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1.
在开放式大气CO2浓度升高平台上(Free-Air CO2 Enrichment,简称FACE),采用盆栽实验,研究了不同浓度Cu污染胁迫条件下,稻麦轮作土壤中土壤酶活性及土壤微生物多样性对大气CO2浓度升高的响应。结果表明,大气CO2浓度升高显著诱导了清洁土壤中蛋白酶、脲酶、尿酸酶活性以及微生物多样性;正常大气和大气CO2浓度升高条件下,3种酶活性都随着土壤Cu污染胁迫的增加而逐渐降低;低浓度Cu污染胁迫条件下(50 mg.kg^-1),FACE圈中的土壤脲酶和蛋白酶活性显著高于正常大气(Ambience)圈,尿酸酶活性无显著变化;高浓度Cu污染胁迫条件下(400 mg.kg^-1),土壤脲酶与蛋白酶活性无显著变化,尿酸酶活性则显著降低,其原因可能与不同酶系对铜污染胁迫的敏感差异性以及大气CO2浓度升高对土壤中铜的活化作用有关。与清洁土壤相比,低浓度Cu污染(50 mg.kg^-1)对微生物生长具有一定的刺激作用,Ambience圈和FACE圈土壤微生物多样性都有所增加,FACE圈中这种现象更为明显;高浓度Cu污染胁迫(400 mg.kg^-1)对土壤微生物表现出了明显的毒害作用,微生物多样性有所降低,但在FACE圈中土壤微生物多样性的降低程度要低于Ambience圈,其影响机制有待进一步研究。  相似文献   

2.
用碳素利用法对浙江省天台铅锌银尾矿区重金属复合污染土壤微生物群落功能多样性动力学特征进行了初步探讨。研究结果表明 ,矿区重金属复合污染降低了供试土壤的微生物群落代谢剖面 ,且群落代谢剖面值与培养时间之间呈非线性关系 ,其变化过程符合微生物种群生长动态模型 (S形 )。随着重金属复合污染程度的加剧 ,土壤微生物群落功能多样性动力学参数K和r值越低 ,参数s值所需的时间则越长。上述动力学参数与群落代谢剖面各自的主成分分析结果显示 ,微生物群落功能多样性的动力学参数K值和s值能够很好地区分矿区土壤重金属污染程度 ,并且其区分效果比微生物群落代谢剖面值好。土壤微生物群落功能多样性动力学特征的变化可以较好地显示矿区重金属复合污染土壤微生物群落对碳源利用模式的差异 ,反映矿区特定生境土壤微生物群落功能多样性的变化 ,在一定程度上揭示重金属胁迫下环境微生物种群作用机理。  相似文献   

3.
为了解随粪肥进入农田中的土霉素对土壤生物化学性质产生的可能影响,采用实验模拟方法研究了土霉素污染对土壤微生物生物量碳、土壤酶活性及微生物组成的影响。结果表明,土霉素污染对土壤细菌、放线菌数量和微生物总量均有一定的抑制作用,随土霉素污染程度的提高抑制作用也有所增强;但土霉素污染对真菌的作用较为复杂,一般是低浓度时有促进作用,高浓度时有抑制作用。低量土霉素污染对土壤脲酶和中性磷酸酶活性均无明显的影响,但高量的土霉素污染对土壤脲酶活性起抑制作用。土霉素对土壤微生物生物量碳的影响因土壤类型、土霉素加入量和培养时间不同有所差异。土霉素污染对土壤生物化学性质的影响主要发生在土霉素进入土壤的初期,随着时间的增加,影响逐渐减弱和消失;  相似文献   

4.
当前土壤环境中重金属和抗生素的广泛共存及二者复合存在所诱导出的细菌抗性,与单一物质的污染相比,均能够加剧对土壤质量和作物安全的破坏。在在实验室模拟培养条件下,向土壤中加入不同浓度的重金属(铜)和抗生素(强力霉素),探讨抗生素和重金属复合污染对土壤微生物呼吸、脲酶、蔗糖酶和过氧化氢酶活性和四环素抗性基因的丰度等土壤微生物指标的影响。结果显示,在整个培养期(30 d)内,铜和强力霉素单一及复合污染均会显著抑制土壤微生物呼吸强度,对脲酶活性主要表现为促进作用,对蔗糖酶、过氧化氢酶活性主要为抑制作用,对过氧化氢酶活性的抑制强度明显大于蔗糖酶。综合而言,铜和强力霉素的复合污染相对于单一污染对上述微生物指标的影响较大,强力霉素的加入可以促进铜对微生物呼吸或酶活性的初始影响。此外,该研究还表明添加为400mg·kg~(–1)铜可以提高强力霉素在土壤培养中后期诱导的抗性基因相对丰度的能力水平。本研究从微生物角度定量探讨铜与强力霉素单一及复合污染对土壤微生物指标的影响程度,以期为重金属与抗生素协同污染的土壤构建微生物预警体系,并为土壤修复和风险评估工作提供理论依据。  相似文献   

5.
采用玉米草及海藻寡糖联合修复技术研究了石油污染土壤的修复效果,对修复过程中酶活性变化进行了测定,并采用变性梯度凝胶电泳(DGGE)技术测定了土壤中微生物群落的变化。结果表明,种植玉米草可以有效提高土壤中石油烃的降解,与对照相比石油烃降解率增加了11%;加入不同浓度海藻寡糖进一步增加了石油烃的降解效果,降解率最高达到28.6%。种植植物及加入海藻寡糖可以有效提高多酚氧化酶、脱氢酶及尿酶的活性。PCR-DGGE结果表明植物种植及海藻寡糖的加入增加了土壤中微生物数量,其微生物群落结构与未种植植物及修复前土壤相比发生了较大的变化。  相似文献   

6.
多环芳烃胁迫下稻田土壤细菌及分支杆菌种群多样性研究   总被引:1,自引:0,他引:1  
多环芳烃(PAHs)污染对土壤微生物群落特别是微生物功能群的影响一直备受关注。本文应用变性梯度凝胶电泳(DGGE)技术,从群落和功能群多样性两个角度,分析了PAHs胁迫条件下稻田土壤细菌遗传多样性及与PAHs降解有密切关系的分支杆菌种群多样性变化。结果表明,PAHs污染对稻田土壤细菌群落多样性指数无显著影响,但造成土壤细菌群落结构的改变,重度PAHs造成一些对污染敏感的细菌种类消失,而使一些与PAHs降解有关的细菌种类丰度增加。而对与多环芳烃降解有密切关系的分支杆菌而言,中度PAHs污染稻田土壤分支杆菌种群多样性指数较重度和轻度PAHs污染土壤的略高,不同PAHs污染程度稻田土壤的优势分支杆菌种类不尽相同,PAHs污染造成稻田土壤1种或几种分支杆菌得到富集。长期PAHs污染造成土壤细菌群落和分支杆菌种群结构的变化,将直接影响土壤生态系统功能的发挥,间接改变土壤质量。  相似文献   

7.
随着工业和农业生产的发展,重金属、有毒有机物及其复合污染土壤日益增多,其修复问题在世界范围内是一项具有挑战性的任务。以砂质壤土为试验对象,模拟石油-铅-镉复合污染,共设置4个处理:(1)对照处理,复合污染土壤(CK);(2)不施肥处理,复合污染土壤+黑麦草(NF);(3)施氮肥处理,复合污染土壤+黑麦草+氮肥(F1);(4)施氮肥和磷肥处理,复合污染土壤+黑麦草+氮肥+磷肥(F2),研究种植黑麦草与施肥处理对石油-铅-镉复合污染土壤微生物活性的影响,以期为污染土壤修复及环境影响评价提供初步的理论基础。结果表明:黑麦草与施肥对复合污染土壤基础呼吸、微生物量碳均有促进作用,处理NF、F1和F2的土壤基础呼吸强度比对照处理CK最高分别增加约20.94%,24.41%,42.69%,其中施加氮、磷肥(F2)对土壤基础呼吸影响最显著;第10天时,处理NF、F1和F2土壤微生物量碳含量与对照相比分别增加约26.92%,127.43%,181.89%,施肥处理土壤微生物量碳含量显著高于不施肥处理;不同种类的酶活性对黑麦草与施肥的响应不尽相同,其中种植黑麦草与施肥均会抑制石油-铅-镉复合污染土壤中FDA水解酶活性,施加氮肥在一定时间内能较好地维持石油-铅-镉复合污染土壤内的脱氢酶活性,施肥能有效地提高并维持复合污染土壤中脲酶活性,而过氧化氢酶活性受黑麦草与施肥影响不显著。研究表明种植黑麦草配施氮、磷肥对土壤微生物基础呼吸、土壤微生物量碳及相关土壤酶活性均有增强作用,进而有利于促进土壤污染物的去除及土壤质量的改善。  相似文献   

8.
通过根际袋土培试验,研究了磺胺间甲氧嘧啶(SMM)对玉米根际与非根际土壤酶活性和土壤呼吸强度的影响。结果表明,无论是否有根系作用,过氧化氢酶对磺胺间甲氧嘧啶胁迫的反应均不敏感。但在高浓度磺胺间甲氧嘧啶(50 mg.kg-1)作用下,试验初期对脲酶有明显的抑制作用,而在试验后期则表现出一定的促进作用,且在SMM胁迫下根际效应表现得更为明显。SMM胁迫下对根际与非根际土壤呼吸均有抑制作用,随浓度的增大,抑制作用越明显。且由于玉米根际作用,一定程度上缓解了SMM污染对根际微生物的毒害。一般情况下,根际土壤脲酶和过氧化氢酶活性及土壤呼吸强度均要大于非根际土壤,根际效应明显。  相似文献   

9.
国内首次运用FastPrep○R 核酸快速提取系统提取了重金属复合污染农田土壤的DNA ,并对其进行了聚合酶链反应—变性梯度凝胶电泳 (PCR DGGE)分析。结果表明 ,FastPrep○R核酸提取仪与相应的FastD NASPINKitforSoil试剂盒联用时 ,能有效地分离到纯度较高的重金属污染农田土壤的DNA。PCR DGGE电泳图谱表明 ,PCR产物经DGGE检测后得到的电泳条带清晰且分离效果好 ,可以明显反映出重金属复合污染导致了农田土壤微生物在基因上的损伤 ,影响到农田土壤生态系统的细菌丰富度 ,改变了土壤环境的优势菌群 ,从而使农田土壤微生物群落结构多样性发生变化。可见 ,FastPrep○R核酸提取系统同样适用于重金属污染农田土壤环境中微生物基因组DNA的快速分离和纯化 ,得到的DNA可直接用于PCR DGGE分析。  相似文献   

10.
随着畜禽养殖业规模化发展以及畜禽粪便的大量农用,由此导致的土壤重金属和抗生素复合污染问题日益突出。本文以畜禽养殖常用的饲料添加剂铜(Cu)和磺胺嘧啶(Sulfadiazine, SDZ)为目标污染物,分别采用土壤酶试剂盒(微量法)和Biolog-ECO盘法,研究二者复合污染对土壤酶活性和土壤微生物群落功能多样性的影响。结果发现,低浓度Cu(200 mg/kg)复合污染可缓解SDZ对土壤脱氢酶和β-葡萄糖苷酶的抑制作用,表现为拮抗作用;但加重SDZ对磷酸酶的抑制作用,表现为协同抑制作用。高浓度Cu(500 mg/kg)和SDZ复合对所测试土壤酶均表现为协同抑制作用。Biolog分析表明,10 mg/kg SDZ添加对土壤微生物活性具有促进作用,而Cu和SDZ复合污染处理则可显著抑制土壤微生物代谢活性。多样性指数分析表明Cu和SDZ复合污染对微生物群落多性影响较小,但可显著改变微生物种群的均一性;且主成分分析也表明二者复合污染可显著改变土壤微生物群落结构组成。本研究结论如下:1)Cu和SDZ复合污染对土壤酶活性的影响与土壤酶的种类及复合浓度有关,且土壤脱氢酶是反映SDZ及其与Cu复合污染较为敏感的指标;2)Cu和SDZ复合污染可显著抑制土壤微生物群落的整体代谢活性,并改变代谢功能,进而促使土壤微生物结构的发生变化。  相似文献   

11.
We studied the distribution of the indigenous bacterial and fungal communities in a forest soil profile. The composition of bacterial and fungal communities was assessed by denaturing gradient gel electrophoresis (DGGE) of total and extracellular DNA extracted from all the soil horizons. Microbial biomass C and basal respiration were also measured to assess changes in both microbial biomass and activity throughout the soil profile. The 16S rDNA-DGGE revealed composite banding patterns reflecting the high bacterial diversity as expected for a forest soil, whereas 18S rDNA-DGGE analysis showed a certain stability and a lower diversity in the fungal communities. The banding patterns of the different horizons reflected changes in the microbial community structure with increasing depth. In particular, the DGGE analysis evidenced complex banding patterns for the upper A1 and A2 horizons, and a less diverse microflora in the deeper horizons. The low diversity and the presence of specific microbial communities in the B horizons, and in particular in the deeper ones, can be attributed to the selective environment represented by this portion of the soil profile. The eubacterial profiles obtained from the extracellular DNA revealed the presence of some bands not present in the total DNA patterns. This could be interpreted as the remainders of bacteria not any more present in the soil because of changes of edaphic conditions and consequent shifting in the microbial composition. These characteristic bands, present in all the horizons with the exception of the A1, should support the concept that the extracellular DNA is able to persist within the soil. Furthermore, the comparison between the total and extracellular 16S rDNA-DGGE profiles suggested a downwards movement of the extracellular DNA.  相似文献   

12.
Soil microbial communities are very sensitive to changes in land use and are often used as indicators of soil fertility. We evaluated the microbial communities in the soils of four types of vegetation (cropland (CP), natural grassland (NG), broadleaf forest (BF) and coniferous forest (CF)) at depths of 0–10 and 10–20 cm on the Loess Plateau in China using phospholipid fatty acid (PLFA) profiling and denaturing gradient gel electrophoresis (DGGE) of DNA amplicons from polymerase chain reactions. The soil microbial communities were affected more by vegetation type than by soil depth. Total organic carbon, total nitrogen, soil-water content, pH, bulk density (BD) and C:N ratio were all significantly associated with the composition of the communities. Total PLFA, bacterial PLFA and fungal PLFA were significantly higher in the BF than the CP. The DGGE analyses showed that NG had the most diverse bacterial and fungal communities. These results confirmed the significant effect of vegetation type on soil microbial communities. BFs and natural grass were better than the CFs for the restoration of vegetation on the Loess Plateau.  相似文献   

13.
Soil microbial biomass and community structures are commonly used as indicators for soil quality and fertility. A investigation was performed to study the effects of long-term natural restoration, cropping, and bare fallow managements on the soil microbial biomass and bacterial community structures in depths of 0--10, 20--30, and 40--50 cm in a black soil (Mollisol). Microbial biomass was estimated from chloroform fumigation-extraction, and bacterial community structures were determined by analysis of 16S rDNA using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Experimental results showed that microbial biomass significantly declined with soil depth in the managements of restoration and cropping, but not in the bare fallow. DGGE profiles indicated that the band number in top 0--10 cm soils was less than that in depth of 20--30 or 40--50 cm. These suggested that the microbial population was high but the bacterial community structure was simple in the topsoil. Cluster and principle component analysis based on DGGE banding patterns showed that the bacterial community structure was affected by soil depth more primarily than by managements, and the succession of bacterial community as increase of soil depth has a similar tendency in the three managements. Fourteen predominating DGGE bands were excised and sequenced, in which 6 bands were identified as the taxa of Verrucomicrobia, 2 bands as Actinobacteria, 2 bands as α-Proteobacteria, and the other 4 bands as δ-Proteobacteria, Acidobacteria, Nitrospira, and unclassified bacteria. In addition, the sequences of 11 DGGE bands were closely related to uncultured bacteria. Thus, the bacterial community structure in black soil was stable, and the predominating bacterial groups were uncultured.  相似文献   

14.
In gold mining regions, the risk of soil pollution by mercury is a major environmental hazard, especially in tropical areas where soil microflora plays a major part in soil functioning, major bio-geochemical cycles and carbon turn-over. The impact of mercury pollution on soil microflora should thus be carefully assessed in such environments while taking into consideration the specificities of tropical soils. The aim of this study was to compare the effects of mercury (0, 1 and 20 μg of inorganic mercury per gram of soil) on the functional diversity and genetic structure of microbial communities in a tropical soil. We investigated the effects of mercury on tropical soil microflora using soil microcosms spiked with mercury and incubated at 28 °C for 1 month. Microcosm flora, its biomass and its activity, as well as its functional and genetic structure, were followed by cultural methods, measures of respiration, ECOLOG plates, and DGGE (denaturing gel gradient electrophoresis), respectively. Fate of total and bioavailable mercury was estimated by CVAFS (cold vapor atomic fluorescence spectrometry). Results obtained for the microcosms enriched with only 1 μg g?1 mercury were indistinguishable from controls. Conversely, in the presence of high mercury contents (20 μg g(1), an immediate effect was measured on soil respiration, functional diversity (ECOLOG plates) and genetic structure (DGGE), although no significant effect was observed on plate counts or microbial biomass. In addition, whereas microbial activities (respiration and functional diversity) rapidly regained control values, a lasting effect of the high mercury concentration was observed on the genetic structure of the soil microbial community. These modifications took place during the first week of incubation when total mercury concentration was declining and bioavailable mercury was at its highest.This multiple approach study is one of the first attempts at investigating the effects of mercury on soil microbial communities in tropical soils. Our results demonstrate that in the tropical soil under study, mercury affects the soil microbial communities in a different manner than was previously reported in temperate soils. Furthermore, mercury toxicity on soil microbes may be modulated by typical tropical soil characteristics.  相似文献   

15.
土壤微生物多样性研究的DGGE/TGGE技术   总被引:1,自引:0,他引:1  
《核农学报》2009,23(4):721-727
分子生物学技术比传统的培养方法可得到土壤微生物种群多样性更全面的信息。变性梯度凝胶电泳(Denaturing gradient gel electrophoresis, DGGE)和温度梯度凝胶电泳(Temperature gradient gel electrophoresis, TGGE)可分离PCR扩增的DNA片段,已成为研究土壤微生物群落多样性的重要手段。本文综述了DGGE/TGGE技术在土壤微生物多样性研究中的应用进展,分析了该方法的主要影响因素及其优点和存在的问题。  相似文献   

16.
铜污染土壤线虫多样性的PCR-DGGE分析   总被引:11,自引:1,他引:10  
A wheat pot experiment was conducted under greenhouse conditions to assess the effect of copper contamination on soil nematode diversity by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method and morphological analysis. The soil was treated with CuSO4.5H2O at the following concentrations: 0, 50, 100, 200, 400, and 800 mg kg^-1 dry soil, and the soil samples were collected at wheat jointing and ripening stages. Nematode diversity index (H′) from morphological analysis showed no difference between the control and the treated samples in either of the sampling dates. At the wheat ripening stage, nematode diversity obtained by the PCR-DGGE method decreased noticeably in the Cu800 treatment in comparison with the control. With optimization of the method of nematode DNA extraction, PCR-DGGE could give more information on nematode genera, and the intensity of the bands could reflect the abundance of nematode genera in the assemblage. The PCR-DGGE method proved promising in distinguishing nematode diversity in heavy metal coritaminated soil.  相似文献   

17.
To gain insight into microbial function following increased atmospheric CO2 concentration, we investigated the influence of 9 years of enriched CO2 (600 μl litre−1) on the function and structural diversity of soil microorganisms in a grassland ecosystem under free air carbon dioxide enrichment (FACE), as affected by plant species (Trifolium repens L. and Lolium perenne L. in monocultures and mixed culture) and nitrogen (N) supply. We measured biomass and activities of enzymes covering cycles of the most important elements (C, N and P). The microbial community was profiled by molecular techniques of phospholipid fatty acid (PLFA) and denaturing gradient gel electrophoresis (DGGE) analysis. The enrichment in CO2 increased soil microbial biomass (+48.1%) as well as activities of invertase (+36.2%), xylanase (+22.9%), urease (+23.8%), protease (+40.2%) and alkaline phosphomonoesterase (+54.1%) in spring 2002. In autumn, the stimulation of microbial biomass was 25% less and that of enzymes 3–12% less than in spring. Strong correlations between activities of invertase, protease, urease and alkaline phosphomonoesterase and microbial biomass were found. The stimulation of microbial activity in the enriched atmosphere was probably caused by changes in the quantity and kind of root litter and rhizodeposition. The response of soil microorganisms to enriched CO2 was most pronounced under Trifolium monoculture and under greater N supply. The PLFA analysis revealed that total PLFA contents were greater by 24.7% on average, whereby the proportion of bioindicators representative of Gram‐negative bacteria increased significantly in the enriched CO2 under less N‐fertilized Lolium culture. Discriminant analysis showed marked differences between the PLFA profiles of the three plant communities. Shannon diversity indices calculated from DGGE patterns were greater (+12.5%) in the enriched CO2, indicating increased soil bacterial diversity. We conclude that greater microbial biomass and enzyme activity buffer the potential increase in C sequestration occurring from greater C addition in enriched CO2 due to greater mineralization of soil organic matter.  相似文献   

18.
Application of phosphate-solubilizing microorganisms (PSMs) has been reported to increase P uptake and plant growth. However, no information is available regarding the ecological consequences of the inoculation with PSMs. The effect of inoculation with phosphate-solubilizing fungal (PSF) isolates Aspergillus niger P39 and Penicillium oxalicum P66 on the bacterial communities in the rhizospheres of maize (Zea mays L. ‘Haiyu 6') and soybean (Glycine max Merr. ‘Heinong 35') was examined using culture-dependent methods as well as a culture-independent method, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Compared with the control, the number of culturable microbes for soybean was significantly greater with P39, whereas for maize, the same was significantly greater with P66. In addition, a greater number of microbes were found in the rhizosphere of maize compared with soybean. The fingerprint of DGGE for 16S rDNA indicated that inoculation with PSF also increased bacterial communities, with the P66 treatment having higher numbers of DGGE bands and a higher Shannon-Weaver diversity index compared with P39; the composition of the microbial community was also more complex with the P66 treatment. Overall, complex interactions between plant species and exotic PSMs affected the structure of the bacterial community in the rhizosphere, but plant species were more important in determining the bacterial community structure than the introduction of exotic microorganisms.  相似文献   

19.
Abstract

We studied the effects of the application of organic matter (OM) and chemical fertilizer (CF) on soil alkaline phosphatase (ALP) activity and ALP-harboring bacterial communities in the rhizosphere and bulk soil in an experimental lettuce field in Hokkaido, Japan. The ALP activity was higher in soils with OM than in soils with CF, and activity was higher in the rhizosphere for OM than in the bulk soil. Biomass P and available P in the soil were positively related to the ALP activity of the soil. As a result, the P concentration of lettuce was higher in OM soil than in CF soil. We analyzed the ALP-harboring bacterial communities using polymerase chain reaction based denaturing gradient gel electrophoresis (DGGE) on the ALP genes. Numerous ALP genes were detected in the DGGE profile, regardless of sampling time, fertilizer treatment or sampled soil area, which indicated a large diversity in ALP-harboring bacteria in the soil. Several ALP gene fragments were closely related to the ALP genes of Mesorhizobium loti and Pseudomonas fluorescens. The community structures of the ALP-harboring bacteria were assessed using principal component analysis of the DGGE profiles. Fertilizer treatment and sampled soil area significantly affected the community structures of ALP-harboring bacteria. As the DGGE bands contributing to the principal component were different from sampling time, it is suggested that the major bacteria harboring the ALP gene shifted. Furthermore, there was, in part, a significant correlation between ALP activity and the community structure of the ALP-harboring bacteria. These results raise the possibility that different ALP-harboring bacteria release different amounts and/or activity of ALP, and that the structure of ALP-harboring bacterial communities may play a major role in determining overall soil ALP activity.  相似文献   

20.
土壤微生物多样性研究的DGGE/TGGE技术进展   总被引:2,自引:0,他引:2  
分子生物学技术比传统的培养方法可得到土壤微生物种群多样性更全面的信息。变性梯度凝胶电泳(Denaturing gradient gel electrophoresis,DGGE)和温度梯度凝胶电泳(Temperature gradient gel electrophoresis,TGGE)可分离PCR扩增的DNA片段,已成为研究土壤微生物群落多样性的重要手段。本文综述了DGGE/TGGE技术在土壤微生物多样性研究中的应用进展,分析了该方法的主要影响因素及其优点和存在的问题。  相似文献   

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