WO2021248974A1 - 具有降低蔬菜重金属含量、提高蔬菜质量的功能菌株及应用 - Google Patents

具有降低蔬菜重金属含量、提高蔬菜质量的功能菌株及应用 Download PDF

Info

Publication number
WO2021248974A1
WO2021248974A1 PCT/CN2021/082435 CN2021082435W WO2021248974A1 WO 2021248974 A1 WO2021248974 A1 WO 2021248974A1 CN 2021082435 W CN2021082435 W CN 2021082435W WO 2021248974 A1 WO2021248974 A1 WO 2021248974A1
Authority
WO
WIPO (PCT)
Prior art keywords
strain
soil
content
vegetables
heavy metal
Prior art date
Application number
PCT/CN2021/082435
Other languages
English (en)
French (fr)
Inventor
夏涛
王晓菡
Original Assignee
齐鲁工业大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 齐鲁工业大学 filed Critical 齐鲁工业大学
Publication of WO2021248974A1 publication Critical patent/WO2021248974A1/zh
Priority to US18/060,251 priority Critical patent/US20230114795A1/en

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01B—SOIL WORKING IN AGRICULTURE OR FORESTRY; PARTS, DETAILS, OR ACCESSORIES OF AGRICULTURAL MACHINES OR IMPLEMENTS, IN GENERAL
    • A01B79/00—Methods for working soil
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G22/00—Cultivation of specific crops or plants not otherwise provided for
    • A01G22/15—Leaf crops, e.g. lettuce or spinach 
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
    • A01N63/22—Bacillus
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P15/00—Biocides for specific purposes not provided for in groups A01P1/00 - A01P13/00
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus
    • C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
    • Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures

Definitions

  • the invention belongs to the technical field of agricultural microbial application, and specifically relates to functional strains and applications that can reduce the content of heavy metals in vegetables and improve the quality of vegetables.
  • Cadmium (Cd) is one of the most toxic heavy metals, which is extremely harmful to human health.
  • the excessive use of chemical fertilizers and pesticides, cadmium pollution of farmland soil caused by sewage irrigation and mining activities have become a concern of all countries in the world.
  • Cd pollution is the most serious heavy metal pollution in my country, accounting for about 7% of farmland pollution.
  • Vegetables, crops, fruit trees, pastures and other plants that grow in mildly and moderately cadmium-polluted soils accumulate cadmium through the food chain, causing harm to human and animal health.
  • Fixing Cd in situ is an effective measure to reduce the transfer of Cd from the soil and ensure food safety.
  • Soil heavy metal fixatives reduce the bioavailability of Cd and reduce the transfer of Cd from soil to plants through adsorption, complexation and precipitation.
  • continuous addition of these organic and inorganic fixatives may have harmful effects on soil properties, structure and ecosystems.
  • Chinese patent document CN110846250A (application number: 201911142617.6) discloses a high-yield ⁇ -PGA Bacillus subtilis and its application, and discloses that the combination of Bacillus subtilis and biochar can effectively improve the growth performance of plants and reduce the heavy metal Pb of the soil. Absorption.
  • the present invention provides two functional strains and applications that can reduce the content of heavy metals in vegetables and improve the quality of vegetables.
  • the strains of the present invention also have the ability to reduce the content of heavy metals in plants, especially the content of cadmium (Cd), promote plant growth, increase the Vc content of plants, increase the content of plant soluble protein, increase soil microbial abundance, and increase soil urease Various functions such as activity; using plant growth-promoting bacteria to reduce the content of heavy metals in plants, improve soil activity, reduce the transport of heavy metals to plants, and ensure the safety of agricultural products. It is a safe, ecological, economic, and environmentally friendly effective method.
  • Cd cadmium
  • the method for culturing the strain W7 includes the following steps:
  • Strain W7 was inoculated on a solid medium, cultured at 28-32°C for 2-3 days, and then colonies were picked to inoculate in liquid medium, and cultured at 35-37°C at 150-200 rpm for 16-20 hours to obtain the fermentation broth of strain W7;
  • the solid medium is: sucrose 10.0g/L, (NH 4 ) 2 SO 4 0.5g/L, Na 2 HPO 4 2.0g/L, NaCl 0.1g/L, KCl 0.191g/L, MgSO 4 ⁇ 7H 2 O 0.5g/L, agar 20g/L, balance water;
  • the liquid culture medium is: sucrose 10.0g/L, (NH 4 ) 2 SO 4 0.5g/L, Na 2 HPO 4 2.0g/L, NaCl 0.1g/L, KCl 0.191g/L, MgSO 4 ⁇ 7H 2 O 0.5g/L, the balance water.
  • the strain W7 is used for the production of biological agents.
  • the biological agent is a liquid formulation comprising an effective number of viable cells of the strain W7 is not less than 108 / ml.
  • the strain W7 is used for crop cultivation.
  • the strain W7 is preferably used to reduce the content of heavy metal cadmium in vegetables.
  • the strain W7 is preferably used to increase the content of vitamin C and soluble protein in vegetables.
  • the vegetable is lettuce.
  • the strain is used for soil improvement.
  • the strain W7 is preferably used to fix the heavy metal cadmium in the soil.
  • the strain W7 is preferably used to promote the proliferation of microorganisms in the soil.
  • the strain W7 is used to promote the proteobacteria, Firmicutes, Sphingomonas, and Aneurinibacillus in soil contaminated by heavy metal cadmium. Proliferation of one or more than two.
  • Bacillus W25 taxonomically named Bacillus amyloliquefaciens, deposited in the General Microbiology Center of China Microbial Culture Collection Management Committee, preservation address: No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing, and the preservation date is 2020 On June 8, 2010, the collection number of the strain was CGMCC No. 20042.
  • the method for culturing the strain W25 includes the following steps:
  • Strain W25 was inoculated on a solid medium, cultured at 28-32°C for 2-3 days, and then colonies were picked to inoculate in liquid medium, and cultured at 35-37°C at 150-200 rpm for 16-20 hours to obtain a fermentation broth of strain W7;
  • the solid medium is: sucrose 10.0g/L, (NH 4 ) 2 SO 4 0.5g/L, Na 2 HPO 4 2.0g/L, NaCl 0.1g/L, KCl 0.191g/L, MgSO 4 ⁇ 7H 2 O 0.5g/L, agar 20g/L, balance water;
  • the liquid culture medium is: sucrose 10.0g/L, (NH 4 ) 2 SO 4 0.5g/L, Na 2 HPO 4 2.0g/L, NaCl 0.1g/L, KCl 0.191g/L, MgSO 4 ⁇ 7H 2 O 0.5g/L, the balance water.
  • the strain W25 is used for the production of biological agents.
  • the biological agent is a liquid formulation comprising an effective number of viable cells of the strain W25 is not less than 108 / ml.
  • the strain W25 is used for crop cultivation.
  • the strain W25 is preferably used to reduce the content of heavy metal cadmium in vegetables.
  • the strain W25 is preferably used to increase the content of vitamin C and soluble protein in vegetables.
  • the vegetable is lettuce.
  • the strain W25 is used for soil improvement.
  • the strain W25 is preferably used to fix the heavy metal cadmium in the soil.
  • the strain W25 is preferably used to promote the proliferation of microorganisms in the soil.
  • the strain W25 is used to promote the proteobacteria, Firmicutes, Sphingomonas, and Aneurinibacillus in soil contaminated by heavy metal cadmium. Proliferation of one or more than two.
  • the present invention provides two strains W7 and W25 that have multiple functions at the same time.
  • the strain provided by the present invention can also reduce the content of heavy metals in plants, especially the content of cadmium (Cd), promote plant growth, increase plant Vc content, increase plant soluble protein content, increase soil microbial abundance, and increase Various functions such as soil urease activity.
  • Cd cadmium
  • the strain involved in the present invention can produce ⁇ -PGA.
  • Figure 1 is a diagram showing the detection results of Cd content in the edible tissue and roots of lettuce in the pot experiment of Example 3;
  • Figure 2 is a graph showing the detection results of the ⁇ -PGA content of the culture solution corresponding to different incubation times in Example 4;
  • FIG. 3 is a diagram showing the detection results of Cd content in the culture medium with an initial Cd content of 3 mg ⁇ L -1 in Example 4 corresponding to different incubation times;
  • Example 4 is a diagram showing the detection results of the Cd content of the culture medium with an initial Cd content of 6 mg ⁇ L -1 in Example 4 at different incubation times;
  • FIG. 5 is a diagram showing the detection results of OD 600 corresponding to different incubation times of the culture solution in Example 4; FIG.
  • FIG. 6 is a graph of pH detection results corresponding to different incubation times of the culture solution in Example 4.
  • FIG. 7 is a diagram showing the detection result of urease activity of the soil collected in Example 5.
  • Example 8 is a diagram showing the detection result of Proteobacteria in the soil collected in Example 5;
  • Figure 9 is a diagram showing the detection results of Firmicutes in the soil collected in Example 5.
  • Example 10 is a diagram showing the detection result of Sphingomonas in the soil collected in Example 5;
  • FIG. 11 is a diagram showing the detection result of Bacillus in the soil collected in Example 5.
  • FIG. 11 is a diagram showing the detection result of Bacillus in the soil collected in Example 5.
  • Bacillus W25 taxonomically named Bacillus amyloliquefaciens, deposited in the General Microbiology Center of China Microbial Culture Collection Management Committee, preservation address: No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing, and the preservation date is 2020 On June 8, 2010, the collection number of the strain was CGMCC No. 20042.
  • Soil samples were collected from the rhizosphere soil of lettuce grown in Cd-contaminated farmland (Cd content in the soil is 1.35 mg ⁇ kg -1) in the suburbs of Jinan. Suspend 10 g of the sample in 90 ml of sterile distilled water, boil for 5 min, and spread the diluted suspension on the separation plate.
  • the separation medium is: glucose 10g/L; yeast extract 5g/L, sodium L-glutamate 5g/L, KH 2 PO 4 0.5g/L, K 2 HPO 4 0.5g/L, MgSO 4 ⁇ 7H 2 O 0.1g/L, neutral red 0.06g/L, agar 15g/L, the balance water. Adjust the initial pH of the medium to 7.2 ⁇ 0.1.
  • the colonies that interact with the dye (color changes from red to yellow) to form a specific concentric zone are considered to be ⁇ -PGA-producing bacteria, and the ⁇ -PGA-producing strains are numbered W7 and W25. .
  • the slant strain of strain W7 was inoculated on a solid medium and cultured at 30°C for 3 days. Then select the full and viscous W7 colony to inoculate the liquid medium, and culture it with shaking at 150rpm at 37°C for 20h.
  • the fermentation broth was transferred to a sterile centrifuge bottle, centrifuged at 5000 rpm for 5 minutes to collect the bacteria, washed with sterile deionized water, and resuspended so that the number of cells reached more than 500 million CFU/mL; the solid medium was: sucrose 10.0g /L, (NH 4 ) 2 SO 4 0.5g/L, Na 2 HPO 4 2.0g/L, NaCl 0.1g/L, KCl 0.191g/L, MgSO 4 ⁇ 7H 2 O 0.5g/L, agar 20g/ L.
  • the liquid culture medium is: sucrose 10.0g/L, (NH 4 ) 2 SO 4 0.5g/L, Na 2 HPO 4 2.0g/L, NaCl 0.1g/L, KCl 0.191g/ L, MgSO 4 ⁇ 7H 2 O 0.5g/L, the balance water.
  • strain W25 The method of activation of strain W25 and preparation of bacterial suspension is the same as that of strain W7.
  • each pot is 28 cm diameter ⁇ 35 cm high, containing 4.8 kg of soil, plus cadmium (CdCl 2 ⁇ 2.5H 2 O), until the final concentration of Cd in the soil is 0 , 0.5 and 1 mg/kg, fully mixed with soil and equilibrate for 45 days; each group is treated with 3 repetitions.
  • the collected surface-sterilized lettuce seeds were sown in each pot, and the seedlings were thinned into 15 plants/pot after germination; 3 pots were treated in parallel for each treatment; the soil was regularly irrigated to keep it moist. Before inoculation, the bacteria prepared in Example 2 were selected.
  • the edible tissues and roots of each pot of lettuce were collected for subsequent analysis; the roots and edible tissues of the lettuce were washed with 0.01M ethylenediaminetetraacetic acid (EDTA) and distilled water, and divided into two evenly. One part was inactivated at 105°C for 30 minutes, dried to constant weight at 65°C, and the weight after drying was recorded; the dried edible tissues and roots were grinded and digested to determine the Cd content.
  • the content of vitamin C (Vc) and soluble protein in fresh edible tissues was determined by standard methods.
  • the pot experiment method of strain W25 is the same as that of strain W7.
  • Method for determining Cd content refer to Gao Yuanyuan, Peng Zhaofeng, Qiu Haiou, Gu Yansheng and Cheng Wei. IPC-OES determination of heavy metal elements in dominant plants in gold mine tailings[J].Analysis Laboratory,2016,35(5):521 -525. According to the published method, 0.1g plant samples are accurately weighed for microwave digestion, the volume is fixed with 5% HNO3, and the Cd content in the digestion solution is determined by ICP-OES.
  • the strain W7 reduces the Cd content in lettuce edible tissues by 17-33%, increases the biomass of lettuce edible tissues by 41-47%, and increases the Vc content of lettuce edible tissues by 29-40%. , Increase the soluble protein content of the edible tissues of lettuce by 9-43%.
  • Strain W25 reduces the Cd content in lettuce edible tissues by 30-41%, increases the biomass of lettuce edible tissues by 61-85%, increases the Vc content of lettuce edible tissues by 38-73%, and improves the edible tissues of lettuce.
  • the soluble protein content is 37-43%.
  • ⁇ -PGA content detection method refer to the method of Wei Zeng et al. (2013). (An integrated high-throughput strategy for rapid screening of poly( ⁇ -glutamic acid)-producing bacteria.Appl Microbiol Biotechnol(2013)97:2163-2172)
  • Cd content detection method Determine the cadmium concentration by inductively coupled plasma emission spectrometer (ICP-OES) (Optima 2100DV, Perkin-Elmer); refer to Gao Yuanyuan, Peng Zhaofeng, Qiu Haiou, Gu Yansheng and Cheng Wei. IPC-OES for gold ore determination Heavy metal elements in dominant plants in the tailings area[J].Analysis Laboratory,2016,35(5):521-525. Public methods.
  • ICP-OES inductively coupled plasma emission spectrometer
  • strain W25 The detection method of strain W25 is the same as that of strain W7.
  • the content of ⁇ -PGA in the culture fluid varies greatly at different culture times.
  • strain W7 at the same culture time, the higher the Cd content in the culture fluid, the greater the amount of ⁇ -PGA produced; the same in strain W25 Phenomenon:
  • the amount of ⁇ -PGA produced by the strain W25 was higher than that of the strain W7.
  • Strains W7 and W25 respectively increase the detection of rhizosphere soil enzyme activity, and increase the rhizosphere soil protein bacteria (Proteobacteria), Bacteroides (Firmicutes), Sphingomonas (Sphingomonas), Bacillus (Bacillus) Relative abundance detection
  • strains W7 and W25 have a certain effect on improving the urease activity of the soil.
  • the strains W7 and W25 have an enhanced effect on the urease activity of the soil.
  • strains W7 and W25 have a certain proliferation of Proteobacteria, Firmicutes, Sphingomonas, and Bacillus in the soil. It can be seen from Figure 10 that the Cd content in the soil is higher, and the strains W7 and W25 have more significant effects on the proliferation of Sphingomonas. Compared with W7, the strain W25 is more effective against Sphingomonas. (Sphingomonas) has better proliferation effect.
  • strains W7 and W25 involved in the present invention can simultaneously reduce the content of heavy metals in plants, especially the content of cadmium (Cd), promote plant growth, increase plant Vc content, increase plant soluble protein content, and increase soil Microbial abundance, increase soil urease activity and other functions, can be widely used in soil improvement, crop biomass improvement, soil heavy metal remediation and other fields.
  • Cd cadmium

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Virology (AREA)
  • Environmental Sciences (AREA)
  • Biochemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • General Engineering & Computer Science (AREA)
  • Botany (AREA)
  • Pest Control & Pesticides (AREA)
  • Plant Pathology (AREA)
  • Dentistry (AREA)
  • Agronomy & Crop Science (AREA)
  • Mechanical Engineering (AREA)
  • Soil Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Processing Of Solid Wastes (AREA)

Abstract

提供了具有降低蔬菜重金属含量、提高蔬菜质量的功能菌种及应用，具体为菌株W7和W25，具有降低植物中重金属的含量，尤其是镉(Cd)的含量，促进植物生长，增加植物的Vc含量，增加植物可溶性蛋白的含量，增加土壤微生物丰度，增加土脲酶活性等多种功能，可广泛应用于土壤改良、农作物生物量提升、土壤重金属修复等领域。

Description

具有降低蔬菜重金属含量、提高蔬菜质量的功能菌株及应用 技术领域
本发明属于农业微生物应用技术领域，具体涉及具有降低蔬菜重金属含量、提高蔬菜质量的功能菌株及应用。
背景技术
镉(Cd)是最具毒性的重金属之一，对人体健康危害极大。化肥、农药的过度使用、污水灌溉和采矿活动引起的农田土壤镉污染已成为世界各国关注的问题。Cd污染是我国最严重的重金属污染，约占农田污染的7％。在轻度和中度镉污染土壤中生长的蔬菜、农作物、果树、牧草等植物通过食物链积累镉，对人畜健康造成危害。
原位固定Cd是减少Cd从土壤中转移，保证食品安全的有效措施。土壤重金属固定剂通过吸附、络合和沉淀作用，降低Cd的生物有效性，减少Cd从土壤向植物的转移。然而，持续添加这些有机、无机固定剂可能会对土壤性质、结构和生态系统产生有害影响。
中国专利文献CN110846250A(申请号：201911142617.6)公开了一株高产γ-PGA的枯草芽孢杆菌及其应用，公开了该枯草芽孢杆菌与生物炭结合，有效提高植物的生长性能，降低植物对土壤重金属Pb的吸收。
现有技术中关于应用微生物菌种用于重金属降解、促进植物生长的相关报道很多，但是大部分是多种微生物相结合，或者微生物与其它物质相结合，才可以起到多种功能的作用，但是就一株菌而言，对于农作物的种植以及土壤改良同时起到多种功能作用的相关报道极少，对于多功能微生物菌种的开发，也是目前农业微生物技术领域急需解决的问题。
发明内容
针对现有技术的不足，本发明提供两株具有降低蔬菜重金属含量、提高蔬菜质量的功能菌株及应用。
本发明涉及的菌种同时具有降低植物中重金属的含量，尤其是镉(Cd)的含量，促进植物生长，增加植物的Vc含量，增加植物可溶性蛋白的含量，增加土壤微生物丰度，增加土脲酶活性等多种功能；利用植物促生细菌来降低植物中重金属含量，改善土壤活性，降低重金属向植物中的转运，保障农产品安全，是一种安全、生态、经济、环保的有效方法。
一株芽孢杆菌W7，分类学命名为枯草芽孢杆菌Bacillus subtilis，保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏地址：北京市朝阳区北辰西路1号院3号，保藏日期为2020年6月8日，菌种保藏号为CGMCC No.20043。
所述菌株W7的培养方法，包括如下步骤：
将菌株W7接种于固体培养基上，28-32℃培养2-3天，然后挑取菌落接种于液体培养基中，35-37℃，150-200rpm培养16-20h，获得菌株W7发酵液；所述固体培养基为：蔗糖10.0g/L、(NH 4) 2SO 4 0.5g/L、Na 2HPO 4 2.0g/L、NaCl 0.1g/L、KCl 0.191g/L、MgSO 4·7H 2O 0.5g/L、琼脂20g/L、余量水；所述液体培养基为：蔗糖10.0g/L、(NH 4) 2SO 4 0.5g/L、Na 2HPO 4 2.0g/L、NaCl 0.1g/L、KCl 0.191g/L、MgSO 4·7H 2O 0.5g/L、余量水。
所述菌株W7用于生产生物制剂。
根据本发明优选的，所述生物制剂为液体制剂，含有所述菌株W7的有效活菌数为10 8个/毫升以上。
所述菌株W7用于农作物种植。
根据本发明优选的，所述菌株W7用于降低蔬菜中重金属镉的含量。
根据本发明优选的，所述菌株W7用于增加蔬菜中维生素C和可溶性蛋白的含量。
进一步优选的，所述蔬菜为生菜。
所述菌株用于土壤改良。
根据本发明优选的，所述菌株W7用于固定土壤中的重金属镉。
根据本发明优选的，所述菌株W7用于促进土壤让中微生物的增殖。
进一步优选的，所述菌株W7用于促进重金属镉污染的土壤中蛋白菌(Proteobacteria)、拟杆菌门(Firmicutes)、鞘氨醇单胞菌(Sphingomonas)、硫胺素芽孢杆菌属(Aneurinibacillus)之一或二者以上的增殖。
一种芽孢杆菌W25，分类学命名为解淀粉芽孢杆菌Bacillus amyloliquefaciens，保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏地址：北京市朝阳区北辰西路1号院3号，保藏日期为2020年6月8日，菌种保藏号为CGMCC No.20042。
所述菌株W25的培养方法，包括如下步骤：
将菌株W25接种于固体培养基上，28-32℃培养2-3天，然后挑取菌落接种于液体培养基中，35-37℃，150-200rpm培养16-20h，获得菌株W7发酵液；所述固体培养基为：蔗糖10.0g/L、(NH 4) 2SO 4 0.5g/L、Na 2HPO 4 2.0g/L、NaCl 0.1g/L、KCl 0.191g/L、MgSO 4·7H 2O 0.5g/L、琼脂20g/L、余量水；所述液体培养基为：蔗糖10.0g/L、(NH 4) 2SO 4 0.5g/L、Na 2HPO 4 2.0g/L、NaCl 0.1g/L、KCl 0.191g/L、MgSO 4·7H 2O 0.5g/L、余量水。
所述菌株W25用于生产生物制剂。
根据本发明优选的，所述生物制剂为液体制剂，含有所述菌株W25的有效活菌数为10 8个/毫升以上。
所述菌株W25用于农作物种植。
根据本发明优选的，所述菌株W25用于降低蔬菜中重金属镉的含量。
根据本发明优选的，所述菌株W25用于增加蔬菜中维生素C和可溶性蛋白的含量。
进一步优选的，所述蔬菜为生菜。
所述菌株W25用于土壤改良。
根据本发明优选的，所述菌株W25用于固定土壤中的重金属镉。
根据本发明优选的，所述菌株W25用于促进土壤让中微生物的增殖。
进一步优选的，所述菌株W25用于促进重金属镉污染的土壤中蛋白菌(Proteobacteria)、拟杆菌门(Firmicutes)、鞘氨醇单胞菌(Sphingomonas)、硫胺素芽孢杆菌属(Aneurinibacillus)之一或二者以上的增殖。
本发明的有益效果
1、本发明提供了两株同时具有多种功能的菌株W7和W25。
2、本发明提供的菌种同时具有降低植物中重金属的含量，尤其是镉(Cd)的含量，促进植物生长，增加植物的Vc含量，增加植物可溶性蛋白的含量，增加土壤微生物丰度，增加土脲酶活性等多种功能。
3、本发明涉及的菌株能够产生γ-PGA。
附图说明
图1为实施例3盆栽实验中生菜食用组织和根部的Cd含量检测结果图；
图2为实施例4中的培养液不同培养时间对应的γ-PGA含量检测结果图；
图3为实施例4中初始Cd含量为3mg·L -1的培养液不同培养时间对应的Cd含量检测结果图；
图4为实施例4中初始Cd含量为6mg·L -1的培养液不同培养时间对应的Cd含量检测结果图；
图5为实施例4中的培养液不同培养时间对应的OD 600检测结果图；
图6为实施例4中的培养液不同培养时间对应的pH检测结果图；
图7为实施例5中采集土壤的脲酶活性检测结果图；
图8为实施例5中采集土壤中蛋白菌(Proteobacteria)检测结果图；
图9为实施例5中采集土壤中拟杆菌门(Firmicutes)检测结果图；
图10为实施例5中采集土壤中鞘氨醇单胞菌(Sphingomonas)检测结果图；
图11为实施例5中采集土壤中芽孢杆菌属(Bacillus)检测结果图。
具体实施方式
下面结合具体实施方式，对本发明做进一步阐述，但保护范围不限于此。
以下实施例中除特例说明外，均为本领域内的常规实验方法和操作步骤。
生物样品保藏信息
一株芽孢杆菌W7，分类学命名为枯草芽孢杆菌Bacillus subtilis，保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏地址：北京市朝阳区北辰西路1号院3号，保藏日期为2020年6月8日，菌种保藏号为CGMCC No.20043。
一种芽孢杆菌W25，分类学命名为解淀粉芽孢杆菌Bacillus amyloliquefaciens，保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏地址：北京市朝阳区北辰西路1号院3号，保藏日期为2020年6月8日，菌种保藏号为CGMCC No.20042。
实施例1
菌株W7和W25的筛选
从济南郊区Cd污染农田(土壤中Cd含量为1.35mg·kg -1)中生长的莴苣根际土壤中采集土壤样品。将10g样品悬浮于90ml无菌蒸馏水中，煮沸5min，稀释后的悬浮液涂在分离板上。分离培养基为：葡萄糖10g/L；酵母提取物5g/L、L-谷氨酸钠5g/L、KH 2PO 4 0.5g/L、K 2HPO 40.5g/L、MgSO 4·7H 2O 0.1g/L、中性红0.06g/L、琼脂15g/L、余量水。将培养基的初始pH值调整为7.2±0.1。在37℃黑暗中孵育48h后，与染料(颜色由红色变为黄色)相互作用形成一个特定同心区的菌落被认为是γ-PGA产生菌，筛得产γ-PGA的菌株编号为W7和W25。
实施例2
菌株W7、W25分别活化和菌悬液制备
将菌株W7斜面菌种接种于固体培养基上，30℃培养3d。然后选取饱满、粘稠的W7菌落接种于液体培养基，37℃150rpm振荡培养20h。将发酵液转移至无菌离心瓶中，5000rpm离心5min收集菌体，用无菌去离子水洗涤，重新悬浮，使细胞数量达到5亿CFU/mL以上；所述固体培养基为：蔗糖10.0g/L、(NH 4) 2SO 4 0.5g/L、Na 2HPO 4 2.0g/L、NaCl 0.1g/L、KCl0.191g/L、MgSO 4·7H 2O 0.5g/L、琼脂20g/L、余量水；所述液体培养基为：蔗糖10.0g/L、(NH 4) 2SO 4 0.5g/L、Na 2HPO 4 2.0g/L、NaCl 0.1g/L、KCl 0.191g/L、MgSO 4·7H 2O 0.5g/L、余量水。
菌株W25的活化和菌悬液制备方法与菌株W7相同。
实施例3
菌株W7、W25分别应用于生菜种植
W7应用于生菜盆栽实验，按下述方法进行：每盆28厘米直径×35厘米高，含4.8公斤土壤，外加镉(CdCl 2·2.5H 2O)，至土壤中Cd含量的终浓度为0、0.5和1毫克/公斤，与土壤 充分混合后平衡45天；每组处理3个重复。将采集的表面灭菌的生菜种子，每盆播种，发芽后疏苗成15株/盆；每个处理3盆平行；土壤定期灌溉以保持湿润，接种前，将实施例2制备的菌选用，用无菌去离子水稀释至1×10 8个/mL。以无菌去离子水为对照，在生菜第三叶期，在根部周围挖沟(1-2cm深)，在沟中加入菌悬液，加液量为90ml/盆；盆栽于温室(温度10-22℃，相对湿度30-45％，光照正常)中进行栽培，整个栽培周期为45天。
栽培结束后，采集每盆生菜的食用组织和根进行后续分析；用0.01M乙二胺四乙酸(EDTA)和蒸馏水清洗生菜根和食用组织，并平均分成两份。其中一份在105℃失活30min，在65℃干燥至恒重，记录干燥后的重量；分别将干燥的食用组织和根磨碎并消化以测定Cd含量。新鲜食用组织用标准方法测定维生素C(Vc)、可溶性蛋白质含量。
菌株W25的盆栽实验方法与菌株W7盆栽实验方法相同。
测定Cd含量的方法：参照高媛媛,彭兆丰,邱海鸥,顾延生和程伟.IPC-OES测定金矿尾矿区优势植物中的重金属元素[J].分析试验室,2016,35(5):521-525.公开的方法，准确称取0.1g植物样品进行微波消解，用5％HNO3定容，采用ICP-OES测定消解液中Cd含量。
测定维生素C的方法：参照赵晓梅,江英,吴玉鹏,刘宽和张志强.果疏中VC含量测定方法的研究[J].食品科学,2006,27(3):197-199.公开的方法测定蔬菜可食用组织维生素C的含量。
测定可溶性蛋白质含量的方法：参照赵英永,戴云,崔秀明,张文斌和马妮.考马斯亮蓝G-250染色法测定草乌中可溶性蛋白质含量[J].云南民族大学学报:自然科学版,2006,15(3):235-237.公开的方法测定蔬菜可食用组织的可溶性蛋白含量。
实验结果见图1和表1：
表1
Figure PCTCN2021082435-appb-000001
Figure PCTCN2021082435-appb-000002
由上述结果可以得出，菌株W7降低生菜可食用组织中Cd含量达17-33％、提高生菜可食用组织的生物量达41-47％、提高生菜可食用组织的Vc含量达29-40％、提高生菜可食用组织的可溶性蛋白含量达9-43％。
菌株W25降低生菜可食用组织中Cd含量达30-41％、提高生菜可食用组织的生物量达61-85％、提高生菜可食用组织的Vc含量达38-73％、提高生菜可食用组织的可溶性蛋白含量达37-43％。
实施例4
菌株W7、W25分别接种土壤滤液中对水溶性Cd的作用以及γ-PGA的含量
将2.5kg土壤加入10L去离子水，150rpm振荡48h，5000rpm离心15min，收集上清液，通过微孔滤膜(0.45μm孔径)过滤，灭菌；然后与无菌基本发酵培养基按体积比4：1的比例混合均匀，制得无菌混合液，所述无菌基本发酵培养基各种组分的质量浓度为：3％葡萄糖、0.25％酵母抽提物、2％谷氨酸盐、0.05％磷酸二氢钾、0.05％K 2HPO 4、0.01％MgSO 4·7H2O、余量水，pH7.2±0.1。向上述制备的无菌混合液中添加不同浓度的Cd 2+(CdCl 2·2.5H 2O)，使无菌混合液中Cd 2+的终浓度分别为0、3和6mg·L -1；取100mL所述混合液加入到三角瓶中，接种实施例2制备的菌悬液，接种量为1％(v:v)，每个浓度6个平行，以Cd 2+终浓度为0的混合液作为对照。
在37℃下，以150转/分的速度培养；分别在0、24、48、72和96小时，测量培养液中的γ-PGA含量、Cd含量、OD 600值和pH值。
通过测量OD 600监测细菌生长，并用pH计测定pH值，
γ-PGA含量的检测方法：参照Wei Zeng等(2013)的方法。(An integrated high-throughput strategy for rapid screening of poly(γ-glutamic acid)-producing bacteria.Appl Microbiol Biotechnol(2013)97:2163–2172)
Cd含量的检测方法：通过电感耦合等离子体发射光谱仪(ICP-OES)(Optima 2100DV，Perkin-Elmer)测定镉浓度；参照高媛媛,彭兆丰,邱海鸥,顾延生和程伟.IPC-OES测定金矿尾矿区优势植物中的重金属元素[J].分析试验室,2016,35(5):521-525.公开的方法。
菌株W25的检测方法与菌株W7的方法相同。
实验结果见图2、图3、图4、图5、图6所示。
由图2所示，不同培养时间，培养液中γ-PGA含量变化较大，菌株W7在相同培养时间，培养液中Cd含量越高，产生γ-PGA的量越多；菌株W25出现同样的现象；在相同培养时间，相同Cd含量培养液，菌株W25产生γ-PGA的量高于菌株W7。
由图3、图4可以看出，随着培养时间的延长，培养液中Cd含量下降显著，就图4中培养96小时的柱状图显示，菌株W7对应的培养液中Cd含量降低了50％；菌株W25对应的培养液中Cd含量降低了60％以上。
由图5可以看出，随着培养时间的延长，培养液中OD 600值在不断增加，在培养24h时，各培养液的OD 600值还有所不同，在培养48h时，各培养液的OD 600值几乎相同，即菌体浓度基本相同，通过72h和96h时的OD 600值可以看出，培养后期各培养液中的菌体浓度相差不大，基本相同。
由图6可以看出，随着培养时间的延长，培养液中pH在不断增加，在培养后期几乎不变。
实施例5
菌株W7、W25分别增加根际土壤酶活性的检测，以及增加根际土壤中蛋白菌(Proteobacteria)、拟杆菌门(Firmicutes)、鞘氨醇单胞菌(Sphingomonas)、芽孢杆菌属(Bacillus)的相对丰度的检测
采集实施例3盆栽实验根系紧密结合的土壤(作为根际土壤)进行后续分析；测定采集土壤的脲酶活性(Soil Sci Ch.Acad，1980；S.Guan等，1986)。
用快速DNA提取试剂盒(MP Biomedicals，Santa Ana，CA)提取采集土壤样品的细菌基因组DNA，并在-20℃下保存，然后进行进一步分析；分别用分光光度计(NanoDrop 1000，Thermo Scientific，USA)和凝胶电泳法测定提取DNA的数量和质量。提取的DNA经338F(5'-actcctagggggcagca-3')和806R(5'-GGACTACHVGGGTWTCTAAT-3')引物扩增，其靶向为细菌16s rRNA的V4区。高通量测序在Illumina Hiseq 2000(Illumina Inc.，美国，圣地亚哥)完成。
实验结果见图7、图8、图9、图10、图11所示。
由图7可以看出，菌株W7和W25对土壤的脲酶活性有一定的提高作用，在土壤含有Cd时，菌株W7和W25对土壤的脲酶活性提高作用增强。
由图8-11可以看出，菌株W7和W25对土壤中蛋白菌(Proteobacteria)、拟杆菌门(Firmicutes)、鞘氨醇单胞菌(Sphingomonas)、芽孢杆菌属(Bacillus)的有一定的增殖作用， 由图10可以看出，在土壤中Cd含量较高，菌株W7和W25对鞘氨醇单胞菌(Sphingomonas)的增殖效果更显著，菌株W25相对于W7，对鞘氨醇单胞菌(Sphingomonas)的增殖效果更优。
综上所示，本发明涉及的菌株W7和W25同时具有降低植物中重金属的含量，尤其是镉(Cd)的含量，促进植物生长，增加植物的Vc含量，增加植物可溶性蛋白的含量，增加土壤微生物丰度，增加土脲酶活性等多种功能，可广泛应用于土壤改良、农作物生物量提升、土壤重金属修复等领域。

Claims (10)

  1. 一株芽孢杆菌W7，分类学命名为枯草芽孢杆菌Bacillus subtilis，保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏地址：北京市朝阳区北辰西路1号院3号，保藏日期为2020年6月8日，菌种保藏号为CGMCC No.20043。
  2. 权利要求1所述菌株W7的培养方法，其特征在于，包括如下步骤：
    将菌株W7接种于固体培养基上，28-32℃培养2-3天，然后挑取菌落接种于液体培养基中，35-37℃，150-200rpm培养16-20h，获得菌株W7发酵液；所述固体培养基为：蔗糖10.0g/L、(NH 4) 2SO 40.5g/L、Na 2HPO 42.0g/L、NaCl 0.1g/L、KCl 0.191g/L、MgSO 4·7H 2O 0.5g/L、琼脂20g/L、余量水；所述液体培养基为：蔗糖10.0g/L、(NH 4) 2SO 40.5g/L、Na 2HPO 42.0g/L、NaCl 0.1g/L、KCl 0.191g/L、MgSO 4·7H 2O 0.5g/L、余量水。
  3. 权利要求1所述菌株W7用于生产生物制剂；
    优选的，所述生物制剂为液体制剂，含有所述菌株W7的有效活菌数为10 8个/毫升以上。
  4. 权利要求1所述菌株W7用于农作物种植；
    优选的，所述菌株W7用于降低蔬菜中重金属镉的含量；
    优选的，所述菌株W7用于增加蔬菜中维生素C和可溶性蛋白的含量；
    进一步优选的，所述蔬菜为生菜。
  5. 权利要求1菌株用于土壤改良；
    优选的，所述菌株W7用于去除土壤中的重金属镉；
    优选的，所述菌株W7用于促进土壤让中微生物的增殖；
    进一步优选的，所述菌株W7用于促进重金属镉污染的土壤中蛋白菌(Proteobacteria)、拟杆菌门(Firmicutes)、鞘氨醇单胞菌(Sphingomonas)、硫胺素芽孢杆菌属(Aneurinibacillus)之一或二者以上的增殖。
  6. 一种芽孢杆菌W25，分类学命名为解淀粉芽孢杆菌Bacillus amyloliquefaciens，保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏地址：北京市朝阳区北辰西路1号院3号，保藏日期为2020年6月8日，菌种保藏号为CGMCC No.20042。
  7. 权利要求6所述菌株W25的培养方法，其特征在于，包括如下步骤：
    将菌株W25接种于固体培养基上，28-32℃培养2-3天，然后挑取菌落接种于液体培养基中，35-37℃，150-200rpm培养16-20h，获得菌株W7发酵液；所述固体培养基为：蔗糖10.0g/L、(NH 4) 2SO 40.5g/L、Na 2HPO 42.0g/L、NaCl 0.1g/L、KCl 0.191g/L、MgSO 4·7H 2O 0.5g/L、琼脂20g/L、余量水；所述液体培养基为：蔗糖10.0g/L、(NH 4) 2SO 40.5g/L、Na 2HPO 42.0g/L、NaCl 0.1g/L、KCl 0.191g/L、MgSO 4·7H 2O 0.5g/L、余量水。
  8. 权利要求6所述菌株W25用于生产生物制剂；
    优选的，所述生物制剂为液体制剂，含有所述菌株W25的有效活菌数为10 8个/毫升以上。
  9. 权利要求6所述菌株W25用于农作物种植；
    优选的，所述菌株W25用于降低蔬菜中重金属镉的含量；
    优选的，所述菌株W25用于增加蔬菜中维生素C和可溶性蛋白的含量；
    进一步优选的，所述蔬菜为生菜。
  10. 权利要求6所述菌株W25用于土壤改良；
    优选的，所述菌株W25用于去除土壤中的重金属镉；
    优选的，所述菌株W25用于促进土壤让中微生物的增殖；
    进一步优选的，所述菌株W25用于促进重金属镉污染的土壤中蛋白菌(Proteobacteria)、拟杆菌门(Firmicutes)、鞘氨醇单胞菌(Sphingomonas)、硫胺素芽孢杆菌属(Aneurinibacillus)之一或二者以上的增殖。
PCT/CN2021/082435 2020-06-11 2021-03-23 具有降低蔬菜重金属含量、提高蔬菜质量的功能菌株及应用 WO2021248974A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US18/060,251 US20230114795A1 (en) 2020-06-11 2022-11-30 Strains capable of reducing heavy metal contents in vegetables and improving quality of vegetables and application thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202010531482.9 2020-06-11
CN202010531482.9A CN111718870B (zh) 2020-06-11 2020-06-11 具有降低蔬菜重金属含量、提高蔬菜质量的功能菌株及应用

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US18/060,251 Continuation US20230114795A1 (en) 2020-06-11 2022-11-30 Strains capable of reducing heavy metal contents in vegetables and improving quality of vegetables and application thereof

Publications (1)

Publication Number Publication Date
WO2021248974A1 true WO2021248974A1 (zh) 2021-12-16

Family

ID=72568021

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/082435 WO2021248974A1 (zh) 2020-06-11 2021-03-23 具有降低蔬菜重金属含量、提高蔬菜质量的功能菌株及应用

Country Status (3)

Country Link
US (1) US20230114795A1 (zh)
CN (2) CN111718870B (zh)
WO (1) WO2021248974A1 (zh)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114317373A (zh) * 2022-01-14 2022-04-12 湘潭大学 一种鞘氨醇单胞菌pah02、微生物制剂及其作为农作物降镉富硒功能调理剂的应用
CN114907987A (zh) * 2022-04-28 2022-08-16 江西师范大学 一株耐镉且能吸附镉的弯孢菌菌株及其应用
CN114940909A (zh) * 2022-05-11 2022-08-26 贵州大学 一种喀斯特地区铅镉污染土壤调理剂及其制备方法和应用

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111718870B (zh) * 2020-06-11 2021-07-06 齐鲁工业大学 具有降低蔬菜重金属含量、提高蔬菜质量的功能菌株及应用
CN112625967B (zh) * 2020-12-31 2022-05-10 江南大学 一种可去除镉离子的菌株及其应用
CN113278558A (zh) * 2021-05-26 2021-08-20 南宁市拜欧生物工程有限责任公司 一种高产脲酶的枯草芽孢杆菌及其应用方法
CN117402767A (zh) * 2023-02-01 2024-01-16 齐鲁工业大学(山东省科学院) 一种复合微生物菌剂及在农作物种植和土壤改良中的应用
CN117063943B (zh) * 2023-09-07 2024-03-15 齐鲁工业大学(山东省科学院) 芽孢杆菌w25在提高植物耐盐性和盐渍化土壤改良中的应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104560804A (zh) * 2014-12-25 2015-04-29 洪湖市添瑞生物制品厂 一种土壤修复的微生物复合菌剂及制备方法和用途
CN105861351A (zh) * 2015-01-22 2016-08-17 北京禾和润生科技有限公司 具有钝化重金属功能的解淀粉芽孢杆菌及其应用
CN110846250A (zh) * 2019-11-20 2020-02-28 中国科学院成都生物研究所 一株高产γ-PGA的枯草芽孢杆菌及其应用
CN111718870A (zh) * 2020-06-11 2020-09-29 齐鲁工业大学 具有降低蔬菜重金属含量、提高蔬菜质量的功能菌株及应用

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101517326B1 (ko) * 2014-01-28 2015-05-04 씨제이제일제당 (주) 발효 대두박 생산능이 향상된 바실러스 속 균주 및 이를 이용하여 발효 대두박을 제조하는 방법
CN108865923B (zh) * 2018-05-24 2021-10-01 苏州逸凡特环境修复有限公司 用于重金属污染土壤处理的微生物菌株及其筛选方法与应用
CN108976078A (zh) * 2018-08-01 2018-12-11 湖南工业大学 一种可吸附土壤中镉的生物炭有机肥及其制备方法

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104560804A (zh) * 2014-12-25 2015-04-29 洪湖市添瑞生物制品厂 一种土壤修复的微生物复合菌剂及制备方法和用途
CN105861351A (zh) * 2015-01-22 2016-08-17 北京禾和润生科技有限公司 具有钝化重金属功能的解淀粉芽孢杆菌及其应用
CN110846250A (zh) * 2019-11-20 2020-02-28 中国科学院成都生物研究所 一株高产γ-PGA的枯草芽孢杆菌及其应用
CN111718870A (zh) * 2020-06-11 2020-09-29 齐鲁工业大学 具有降低蔬菜重金属含量、提高蔬菜质量的功能菌株及应用

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
WANG XIAOHAN, GEYU DONG, XINGWANG LIU, SHENGKUI ZHANG, CAN LI, XIANQIN LU, TAO XIA: "Poly-γ-glutamic acid-producing bacteria reduced Cd uptake and effected the rhizosphere microbial communities of lettuce", JOURNAL OF HAZARDOUS MATERIALS, vol. 398, 20 June 2020 (2020-06-20), XP055878921, DOI: 10.1016/j.jhazmat.2020.123146 *
WANG, Q. ET AL.: "Increased biomass and quality and reduced heavy metal accumulation of edible tissues of vegetables in the presence of Cd-tolerant and immobilizing Bacillus megaterium H3", ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY, vol. 148, 6 November 2017 (2017-11-06) *
ZHANG RANRAN: "Aba Generating-Bacteria Reduces Cadmium Accumulation in Plants Grown in Cd-Contaminated Soil:Mechanisms and Applications", CHINESE MASTER’S THESES FULL-TEXT DATABASE, ENGINEERING SCIENCE AND TECHNOLOGY I, 15 June 2017 (2017-06-15), XP055878916 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114317373A (zh) * 2022-01-14 2022-04-12 湘潭大学 一种鞘氨醇单胞菌pah02、微生物制剂及其作为农作物降镉富硒功能调理剂的应用
CN114317373B (zh) * 2022-01-14 2023-05-02 湘潭大学 一种鞘氨醇单胞菌pah02、微生物制剂及其作为农作物降镉富硒功能调理剂的应用
CN114907987A (zh) * 2022-04-28 2022-08-16 江西师范大学 一株耐镉且能吸附镉的弯孢菌菌株及其应用
CN114907987B (zh) * 2022-04-28 2024-03-22 江西师范大学 一株耐镉且能吸附镉的弯孢菌菌株及其应用
CN114940909A (zh) * 2022-05-11 2022-08-26 贵州大学 一种喀斯特地区铅镉污染土壤调理剂及其制备方法和应用
CN114940909B (zh) * 2022-05-11 2023-05-05 贵州大学 一种喀斯特地区铅镉污染土壤调理剂及其制备方法和应用

Also Published As

Publication number Publication date
CN111718870B (zh) 2021-07-06
CN113337418A (zh) 2021-09-03
CN113337418B (zh) 2022-05-31
US20230114795A1 (en) 2023-04-13
CN111718870A (zh) 2020-09-29

Similar Documents

Publication Publication Date Title
WO2021248974A1 (zh) 具有降低蔬菜重金属含量、提高蔬菜质量的功能菌株及应用
CN102936574B (zh) 一种耐重金属的根瘤菌及其促进尾矿区植物修复的方法
CN109652329A (zh) 一种芽孢杆菌属固体微生物菌剂的制备及应用
CN102409014B (zh) 一株冬枣根际促生枯草芽孢杆菌及其应用
CN108624528B (zh) 一种对豆科植物具有促生增产作用的复合菌剂及其应用
CN110846249B (zh) 一种可用于改良盐碱土壤的微生物、微生物菌剂及盐碱地土壤微生物改良剂
CN109504622A (zh) 一种赖氨酸芽孢杆菌和枯草芽孢杆菌的复合菌剂及其制备方法
CN111187737B (zh) 一种耐盐碱促生菌及其在盐碱地改良中的应用
CN110846250B (zh) 一株高产γ-PGA的枯草芽孢杆菌及其应用
CN104673715A (zh) 对镉具有固定效应并能促进植物生长的肠杆菌及其应用
CN109735475B (zh) 一株耐酸产乙偶姻的解淀粉芽孢杆菌及其应用
CN109576177B (zh) 一种中华微杆菌株sm8及其在耐盐促生中的应用
CN109868242B (zh) 一株耐盐产乙偶姻的枯草芽孢杆菌及其应用
CN113528398B (zh) 一种具有解磷解钾作用的枯草芽孢杆菌、微生物菌剂及其应用
CN110076193A (zh) 黎巴嫩假单胞菌株my及其在重金属污染盐渍土壤修复中的应用
CN117106614B (zh) 一株根际细菌青岛假单胞菌yim b08402和其微生物菌剂及应用
CN102911900B (zh) 紫金牛叶杆菌RC6b及其在土壤修复中的应用
CN109762766B (zh) 一株具有吸附重金属、解磷及植物益生的细菌及其应用
CN108097711A (zh) 一种利用微紫青霉菌联合印度芥菜修复锌污染土壤的方法
CN117050913A (zh) 类芽孢杆菌cbp-2及其应用
CN110713953A (zh) 一种具有解磷特性的中间根瘤菌属菌株及其应用
CN112779187B (zh) 一株五大连池芽孢杆菌及其应用
CN107937302A (zh) 一种提高重金属有效性的细菌d10及其应用
CN103044094A (zh) 一种用水葫芦渣和复合微生物菌种制备的微生物活性磷肥
CN114958678B (zh) 一种肠杆菌y1-10及其应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21821846

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 15.05.2023)

122 Ep: pct application non-entry in european phase

Ref document number: 21821846

Country of ref document: EP

Kind code of ref document: A1