焦化厂地肤根内解芘细菌的筛选及促生潜力
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摘要
从多环芳烃耐受植物根内分离具多环芳烃降解功能的内生细菌并研究其促生特性,为内生菌协同宿主植物修复多环芳烃污染土壤提供基础.以长期受多环芳烃污染的焦化厂区生长的地肤为材料,从其根内分离出以芘和1-氨基环丙烷-1-羧酸(ACC)为唯一碳源和氮源的内生细菌8株.通过芘降解试验,筛选得到3株高效芘降解内生细菌KSE4、KSE7和KSE8,经鉴定分别为芽孢杆菌属、假单胞菌属和鞘氨醇菌属.通过液体培养试验,研究了3株菌在芘胁迫下产ACC脱氨酶的能力和对地肤种子萌发的影响.结果表明:随着芘浓度(0～15 mg·L~(-1))的升高,ACC脱氨酶活性降低,其中KSE7的效果最好,在芘浓度为15 mg·L~(-1)时,地肤发芽率和芽长分别比对照提高了44.8%和61.1%,在地肤-微生物修复焦化厂污染土壤的修复中具有一定的应用潜力.
The endophytic bacteria were isolated from the roots of polycyclic aromatic hydrocarbon(PAHs)-tolerant plant. We investigated their ability of PAHs degradation and plant growth promo-ting, with the aim to provide theoretical support for bacterial-plant cooperative soil remediation. Kochia scoparia living in coking plant area were selected for strains isolation. Eight endophytic bacteria strains, which used pyrene and 1-aminocyclopropane-1-carboxylic acid(ACC) as sole carbon and nitrogen source, were isolated from the roots of K. scoparia. Three endophytic bacteria, KSE4, KSE7 and KSE8, displayed high degradation efficiency in pyrene degradation experiment. They were identified as Bacillus sp., Pseudomonas sp., and Sphingobacterium sp., respectively. The abili-ties of those three strains to produce ACC deaminase and their effects on seed germination of K. scoparia were examined under pyrene stress through liquid culture tests. The results showed that the activity of ACC deaminase decreased with increasing pyrene concentration(0-15 mg·L~(-1)). KSE7 had the strongest promotion effect. When pyrene concentration reached to 15 mg·L~(-1), the germination rate and bud length of K. scoparia increased by 44.8% and 61.1%, respectively. Our results indicated that KSE7 is a promising bacterial strain for soil remediation in coking plant area.
The endophytic bacteria were isolated from the roots of polycyclic aromatic hydrocarbon(PAHs)-tolerant plant. We investigated their ability of PAHs degradation and plant growth promo-ting, with the aim to provide theoretical support for bacterial-plant cooperative soil remediation. Kochia scoparia living in coking plant area were selected for strains isolation. Eight endophytic bacteria strains, which used pyrene and 1-aminocyclopropane-1-carboxylic acid(ACC) as sole carbon and nitrogen source, were isolated from the roots of K. scoparia. Three endophytic bacteria, KSE4, KSE7 and KSE8, displayed high degradation efficiency in pyrene degradation experiment. They were identified as Bacillus sp., Pseudomonas sp., and Sphingobacterium sp., respectively. The abili-ties of those three strains to produce ACC deaminase and their effects on seed germination of K. scoparia were examined under pyrene stress through liquid culture tests. The results showed that the activity of ACC deaminase decreased with increasing pyrene concentration(0-15 mg·L~(-1)). KSE7 had the strongest promotion effect. When pyrene concentration reached to 15 mg·L~(-1), the germination rate and bud length of K. scoparia increased by 44.8% and 61.1%, respectively. Our results indicated that KSE7 is a promising bacterial strain for soil remediation in coking plant area.
引文
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[14] Song L-C (宋立超),Li P-J (李培军),Liu W (刘宛),et al.Isolation,identification and degradation characteristics of a PAHs-degrading bacteria from salt-alkaline soil.Microbiology China (微生物学通报),2011,38(2):282-287 (in Chinese)
[15] Penrose DM,Glick BR.Methods for isolating and chara-cterizing ACC deaminase-containing plant growth-promoting rhizobacteria.Physiologia Plantarum,2003,118:10-15
[16] Zhang G-Z (张国壮),Li H-C (李海超),Sun Y-L (孙永林),et al.Isolation and identification of rhizobacteria producing 1-aminocyclopropane-1-carboxylate (ACC) deaminase.Journal of Northwest A&F University (Natural Science) (西北农林科技大学学报:自然科学版),2014,42(6):89-196 (in Chinese)
[17] Glickmann E,Dessaux Y.A critical examination of the specificity of the Salkowski reagent for indole compounds produced by phytopathogenic bacteria.Applied and Environmental Microbiology,1995,61:793-796
[18] Liu WX,Sun JY,Ding LL,et al.Rhizobacteria(Pseudo-monas sp.SB) assist phytoremediation of oily-sludge-contaminated soil by tall fescue (Testuca arundinacea L.).Plant and Soil,2013,371:533-542
[19] Zeng J,Zhu Q,Wu YC,et al.Oxidation of polycyclic aromatic hydrocarbons using Bacillus subtilis CotA with high laccase activity and copper independence.Chemosphere,2016,148:1-7
[20] Wang R-T (汪如婷),Luo X-F (雒晓芳),Tian D-N (田丹妮),et al.Research on the degradation characteristics of naphthalene and phenanthrene by Pseudomonas aeruginosa.China Brewing (中国酿造),2015,34(10):82-85 (in Chinese)
[21] Pei XH,Hang XH,Wang SM,et al.Effects of a biosurfactant and a synthetic surfactant on phenanthrene degradation by a sphingomonas strain.Pedosphere,2010,20:771-779
[22] Liu J-L (刘佳莉),Fang F (方芳),Shi X-H (史煦涵),et al.Isolation and characterization of PGPR from the rhizosphere of the Avena sativa in saline-alkali soil.Acta Prataculturae Sinica (草业学报),2013,22(2):132-139 (in Chinese)
[23] Zhang Y-Q (张艳群),Lai H-X (来航线),Wei X-M (韦小敏),et al.Screening and identification of two promoting plant growth bacillus strains.Acta Agriculturae Boreali-Occidentalis Sinica (西北农业学报),2014,23(3):193-200 (in Chinese)
[24] Zhang H (张晖),Song Y-Y (宋圆圆),Lyu S (吕顺),et al.The antifungal activity and crop growth stimu-lation of growth-promoting rhizobacteria from banana rhizosphere soil.Journal of South China Agricultural University (华南农业大学学报),2015,36(3):65-70 (in Chinese)
[25] Wang J,Liu J,Wang L,et al.Composite of PAH-degrading endophytic bacteria reduces contamination and health risks caused by PAHs in vegetables.Science of the Total Environment,2017,598:471-478
[26] Li-S (李爽),Zuo S-W (左尚武),Wang W-Q (王万清),et al.Determining the effects of Serratia sp.PW7 on pyrene removal and the endophytic bacterial community in ryegrass (Lolium multiflorum L.) via different inoculation methods.Journal of Agro-Environment Science (农业环境科学学报),2018,37(12):2755-2764 (in Chinese)
[27] Ho YN,Shih CH,Hsiao SC,et al.A novel endophytic bacterium,Achromobacter xylosoxidans,helps plants against pollutant stress and improves phytoremediation.Journal of Bioscience and Bioengineering,2009,108:S75-S95
[28] Bezza FA,Nkhalambayausi CEM.The role of lipopeptide biosurfactant on microbial remediation of aged polycyclic aromatic hydrocarbons (PAHs)-contaminated soil.Chemical Engineering Journal,2017,309:563-576
[29] Yang X (杨轩),Zhang W (张威),Li S-W (李师翁),et al.Study on isolation,identification and physiological characteristics of PAHs-degrading bacteria.Acta Scientiae Circumstantiae (环境科学学报),2012,32(5):1033-1040 (in Chinese)
[30] He L-Q (何丽娟),Li Z-H (李正华),Hong Q (洪青),et al.Characterization of a phenanthrene-degrading strain and cloning of degradation-related gene.Chinese Journal of Applied & Environmental Biology (应用与环境生物学报),2009,15(5):682-685 (in Chinese)
[2] Sun K (孙凯),Liu J (刘娟),Li X (李欣),et al.Isolation,identification,and performance of two pyrene-degrading endophytic bacteria.Acta Ecologica Sinica (生态学报),2014,34(4):853-861 (in Chinese)
[3] Wang RW,Yousaf B,Sun RY,et al.Emission characterization and δ13C values of parent PAHs and nitro-PAHs in size-segregated particulate matters from coal-fired power plants.Journal of Hazardous Materials,2016,318:487-496
[4] Liu (刘庚),Bi R-T (毕如田),Wang S-J (王世杰),et al.Statistical characteristic analysis of soil PAHs in a coking contaminated site of China.Chinese Journal of Applied Ecology (应用生态学报),2013,24(6):1722-1728 (in Chinese)
[5] Gan S,Lau EV,Ng HK.Remediation of soils contaminated with polycyclic aromatic hydrocarbons (PAHs).Journal of Hazardous Materials,2009,172:532-549
[6] Teng Y,Shen YY,Luo YM,et al.Influence of rhizobium meliloti on phytoremediation of polycyclic aromatic hydrocarbons by alfalfa in an aged contaminated soil.Journal of Hazardous Materials,2011,186:1271-1276
[7] Liu W-W (刘魏魏),Yin R (尹睿),Lin X-G (林先贵),et al.Interaction of phytoremediation-microorganism to remediation of aged polycyclic aromatic hydrocarbons (PAHs) polluted soils.Soils (土壤),2010,42(5):800-806 (in Chinese)
[8] Wang Y,Dai CC.Endophytes:A potential resource for biosynthesis,biotransformation,and biodegradation.Annals of Microbiology,2011,61:207-215
[9] Kukla M,P?ociniczak T,Piotrowska-Seget Z.Diversity of endophytic bacteria in Lolium perenne and their potential to degrade petroleum hydrocarbons and promote plant growth.Chemosphere,2014,117:40-46
[10] Song L-C (宋立超),Zhang W (张薇),Niu X-G (钮旭光),et al.Screening of pyrene-degrading bacteria from Kochia scoparia rhizosphere in coking plant and study of plant growth-promoting ability.Journal of Agro-Environment Science (农业环境科学学报),2017,36(11):2275-2280 (in Chinese)
[11] Chen X-B (陈小兵),Sheng X-F (盛下放),He L-Y (何琳燕),et al.Isolation and characteristics of a plant growth-promoting,phenanthrene-degrading endophytic bacterium from plants.Acta Scientiae Circumstantiae (环境科学学报),2008,28(7):1308-1313 (in Chinese)
[12] Hegazy AK.Phytomonitoring and management of tar piles on the qatari coastal marshes,arabian gulf.Enviromental Pollution,1995,90:187-190
[13] Moubasher HA,Hegazy AK,Mohamed NH,et al.Phytoremediation of soils polluted with crude petroleum oil using Bassia scoparia and its associated rhizosphere microorganisms.International Biodeterioration & Biode-gradation,2015,98:113-120
[14] Song L-C (宋立超),Li P-J (李培军),Liu W (刘宛),et al.Isolation,identification and degradation characteristics of a PAHs-degrading bacteria from salt-alkaline soil.Microbiology China (微生物学通报),2011,38(2):282-287 (in Chinese)
[15] Penrose DM,Glick BR.Methods for isolating and chara-cterizing ACC deaminase-containing plant growth-promoting rhizobacteria.Physiologia Plantarum,2003,118:10-15
[16] Zhang G-Z (张国壮),Li H-C (李海超),Sun Y-L (孙永林),et al.Isolation and identification of rhizobacteria producing 1-aminocyclopropane-1-carboxylate (ACC) deaminase.Journal of Northwest A&F University (Natural Science) (西北农林科技大学学报:自然科学版),2014,42(6):89-196 (in Chinese)
[17] Glickmann E,Dessaux Y.A critical examination of the specificity of the Salkowski reagent for indole compounds produced by phytopathogenic bacteria.Applied and Environmental Microbiology,1995,61:793-796
[18] Liu WX,Sun JY,Ding LL,et al.Rhizobacteria(Pseudo-monas sp.SB) assist phytoremediation of oily-sludge-contaminated soil by tall fescue (Testuca arundinacea L.).Plant and Soil,2013,371:533-542
[19] Zeng J,Zhu Q,Wu YC,et al.Oxidation of polycyclic aromatic hydrocarbons using Bacillus subtilis CotA with high laccase activity and copper independence.Chemosphere,2016,148:1-7
[20] Wang R-T (汪如婷),Luo X-F (雒晓芳),Tian D-N (田丹妮),et al.Research on the degradation characteristics of naphthalene and phenanthrene by Pseudomonas aeruginosa.China Brewing (中国酿造),2015,34(10):82-85 (in Chinese)
[21] Pei XH,Hang XH,Wang SM,et al.Effects of a biosurfactant and a synthetic surfactant on phenanthrene degradation by a sphingomonas strain.Pedosphere,2010,20:771-779
[22] Liu J-L (刘佳莉),Fang F (方芳),Shi X-H (史煦涵),et al.Isolation and characterization of PGPR from the rhizosphere of the Avena sativa in saline-alkali soil.Acta Prataculturae Sinica (草业学报),2013,22(2):132-139 (in Chinese)
[23] Zhang Y-Q (张艳群),Lai H-X (来航线),Wei X-M (韦小敏),et al.Screening and identification of two promoting plant growth bacillus strains.Acta Agriculturae Boreali-Occidentalis Sinica (西北农业学报),2014,23(3):193-200 (in Chinese)
[24] Zhang H (张晖),Song Y-Y (宋圆圆),Lyu S (吕顺),et al.The antifungal activity and crop growth stimu-lation of growth-promoting rhizobacteria from banana rhizosphere soil.Journal of South China Agricultural University (华南农业大学学报),2015,36(3):65-70 (in Chinese)
[25] Wang J,Liu J,Wang L,et al.Composite of PAH-degrading endophytic bacteria reduces contamination and health risks caused by PAHs in vegetables.Science of the Total Environment,2017,598:471-478
[26] Li-S (李爽),Zuo S-W (左尚武),Wang W-Q (王万清),et al.Determining the effects of Serratia sp.PW7 on pyrene removal and the endophytic bacterial community in ryegrass (Lolium multiflorum L.) via different inoculation methods.Journal of Agro-Environment Science (农业环境科学学报),2018,37(12):2755-2764 (in Chinese)
[27] Ho YN,Shih CH,Hsiao SC,et al.A novel endophytic bacterium,Achromobacter xylosoxidans,helps plants against pollutant stress and improves phytoremediation.Journal of Bioscience and Bioengineering,2009,108:S75-S95
[28] Bezza FA,Nkhalambayausi CEM.The role of lipopeptide biosurfactant on microbial remediation of aged polycyclic aromatic hydrocarbons (PAHs)-contaminated soil.Chemical Engineering Journal,2017,309:563-576
[29] Yang X (杨轩),Zhang W (张威),Li S-W (李师翁),et al.Study on isolation,identification and physiological characteristics of PAHs-degrading bacteria.Acta Scientiae Circumstantiae (环境科学学报),2012,32(5):1033-1040 (in Chinese)
[30] He L-Q (何丽娟),Li Z-H (李正华),Hong Q (洪青),et al.Characterization of a phenanthrene-degrading strain and cloning of degradation-related gene.Chinese Journal of Applied & Environmental Biology (应用与环境生物学报),2009,15(5):682-685 (in Chinese)