CN115058252B - Microbial soil conditioner for restoring phthalate polluted soil and application thereof - Google Patents

Microbial soil conditioner for restoring phthalate polluted soil and application thereof Download PDF

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CN115058252B
CN115058252B CN202210049855.8A CN202210049855A CN115058252B CN 115058252 B CN115058252 B CN 115058252B CN 202210049855 A CN202210049855 A CN 202210049855A CN 115058252 B CN115058252 B CN 115058252B
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葛静
王亚
余向阳
黄博闻
张猛
宋立晓
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Jiangsu Yanjiang Agricultural Science Research Institute
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Abstract

本申请提供了一种用于酞酸酯污染土壤修复的微生物土壤调理剂,该土壤调理剂包括微生物搭载生物炭、有机质、糖碳源和养分,所述微生物搭生物炭由由鞘氨醇单胞菌、枯草芽孢杆菌和生物炭组成;该调理剂内生菌还可以定殖植物内部,促进植物内部污染物(PAEs)降解,达到内外同时修复的效果,具修复与促生为一体,能在修复去除塑化剂污染的同时促进植物生长,调节土壤微生态,减少化肥的使用,有利于高质量农田建设和农业的可持续发展。

The application provides a microbial soil conditioner for the remediation of phthalate-contaminated soil, the soil conditioner comprises microorganisms carrying biochar, organic matter, sugar carbon sources and nutrients, and the microorganisms and biochar are composed of sphingosine mono bacteria, Bacillus subtilis and biochar; the endophytic bacteria of the conditioner can also colonize the interior of plants, promote the degradation of plant internal pollutants (PAEs), achieve the effect of internal and external repair at the same time, with repair and growth promotion as a whole, can While repairing and removing plasticizer pollution, it promotes plant growth, regulates soil microecology, reduces the use of chemical fertilizers, and is conducive to the construction of high-quality farmland and the sustainable development of agriculture.

Description

用于酞酸酯污染土壤修复的微生物土壤调理剂及其应用Microbial soil conditioner and its application for remediation of phthalate-contaminated soil

技术领域technical field

本发明涉及肥料技术领域,特别是有毒有机物污染土壤修复微生物土壤调理剂的制备方法及其在酞酸酯降解中的应用。The invention relates to the technical field of fertilizers, in particular to a preparation method of a microbial soil conditioner for remediating soil contaminated by toxic organic substances and its application in the degradation of phthalate esters.

技术背景technical background

酞酸酯也称作塑化剂(PAEs),是邻苯二甲酸酯类化合物,被广泛用作增塑剂,应用于玩具、化妆品、建筑材料、农药载体等。有些PAEs是内分泌干扰物质,对动物和人类具有潜在致癌、致畸和致突变作邻苯二甲酸正丁酯(DBP)、领苯二甲酸二(2-乙基己基)酯(DEHP)等6种塑化剂被美国国家环保局列为优先控制污染物,邻苯二甲酸二甲酯等3种PAEs也被中国环境保护部认定为环境优先控制污染物。由于塑化剂的广泛使用,导致其在空气、土壤、水等环境中残留较为普遍。在农产品产地中,设施和覆膜蔬菜产地土壤和蔬菜中塑化剂检出浓度和检出率要远远高于常规产地。环境中的塑化剂容易被作物吸收和富集,不仅影响作物品质,还将对人体造成潜在健康风险。Phthalates, also known as plasticizers (PAEs), are phthalates and are widely used as plasticizers in toys, cosmetics, construction materials, and pesticide carriers. Some PAEs are endocrine disrupting substances, which are potentially carcinogenic, teratogenic and mutagenic to animals and humans, such as n-butyl phthalate (DBP), bis(2-ethylhexyl) phthalate (DEHP), etc. 6 A plasticizer is listed as a priority pollutant by the US National Environmental Protection Agency, and three PAEs including dimethyl phthalate are also recognized as a priority pollutant by the Ministry of Environmental Protection of China. Due to the wide use of plasticizers, their residues in air, soil, water and other environments are more common. In the production areas of agricultural products, the detection concentration and detection rate of plasticizers in the soil and vegetables of facilities and film-covered vegetable production areas are much higher than those in conventional production areas. Plasticizers in the environment are easily absorbed and enriched by crops, which not only affects the quality of crops, but also poses potential health risks to humans.

塑化剂修复技术有生物修复和非生物修复两种形式,生物修复包括微生物修复和植物修复;而非生物修复技术包括水解技术、物理技术和高级氧化技术等。相比于非生物修复,生物修复较适合在农田中使用,但微生物在环境中稳定性差,导致微生物修复技术很难大面积推广;而植物修复又会破坏正常的农耕节奏。低成本、高效、快速、又不引入新污染物的农田塑化剂污染修复技术仍然很缺乏。目前,具有塑化剂修复及植物促生功能的微生物土壤调理剂的应用尚未见报道。Plasticizer remediation technology has two forms: bioremediation and non-biological remediation. Bioremediation includes microbial remediation and phytoremediation; non-biological remediation technology includes hydrolysis technology, physical technology and advanced oxidation technology. Compared with non-biological remediation, bioremediation is more suitable for use in farmland, but the stability of microorganisms in the environment is poor, making it difficult to promote microbial remediation technology on a large scale; and phytoremediation will disrupt the normal farming rhythm. Low-cost, high-efficiency, fast, and non-introducing new pollutants in farmland plasticizer pollution remediation technologies are still lacking. At present, the application of microbial soil conditioners with plasticizer repair and plant growth promotion functions has not been reported.

发明内容Contents of the invention

针对上述问题,本发明提供了一种具有塑化剂降解功能的微生物土壤调理剂。该土壤调理剂能促进蔬菜-土壤种植体系中塑化剂的降解,并同时具有促进植物生长的作用,显著提高蔬菜生物量,同时改善土壤微生物群落结构,提高了耕地质量,大幅度减少了化肥使用。In view of the above problems, the present invention provides a microbial soil conditioner with plasticizer degradation function. The soil conditioner can promote the degradation of plasticizers in the vegetable-soil planting system, and at the same time has the effect of promoting plant growth, significantly increasing the biomass of vegetables, improving the structure of soil microbial communities, improving the quality of cultivated land, and greatly reducing the use of chemical fertilizers. use.

具体来说,本发明通过如下技术方案实现:Specifically, the present invention is realized through the following technical solutions:

首先,本申请提供了一种用于酞酸酯污染土壤修复的微生物土壤调理剂,该微生物土壤调理剂包括微生物搭载生物炭、有机质、糖碳源、养分。所述微生物搭载生物炭内由生物炭和内生菌组成,微生物土壤调理剂中,内生菌的含量≧1.5×108cfu/g,该内生菌由鞘氨醇单胞菌和枯草芽孢杆菌混合后获得。First of all, the application provides a microbial soil conditioner for remediation of phthalate-contaminated soil, the microbial soil conditioner includes microorganism-carrying biochar, organic matter, sugar carbon source, and nutrients. The microorganism-carrying biochar is composed of biochar and endophytic bacteria. In the microbial soil conditioner, the content of endophytic bacteria is ≧1.5×10 8 cfu/g, and the endophytic bacteria are composed of Sphingomonas and Bacillus subtilis Bacteria obtained after mixing.

优选的,上述内生菌中,鞘氨醇单胞菌和枯草芽孢杆菌的含菌量比为1:1。Preferably, among the above-mentioned endophytes, the bacteria content ratio of Sphingomonas and Bacillus subtilis is 1:1.

优选的,有机质、糖碳源、养分占微生物土壤调理剂质量百分比依次为65%,5%,5%,余量为微生物搭载有机质。Preferably, the organic matter, sugar carbon source, and nutrient account for 65%, 5%, and 5% of the mass percentage of the microbial soil conditioner in sequence, and the balance is organic matter carried by microorganisms.

优选的,上述养分包括尿素、过磷酸钙、氯化钾中的至少一种。Preferably, the above nutrients include at least one of urea, superphosphate and potassium chloride.

优选的,有机质为豆粕和菜籽粕中的一种或两种;糖碳源优选D-纤维二糖或其他多聚寡糖;Preferably, the organic matter is one or both of soybean meal and rapeseed meal; the sugar carbon source is preferably D-cellobiose or other polyoligosaccharides;

优选的,生物炭为稻壳生物炭或秸秆(优选玉米秸秆)生物炭。Preferably, the biochar is rice husk biochar or straw (preferably corn stalk) biochar.

其次,本申请还提供了上述微生物土壤调理剂在修复酞酸酯污染土壤中的应用。具体而言,将该微生物土壤调理剂作为底肥在蔬菜播种前一周撒施,然后使调理剂与大田表层土壤(约15cm深)翻拌均匀,使用量为110kg/亩。Secondly, the application also provides the application of the above-mentioned microbial soil conditioner in repairing phthalate-contaminated soil. Specifically, the microbial soil conditioner is used as a base fertilizer and spread one week before vegetable sowing, and then the conditioner is evenly mixed with the field surface soil (about 15 cm deep), and the usage rate is 110 kg/mu.

本申请构建的用于有机污染修复用微生物土壤调理剂的原理如图1所示,该调理剂以有益内生菌为核心构建,利用生物炭搭载内生菌,提高内生菌在环境中的稳定性;生物炭还可吸附土壤环境中有机污染物,阻断植物对有机污染物的吸收,同时给内生菌提供集中降解污染物的环境;辅助添加糖碳源,提高内生菌的降解能力;添加有机质,强化根际微生物群落;同时辅以添加少许养分,以保证作物前期生长所需。The principle of the microbial soil conditioner for organic pollution remediation constructed by this application is shown in Figure 1. The conditioner is constructed with beneficial endophytes as the core, and uses biochar to carry endophytes to improve the stability of endophytes in the environment. Stability; biochar can also adsorb organic pollutants in the soil environment, block the absorption of organic pollutants by plants, and at the same time provide an environment for endophytes to degrade pollutants; assist in adding sugar carbon sources to improve the degradation of endophytes ability; add organic matter to strengthen the rhizosphere microbial community; at the same time add a little nutrient to ensure the growth of crops in the early stage.

本申请提供的土壤调理剂不仅可以修复土壤中污染物,其内生菌还可以定殖植物内部,促进植物内部污染物(PAEs)降解,达到内外同时修复的效果。该微生物调剂具修复与促生为一体,能在修复去除塑化剂污染的同时促进植物生长,调节土壤微生态,减少化肥的使用,有利于高质量农田建设和农业的可持续发展。The soil conditioner provided by this application can not only repair the pollutants in the soil, but also its endophytes can colonize the inside of the plant, promote the degradation of the pollutants (PAEs) inside the plant, and achieve the effect of repairing the inside and outside at the same time. The microbial adjustment integrates repair and growth promotion, can promote plant growth while repairing and removing plasticizer pollution, regulate soil microecology, reduce the use of chemical fertilizers, and is conducive to high-quality farmland construction and sustainable development of agriculture.

附图说明Description of drawings

图1为本申请有机污染修复用微生物土壤调理剂构建原理示意图。Figure 1 is a schematic diagram of the construction principle of the microbial soil conditioner for organic pollution remediation of the present application.

图2为实施例糖碳源的添加显著提高内生菌降解塑化剂的能力示意图。Fig. 2 is a schematic diagram showing that the addition of sugar carbon sources significantly improves the ability of endophytes to degrade plasticizers in the example.

图3为不同处理对蔬菜(a)和土壤(b)中PAEs浓度影响示意图;Figure 3 is a schematic diagram of the influence of different treatments on the concentration of PAEs in vegetables (a) and soil (b);

图3中不同字母表示二者在α=0.05水平上差异显著,下同。Different letters in Figure 3 indicate significant difference at the level of α=0.05, the same below.

图4为不同处理对蔬菜生物量的影响示意图。Figure 4 is a schematic diagram of the effect of different treatments on vegetable biomass.

图5为不同处理对土壤脲酶(a)、蔗糖酶(b)、酸性磷酸酶(c)酶活的影响示意图。Figure 5 is a schematic diagram of the effects of different treatments on the enzyme activities of soil urease (a), invertase (b), and acid phosphatase (c).

图6为不同处理对青菜根际微生物群落的影响示意图。Figure 6 is a schematic diagram of the effect of different treatments on the microbial community of the cabbage rhizosphere.

图7为小区试验蔬菜茎叶(a)、蔬菜根部(b)和土壤(c)中PAEs浓度示意图。Fig. 7 is a schematic diagram of PAEs concentrations in vegetable stems and leaves (a), vegetable roots (b) and soil (c) in plot experiments.

图8为不同处理蔬菜叶绿素和维生素C含量示意图。Fig. 8 is a schematic diagram of chlorophyll and vitamin C contents in vegetables with different treatments.

具体实施方式Detailed ways

下述实例中鞘氨醇单胞菌和枯草芽孢杆菌由江苏省农业科学院农产品产地有机污染减控创新团队从植物中分离,纯化、鉴定、保存。其中,枯草芽孢杆菌已由中国专利201910079682.2(公开号109666612A,其保藏号为CGMCC NO.16233),鞘氨醇单胞菌已由中国专利ZL201510775953.X(公开号105274031B,其保藏号为CGMCCNo.11031)。In the following examples, Sphingomonas and Bacillus subtilis were isolated, purified, identified, and preserved from plants by the innovative team of organic pollution reduction and control in agricultural product origins of Jiangsu Academy of Agricultural Sciences. Among them, Bacillus subtilis has been obtained by Chinese patent 201910079682.2 (publication number 109666612A, its preservation number is CGMCC No.16233), and Sphingomonas has been obtained by Chinese patent ZL201510775953.X (publication number 105274031B, its preservation number is CGMCC No.11031 ).

实例中涉及的原料及培养基:Raw material and culture medium involved in the example:

LB液体培养基:10g/L胰蛋白胨,5g/L酵母膏,10g/L氯化钠,pH7.0;LB liquid medium: 10g/L tryptone, 5g/L yeast extract, 10g/L sodium chloride, pH7.0;

无机盐液体培养基MSM:MgSO4·7H2O(0.4g),FeSO4·7H2O(0.2g),K2HPO4(0.2g),(NH4)2SO4(0.2g)和CaSO4(0.08g)去离子水1L,pH7.0~7.2;Inorganic salt liquid medium MSM: MgSO 4 7H 2 O (0.4g), FeSO 4 7H 2 O (0.2g), K 2 HPO 4 (0.2g), (NH 4 ) 2 SO 4 (0.2g) and CaSO 4 (0.08g) 1L deionized water, pH7.0~7.2;

种子培养基:K2HPO4(4.8g),KH2PO4(3.5g),(NH4)2SO4(2g),MgCl2(0.16g),CaCl2(0.02g),NaMoO4.2H2O(0.0024g),FeCl3(0.0018g),MnCl2.2H2O(0.0015g),PH=7.0,加水至1L。Seed medium: K 2 HPO 4 (4.8g), KH 2 PO 4 (3.5g), (NH 4 ) 2 SO 4 (2g), MgCl 2 (0.16g), CaCl 2 (0.02g), NaMoO 4 . 2H 2 O (0.0024g), FeCl 3 (0.0018g), MnCl 2 .2H 2 O (0.0015g), pH=7.0, add water to 1L.

无机盐固体培养基:无机盐液体培养基中添加终浓度为20g/L的琼脂。Inorganic salt solid medium: Add agar with a final concentration of 20 g/L to the inorganic salt liquid medium.

高温开启式炉购自中国科学院上海光学精密机械研究所,型号SG-GL1100K。The high-temperature open-type furnace was purchased from Shanghai Institute of Optics and Fine Mechanics, Chinese Academy of Sciences, model SG-GL1100K.

其他实施例所涉及的仪器及原料,除非特别说明,均为商业渠道购买。The instruments and raw materials involved in other embodiments were purchased from commercial channels unless otherwise specified.

实施例1制备土壤调理剂Embodiment 1 prepares soil conditioner

取纯化后的内生菌(鞘氨醇单胞菌和枯草芽孢杆菌),分别接到LB液体培养基中,32±1℃培养24h,获得内生菌发酵液,菌含量均为1010左右,备用。Take the purified endophytic bacteria (Sphingomonas and Bacillus subtilis), respectively put them into LB liquid medium, and culture them at 32±1°C for 24 hours to obtain the endophytic fermentation liquid, the bacterial content is about 10 10 ,spare.

以稻壳为原料,采用高温开启式管式炉450℃厌氧条件下制得生物质炭,粉碎过100目筛,在121℃灭菌锅内灭菌1h后取出,获得稻壳生物炭备用。本实施例采用450℃厌氧烧制而成的生物炭具有相对高的吸附能力和电子转移能力,较适合用作土壤有机污染修复。Using rice husk as raw material, biochar was prepared under anaerobic conditions at 450°C in a high-temperature open-type tube furnace, crushed through a 100-mesh sieve, sterilized in a sterilizing pot at 121°C for 1 hour, and then taken out to obtain rice husk biochar for future use. . In this example, the biochar produced by anaerobic firing at 450°C has relatively high adsorption capacity and electron transfer capacity, and is more suitable for remediation of soil organic pollution.

将豆粕在121℃灭菌并磨碎至1mm左右,即为有机质,备用;Sterilize the soybean meal at 121°C and grind it to about 1mm, which is the organic matter and set aside;

将鞘氨醇单胞菌和枯草芽孢杆菌按照含菌量1:1混合后获得内生菌液,接种到生物炭中,内生菌液与生物炭质量比1:4;培养2天,风干至含水率<25%,获得微生物搭载生物炭,生物炭中内生菌含量1010cfu/g左右。Sphingomonas and Bacillus subtilis were mixed according to the bacterial content of 1:1 to obtain the endophyte liquid, inoculated into biochar, the mass ratio of endophyte liquid to biochar was 1:4; cultured for 2 days, air-dried When the water content is less than 25%, the microorganism-loaded biochar is obtained, and the content of endophytic bacteria in the biochar is about 10 10 cfu/g.

将豆粕、D-纤维二糖、养分(以质量份计,尿素:过磷酸钙:氯化钾=2:3:1)、微生物搭载生物炭混合均匀,获得微生物土壤调理剂,其中,有机质豆粕、糖碳源D-纤维二糖、养分分别占微生物土壤调理剂质量百分数65%,5%,5%,余量为微生物搭载生物炭;所获得的调理剂中,内生菌的含量≧1.5×108cfu/g。Mix soybean meal, D-cellobiose, nutrients (in parts by mass, urea: superphosphate: potassium chloride = 2:3:1), and microorganism-carrying biochar to obtain a microbial soil conditioner, wherein organic soybean meal , sugar carbon source D-cellobiose, and nutrients account for 65%, 5%, and 5% of the mass percentage of the microbial soil conditioner, respectively, and the balance is biochar carried by microorganisms; in the obtained conditioner, the content of endophytes ≧1.5 ×10 8 cfu/g.

在具体实施过程中,可根据修复目标污染物选择具有相应降解能力内生菌及具促生能力内生菌中的一种或多种,选择合适的糖碳源,制成土壤调理剂。In the specific implementation process, one or more endophytic bacteria with corresponding degradative ability and endophytic bacteria with growth-promoting ability can be selected according to the remediation target pollutants, and a suitable sugar carbon source can be selected to make a soil conditioner.

糖碳源对比实验:将多种塑化剂(DMP、DEP、DBP、BBP、DEHP、DnOP)分别加入LB液体培养基中,塑化剂添加终浓度为40mg/kg;分别将菌液浓度OD600=1.0的内生菌液1mL接种到50mL培养基中,然后在实验组1(菌+糖)培养基中加入浓度为10mmol的D-纤维二糖,实验组2(菌)中不加入糖,同时设立不接菌不加糖的对照组。7d后检测培养皿中残余塑化剂浓度,检测结果如图2所示。由图2可见,糖碳源的加入明显提高塑化剂的降解量。Sugar carbon source comparison experiment: various plasticizers (DMP, DEP, DBP, BBP, DEHP, DnOP) were added to LB liquid medium respectively, and the final concentration of plasticizer added was 40mg/kg; 600 = 1.0 of the endophytic bacteria liquid 1mL was inoculated into 50mL medium, and then in the experimental group 1 (bacteria + sugar) medium, a concentration of 10mmol of D-cellobiose was added, and in the experimental group 2 (bacteria), no sugar was added. , and set up a control group without inoculation and no sugar at the same time. After 7 days, the residual plasticizer concentration in the petri dish was detected, and the detection results are shown in Figure 2. It can be seen from Figure 2 that the addition of sugar carbon sources significantly increases the degradation of plasticizers.

实施例2盆钵试验Embodiment 2 pot test

将实施例1获得微生物土壤调理剂与人工污染的土壤(向无污染土壤中同时添加终浓度均为20mg/kg的DBP和DEHP),混匀,待溶剂挥发干,将污染后的土壤装入盆钵,每钵装土量2kg。The microbial soil conditioning agent obtained in Example 1 and the artificially polluted soil (add DBP and DEHP that the final concentration is 20mg/kg simultaneously to the non-polluted soil), mix evenly, and wait for the solvent to volatilize and dry, and the polluted soil is packed into For pots and bowls, the amount of soil in each pot is 2kg.

试验中所使用的土壤分别采集于江苏南京和四川眉山。The soil used in the experiment was collected in Nanjing, Jiangsu and Meishan, Sichuan respectively.

试验作物为小白菜,试验共设4个处理,NF:不施肥;CF:施用尿素;OF:施用不添加菌土壤调理剂(除不加入内生菌,其制备方法与实施例1微生物土壤调理剂制备方法相同);EOF:施用实施例1获得的微生物土壤调理剂;除NF外,CF和OF处理组保持与EOF处理组养分(NPK)相同;EOF处理组土壤调理剂使用量为6g/钵。The test crop is Chinese cabbage, and 4 treatments are established in the test, NF: do not apply fertilizer; CF: apply urea; OF: apply without adding bacterial soil conditioner (except that endophytic bacteria are not added, its preparation method is the same as in Example 1 microbial soil conditioning EOF: use the microbial soil conditioner obtained in Example 1; except NF, the CF and OF treatment groups remain the same as the EOF treatment group nutrient (NPK); the EOF treatment group soil conditioner consumption is 6g/ Bowl.

上述各处理除施用肥料品种不同外,其余管理措施均一致,每个处理3个重复,试验于2020年12月10日播种,2021年1月22日收获,收获时采集蔬菜和土壤样品,分析蔬菜和土壤中PAEs浓度、蔬菜生物量、土壤培肥效果和微生物多样性,检测结果如下:Except for the different types of fertilizers, the above-mentioned treatments have the same management measures. Each treatment has 3 replicates. The experiment was sown on December 10, 2020, and harvested on January 22, 2021. Vegetable and soil samples were collected at the time of harvest and analyzed. The concentration of PAEs in vegetables and soil, vegetable biomass, soil fertilization effect and microbial diversity, the test results are as follows:

1)不同处理对蔬菜和土壤中PAEs浓度的影响1) Effects of different treatments on the concentration of PAEs in vegetables and soil

实验结果如图3所示,图3中,(a)为蔬菜中PAEs浓度检测结果,(b)为土壤中PAEs浓度检测结果。由图3可见,施用含有枯草芽孢杆菌的微生物土壤调理剂EOF处理组中,青菜根和茎叶中典型塑化剂DBP和DEHP的浓度显著低于其他处理组;与对照相比,EOF处理组中小白菜根部和茎叶部DBP和DEHP浓度均下降了70%左右;与OF组相比,也有较显著的降低。结果表明,添加枯草芽孢杆菌的土壤调理剂能有效的去除青菜中DBP和DEHP。DBP和DEHP是环境中检出较多且是优先控制的塑化剂种类,毒性相对较大,因此用来作为塑化剂污染修复的效果的指示。The experimental results are shown in Figure 3. In Figure 3, (a) is the detection result of PAEs concentration in vegetables, and (b) is the detection result of PAEs concentration in soil. As can be seen from Figure 3, in the EOF treatment group using the microbial soil conditioner containing Bacillus subtilis, the concentrations of typical plasticizers DBP and DEHP in the cabbage roots and stems and leaves were significantly lower than other treatment groups; compared with the control, the EOF treatment group The concentrations of DBP and DEHP in the roots, stems and leaves of medium and small Chinese cabbage decreased by about 70%; compared with the OF group, there was also a significant decrease. The results showed that the soil conditioner added with Bacillus subtilis could effectively remove DBP and DEHP from green vegetables. DBP and DEHP are plasticizers that are more detected in the environment and are prioritized for control. They are relatively toxic, so they are used as indicators of the effect of plasticizer pollution remediation.

总体而言,含有两种内生菌的土壤调理剂显著的降低了土壤中PAEs的浓度,与对照相比,DBP浓度降低了42%左右,DEHP浓度降低了48%左右。Overall, the soil conditioner containing the two endophytes significantly reduced the concentration of PAEs in the soil, with DBP concentration reduced by about 42% and DEHP concentration reduced by about 48% compared with the control.

2)不同处理对青菜生物量的影响实验2) Experiments on the influence of different treatments on the biomass of green vegetables

实验结果如图4所示。由图4可见,实施例1制备的土壤调理剂在20mg/kg塑化剂污染的土壤中能显著提高青菜生物量。EOF与NF、CF、OF处理组相比,青菜总生物量分别提高了约150%、30%、12%。EOF对青菜茎叶和根部促生效果都较显著。The experimental results are shown in Figure 4. As can be seen from Figure 4, the soil conditioner prepared in Example 1 can significantly increase the biomass of green vegetables in soil polluted by 20mg/kg plasticizer. Compared with NF, CF, OF treatment groups, EOF increased the total biomass of green vegetables by about 150%, 30%, 12%, respectively. EOF has a more significant effect on promoting the growth of stems, leaves and roots of Chinese cabbage.

3)不同处理对土壤性质的改变检测结果如下表1、2和图5所示3) The detection results of different treatments on the change of soil properties are shown in Tables 1, 2 and Figure 5 below

表1.不同处理下南京土壤肥力情况Table 1. Soil fertility in Nanjing under different treatments

表2.不同处理下四川山土肥力情况Table 2. Mountain soil fertility in Sichuan under different treatments

表1、2结果表明,与NF、CF相比,EOF对不同性质的土壤都具有一定的改善效果,提高了土壤有机质、有效磷等的含量。The results in Tables 1 and 2 show that compared with NF and CF, EOF has certain improvement effects on soils with different properties, and increases the content of soil organic matter and available phosphorus.

图5中,(a)、(b)、(c)分别为不同处理对土壤脲酶、蔗糖酶、酸性磷酸酶酶活的影响结果,可见,与对照及施用化肥组相比,EOF显著的增强了土壤脲酶、蔗糖酶和酸性磷酸酶的活性,对塑化剂污染土壤性质改良较为明显。4)土壤调理剂对青菜根际微生物群落的影响In Figure 5, (a), (b), and (c) are the effects of different treatments on the activity of soil urease, invertase, and acid phosphatase, respectively. It can be seen that compared with the control group and the chemical fertilizer application group, EOF is significantly enhanced The activities of soil urease, sucrase and acid phosphatase were significantly improved, and the properties of plasticizer-contaminated soil were significantly improved. 4) Effect of soil conditioner on vegetable rhizosphere microbial community

影响结果如图6结果所示,施用EOF显著增加了鞘氨醇单胞菌、黄杆菌、溶杆菌和黄色土源菌属等有益菌属的丰度,其中鞘氨醇单胞菌属和黄杆菌属中多株菌被报道具有塑化剂降解功能,溶杆菌属具有抵抗植物病原菌、固氮和降解有机物功能,而黄色土源菌属具有根际促生功能。The impact results are shown in Figure 6. The application of EOF significantly increased the abundance of beneficial bacterial genera such as Sphingomonas, Flavobacterium, Lysobacterium, and Geophylla, among which Sphingomonas and Flavobacterium Many strains of Bacillus have been reported to have the function of degrading plasticizers, Lysobacter has the function of resisting plant pathogenic bacteria, nitrogen fixation and degrading organic matter, and the genus Soilbacillus has the function of promoting rhizosphere growth.

实施例3田间试验Embodiment 3 field test

田间小区试验:将DBP和DEHP配制成一定浓度的溶液,同时喷洒于试验小区内,并翻耕(表层土壤15cm)混匀。土壤中,DBP和DEHP的污染浓度均为20mg/kg(以表层15cm土层厚度计)。人工污染土壤后,将试验小区晾置2天。2天后进行施肥或土壤调理剂处理,将肥料或土壤调理剂均匀撒施与土壤表层,翻拌均匀后撒播一定量蔬菜种子。Field plot test: DBP and DEHP were prepared into a solution of a certain concentration, sprayed in the test plot at the same time, and plowed (surface soil 15cm) to mix. In the soil, the pollution concentrations of DBP and DEHP are both 20mg/kg (based on the thickness of the surface 15cm soil layer). After artificially polluting the soil, the test plot was left to air for 2 days. After 2 days, fertilization or soil conditioner treatment is carried out, and the fertilizer or soil conditioner is evenly spread on the soil surface, and a certain amount of vegetable seeds are spread after mixing evenly.

试验作物为小白菜,试验共设4个处理,NF:不施肥,CF:施用化肥,EOF:施用实施例1获得的土壤调理剂;除NF外,CF和EOF处理组养分(NPK)相同;EOF处理组土壤调理剂使用量为110kg/亩。The test crop is Chinese cabbage, and the test has 4 treatments, NF: no fertilization, CF: application of chemical fertilizers, EOF: application of the soil conditioner obtained in Example 1; except for NF, the nutrients (NPK) of the CF and EOF treatment groups are the same; The amount of soil conditioner used in the EOF treatment group was 110kg/mu.

上述各处理除施用肥料品种不同外,其余管理措施均一致,每个小区3个重复,蔬菜种植后45天收获,收获时采集蔬菜和土壤样品,分析蔬菜和土壤中PAEs浓度、蔬菜叶绿素及VC含量。Except for the different types of fertilizers, the management measures of the above treatments were the same. Each plot was repeated 3 times. Vegetables were harvested 45 days after planting. Vegetable and soil samples were collected at the time of harvest, and the concentrations of PAEs, vegetable chlorophyll and VC in vegetables and soil were analyzed. content.

试验结果如下:The test results are as follows:

1)蔬菜和土壤中PAEs(DBP和DEHP)浓度检测1) Detection of PAEs (DBP and DEHP) concentration in vegetables and soil

检测结果如图7所示,图7中a、b分别为小区试验青菜茎叶、青菜根部、PAEs(DBP和DEHP)浓度的检测结果。可见,使用添加枯草芽孢杆菌的土壤调理剂组蔬菜茎叶和根中PAEs浓度均显著低于不施肥、使用化肥组,与不施肥组相比,蔬菜茎叶部DBP和DEHP浓度分别降低15%和31%;蔬菜根部DBP和DEHP浓度降分别低46%和60%。The detection results are shown in Figure 7, where a and b in Figure 7 are the detection results of the concentration of green vegetable stems and leaves, green vegetable roots, and PAEs (DBP and DEHP) in the plot test, respectively. It can be seen that the concentration of PAEs in vegetable stems, leaves and roots of the soil conditioner group added with Bacillus subtilis was significantly lower than that of the non-fertilization and chemical fertilizer groups. Compared with the non-fertilization group, the concentrations of DBP and DEHP in vegetable stems and leaves were respectively reduced by 15%. and 31%; the concentration of DBP and DEHP in vegetable roots decreased by 46% and 60%, respectively.

图7中,c为土壤中PAEs(DBP和DEHP)浓度的检测结果,可见,土壤调理剂处理组土壤中PAEs浓度显著低于不施肥和施化肥组,与不施肥组相比,土壤调理剂组土壤中DBP和DEHP浓度分别降低56%和61%。Among Fig. 7, c is the detection result of the concentration of PAEs (DBP and DEHP) in the soil, it can be seen that the concentration of PAEs in the soil of the soil conditioner treatment group is significantly lower than that of the non-fertilization and chemical fertilizer group, compared with the non-fertilization group, the soil conditioner The concentrations of DBP and DEHP in the soil of the group decreased by 56% and 61%, respectively.

3)不同处理蔬菜叶绿素和VC含量检测结果3) Detection results of chlorophyll and VC content in vegetables with different treatments

检测结果如图8所示,图8中,a、b分别为青菜叶绿素和维生素C含量检测结果。可见,土壤处理剂组青菜叶绿素和维生素C含量显著高于不施肥或施用化肥组,表明土壤调理有效的提高了蔬菜的品质指标。The test results are shown in Figure 8. In Figure 8, a and b are the test results of the chlorophyll and vitamin C contents of vegetables, respectively. It can be seen that the chlorophyll and vitamin C content of vegetables in the soil treatment agent group were significantly higher than those in the no fertilizer or chemical fertilizer application group, indicating that soil conditioning effectively improved the quality indicators of vegetables.

综上结果表明,该发明构建的土壤调理剂具减毒和促生多种功能,能有效的降低蔬菜和土壤中塑化剂污染,改良土壤,改善土壤微生物群落,促进蔬菜生长,大幅度减少化肥使用,提高农产品产地品质,促进农业可持续发展。The above results show that the soil conditioner constructed by the invention has multiple functions of attenuating and promoting growth, can effectively reduce plasticizer pollution in vegetables and soil, improve soil, improve soil microbial community, promote vegetable growth, and greatly reduce The use of chemical fertilizers can improve the quality of agricultural products and promote the sustainable development of agriculture.

以上所述仅是本发明的优选实施方式,应当指出,由于文字表达的有限性,而客观上存在无限的具体结构,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进、润饰或变化,也可以将上述技术特征以适当的方式进行组合;这些改进润饰、变化或组合,或未经改进将发明的构思和技术方案直接应用于其他场合的,均应视为本发明的保护范围。The above is only a preferred embodiment of the present invention. It should be pointed out that due to the limitation of literal expression, there are objectively unlimited specific structures. Under these conditions, several improvements, modifications or changes can also be made, and the above technical features can also be combined in an appropriate manner; these improvements, modifications, or combinations, or the idea and technical solution of the invention can be directly applied to other occasions without improvement should be regarded as the protection scope of the present invention.

Claims (5)

1. The microbial soil conditioner for repairing phthalate polluted soil is characterized by comprising microbial loaded biochar, organic matters, sugar carbon sources and nutrients;
the nutrient comprises at least one of urea, superphosphate and potassium chloride;
the organic matter is at least one of soybean meal and rapeseed meal;
the microorganism-carried biochar consists of biochar and endophytes, wherein the content of endophytes in the microorganism soil conditioner is not less than 1.5X10 8 cfu/g, wherein the endophyte is obtained by mixing Sphingomonas and bacillus subtilis;
the sugar carbon source is D-cellobiose;
the organic matters, the sugar carbon source and the nutrients account for 65% of the microbial soil conditioner in percentage by mass, 5% of the microbial soil conditioner, and the balance is the microbial loaded organic matters.
2. The microbial soil conditioner for phthalate contaminated soil restoration according to claim 1 wherein said endophyte has a bacterial content ratio of Sphingomonas and Bacillus subtilis of 1:1.
3. The microbial soil conditioner for phthalate contaminated soil restoration according to claim 1 wherein said biochar is rice hull biochar or straw biochar.
4. Use of the microbial soil conditioner for phthalate contaminated soil restoration according to any of claims 1 to 3 for restoring phthalate contaminated soil.
5. The use according to claim 4, wherein the microbial soil conditioner is applied as a base fertilizer for one week before vegetable sowing at an amount of 110 kg/mu.
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