除草剂异丙隆降解菌株的筛选鉴定及降解特性和应用研究
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摘要
随着城市化进程和农村劳动力转移的不断加快,为提高农业生产效率保证农业生产的正常进行,除草剂被大量广泛使用,取代脲类除草剂就是其中的一种。因取代脲类除草剂的大量使用而造成的除草剂残留问题已经引起人们的广泛关注,对污染环境修复问题已成为迫在眉睫的重要课题。生物修复因具有经济、安全、方便、高效等优点,成为处理有机物面源污染的重要手段。本研究将对异丙隆(取代脲类除草剂的一种)降解菌株的分离过程进行介绍,对降解菌株的生长和降解性状进行研究,在此基础上完成对菌株的修复取代脲类除草剂污染能力的评估,以期为人们认识取代脲类除草剂在环境中的生物降解过程提供帮助,并为解决取代脲类除草剂的面源污染提供一种安全高效的方法。
采用富集培养的方式从长期受除草剂污染土壤中分离了五株细菌,分别命名为Y57、YBL1、YBL2、YBL3和CA6。其中菌株Y57、YBL1、YBL2和YBL3降解取代脲类除草剂异丙隆,菌株CA6可以降解取代脲类除草剂降解过程中产生的苯胺化合物。根据生理生化特性和16SrRNA基因序列比对,将其中的一株细菌Y57鉴定为鞘氨醇单胞菌属(Sphingomonas sp.)细菌。将细菌YBL1、YBL2和YBL3鉴定为鞘氨醇杆菌属(Sphingobium sp.)细菌。
苯胺降解菌株CA6通过16S rRNA基因序列分析鉴定为固氮螺菌属细菌,但与目前该属内已发表的种存在较大的差异。经过生理生化试验发现,菌株CA6培养1d后,在NFb固体培养基平板上会出现肉眼可见的菌落,菌落呈圆形,光滑,凸起,微小,3d后菌落在NFb培养基平板上的单菌落逐渐长大,菌落的形态发生变化,菌落的表面变粗燥,突起,与固体培养基平板牢固的结合；CA6菌体大小约为0.7-0.9μm×1.5-2.4μm,菌体呈杆状,稍有螺旋,单极生鞭毛；革兰氏染色为阴性,不产芽孢,严格好氧,接触酶阳性,氧化酶阳性,不液化明胶,脲酶阳性,不水解淀粉,最适生长温度为37℃,但菌株可以在20-40℃广泛的温度范围内生长,最适生长pH值为6.0-7.0。不能发酵葡萄糖、乳糖、蔗糖产酸,菌株为化能异养,菌株可以在含有3%的氯化钠的NFb液体培养基中生长,柠檬酸盐还原、硝酸还原和亚硝酸还原反应阴性；不产生吲哚；H2S产生反应阴性；水解磷酸盐阴性。16SrRNA基因序列(GenBank No.EU573183)与菌株Azospirillum rugosum AFH-6T, Azospirillum brasilense DSM1690T和Azospirillum lipoferum DSM1691T的序列同源性达到96.6%、96.3%、96.2%；CA6的成分如下：Feature2(4.92%),13:1AT12-13(0.45%), unknown14.959(0.31%), Sum In Feature3(4.72%),16:1ω5c(2.59%),16:0(2.67%),16:03OH (2.77%), Sum In Feature5(0.36%),18:1ω9c (4.55%),18:1ω7c (66.39%),18:0(1.19%),19:0CYCLOω8c (4.61%),18:12OH(3.75%),18:03OH (0.74%).;菌株可以利用环糊精、糊精、吐温40、N-乙酰基-D半乳糖胺、N-乙酰基-D-葡萄糖胺、侧金盏花醇、D-果糖、a-D-葡萄糖、甲基丙酮酸、麦芽糖、D-甘露糖、D-棉子糖、L-棉子糖、蔗糖、单甲基琥珀酸、甲酸、L-丙氨酸胺、D-丙氨酸、L-丙氨酸、L-丙氨酰、甘氨酸、L-脯氨酸、p-羟基丁酸、丙酸、琥珀酸、溴丁二酸、D,L-乳酸等为唯一碳源生长。其中菌株对糊精、蔗糖、单甲基琥珀酸、p-羟基丁酸、L-丙氨酰甘氨酸的利用最好。通过与现有菌株的特性比较后初步将该菌鉴定为固氮螺菌属的一个新种,命名为宪武固氮螺菌(Azospirillum xianwuii),模式菌株为Azospirillum xianwuii sp. nov. CA6(CCTCC AB208097=JCM15509).
菌株降解异丙隆的实验表明,在接种量为1.0%时,四株细菌均可以在36h之内将30mg·1-1的异丙隆完全降解,最终降解率达99%以上。在降解异丙隆的同时,四株细菌利用异丙隆提供的碳氮源进行生长。菌株Y57、YBL1、YBL2和YBL3降解异丙隆的最适pH分别为7.0,培养基中异丙隆的初始浓度对菌株的降解速度的影响较小,在培养基中添加不同的金属离子对菌株的降解能力和降解速度产生不同的影响,Mg2+对菌株的降解产生促进作用,1mmol·l-1的Cu2+或Ni2+对菌株的降解有明显地抑制作用；在培养基中添加葡萄糖对菌株的降解有促进作用；菌株YBL1、YBL2和YBL3降解异丙隆的最适温度为30℃,菌株Y57的最适降解温度为35℃。菌株YBL1、YBL2、 YBL3和Y57均具有较宽的降解谱,四株细菌均可以降解异丙隆、绿麦隆、敌草隆、氟草隆等N,N.二甲基取代脲类除草剂,而YBL1菌株还可以降解甲氧基取代脲类除草剂中的利谷隆；菌株Y57不但可以降解取代脲类除草剂,还可以降解酰胺类除草剂中的敌稗,降解率达到80%以上。
菌株YBL1、YBL2、YBL3可以迅速降解马肝土中的异丙隆、绿麦隆、敌草隆等除草剂；但是菌株对红壤中的除草剂的降解效果明显差于马肝土中的降解,这可能是红壤的pH过低所致；在土壤中的接种量较低时,菌株对土壤中的异丙隆的降解效果较差,只有当菌株在土壤中的浓度超过一定的数值,菌株才能对异丙隆残留有较好的降解效果。菌株在灭土壤中降解异丙隆的速率略快于新鲜土壤中的降解速度。在土壤中添加碳源、土壤的含水量、土壤中的土著微生物等对菌株降解能力产生明显地影响,其中土壤pH的影响最大。
菌株YBL2降解苯脲基类化合物时,首先作用于苯环侧链,逐一脱去异丙隆N原子上所连的两个甲基基团,通过进一步的酶促反应生成相应的苯胺化合物。菌株YBL1、YBL2和YBL3降解苯胺化合物,降解苯胺的过程中生成了邻苯二酚,再通过1,2位双加氧酶降解邻苯二酚生成顺,顺-粘糠酸。其中YBL1可以降解以异丙隆为代表的双甲基取代脲类除草剂和以利谷隆为代表的N-甲基-N-甲氧基取代脲类除草剂,并且可以降解苯胺类化合物,为首次报道。菌株YBL2可以降解4-甲基邻苯二酚,但不能降解3-甲基邻苯二酚,苯胺的间位上存在的取代基团对菌株的降解产生明显地影响,使得菌株在对间位上含有取代基团的苯胺进行双加氧反应时,只有一个方向可以进行。说明间位上存在取代基团会影响到间位含有取代基团的苯胺生成的化合物的种类。
通过试验发现菌株可以降低因加入绿麦隆而引起的药害,降低因加入异丙隆而对油菜的药害。将所分离的菌株发酵成菌剂投加到同时含有绿麦隆和绿黄隆残留的泰兴某处农田中,经测定菌株可以在一周之内将土壤中的0.1mg·kg-1的绿麦隆残留完全降解,直至检测不出,但在加入绿黄隆降解菌株SW3时,可使降解绿黄隆残留但降解率较低。
Five strains of bacteria (designated as Y57, YBL1, YBL2, YBL3, CA6respectively) capable of degrading isoproturon,3-(4-isopropylphenyl)-1,1-dimethylurea, or capable of degrading aniline which was thought as the critical metabolite in the degradation of isoproturon in environment, were isolated from the soils of three herbicide plants. Strain Y57was isolated from the soil collected from Changzhou, Strain YBL1was isolated from the soil collected from Nanjing and other two strains were isolated from the soil collected from Suzhou. Based on the comparative analysis of16S rRNA gene, phenotypic and biochemical characterization, strain Y57was identified as Sphingomonas sp., other three strains were identified as Sphingobium sp.. The optimum conditions for strains YBL1, YBL2and YBL3growth were pH6.0, and temperature30℃, while the optimum condition for strain Y57growth was pH7.0and temperature35℃. All these strains can not tolerate2.5%NaCl. Growth rate was related positively to oxygen availability.
Strain CA6was isolated from soil which was collected from Suzhou. On the basis of polyphasic taxonomy, morphological characterization, Biolog analysis, Cellular fatty acid and16S rRNA gene sequence analysis, the isolate represent a new species within the genus Azospirillum. Its closest phylogenetic neighbors, as deduced by16S rRNA gene analysis, are Azospirillum rugosum AFH-6T, Azospirillum brasilense DSM1690T and Azospirillum Hpoferum DSM1691r with96.6,96.3and96.2%sequence similarity respectively. The chemotaxonomic characteristics of CA6(G+C content is70.5mol%and18:1ω7c as the major cellular fatty acid, and14:03-OH and16:03-OH as the major hydroxy fatty acids) were consistent with the genus Azospirillum. Cells of CA6are rods and curve,0.7-0.9μm in width and1.5-2.4μm in length, Gram-negative and motile with single polar flagellum. Optimum growth occurs at37℃and at pH values between6.0and7.0. They have a respiratory type of metabolism, grow well on Dextrin, a-D-Glucose, Maltose, Mono- Methyl-Succinate, β-Hydroxy Butyric Acid, L-Alanyl-glycine. For this species, the name Azospirillum xianwuii sp. nov. is suggested, with strain CA6(=CCTCC AB208097=JCM15509) as the type strain.
In the degradation test, more than99%of the initial isoproturon (50mg·l-1, Y57was30mg·l-), added to the medium as the sole carbon source, were degraded by these strains at the inoculation rate of107cells per mili liter within20hours. Strains showed optimum degradation activity at pH7.0, which dropped sharply above pH9.0or below pH6.0, and the strain lost the ability to degrade isoproturon at pH5.0; the optimum temperature for isoproturon degradation by these strains was30℃, however these strains can degrade isoproturon between20℃to35℃as well. Supplementing the medium with glucose considerably enhanced isoproturon degradation rate; but slow down degradation rate by strain Y57in MSM medium, the addition of Cu2+, Ni2+at the rate of1mmol·l-1inhibited the capability of these strains to degrade isoproturon, while the Mg2+in the same quantity enhanced the isoproturon degradation rate under the same conditions. These four strains also showed the ability to remove the residues of other phenylurea herbicides such as chlorotoluron, diuron and fluometuron in mineral salt culture medium. Strain YBL1was found capable of degrading both, dimethylurea-substituted herbicides and methoxy methyl phenylurea herbicide i.e. linuron (3-(3,4-dichlorophenyl)-1-methoxy-1-methylurea). Strain Y57was capable of degrading propail in addition to isoproturon and chlorotoluron. The concentration of isoproturon (within the chemical solubility in water) had no apparent effect on the degradation rate.
Strains YBL1, YBL2and YBL3degraded isoproturon significantly (>80%) in Magan soil, and Flvo-Aquic soil within two days, while in Red soil the degradation activity was very poor. The degradation rate of other phenylurea herbicides, such as chlorotoluron and diuron was also observed higher in Magan soil. These three strains also have ability to degrade isoproturon in fresh soil (non-sterilized), but the degradation rate was slower in fresh soil than in sterilized soil, and the degradation rate of isoproturon by these strain was related positively to the soil moisture (10%-40%, v:v). The most adaptable temperature for these strains degrading isoproturon in soil was35℃, which was higher than the optimum temperature for isoproturon degradation in MSM The degradation rate was related positively to inoculation quantity, when the inoculum was below10CFU per gram soil, these strains were unable to degrade the isoproturon after two days inoculation. Glucose suplementation can enhance the degradation rate of isoproturon in soil, while the addition of NH4NO3showed the reverse results.
MDBPU (3-(4-isopropylphenyl)-1-methylurea) was detected in the catabolic products of isoproturon, and the chemical3-(3-chloro-4-methylphenyl)-1-methylurea, was detected in the degradation of chlorotoluron by YBL2, analysed by HPLC-MS. The catechol, which was thought as the critical metabolite in the process of phenyl-ring cleavage of most aromatic hydrocarbon, was detected in the beginning of aniline degradation by these strains. Moreover the cis, cis-muconic acid, the first metabolite after phenyl-ring cleavage, was also detected in the degradation products of aniline by HPLC.
Additon of strain Y57can reduce the effect of chlorotoluron residue on the growth of maize, and addition of strains YBL1, YBL2and YBL3respectively can clean up the isoproturon residue in laboratory experiments, and protected the rape from prototoxitity of isoproturon. Using culture of YBL3in a field, which was contaminated by chlorotoluron as a result of repeated use located at Taixing, can clean up the residue of chlorotoluron in soil. All these results indicate that these strains can be used for bioremediation of phenylurea herbicides contamination.
采用富集培养的方式从长期受除草剂污染土壤中分离了五株细菌,分别命名为Y57、YBL1、YBL2、YBL3和CA6。其中菌株Y57、YBL1、YBL2和YBL3降解取代脲类除草剂异丙隆,菌株CA6可以降解取代脲类除草剂降解过程中产生的苯胺化合物。根据生理生化特性和16SrRNA基因序列比对,将其中的一株细菌Y57鉴定为鞘氨醇单胞菌属(Sphingomonas sp.)细菌。将细菌YBL1、YBL2和YBL3鉴定为鞘氨醇杆菌属(Sphingobium sp.)细菌。
苯胺降解菌株CA6通过16S rRNA基因序列分析鉴定为固氮螺菌属细菌,但与目前该属内已发表的种存在较大的差异。经过生理生化试验发现,菌株CA6培养1d后,在NFb固体培养基平板上会出现肉眼可见的菌落,菌落呈圆形,光滑,凸起,微小,3d后菌落在NFb培养基平板上的单菌落逐渐长大,菌落的形态发生变化,菌落的表面变粗燥,突起,与固体培养基平板牢固的结合；CA6菌体大小约为0.7-0.9μm×1.5-2.4μm,菌体呈杆状,稍有螺旋,单极生鞭毛；革兰氏染色为阴性,不产芽孢,严格好氧,接触酶阳性,氧化酶阳性,不液化明胶,脲酶阳性,不水解淀粉,最适生长温度为37℃,但菌株可以在20-40℃广泛的温度范围内生长,最适生长pH值为6.0-7.0。不能发酵葡萄糖、乳糖、蔗糖产酸,菌株为化能异养,菌株可以在含有3%的氯化钠的NFb液体培养基中生长,柠檬酸盐还原、硝酸还原和亚硝酸还原反应阴性；不产生吲哚；H2S产生反应阴性；水解磷酸盐阴性。16SrRNA基因序列(GenBank No.EU573183)与菌株Azospirillum rugosum AFH-6T, Azospirillum brasilense DSM1690T和Azospirillum lipoferum DSM1691T的序列同源性达到96.6%、96.3%、96.2%；CA6的成分如下：Feature2(4.92%),13:1AT12-13(0.45%), unknown14.959(0.31%), Sum In Feature3(4.72%),16:1ω5c(2.59%),16:0(2.67%),16:03OH (2.77%), Sum In Feature5(0.36%),18:1ω9c (4.55%),18:1ω7c (66.39%),18:0(1.19%),19:0CYCLOω8c (4.61%),18:12OH(3.75%),18:03OH (0.74%).;菌株可以利用环糊精、糊精、吐温40、N-乙酰基-D半乳糖胺、N-乙酰基-D-葡萄糖胺、侧金盏花醇、D-果糖、a-D-葡萄糖、甲基丙酮酸、麦芽糖、D-甘露糖、D-棉子糖、L-棉子糖、蔗糖、单甲基琥珀酸、甲酸、L-丙氨酸胺、D-丙氨酸、L-丙氨酸、L-丙氨酰、甘氨酸、L-脯氨酸、p-羟基丁酸、丙酸、琥珀酸、溴丁二酸、D,L-乳酸等为唯一碳源生长。其中菌株对糊精、蔗糖、单甲基琥珀酸、p-羟基丁酸、L-丙氨酰甘氨酸的利用最好。通过与现有菌株的特性比较后初步将该菌鉴定为固氮螺菌属的一个新种,命名为宪武固氮螺菌(Azospirillum xianwuii),模式菌株为Azospirillum xianwuii sp. nov. CA6(CCTCC AB208097=JCM15509).
菌株降解异丙隆的实验表明,在接种量为1.0%时,四株细菌均可以在36h之内将30mg·1-1的异丙隆完全降解,最终降解率达99%以上。在降解异丙隆的同时,四株细菌利用异丙隆提供的碳氮源进行生长。菌株Y57、YBL1、YBL2和YBL3降解异丙隆的最适pH分别为7.0,培养基中异丙隆的初始浓度对菌株的降解速度的影响较小,在培养基中添加不同的金属离子对菌株的降解能力和降解速度产生不同的影响,Mg2+对菌株的降解产生促进作用,1mmol·l-1的Cu2+或Ni2+对菌株的降解有明显地抑制作用；在培养基中添加葡萄糖对菌株的降解有促进作用；菌株YBL1、YBL2和YBL3降解异丙隆的最适温度为30℃,菌株Y57的最适降解温度为35℃。菌株YBL1、YBL2、 YBL3和Y57均具有较宽的降解谱,四株细菌均可以降解异丙隆、绿麦隆、敌草隆、氟草隆等N,N.二甲基取代脲类除草剂,而YBL1菌株还可以降解甲氧基取代脲类除草剂中的利谷隆；菌株Y57不但可以降解取代脲类除草剂,还可以降解酰胺类除草剂中的敌稗,降解率达到80%以上。
菌株YBL1、YBL2、YBL3可以迅速降解马肝土中的异丙隆、绿麦隆、敌草隆等除草剂；但是菌株对红壤中的除草剂的降解效果明显差于马肝土中的降解,这可能是红壤的pH过低所致；在土壤中的接种量较低时,菌株对土壤中的异丙隆的降解效果较差,只有当菌株在土壤中的浓度超过一定的数值,菌株才能对异丙隆残留有较好的降解效果。菌株在灭土壤中降解异丙隆的速率略快于新鲜土壤中的降解速度。在土壤中添加碳源、土壤的含水量、土壤中的土著微生物等对菌株降解能力产生明显地影响,其中土壤pH的影响最大。
菌株YBL2降解苯脲基类化合物时,首先作用于苯环侧链,逐一脱去异丙隆N原子上所连的两个甲基基团,通过进一步的酶促反应生成相应的苯胺化合物。菌株YBL1、YBL2和YBL3降解苯胺化合物,降解苯胺的过程中生成了邻苯二酚,再通过1,2位双加氧酶降解邻苯二酚生成顺,顺-粘糠酸。其中YBL1可以降解以异丙隆为代表的双甲基取代脲类除草剂和以利谷隆为代表的N-甲基-N-甲氧基取代脲类除草剂,并且可以降解苯胺类化合物,为首次报道。菌株YBL2可以降解4-甲基邻苯二酚,但不能降解3-甲基邻苯二酚,苯胺的间位上存在的取代基团对菌株的降解产生明显地影响,使得菌株在对间位上含有取代基团的苯胺进行双加氧反应时,只有一个方向可以进行。说明间位上存在取代基团会影响到间位含有取代基团的苯胺生成的化合物的种类。
通过试验发现菌株可以降低因加入绿麦隆而引起的药害,降低因加入异丙隆而对油菜的药害。将所分离的菌株发酵成菌剂投加到同时含有绿麦隆和绿黄隆残留的泰兴某处农田中,经测定菌株可以在一周之内将土壤中的0.1mg·kg-1的绿麦隆残留完全降解,直至检测不出,但在加入绿黄隆降解菌株SW3时,可使降解绿黄隆残留但降解率较低。
Five strains of bacteria (designated as Y57, YBL1, YBL2, YBL3, CA6respectively) capable of degrading isoproturon,3-(4-isopropylphenyl)-1,1-dimethylurea, or capable of degrading aniline which was thought as the critical metabolite in the degradation of isoproturon in environment, were isolated from the soils of three herbicide plants. Strain Y57was isolated from the soil collected from Changzhou, Strain YBL1was isolated from the soil collected from Nanjing and other two strains were isolated from the soil collected from Suzhou. Based on the comparative analysis of16S rRNA gene, phenotypic and biochemical characterization, strain Y57was identified as Sphingomonas sp., other three strains were identified as Sphingobium sp.. The optimum conditions for strains YBL1, YBL2and YBL3growth were pH6.0, and temperature30℃, while the optimum condition for strain Y57growth was pH7.0and temperature35℃. All these strains can not tolerate2.5%NaCl. Growth rate was related positively to oxygen availability.
Strain CA6was isolated from soil which was collected from Suzhou. On the basis of polyphasic taxonomy, morphological characterization, Biolog analysis, Cellular fatty acid and16S rRNA gene sequence analysis, the isolate represent a new species within the genus Azospirillum. Its closest phylogenetic neighbors, as deduced by16S rRNA gene analysis, are Azospirillum rugosum AFH-6T, Azospirillum brasilense DSM1690T and Azospirillum Hpoferum DSM1691r with96.6,96.3and96.2%sequence similarity respectively. The chemotaxonomic characteristics of CA6(G+C content is70.5mol%and18:1ω7c as the major cellular fatty acid, and14:03-OH and16:03-OH as the major hydroxy fatty acids) were consistent with the genus Azospirillum. Cells of CA6are rods and curve,0.7-0.9μm in width and1.5-2.4μm in length, Gram-negative and motile with single polar flagellum. Optimum growth occurs at37℃and at pH values between6.0and7.0. They have a respiratory type of metabolism, grow well on Dextrin, a-D-Glucose, Maltose, Mono- Methyl-Succinate, β-Hydroxy Butyric Acid, L-Alanyl-glycine. For this species, the name Azospirillum xianwuii sp. nov. is suggested, with strain CA6(=CCTCC AB208097=JCM15509) as the type strain.
In the degradation test, more than99%of the initial isoproturon (50mg·l-1, Y57was30mg·l-), added to the medium as the sole carbon source, were degraded by these strains at the inoculation rate of107cells per mili liter within20hours. Strains showed optimum degradation activity at pH7.0, which dropped sharply above pH9.0or below pH6.0, and the strain lost the ability to degrade isoproturon at pH5.0; the optimum temperature for isoproturon degradation by these strains was30℃, however these strains can degrade isoproturon between20℃to35℃as well. Supplementing the medium with glucose considerably enhanced isoproturon degradation rate; but slow down degradation rate by strain Y57in MSM medium, the addition of Cu2+, Ni2+at the rate of1mmol·l-1inhibited the capability of these strains to degrade isoproturon, while the Mg2+in the same quantity enhanced the isoproturon degradation rate under the same conditions. These four strains also showed the ability to remove the residues of other phenylurea herbicides such as chlorotoluron, diuron and fluometuron in mineral salt culture medium. Strain YBL1was found capable of degrading both, dimethylurea-substituted herbicides and methoxy methyl phenylurea herbicide i.e. linuron (3-(3,4-dichlorophenyl)-1-methoxy-1-methylurea). Strain Y57was capable of degrading propail in addition to isoproturon and chlorotoluron. The concentration of isoproturon (within the chemical solubility in water) had no apparent effect on the degradation rate.
Strains YBL1, YBL2and YBL3degraded isoproturon significantly (>80%) in Magan soil, and Flvo-Aquic soil within two days, while in Red soil the degradation activity was very poor. The degradation rate of other phenylurea herbicides, such as chlorotoluron and diuron was also observed higher in Magan soil. These three strains also have ability to degrade isoproturon in fresh soil (non-sterilized), but the degradation rate was slower in fresh soil than in sterilized soil, and the degradation rate of isoproturon by these strain was related positively to the soil moisture (10%-40%, v:v). The most adaptable temperature for these strains degrading isoproturon in soil was35℃, which was higher than the optimum temperature for isoproturon degradation in MSM The degradation rate was related positively to inoculation quantity, when the inoculum was below10CFU per gram soil, these strains were unable to degrade the isoproturon after two days inoculation. Glucose suplementation can enhance the degradation rate of isoproturon in soil, while the addition of NH4NO3showed the reverse results.
MDBPU (3-(4-isopropylphenyl)-1-methylurea) was detected in the catabolic products of isoproturon, and the chemical3-(3-chloro-4-methylphenyl)-1-methylurea, was detected in the degradation of chlorotoluron by YBL2, analysed by HPLC-MS. The catechol, which was thought as the critical metabolite in the process of phenyl-ring cleavage of most aromatic hydrocarbon, was detected in the beginning of aniline degradation by these strains. Moreover the cis, cis-muconic acid, the first metabolite after phenyl-ring cleavage, was also detected in the degradation products of aniline by HPLC.
Additon of strain Y57can reduce the effect of chlorotoluron residue on the growth of maize, and addition of strains YBL1, YBL2and YBL3respectively can clean up the isoproturon residue in laboratory experiments, and protected the rape from prototoxitity of isoproturon. Using culture of YBL3in a field, which was contaminated by chlorotoluron as a result of repeated use located at Taixing, can clean up the residue of chlorotoluron in soil. All these results indicate that these strains can be used for bioremediation of phenylurea herbicides contamination.
引文
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