy metals by plants. Accordingly, an objective of the invention is realized by a process for soil remediation by using graphene and a fermentation broth from rhodoforula mucilaginosa OP11, comprising:
    mixing graphene with soil to be treated at a mixing ratio of 0.01 to 1 % by mass to cultivate a plant, and watering the soil near roots of the plant with a fermentation broth from rhodotorula mucilaginosa OP11. In a preferred embodiment, the fermentation broth has a quantity of living bacterial cells ranging from 8.96x107 to 5.88x10% CFU/ml. In a preferred embodiment, the soil near roots of the plant is watered two times with the fermentation broth from rhodotorula mucilaginosa OP11, with the first watering being performed at day 15 after emergence of the plant and the second being performed at day 30 after the first watering. The amount of the fermentation broth used each time may be 10to 15 % by mass with respect to the mass of the soil to be treated.
    In a preferred embodiment, the plant comprises a plant which tends to accumulate a heavy metal.
    In a further preferred embodiment, the plant, which tends to accumulate a heavy metal, comprises plants of Suaeda.
    Ina preferred embodiment, the soil remediation comprises increasement of the pH level of the coastal soil, and reduction of the active heavy metal content therein and of the heavy metal content in the plant.
    In a preferred embodiment, the heavy metal comprises one or more of mercury (Hg), cadmium (Cd), and lead (Pb).
    The process of the present invention has several advantages. The invention provides a process for soil remediation by using graphene and a fermentation broth from rhodotorula mucilaginosa OP11, comprising mixing graphene with soil to be treated at a mixing ratio of 0.01 to 1 % by mass to cultivate a plant, and watering the soil near roots of the plant with a fermentation broth from rhodotorula mucilaginosa OP11. The process can enable the pH level of and the heavy metal content in the coastal soil to be increased and reduced, respectively, and can reduce the absorption of the heavy metal by the plant. Experiments, as described hereinafter, have shown that, with the process of the invention, the pH value of the soil sample to be treated was increased by 0.08 to
    0.012; the contents of active Hg, Cd, and Pd were decreased by 6.38 to 14.89 %, 20 to 40%, and 13.20 to 38.85 %, respectively; the biomasses of aerial and subterranean parts of the Suaeda plant were increased to 0.53 to 0.57 g/pot and 0.42 to 0.50 g/pot, respectively; and the accumulated amounts of Hg, Cd, and Pd in the aerial part of the Suaeda glauca plant were decreased by 43.14 to 60.39 %, 66.67 to 75.47 %, and 51.98 to 60.45 %, respectively, and the accumulated amounts thereof in the subterranean part of the Suaeda plant were decreased by 37.45 to 50 %, 70.06 to 70.38 %, and 42.88 to
    56.44 %, respectively. These indicate that the absorption of the heavy metals by the Suaeda plant was reduced by the application of the process.
    Name and Address of Depository Institution at Which the Biological Materials Were Deposited Rhodotorula mucilaginosa OP11 was deposited under the accession number CGMCC No. 13540 on 6 January, 2017 at China General Microbiological Culture Collection IO Center (CGMCC). Detailed Description The invention provides a process for soil remediation by using graphene and a fermentation broth from rhodotorula mucilaginosa OP11, comprising: mixing graphene with soil to be treated at a mixing ratio of 0.01 to 1 % by mass, preferably 0.1 to 0.9 % by mass, to cultivate a plant, and watering the soil near roots of the plant with a fermentation broth from rhodotorula mucilaginosa OP11.
    In this description, the graphene used is stock number G100 purchased from SHANDONG JINCHENG GRAPHENE TECHNOLOGY Co., LtD. The fermentation broth from rhodotorula mucilaginosa OP11 preferably has a quantity of living bacterial cells ranging from 8.96x107 to 5.88x103 CFU/ml, more preferably 3.67x10* CFU/ml. According to the process of the invention, the graphene serves to adsorb and deactivate the heavy metal present in the soil to be treated and thus to improve the soil. The fermentation broth from rhodotorula mucilaginosa OP11 serves to dissolve phosphorus (P), secrete auxin and thus promote plant growth, and to reduce absorption of the heavy metal by the plant. The process of the invention enables the pH value of moderately alkaline soil to be increased and the active heavy metal content in the soil to be reduced, thereby inhibiting absorption of the heavy metal by the plant and achieving remediation of the soil contaminated by the heavy metal.
    In this description, the soil sample selected for treatment through the process of the invention had a total Hg content of 22.9 mg/kg, a total Pd content of 46.8 mg/kg, a total Cd content of 0.97 mg/kg, a cation exchange capacity of 16.8 cmol/kg, and a pH value of 5.12. In the examples described hereinafter, flowerpots, having a diameter of 20 cm and a height of 17.5 cm, were used for performing the investigation on soil remediation through the process of the invention, with each pot containing 1500 g of the soil. The mixing and watering are not particularly limited herein and may be performed in any manner well known to a skilled person in the art.
    The watering is preferably performed two times. Preferably, the first watering is performed at day 15 after emergence of the plant, and the second watering is performed at day 30 after the first watering. The amount of the fermentation broth used each time is preferably 10 to 20 % by mass with respect to the mass of the soil to be treated. Further preferably, the amounts of the fermentation broth used by the first and second watering are 10 % and 15 %, respectively, with respect to the mass of the soil to be treated. The plant, as described above, preferably comprises a plant which tends to accumulate a heavy metal. The plant, which tends to accumulate a heavy metal, preferably comprises plants of Suaeda. Species of the plants of Suaeda is not particularly limited herein, and commercially available one may be employed. In the examples as described hereinafter, the Suaeda plant was derived from a river beach at Jihongtan in Qingdao city, Shandong province.
    The preparation of the fermentation broth from rhodotorula mucilaginosa OP11 is not particularly limited, and may be performed in any manner well known to a skilled person in the art. A preferred preparation method comprises: inoculating cells of rhodotorula mucilaginosa OP11 into malt extract agar medium (melt extract: 130 g, agar: 15 g, natural pH) for activation to obtain activated cells of rhodoforula mucilaginosa OP11; and inoculating the activated cells of rhodotorula mucilaginosa OP11 into malt extract medium (melt extract: 130 g, water: 1000 ml, natural pH) for fermentation to obtain the fermentation broth from rhodotorula mucilaginosa OP11. The activation is preferably carried out at 30 °C for 24 hours. The fermentation is preferably carried out at 28 °C for 2 days with an agitation speed of 200 rpm. The fermentation broth from rhodotorula mucilaginosa OP11 can activate the phosphorus of low solubility, secrete auxin and thus promote plant growth, and adsorb heavy metals.
    The soil remediation preferably comprises increasement of the pH level of the coastal soil, and reduction of the active heavy metal content therein and of the heavy metal content in the plant. The heavy metal preferably comprises one or more of Hg, Cd, and Pb.
    The process of the invention can enable the pH level of and the heavy metal contents in the coastal soil to be increased and reduced, respectively, and can reduce the absorption of the heavy metal by the plant.
    Experiments, as described hereinafter, have shown that, with the process of the invention, the biomasses of aerial and subterranean parts of the 5 Suaeda plant were increased to 0.53 to 0.57 g/pot and 0.42 to 0.50 g/pot, respectively; and the accumulated amounts of Hg, Cd, and Pd in the aerial part of the Suaeda plant were decreased by 43.14 to 60.39 %, 66.67 to 75.47 %, and 51.98 to 60.45 %, respectively, and the accumulated amounts thereof in the subterranean part of the Suaeda plant were decreased by 37.45 to 50 %, 70.06 to 70.38 %, and 42.88 to 56.44 %, respectively.
    These indicate that the absorption of the heavy metals by the Suaeda plant was reduced by the application of the process.
    In order to further illustrate the invention, the process for soil remediation by using graphene and a fermentation broth from rhodotorula mucilaginosa OP11 according to the invention will be further described with respect to the following examples, which, however, are not intended to limit the scope of the invention.
    Example 1 Cells of rhodotorula mucilaginosa OP11 preserved on slants were inoculated into malt extract agar medium for activation.
    The activation was carried out at 30 °C for 24 h to obtain activated cells of rhodotorula mucilaginosa OP11. The activated cells of rhodotorula mucilaginosa OP11 were then inoculated into malt extract medium and cultured in a constant temperature incubator at 28 °C for 2 d with an agitation speed of 200 rpm to obtain a fermentation broth from rhodotorula mucilaginosa OP11. The fermentation broth was well mixed for later use.
    It was found that the broth had a quantity of living bacterial cells ranging from 8.96107 to 5.88x105 CFU/ml.
    The fermentation broth from rhodotorula mucilaginosa OP11 prepared in Example 1 was used in the following Application Examples.
    Application Example 1 The example was performed as an experiment in the Chemistry Building at Qingdao Agricultural University, conducted from October, 2017 to April, 2019.
    Flowerpots were divided into 8 experimental groups (3 replicates per group). Various groups are shown in table 1. Table 1 Amounts of graphene and fermentation broth from rhodotorula mucilaginosa OPI] used in various groups ~~ Fermentation broth from rhodotorula Group Graphene mucilaginosa OP11 /(% by mass) /(% by mass) tT 0 2 0.01 0 3 0.10 0 4 1 0 5 0 25 6 0.01 25 7 0.10 25 8 1 25 In particular, after air drying, a soil sample was carefully ground and passed through a 20 mesh screen. 1500 g of the soil sample was then placed into each pot after mixing with graphene. Thereafter, Suaeda seeds were planted in the soil contained in each pot. Physicochemical properties of the soil sample are shown in Table 2.
    During growth of the plants, water content in the soil contained in each pot was maintained at around 65 % of the maximum field capacity. At day 15 after emergence of the Suaeda plants, the soil near roots of the plant in each pot was watered with the fermentation broth in an amount of 10 % with respect to the mass of the soil. At day 30 after watering, the soil near roots of the plant in each pot was then watered again with the fermentation broth in an amount of 15 % with respect to the mass of the soil. After 120 days, all plants were harvested for later experiments. Table 2 Soil physicochemical properties Total Hg TotalPd Total Cd Cation exchange Soil sample pH (mg/kg) /(mg/kg) /(mg/kg) capacity
    | city, Shandong 22.9 46.8 0.97 16.8 5.12 province Application Example 2 The soil, in Application Example 1, contained in each pot after harvest of the plants was subjected to determination of pH. The results are shown in Table 3. Table 3 Soil pH values Group pH T5155 2 5.17 3 5.18 4 5.16 5 5.17 6 5.23 7 5.27 8 5.24 Table 3 shows that there was a significant difference between the groups subj ected to the fermentation broth and the groups not subjected to the fermentation broth, and the pH of the soil subjected to the fermentation broth was increased. In particular, as compared with Group 1, the pH of the soil in Groups 2 to 8 was increased by 0.01 to
    0.09; as compared with Group 2, the pH of the soil in Group 6 was increased by 0.06; as compared with Group 3, the pH of the soil in Group 7 was increased by 0.09; and as compared with Group 4, the pH of the soil in Group 8 was increased by 0.08. Therefore, with the process of the invention, the pH of the soil was increased. Application Example 3 Active Hg, Cd, and Pd were extracted from the soil, in Application Example 1, contained in each pot after harvest of the plants through diethylenetriaminepentaacetic acid
    (DTPA) extraction. The results of measuring the contents of these active heavy metals are shown in Table 4. Table 4 Active heavy metal contents in the soil “Group Hg = ¢€ pd 1 0047 0045 0803 2 0.046 0.038 0.752 3 0.043 0.033 0.623 4 0.041 0.030 0.533
    0.045 0.043 0.761 6 0.044 0.036 0.697 7 0.041 0.029 0.568 8 0.040 0.027 0.491 5 Table 4 shows that there was a significant difference between the groups subjected to the fermentation broth and the groups not subjected to the fermentation broth, and the active heavy metal contents in the soil subjected to the fermentation broth were increased. In particular, as compared with Group 1, the contents of active Hg, Cd, and Pd in the soil in Groups 2 to 8 were decreased by 0.001 to 0.007, 0.002 to 0.018, and IO 0.042 to 0.312 mg/kg, respectively; as compared with Group 2, the contents of active Hg, Cd, and Pd in the soil in Group 6 were decreased by 0.002, 0.002, and 0.055 mg/kg, respectively; as compared with Group 3, the contents of active Hg, Cd, and Pd in the soil in Group 7 were decreased by 0.002, 0.004, and 0.055 mg/kg, respectively; and as compared with Group 4, the contents of active Hg, Cd, and Pd in the soil in Group 8 were decreased by 0.001, 0.03, and 0.042 mg/kg, respectively. Therefore, with the process of the invention, the contents of active heavy metals in the soil were decreased. Application Example 4 Each entire plant after being harvested was carefully rinsed using tap water to remove the soil therefrom. Aerial and subterranean parts of the plants were separated from each other, and were then carefully rinsed using distilled water. These parts were air dried to remove surface moisture, and their fresh weights were then measured. Thereafter, they were placed in a dry, well ventilated place so as to be air dried and thus obtain dried samples. Dry weights of the samples were measured to determinate the biomass thereof. The results are shown in table 5. Table $ Biomass of dried aerial and subterranean parts © Aerialpart Subterranean part Group /(g/pot) /(g/pot) 104723 0363 2 0.4355 0.3553 3 0.2758 0.2472 4 0.0943 0.1321
    0.5061 0.4092 6 0.5775 0.5038 7 0.5496 0.4886 8 0.5382 0.4292 5 Table 5 shows that there was a significant difference between the groups subjected to the fermentation broth and the groups not subjected to the fermentation broth, and the biomass of the plants subjected to the fermentation broth was increased. In particular, as compared with Group 1, the biomasses of the aerial and subterranean parts in Group 5 were increased by 0.0338 and 0.0439 g/pot, respectively, as compared with Group 2, the biomasses of the aerial and subterranean parts in Group 6 were increased by 0.1420 and
    0.1485 g/pot, respectively; as compared with Group 3, the biomasses of the aerial and subterranean parts in Group 7 were increased by 0.2738 and 0.2414 g/pot, respectively; and as compared with Group 4, the biomasses of the aerial and subterranean parts in Group 8 were increased by 0.4439 and 0.2971 g/pot. Therefore, with the process of the invention, the biomass of the Suaeda plants was increased. Application Example 5 The heavy metal contents in the dried Suaeda plants were measured. In particular, the Cd and Pd contents were measured by a wet digestion method and an ICP-MS method, and the Hg content was measured through Atomic Fluorescence Spectrometry (AFS). The results are shown in Tables 6 to 8. Table 6 Hg contents in the Suaeda plants
    Aerial Part Subterranean Part Heavy metal Group / / (mg/kg) (mg/kg) 2255406544 8640344 2 2.24+0.39ab 4.63+0.67a 3 2.05+0.06ab 4.55+0.30a 4 1.33+0.12b 3.30+0.01ab Hy 5 1.53+0.18b 3.30+£0.51ab 6 1.45+0.27ab 3.04+0.09ab 7 1.12+0.06b 2.43+0.65b 8 1.01+0. 10b 2.48+0.50b Note: different letters denote significant differences between the treatments (at 550.05). Table 7 Cd contents in the Szaeda plants Aerial Part Subterranean Part Heavy metal Group (mg/kg) (mg/kg) EE LL5940 46 30 2 1.23+0.30a 1.83+0.65ab 3 1.03+£0.62ab 1.47+0.05ab 4 0.69+0.01ab 1.194£0.01b cd 5 0.76+0.01ab 1.26+0.01b 6 0.53+0.20b 0.94+0.01b 7 0.46+0.09b 0.93+0.22b 8 0.3940. 16b 0.93+£0.35b Table 8 Pd contents in the Suaeda plants Aerial Part Subterrancan Part Heavy metal Group /(mg/kg) (mg/kg) 263241 654 3981£523a 2 24.77+£2.00a 33.0045 00ab Pd 3 16.5944 32ab 33.09+0.61a 4 15.36+3.37ab 32.46£3.99ab 5 12.85+1.06b 24 .09:+0.06b
    Heem Goo Sn eraan (mg/kg) (mg/kg) 6 126420340 22744081b 7 12.31+£3.61b 21.66+£5.15b 8 10.41+2.35b 17.3443.26b Tables 6 to 8 show that there was a significant difference between the groups subjected to the fermentation broth and the groups not subjected to the fermentation broth, and the heavy metal contents in the plants subjected to the fermentation broth was decreased. In particular, as compared with Group 1, the Hg, Cd, and Pd contents in the aerial parts in Groups 2 to 8 were decreased by 12.16 to 60.39 %, 22.64 to 75.47 %, and 5.89 to
    60.45 %, respectively, and the Hg, Cd, and Pd contents in the subterranean parts in Groups 2 to 8 were decreased by 4.73 to 50.00 %, 40.72 to 70.38 %, and 16.88 to
    56.44 %, respectively; as compared with Group 2, the Hg, Cd, and Pd contents in the aerial parts in Group 6 were decreased by 35.27 %, 56.91 %, and 48.97 %, respectively, and the Hg, Cd, and Pd contents in the subterranean parts in Group 6 were decreased by
    34.34 %, 48.63 %, and 31.28 %, respectively; as compared with Group 3, the Hg, Cd, and Pd contents in the aerial parts in Group 7 were decreased by 45.37 %, 55.34 %, and
    25.80%, respectively, and the Hg, Cd, and Pd contents in the subterranean parts in Group 7 were decreased by 46.59 %, 36.73 %, and 34.54 %, respectively; and as compared with Group 4, the Hg, Cd, and Pd contents in the aerial parts in Group 8 were decreased by
    24.06 %, 43.48 %, and 32.23 %, respectively, and the Hg, Cd, and Pd contents in the subterranean parts in Group 8 were decreased by 24.85 %, 21.85 %, and 46.58 %, respectively. Therefore, with the process of the invention, the heavy metal contents in the Swuaeda plants were decreased and thus the absorption of the heavy metals by the Suaeda plants was reduced. While preferred embodiments of the present invention have been described, it should be understood that the present invention is not so limited and various alterations and modifications may be made without departing from the scope and spirit of the present invention. The scope of the invention is to be limited only by the appended claims.
    权利要求:
    Claims (7)
    [1]
    1. Process for soil restoration using graphene and a fermentation infusion of rhodotorula mucilaginosa OP11, which involves mixing graphene with soil to be treated to cultivate a plant, and watering the surrounding soil from roots of the plant with a fermentation infusion of rhodotorula mucilaginosa OP11; wherein the graphene is used in an amount of 0.01 - 1% by mass with respect to the mass of the earth to be treated.
    [2]
    The process of claim 1, wherein the fermentation broth has an amount of live bacterial cells ranging from 8.96x10 7 - 5.88x10 3 CFU/ml.
    [3]
    The process according to claim 1, wherein the soil in the vicinity of roots of the plant is watered twice with the fermentation infusion, the first watering being performed on day 15 after plant emergence and the second watering being performed on day 30 after the first watering; wherein the fermentation infusion is used each time in an amount of 10 to 15 mass% with respect to the mass of the soil to be treated.
    [4]
    The process of claim 1, wherein the plant comprises a plant that tends to accumulate a heavy metal.
    [5]
    The process of claim 4, wherein the plant which tends to accumulate a heavy metal comprises plants of Suaeda.
    [6]
    The process of claim 1, wherein the soil restoration comprises increasing the pH level of coastal soil, and decreasing the active heavy metal content therein and the heavy metal content of the plant.
    [7]
    The process of claim 6, wherein the heavy metal comprises one or more of mercury, cadmium and lead.
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