• <tr id="yyy80"></tr>
  • <sup id="yyy80"></sup>
  • <tfoot id="yyy80"><noscript id="yyy80"></noscript></tfoot>
  • 99热精品在线国产_美女午夜性视频免费_国产精品国产高清国产av_av欧美777_自拍偷自拍亚洲精品老妇_亚洲熟女精品中文字幕_www日本黄色视频网_国产精品野战在线观看 ?

    Lysobacter hymeniacidonis sp. nov., Isolated from a Crude Oil-Contaminated Marine Sponge

    2015-04-01 02:11:34XINYanjuanQUJungeXUJunyiWUPeichunCAOXupengandXUESong
    Journal of Ocean University of China 2015年6期

    XIN Yanjuan, QU Junge, XU Junyi, WU Peichun, CAO Xupeng, and XUE Song, *

    ?

    sp. nov., Isolated from a Crude Oil-Contaminated Marine Sponge

    XIN Yanjuan1), QU Junge2), XU Junyi3), WU Peichun1), CAO Xupeng1), and XUE Song1), *

    1) Marine Bioproducts Engineering Group, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, P.R. China?2)Department of Biology and Pharmacy, Zhejiang Pharmaceutical College, Ningbo315100, P.R. China?3)Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian 116001,P.R. China

    An aerobic, Gram-negative bacterium, strain 2-5T, was isolated from a crude oil-contaminated marine sponge collected near Dalian Bay, China, and subjected to a polyphasic taxonomic investigation. Cells of strain 2-5Twere non-spore forming, non-motile, rods 0.2–0.3μm wide and 1.1–1.2μm long. Strain 2-5Tgrew well on nutrient agar, TSA, R2A agar and LB agar. Colonies of strain 2-5Ton LB agar were circular, smooth with entire margins, non-transparent and pale yellow after 3d of incubation at 30℃. Growth of strain 2-5Toccurred in LN medium with 0–6% NaCl; no growth occurred in the presence of 8.0% NaCl. Strain 2-5Tgrew at 15–42℃ and at pH 6.0–8.0. Comparative 16S rRNA gene sequence analysis showed that strain 2-5Tclustered with the species of the genus. Its closet neighbors were the type strains ofKCTC 12205T(97% similarity),ZS79T(96%), andAPB-9T(96%). The value for DNA-DNA relatedness between strain 2-5TandKCTC 12205Twas 23%. Branched fatty acids iso-C16: 0, iso-C15: 0, iso-C11: 03-OH, iso-C17: 1ω9and iso-C11: 0were found to be predominant. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. Strain 2-5Thad a DNA G+C content of 63.8mol%. On the basis of the phenotypic, chemotaxonomic, DNA-DNA hybridization and phylogenetic data, strain 2-5Trepresents a novel species of the genus, for which the namesp. nov. is proposed. The type strain is 2-5T(=CGMCC 1.12190T= JCM 18137T).

    sp. nov.; taxonomy; phylogenetic analysis

    1 Introduction

    The genuswas first described by Christensen and Cook (1978), and the description was emended by Park. (2008). The genus, grouped in the family, is classified in the class(Christensen and Cook, 1978) based on non-fruiting bodies, lack of flagella, gliding nature and high genomic DNA G+C content (typically ranging between 65.4 and 70.1mol%) (Aslam., 2009). At the time of writing, the genuscomprises 27 species with validly published names (http: //www. bacterio. cict. fr/). Recently, three novel species of the genus,ZS79Tfrom iron-mined soil (Luo., 2012),107-E2Tfrom an Antarctic freshwater lake (Fukuda., 2013) andCJ29Tfrom ginsengsoil (Choi., 2014) have been described. Members of the genus were of great potential for the de-velopment of biocontrol agents against plant fungal pathogens (Islam., 2005; Park., 2008) and antibiotic bioactivities against human pathogens (Ahmed., 2003; Hashizume., 2004). Here we report the characterization of a novel marine bacterium, strain 2-5T, of the genus,which was isolated from a crude oil-contaminated marine sponge.

    2 Materials and Methods

    2.1 Bacterial Strains

    Strain 2-5Twas isolated from a crude oil-contaminated sponge specimen (), collected at the inter-tidal beach of Dalian, on the Chinese Yellow Sea, located in northern China (38?52′N 121?41′E). Freshly collected sponge specimens were rinsed five times in sterile seawater to remove unassociated bacteria and then thoroughly homogenized in a sterile mortar. A 10-fold dilution series of sponge homogenate was made and plated on 2216E plates (Difco, USA) in triplicate. After incubating the plates at 28℃ for 7d, an isolate, designated 2-5T, was picked and sub-cultured on 2216E plates, repeating the above steps until a pure culture was obtained. The novel strain was deposited into the CGM CC (China General Microbiological Culture Collection Center) as CGMCC 1.12190Tand the JCM (Japan Collection of Microorganisms) as JCM 18137T. The reference strain used for the DNA-DNA homology tests wasKCTC 12205T, obtained from the KCTC (Korean Collection for Type Cultures).

    2.2 Morphology and Physiological Characteristics

    To investigate the morphological and physiological characteristics, cell grown on R2A agar plates at 30℃ for 2d. Cell morphology and motility were examined using light microscopy (Olympus; ×1000) and transmission electron microscopy (H-7650; Hitachi, Japan) using cells from an exponentially growing culture. The strain Gram straining reaction was carried out according to the classical procedure described by Doetsch (1981). Gliding motility was determined as described by Bowman (2000). The physiological properties of strain 2-5Twere determined by the CGMCC using established procedures described by Gordon. (1974) and Yokota. (1993). Catalase activity, oxidase activity, enzyme activity and acid production from different carbohydrates were determined by the CGMCC with Biolog GN2, API 20E, API 20NE kits according to the manufacturer’s instructions. The assimilation of single carbon substrates was determined by the CGMCC with Biolog GN2 and API 20NE strips cultured at 28℃ for 24 h. Hydrolysis of casein and chitin was determined using previously described test (Smibert and Krieg, 1994; Brown, 2007). Growth at 4, 10, 15, 25, 30, 37, 42 and 45℃ and at pH 4.0–10.0 (at intervals of 1.0pH unit) was assessed after 5d of incubation on 2216E agar. Growth on nutrient agar, trypticase soy agar (TSA; Difco), R2A agar (Difco, USA) and LB agar (Difco, USA) was also evaluated at 28℃. Salt tolerance was tested in LN medium (LB without NaCl) supplemented with 0–10% NaCl after 10d of incubation.

    2.3 G+C Content and Analysis of Cellular Fatty Acids

    Cell grown on R2A agar plates at 30℃ for 2d were used for the analysis of cellular fatty acid and polar lipid. The fatty acids were extracted, methylated and analysed using the standard Sherlock MIDI (Microbial Identification) system (Sasser, 1990; K?mpfer and Kroppenstedt, 1996). Polar lipids were extracted and analyzed as described by Tindall (1990). A 6.75mL portion of chloroform/methanol/0.3% aqueous NaCl (1: 2: 0.8) was added to 100 mg freeze-dried cell material. The preparation was stirred overnight and cell debris was pelleted by centrifugation. Polar lipids were recovered into the chloroform phase by adjusting the chloroform/methanol/0.3% aqueous NaCl mixture to a ratio of 1: 1: 0.9 and then dried under nitrogen. The dried polar lipids were resuspended in chloroform/methanol (2:1) and separated by two-dimensional TLC.

    The G+C content of the genomic DNA was determined by the CGMCC by thermal denaturation (Mandel and Marmur, 1968), for whichK-12 (CGMCC 1.365) was used as a standard.

    2.4 Phylogenetic Analysis and DNA-DNA Hybridization

    Genomic DNA from strain 2-5Twas extracted and purified according to standard procedures (Sambrook and Russell, 2001). The 16S rRNA gene cloned into pMD- 18T (Takara, Japan) was sequenced using an automated sequencer (Applied Biosystems model 3730). The 16S rRNA gene sequence of strain 2-5Twas compared with known sequences found in the GenBank database using the BLAST program (http://blast.ncbi.nlm.nih.gov/ Blast.cgi). Phylogenetic analysis was performed with MEGA5 program (Tamura., 2011) after multiple alignments of the data via CLUSTAL_X program (Thom- pson., 1997). A distance matrix method (distance options according to the Kimura two-parameter model), including clustering using the neighbor-joining and maximum-likelihood (Kluge and Farris, 1969) method. In each case, bootstrap values were calculated based on 1000 replications (Felsenstein, 1985). The taxonomic relationship between strain 2-5Tand its phylogenetic relative was further examined using DNA-DNA hybridization. DNA–DNA hybridization values between 2-5TandKCTC 12205Twas performed fluorometrically, according to the method developed by Ezaki. (1989) using photobiotin-labelled DNA probes and micro dilution wells. Hybridization was conducted in five replications for each sample. The highest and lowest values obtained for each sample were excluded, and the remaining three values were utilized in the calculation of hybridization values.

    3 Results and Discussion

    3.1 Morphological and Physiological Characteristics

    Cells of strain 2-5Twere Gram-negative, non-spore forming, non-motile (but showing gliding activity), aerobic rods 0.2–0.3μm wide and 1.1–1.2μm long (Supplementary Fig.S1-2). Strain 2-5Tgrew well on nutrient agar, TSA, R2A agar and LB agar. Colonies of strain 2-5Ton LB agar were circular, smooth with entire margins, non- transparent and pale yellow after 3d of incubation at 30℃. Growth of strain 2-5Toccurred in LN medium with 0–6% NaCl; no growth occurred in the presence of 8.0% NaCl. Strain 2-5Tgrew at 15–42℃ (optimal 28–30℃) and at pH 6.0–8.0 (optimum pH 7.0). In the Biolog GN2, API 20E and API 20NE kits, strain 2-5Tassimilated glycogen,-acetyl-D-galactosamine,-acetylglucosamine, ribitol, L-arabinose, D-arabitol, D-cellobiose,D-fructose, alpha-D-glucose, M-inositol,D-galactonic acid lactone,D-galacturonic acid, D, L-lactic acid, malonic acid, propionic acid, quinic acid, D-saccharic acid, succinamic acid, glucuronamide, L-ala- ninamide, L-alanine, L-alanyl-glycine, L-asparagine, L- aspartic acid, glycyl-L-aspartic acid, glycyl-L-glutamic acid, hydroxy-L-proline.The phenotypic characteristics of strain 2-5Tare summarized in the species description and a comparison of selective characteristics with refe- rence strains are summarized in Table 1.

    Table 1 Physiological and biochemical characteristics that differentiate strain 2-5T from related type strains of the genus Lysobacter

    Notes: Strains: 1, 2-5T(data from this study); 2,KCTC 12205T(Bae., 2005); 3,ZS79T(Lue., 2012); 4,DSM 18482T(Yassin., 2007). +, positive; -, negative; ND, no data available.

    3.2 G+C content and Analysis of Cellar Fatty Acids

    Table 2 Cellular fatty acid compositions (%) of strain 2-5T and related type strains of the genus Lysobacter.

    Notes: Strains: sames as those in Table 1. -, <1% or not detected.

    The major cellular fatty acids of strain 2-5Twere iso- C16: 0(15.1%), iso-C15: 0(14.4%), iso-C17: 1ω9(9.9%), iso-C11: 03-OH (8.4%) and iso-C11: 0(7.7%) (Table 2). The presence of branched fatty acids, namely iso-C16:0, iso-C15: 0, iso-C17:1ω19and iso-C17: 0, was consistent with the placement of strain 2-5Twithin the genus(Bae., 2005; Weon., 2006, 2007; Romanenko., 2008). However, strain 2-5Tcontained C16:0ω7/16:1 ω6and C19:0cyclo ω8, which differences with the reference strains. The predominant polar lipid of strain 2-5Twere phosphatidylethanolamine (PE), phosphatidylglycerol (PG) and diphosphatidylglyceroal (DPG) (Supplementary Fig.S3). The DNA G+C content of strain 2-5Twas 63.8 mol%. This value is within the range reported for the genus(Christensen and Cook, 1978; Aslam., 2009).

    3.3 Phylogenetic Analysis and DNA-DNA Hybridization

    An almost-complete 16S rRNA gene sequence of strain 2-5Tconsisting of 1436bp was obtained. The phylogenetic tree shows that strain 2-5Tclusters within the genusin the class Gammaproteobacteria(Fig.1). 16S rRNA gene sequence analysis revealed that strain 2-5Twas related most closely to.Ko07T(97%similarity),.ZS79T(96%), and.APB-9T(96%). Based on the above results, DNA- DNA hybridization experiments were carried out between strain 2-5TandKo07T. The value obtained was 23%, which is significantly below the value of 70% proposed by Wayne. (1987) for species discrimination.

    3.4 Toxonomic Conclusion

    It is clear from the 16S rRNA gene sequence comparison and DNA–DNA hybridization data that strain 2-5Trepresents a novel species of the genus(Wayne., 1987). In addition, strain 2-5Twas positive for L-Arabinose, D-Mannitol, N-Acetylglucosamine, Maltose, and Trisodium citrate, differs from the type strain.Ko07T. Based on the phenotypic, phylogenetic and genomic evidence, strain 2-5Twas identified as a novel species of the genus, for which the name.sp. nov. is proposed.

    3.5 Description ofsp. nov.

    (hy.me.ni.a.ci?do.nis. N.L. gen. n.of, the generic name of the marine sponge, the source of the type strain).

    Cells are Gram-negative, aerobic, non-spore forming, non-motile (but show gliding activity) and rod-shaped,about 0.2–0.3μm wide and 1.1–1.2μmlong. Colonies grown on LB agar are convex, circular, smooth, non- transparent and pale yellow after 3d of incubation at 30℃. Grows at 15–37℃ (optimal 28–30℃); no growth occurs below 4℃ or above 45℃. The pH range for growth is 6.0–8.0 (optimal pH 7.0). Growth occurs in the absence of NaCl and no growth occurs in 8.0% NaCl. Oxidase and catalase-positive. Positive for acid production from glucose, but negative for reduction of nitrates to nitrites. Hydrolyses aesculin, gelatin and casein, but not starch and chitin. Positive for citric acid utilization andacetoin production (Voges–Proskauer reaction). Activities of beta-galactosidase, arginine dihydrolase, lysine decarboxylase, ornithine decarboxylase, urease and tryptophan deaminase are negative (API 20 E).Negative for assimilation of dextrin, tween40, tween80, I-erythritol, L-fruc- tose, D-galactose, dextrinose, D-mannose, D-allulose, sucrose, methyl pyruvate, succinic acid methyl, betaphenylglycollic acid, itaconic acid, alpha-oxo acid, sebacic acid, succinic acid, bromosuccinic acid, D-alanine, L-histidine, D-serine, gamma-amino butyric acid, rocanic acid, thymidine, phenyl ethylamine, putrescine, 2-amino ethanol, 2, 3-butanediol, glycerol, glycerol phosphate, D-glucose-1-phosphate, D-glucose-6-phosphate. Utilizes alpha-Cyclodextrin, glycogen, N-acetylgalactosamine, N- acetylglucosamine, ribitol, L-arabinose, D-arabitol, D- cellobiose, D-fructose, D-glucose, m-inositol,alpha-D- lactose, lactulose, maltose, D-mannitol, D-melibiose, beta-methyl-D-glucoside, D-raffinose, L-rhamnose, D- sorbitol, D-trehalose, turanose, xylitol, acetic acid, cis-aconitic acid, citric acid, formic acid, D-galactose acid lactone, D-galacturonic acid, D-gluconic acid, beta- methyl glucoside, D-glucuronic acid, alpha-hydroxy butyric acid, beta-hydroxy butyric acid, gamma-hydroxy butyric acid,alpha-oxo-butanoic acid,alpha-oxo-penta- nedioic acid,D,L-lactic acid, malonic acid, propionic acid, quinic acid, D-saccharinic acid, succinamic acid, glucuronamide, L-alanyl amine, L-alanine, L-alanylglycine, L-asparagine, glycyl aspartic acid, glycyl glutamic acid, hydroxy proline, L-leucine, L-ornithine, L-phenylalanine, L-proline, L-pyroglutamic acid, L-serine, L-threonine, D, L-carnitine, inosine, uridine(API 20 NE and Biolog GN2).

    Fig.1 Phylogenetic tree of strain 2-5T and the type strains of related taxa based on 16S rRNA gene sequences. The tree wasreconstructed using the neighbor-joining (NJ) and maximum-likelihood (ML) methods and numbers at nodes representbootstrap percentages (NJ/ML, based on 1000 resamplings). Filled circles indicate genetic branches that were present in boththe NJ and ML trees. GenBank accession numbers are given in parentheses. Bootstrap values over 50% are shown at branching points. Bar, 0.005 substitutions per nucleotide position.

    The major cellular fatty acids of strain 2-5Twere iso- C16:0, iso-C15:0, iso-C17:1ω9and iso-C11:03-OH; detailed fatty acid compositions are given in Table 2. The polar lipids consist of phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglyceroal. The genomic DNA G+C content of the type strain is 63.8 mol%. The type strain 2-5T(=CGMCC 1.12190T= JCM 18137T), was isolated from a crude oil-contaminated marine spon- ge in Dalian, China.

    Acknowledgements

    This work was supported by the National Natural Science Foundation of China (31100092), the Hundred Talent Program of the Chinese Academy of Sciences (No. A1097) and the Ningbo Natural Science Foundation of China (2011A610028).

    Supplementary

    Fig.S1 Transmission electron micrograph of strain 2-5.

    Fig.S2 Scanning electron micrograph of strain 2-5.

    Fig.S3 Two-dimensional TLC of polar lipids of strains 2-5Tstained with 5% ethanolic molybdophosphoric acid. PE, phosphatidylethanolamine; PG, phosphatidylglycerol; DPG, diphosphatidylglycerol.

    Ahmed, K., Chohnan, S., Ohashi, H., Hirata, T., Masaki, T., and Sakiyama, F., 2003. Purification, bacteriolytic activity, and specificity of β-lytic protease fromsp. IB-9374., 95: 27-34.

    Aslam, Z., Yasir, M., Jeon, C. O., and Chung, Y. R., 2009.sp. nov., isolated from the rhizosphere of rice (L.)., 59: 675-680.

    Bae, H.-S., Im, W.-T., and Lee, S.-T. 2005.sp. nov., isolated from anaerobic granules in an upflow anaerobic sludge blanket reactor., 55: 1155- 1161.

    Bowman, J. P., 2000. Description ofsp. nov., isolated from the surfaces of Antarctic algae, and reclassification of(ZoBell and Upham 1944) Reichenbach 1989 ascomb. nov., 50: 1861-1868.

    Brown, A. E., 2007.. New York, McGraw-Hill, 20-42.

    Christensen, P., and Cook, F. D., 1978., a new genus of nonfruiting, gliding bacteria with a high base ratio., 28: 367-393.

    Choi, J. H., Seok, J, H., Cha, J. H., and Cha, C. J., 2014.sp. nov., isolated fromginseng soil., 64: 2193-2197

    Doetsch, R. N., 1981. Determinative methods of light microscopy. In:, Gerhardt, P.,., eds., American Society for Microbiology, Washington, DC, 21-33.

    Ezaki, T., Hashimoto, Y., and Yabuuchi, E., 1989. Fluorometric deoxyribonucleic acid-deoxyribonucleic acid hybridization in microdilution wells as an alternative to membrane filter hybridization in which radioisotopes are used to determine genetic relatedness among bacterial strains., 39: 224-229.

    Felsenstein, J., 1985. Confidence limits on phylogenies: an approach using the bootstrap., 39: 783-791.

    Fukuda, W., Kimura, T., Araki, S., Miyoshi, Y., Atomi, H., and Imanaka, T., 2013.sp. nov., isolated from an Antarctic freshwater lake in Antarctica., 63: 3313-3318.

    Gordon, R. E., Barnett, D. A., Handerhan, J. E., and Pang, C. H. N., 1974.,, and the nocardin strain., 24: 54-63.

    K?mpfer, P., and Kroppenstedt, R. M., 1996. Numerical analysis of fatty acid patterns of coryneform bacteria and related taxa., 42: 989-1005.

    Kluge, A. G.,and Farris, F. S., 1969. Quantitative phyletics and theevolution of anurans., 18: 1-32.

    Hashizume, H., Hattori, S., Igarashi, M., and Akamatsu, Y., 2004. Tripropeptin E, a new tripropeptin group antibiotic produced bysp. BMK333-48F3., () 57: 394-399.

    Islam, M. T., Hashidoko, Y., Deora, A., Ito, T. and Tahara, S., 2005. Suppression of damping-off disease in host plants by the rhizoplane bacteriumsp. strain SB-K88 is linked to plant colonization and antibiosis against soilborne peronosporomycetes., 71: 3786-3796.

    Luo, G., Shi, Z.,and Wang, G., 2012.sp. nov.,an arsenite-resistant bacterium isolated from iron-mined soil., 62: 1659-1665.

    Mandel, M., and Marmur, J., 1968. Use of ultraviolet absorbancetemperature profile for determining the guanine plus cytosine content of DNA., 12B: 195-206.

    Park, J. H., Kim, R., Aslam, Z., Jeon, C. O., and Chung, Y. R., 2008.sp. nov., with antimicrobial activity, isolated from the rhizosphere of pepper, and emended description of the genus., 58: 387-392.

    Romanenko, L. A., Uchino, M., Tanaka, N., Frolova, G. M., and Mikhailov, V. V., 2008.sp. nov., isolated from a deep-sea sponge., 58: 370-374.

    Sasser, M., 1990. Identification of bacteria by gas chromatography of cellular fatty acids. Technical Note 101. Newark, DE: MIDI.

    Sambrook, J., and Russell, D. W., 2001.:, 3rd edtion. Cold Spring Harbor, Cold Spring Harbor Laboratory, 12-29.

    Smibert, R. M., and Krieg, N. R., 1994. Phenotypic characterization. In:, Gerhardt, P.,., eds., American Society for Microbiology, Washington, DC, 607-655

    Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., and Kumar,S., 2011. MEGA5: Molecular evolutionary genetics analysis usingmaximum likelihood, evolutionary distance, and maximum parsimonymethods., 28: 2731-2739.

    Tindall, B. J., 1990. A comparative study of the lipid composition offrom various sources., 13: 128-130.

    Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F., and Higgins, D. G., 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools., 25: 4876- 4882.

    Wayne, L. G., Brenner, D. J., Colwell, R. R., Grimont, P. A. D.,Kandler, O., Krichevsky, M. I., Moore, L. H., Moore, W. E. C., Murray,R. G. E., 1987. International Committee onSystematic Bacteriology. Report of the ad hoc committee onreconciliation of approaches to bacterial systematics., 37: 463- 464.

    Weon, H. Y., Kim, B. Y., Baek, Y. K., Yoo, S. H., Kwon, S. W.,Stackebrandt, E., and Go, S. J., 2006. Two novel species,sp. nov. andsp. nov., isolatedfrom Korean greenhouse soils., 56: 947-951.

    Weon, H. Y., Kim, B. Y., Kim, M. K., Yoo, S. H., Kwon, S. W., Go, S. J., and Stackebrandt, E., 2007.sp. nov. andsp. nov., isolated from greenhouse soils inKorea., 57: 548-551.

    Yassin, A. F., Chen, W. M., Hupfer, H., Siering, C., Krop- penstedt,R. M., Arun, A. B., Lai, W. A., Shen, F. T., Rekha, P. D., and Young, C. C., 2007.sp. nov., isolated from municipal solidwaste., 57: 1131-1136.

    Yokota, A., Tamura, T., Hasegawa, T., and Huang, L. H., 1993.gen. nov., sp. nov., nom. rev., a new genus of the order Actinomycetales., 43: 805-812.

    (Edited by Ji Dechun)

    DOI 10.1007/s11802-015-2789-4

    ISSN 1672-5182, 2015 14 (6): 1019-1024

    ? Ocean University of China, Science Press and Springer-Verlag Berlin Heidelberg 2015

    (October 27, 2014; revised August 14, 2015; accepted August 30, 2015)

    * Corresponding author. Tel: 0086-411-84379069 E-mail:xuesong@dicp.ac.cn

    久久久久精品人妻al黑| 老汉色av国产亚洲站长工具| 天堂中文最新版在线下载| 色94色欧美一区二区| 最近中文字幕2019免费版| 一级,二级,三级黄色视频| 久久精品亚洲av国产电影网| 99香蕉大伊视频| 一级片'在线观看视频| 好男人视频免费观看在线| 三级国产精品片| 一个人免费看片子| 国产一区二区激情短视频 | 久久久久久久久免费视频了| 亚洲国产色片| 午夜精品国产一区二区电影| 久久精品aⅴ一区二区三区四区 | 超碰成人久久| 韩国精品一区二区三区| 1024香蕉在线观看| 国产免费现黄频在线看| 国产色婷婷99| 王馨瑶露胸无遮挡在线观看| 国产成人a∨麻豆精品| 亚洲婷婷狠狠爱综合网| av卡一久久| 一区二区三区乱码不卡18| 欧美国产精品va在线观看不卡| 日韩欧美精品免费久久| 一区二区三区四区激情视频| 久久午夜综合久久蜜桃| 亚洲欧洲精品一区二区精品久久久 | 又粗又硬又长又爽又黄的视频| 亚洲精品国产一区二区精华液| 亚洲欧洲精品一区二区精品久久久 | 午夜福利在线免费观看网站| 香蕉丝袜av| 国产有黄有色有爽视频| 精品人妻偷拍中文字幕| 日本色播在线视频| 精品国产超薄肉色丝袜足j| 99久久人妻综合| 性高湖久久久久久久久免费观看| 亚洲天堂av无毛| 观看美女的网站| 亚洲三区欧美一区| 欧美黄色片欧美黄色片| 大香蕉久久成人网| 成人漫画全彩无遮挡| 麻豆乱淫一区二区| 亚洲视频免费观看视频| 这个男人来自地球电影免费观看 | 秋霞在线观看毛片| 日韩在线高清观看一区二区三区| 欧美日韩视频精品一区| 人人妻人人添人人爽欧美一区卜| 蜜桃国产av成人99| 日本黄色日本黄色录像| 免费在线观看黄色视频的| 一区二区三区四区激情视频| 日本91视频免费播放| 国产精品av久久久久免费| 色婷婷av一区二区三区视频| 2021少妇久久久久久久久久久| 精品亚洲成国产av| 黄频高清免费视频| 亚洲综合色网址| 欧美国产精品va在线观看不卡| 欧美在线黄色| 亚洲精品,欧美精品| av一本久久久久| 免费观看性生交大片5| 美女xxoo啪啪120秒动态图| 伦理电影免费视频| 高清不卡的av网站| 亚洲精品日本国产第一区| 青春草国产在线视频| 乱人伦中国视频| 成年女人在线观看亚洲视频| 久久久久国产网址| 欧美日韩一区二区视频在线观看视频在线| 天天躁夜夜躁狠狠久久av| 观看av在线不卡| 亚洲精品国产一区二区精华液| 2021少妇久久久久久久久久久| 国产又爽黄色视频| 久久久久久久久久久久大奶| 搡老乐熟女国产| 国产日韩欧美视频二区| 亚洲精华国产精华液的使用体验| av在线观看视频网站免费| 天堂8中文在线网| 国产精品久久久久久久久免| 人体艺术视频欧美日本| 大码成人一级视频| 国产精品免费视频内射| 亚洲第一青青草原| 日本午夜av视频| 一二三四中文在线观看免费高清| 国产精品麻豆人妻色哟哟久久| 欧美xxⅹ黑人| 久久久久久久久免费视频了| 伊人久久大香线蕉亚洲五| 亚洲人成77777在线视频| 国产成人免费观看mmmm| 亚洲av免费高清在线观看| 一区福利在线观看| 亚洲精品,欧美精品| 最近中文字幕高清免费大全6| 久久久久久久国产电影| 大陆偷拍与自拍| 久久久久视频综合| 日韩 亚洲 欧美在线| 熟女av电影| 欧美人与性动交α欧美精品济南到 | 欧美亚洲日本最大视频资源| 国产有黄有色有爽视频| 在线观看三级黄色| 国产精品一二三区在线看| 亚洲在久久综合| 最新中文字幕久久久久| 高清欧美精品videossex| 国产精品香港三级国产av潘金莲 | 国产成人精品无人区| 一级毛片电影观看| 亚洲精品在线美女| 亚洲经典国产精华液单| 亚洲av.av天堂| 午夜福利视频精品| 97人妻天天添夜夜摸| 日韩大片免费观看网站| 久久青草综合色| 欧美成人精品欧美一级黄| 伦精品一区二区三区| av.在线天堂| 精品国产乱码久久久久久小说| 久久久久国产一级毛片高清牌| 一本久久精品| 一个人免费看片子| 纵有疾风起免费观看全集完整版| 久久久国产一区二区| 下体分泌物呈黄色| 亚洲欧美色中文字幕在线| 午夜福利在线观看免费完整高清在| 18禁裸乳无遮挡动漫免费视频| 色播在线永久视频| 久久久精品区二区三区| 国产av一区二区精品久久| 美女大奶头黄色视频| 熟女少妇亚洲综合色aaa.| av不卡在线播放| 久热久热在线精品观看| 伊人久久大香线蕉亚洲五| 国产在线免费精品| 亚洲成色77777| 制服诱惑二区| 亚洲国产欧美在线一区| 亚洲国产欧美网| 久久人人爽人人片av| 晚上一个人看的免费电影| 亚洲欧美色中文字幕在线| 深夜精品福利| 一级毛片黄色毛片免费观看视频| 丝袜人妻中文字幕| 日韩制服骚丝袜av| 多毛熟女@视频| 热re99久久国产66热| 日日撸夜夜添| 亚洲欧洲日产国产| 国产精品嫩草影院av在线观看| 晚上一个人看的免费电影| 欧美变态另类bdsm刘玥| 国产熟女欧美一区二区| 天天操日日干夜夜撸| 久久精品久久久久久噜噜老黄| 人人澡人人妻人| 亚洲欧美一区二区三区国产| 老熟女久久久| 久久久久精品人妻al黑| 亚洲一区二区三区欧美精品| 国产精品人妻久久久影院| 好男人视频免费观看在线| 18禁观看日本| 人妻一区二区av| 国产精品 国内视频| 18禁裸乳无遮挡动漫免费视频| 国产1区2区3区精品| 老汉色∧v一级毛片| 午夜免费男女啪啪视频观看| 菩萨蛮人人尽说江南好唐韦庄| 欧美精品亚洲一区二区| 国产免费现黄频在线看| √禁漫天堂资源中文www| 狠狠精品人妻久久久久久综合| 亚洲欧美精品自产自拍| 黄片播放在线免费| 国产成人精品一,二区| 欧美亚洲 丝袜 人妻 在线| 9191精品国产免费久久| 久久毛片免费看一区二区三区| 久久99蜜桃精品久久| 亚洲情色 制服丝袜| 欧美日韩精品网址| 丝瓜视频免费看黄片| 人妻一区二区av| 国产成人精品久久二区二区91 | 精品99又大又爽又粗少妇毛片| 秋霞伦理黄片| 黑丝袜美女国产一区| 亚洲精品,欧美精品| 2018国产大陆天天弄谢| 亚洲 欧美一区二区三区| 亚洲精品第二区| 亚洲国产毛片av蜜桃av| 在线观看一区二区三区激情| 99re6热这里在线精品视频| 午夜福利在线观看免费完整高清在| 精品国产一区二区久久| 欧美精品av麻豆av| 一区二区三区四区激情视频| 国产日韩欧美视频二区| 波多野结衣av一区二区av| 伦理电影大哥的女人| 日本av免费视频播放| 丰满乱子伦码专区| 最近2019中文字幕mv第一页| 中文字幕人妻丝袜制服| 校园人妻丝袜中文字幕| 亚洲第一av免费看| 最黄视频免费看| 黄色视频在线播放观看不卡| 久久久久久久久久人人人人人人| 天美传媒精品一区二区| 日韩精品免费视频一区二区三区| 99久久中文字幕三级久久日本| 日韩中文字幕欧美一区二区 | 精品少妇黑人巨大在线播放| 国产精品欧美亚洲77777| 99久久人妻综合| 久久99精品国语久久久| 中文乱码字字幕精品一区二区三区| 国产在线视频一区二区| www.熟女人妻精品国产| 狂野欧美激情性bbbbbb| 免费看不卡的av| 大话2 男鬼变身卡| 搡女人真爽免费视频火全软件| 国产av国产精品国产| 另类精品久久| 久久毛片免费看一区二区三区| 最近最新中文字幕大全免费视频 | 日韩一区二区视频免费看| 午夜福利在线观看免费完整高清在| 国产精品秋霞免费鲁丝片| 91在线精品国自产拍蜜月| 激情视频va一区二区三区| 午夜影院在线不卡| 国产在线视频一区二区| 成人手机av| 黄片播放在线免费| 亚洲国产欧美日韩在线播放| 国产伦理片在线播放av一区| av在线观看视频网站免费| 久久久久久久精品精品| 中文字幕制服av| 国产高清不卡午夜福利| 日韩三级伦理在线观看| 亚洲精品日本国产第一区| 一区福利在线观看| 亚洲精品自拍成人| 国产精品女同一区二区软件| 精品99又大又爽又粗少妇毛片| 久久久国产精品麻豆| 一区二区av电影网| 最近手机中文字幕大全| 成人午夜精彩视频在线观看| 欧美日本中文国产一区发布| 国产又爽黄色视频| 精品人妻偷拍中文字幕| 啦啦啦在线免费观看视频4| 一个人免费看片子| 欧美精品人与动牲交sv欧美| 久久 成人 亚洲| 久久精品aⅴ一区二区三区四区 | 亚洲伊人久久精品综合| 制服人妻中文乱码| 老汉色∧v一级毛片| 蜜桃在线观看..| 久久99蜜桃精品久久| 日韩av免费高清视频| 曰老女人黄片| 高清视频免费观看一区二区| 国产av精品麻豆| 女人被躁到高潮嗷嗷叫费观| 国产一级毛片在线| 日韩一区二区视频免费看| 狠狠婷婷综合久久久久久88av| 欧美人与性动交α欧美精品济南到 | 七月丁香在线播放| 亚洲欧美精品综合一区二区三区 | 夫妻性生交免费视频一级片| 性少妇av在线| av又黄又爽大尺度在线免费看| 黄色视频在线播放观看不卡| 亚洲av在线观看美女高潮| 久久久久久久久久久免费av| 一本—道久久a久久精品蜜桃钙片| 欧美人与性动交α欧美软件| 999久久久国产精品视频| 久久久久久久亚洲中文字幕| 香蕉丝袜av| 777米奇影视久久| 免费日韩欧美在线观看| 午夜福利乱码中文字幕| 成人毛片60女人毛片免费| 国产精品欧美亚洲77777| 亚洲精品成人av观看孕妇| 一区福利在线观看| 亚洲精品久久久久久婷婷小说| 亚洲精品久久午夜乱码| 成年人午夜在线观看视频| 熟女av电影| 日韩三级伦理在线观看| 一边摸一边做爽爽视频免费| 男女无遮挡免费网站观看| 日本91视频免费播放| 久久精品国产亚洲av高清一级| 国产亚洲最大av| 一本大道久久a久久精品| 人妻 亚洲 视频| 国产熟女午夜一区二区三区| 久久韩国三级中文字幕| 自线自在国产av| 在线看a的网站| 大码成人一级视频| 丝袜脚勾引网站| 一本一本久久a久久精品综合妖精 国产伦在线观看视频一区 | 国产精品亚洲av一区麻豆 | 国产成人欧美| 久久女婷五月综合色啪小说| 国产精品免费大片| 亚洲精品中文字幕在线视频| 色网站视频免费| 午夜激情久久久久久久| 国产一级毛片在线| 满18在线观看网站| 中文字幕av电影在线播放| 精品少妇黑人巨大在线播放| xxxhd国产人妻xxx| 黄色配什么色好看| 久久 成人 亚洲| 岛国毛片在线播放| 久久韩国三级中文字幕| 久久女婷五月综合色啪小说| 午夜老司机福利剧场| 国产成人精品在线电影| 久久精品国产综合久久久| 国产免费现黄频在线看| 午夜日韩欧美国产| 色网站视频免费| 亚洲国产最新在线播放| 美女脱内裤让男人舔精品视频| 9191精品国产免费久久| 精品国产乱码久久久久久男人| 咕卡用的链子| 免费观看av网站的网址| 国产精品熟女久久久久浪| 天堂中文最新版在线下载| av不卡在线播放| 寂寞人妻少妇视频99o| 色婷婷久久久亚洲欧美| 亚洲av欧美aⅴ国产| 午夜激情av网站| 日韩人妻精品一区2区三区| 中文乱码字字幕精品一区二区三区| 久久99一区二区三区| 在线 av 中文字幕| 日韩人妻精品一区2区三区| 国产黄色视频一区二区在线观看| 久久久亚洲精品成人影院| 最近的中文字幕免费完整| 亚洲av中文av极速乱| 亚洲欧美日韩另类电影网站| 日本av免费视频播放| 丝袜在线中文字幕| 最新中文字幕久久久久| 男女午夜视频在线观看| 国产男人的电影天堂91| av视频免费观看在线观看| 亚洲精品国产色婷婷电影| 18禁国产床啪视频网站| 色视频在线一区二区三区| 亚洲国产日韩一区二区| 久久午夜福利片| 国产精品av久久久久免费| 一本大道久久a久久精品| 制服丝袜香蕉在线| 777久久人妻少妇嫩草av网站| 91午夜精品亚洲一区二区三区| 一级黄片播放器| 国产精品成人在线| 国产精品久久久久久久久免| 菩萨蛮人人尽说江南好唐韦庄| 男女午夜视频在线观看| 大片电影免费在线观看免费| 午夜免费观看性视频| 欧美另类一区| 亚洲在久久综合| 最近中文字幕高清免费大全6| 捣出白浆h1v1| 亚洲欧美一区二区三区久久| 最近最新中文字幕大全免费视频 | 热re99久久国产66热| 黑丝袜美女国产一区| 免费观看性生交大片5| 日韩精品免费视频一区二区三区| 精品福利永久在线观看| 纵有疾风起免费观看全集完整版| 国产精品 欧美亚洲| 美女视频免费永久观看网站| 满18在线观看网站| 最近中文字幕高清免费大全6| 青春草视频在线免费观看| 免费观看a级毛片全部| 中文字幕精品免费在线观看视频| 人妻一区二区av| 自拍欧美九色日韩亚洲蝌蚪91| 欧美成人午夜免费资源| freevideosex欧美| av不卡在线播放| 看十八女毛片水多多多| 天美传媒精品一区二区| 一区二区日韩欧美中文字幕| 日韩一卡2卡3卡4卡2021年| 999精品在线视频| 一级,二级,三级黄色视频| 一区二区三区乱码不卡18| 精品人妻熟女毛片av久久网站| 夫妻午夜视频| 十八禁高潮呻吟视频| 欧美精品一区二区免费开放| 最近的中文字幕免费完整| 免费高清在线观看视频在线观看| 精品99又大又爽又粗少妇毛片| 日韩制服骚丝袜av| 久久婷婷青草| 久久久久久久久免费视频了| 日韩伦理黄色片| 黄色怎么调成土黄色| 久久精品久久精品一区二区三区| 极品少妇高潮喷水抽搐| 国产一区二区三区av在线| 亚洲国产精品成人久久小说| 亚洲精品美女久久av网站| 2018国产大陆天天弄谢| 女人高潮潮喷娇喘18禁视频| 香蕉国产在线看| 少妇熟女欧美另类| 天天躁夜夜躁狠狠久久av| 国产片特级美女逼逼视频| 叶爱在线成人免费视频播放| 毛片一级片免费看久久久久| 久久精品人人爽人人爽视色| 免费观看性生交大片5| 人妻一区二区av| 人人妻人人添人人爽欧美一区卜| 蜜桃在线观看..| 国产人伦9x9x在线观看 | 69精品国产乱码久久久| 人妻系列 视频| 日产精品乱码卡一卡2卡三| 欧美成人午夜精品| 丝袜人妻中文字幕| 欧美日韩av久久| 亚洲一码二码三码区别大吗| 天天躁日日躁夜夜躁夜夜| 欧美日韩综合久久久久久| 热re99久久国产66热| 精品酒店卫生间| 久久精品久久精品一区二区三区| 免费大片黄手机在线观看| 午夜免费鲁丝| 尾随美女入室| 成人漫画全彩无遮挡| 亚洲国产看品久久| 国产一区亚洲一区在线观看| 巨乳人妻的诱惑在线观看| 精品久久蜜臀av无| av在线老鸭窝| av片东京热男人的天堂| av线在线观看网站| 欧美日韩一级在线毛片| 如何舔出高潮| 日本色播在线视频| 亚洲欧美一区二区三区国产| 1024视频免费在线观看| 日韩 亚洲 欧美在线| 精品人妻偷拍中文字幕| 狂野欧美激情性bbbbbb| 国产成人精品久久久久久| 亚洲 欧美一区二区三区| 亚洲精品美女久久久久99蜜臀 | 精品国产一区二区三区久久久樱花| 9191精品国产免费久久| 免费高清在线观看日韩| 王馨瑶露胸无遮挡在线观看| 欧美日本中文国产一区发布| 综合色丁香网| 亚洲国产看品久久| 国产一区二区三区av在线| 黄色视频在线播放观看不卡| 日韩熟女老妇一区二区性免费视频| 欧美精品av麻豆av| 少妇被粗大的猛进出69影院| 国产精品三级大全| 久热这里只有精品99| www.精华液| 久久久精品免费免费高清| 亚洲精品国产一区二区精华液| 成年动漫av网址| 亚洲欧洲日产国产| 热99久久久久精品小说推荐| 亚洲欧洲日产国产| 国产xxxxx性猛交| 亚洲精品乱久久久久久| 美女中出高潮动态图| 国产一区亚洲一区在线观看| 极品少妇高潮喷水抽搐| 七月丁香在线播放| 午夜福利视频在线观看免费| 久久这里只有精品19| 精品99又大又爽又粗少妇毛片| 最新中文字幕久久久久| 中文字幕亚洲精品专区| 99热国产这里只有精品6| 国产爽快片一区二区三区| 日韩一卡2卡3卡4卡2021年| 男女午夜视频在线观看| 日日撸夜夜添| 国产淫语在线视频| 一级毛片电影观看| 午夜福利乱码中文字幕| 伦理电影免费视频| 熟女av电影| 亚洲国产精品一区三区| 岛国毛片在线播放| 亚洲欧美清纯卡通| 成人亚洲欧美一区二区av| 王馨瑶露胸无遮挡在线观看| 国产精品av久久久久免费| 久久国内精品自在自线图片| 美女脱内裤让男人舔精品视频| 久久精品国产综合久久久| 久久久久精品人妻al黑| 精品少妇内射三级| 国产男女超爽视频在线观看| 亚洲国产最新在线播放| 女性生殖器流出的白浆| 久久午夜福利片| 欧美日韩综合久久久久久| av卡一久久| 丝袜美腿诱惑在线| 久久精品国产综合久久久| 欧美日韩综合久久久久久| 最近手机中文字幕大全| av网站免费在线观看视频| 一边摸一边做爽爽视频免费| 欧美人与性动交α欧美精品济南到 | 国产精品免费视频内射| 男女下面插进去视频免费观看| 男人爽女人下面视频在线观看| 电影成人av| 热re99久久精品国产66热6| 亚洲欧美成人综合另类久久久| 在线观看人妻少妇| 男人操女人黄网站| 99久国产av精品国产电影| 国产精品av久久久久免费| 久久影院123| 女人高潮潮喷娇喘18禁视频| 欧美日韩精品成人综合77777| 国产精品一二三区在线看| 国产极品天堂在线| 黄网站色视频无遮挡免费观看| 日韩一本色道免费dvd| 亚洲欧美精品综合一区二区三区 | 一本色道久久久久久精品综合| 亚洲国产精品国产精品| 日韩一卡2卡3卡4卡2021年| 涩涩av久久男人的天堂| 桃花免费在线播放| 天天影视国产精品| 在线观看三级黄色| 国产av一区二区精品久久| √禁漫天堂资源中文www| 黄片小视频在线播放| 亚洲精品av麻豆狂野| 九草在线视频观看| 久久 成人 亚洲| 熟女电影av网| 午夜激情av网站| 欧美亚洲 丝袜 人妻 在线| 少妇熟女欧美另类| 国产国语露脸激情在线看| 女人高潮潮喷娇喘18禁视频| 久久久久视频综合| av视频免费观看在线观看| 日韩成人av中文字幕在线观看| 久久久久国产网址| 色播在线永久视频| 最近最新中文字幕大全免费视频 | 最新中文字幕久久久久| 91成人精品电影| 2018国产大陆天天弄谢| 欧美人与善性xxx|