首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 640 毫秒
1.
Dioscorin, the tuber storage protein of yam (Dioscorea alata cv. Tainong No. 1), was purified to homogeneity by DE-52 ion-exchange chromatography. This purified dioscorin was shown by spectrophotometric methods to inhibit angiotensin converting enzyme (ACE) in a dose-dependent manner (12.5-750 microg, respectively, 20.83-62.5% inhibitions) using N-[3-(2-furyl)acryloyl]-Phe-Gly-Gly (FAPGG) as substrates. The 50% inhibition (IC(50)) of ACE activity was 6.404 microM dioscorin (250 microg corresponding to 7.81 nmol) compared to that of 0.00781 microM (0.0095 nmol) for captopril. The commercial bovine serum albumin and casein (bovine milk) showed less ACE inhibitory activity. The use of qualitative TLC also showed dioscorin as ACE inhibitors. Dioscorin showed mixed noncompetitive inhibitions against ACE; when 31.25 microg of dioscorin (0.8 microM) was added, the apparent inhibition constant (K(i)) was 2.738 microM. Pepsin was used for dioscorin hydrolysis at 37 degrees C for different times. It was found that the ACE inhibitory activity was increased from 51.32% to about 75% during 32 h hydrolysis. The smaller peptides were increased with increasing pepsin hydrolytic times. Dioscorin and its hydrolysates might be a potential for hypertension control when people consume yam tuber.  相似文献   

2.
Dioscorin, the tuber storage protein of yam (Dioscorea batatas Decne), was purified successively by ammonium sulfate fractionation, DE-52 ion exchange chromatography, and Sephadex G-75 column. Two protein bands (82 and 28 kDa) were found under nonreducing conditions after SDS-PAGE; but only one band (32 kDa) was detected under reducing conditions. The first 21 amino acids in the N-terminal region of the 28 kDa form were VEDEFSYIEGNPNGPENWGNL, which was highly homologous to deductive sequence of dioscorin from cDNA of another yam species (Dioscoreacayenensis Lam) reported by Conlan et al. (Plant Mol. Biol. 1995, 28, 369-380). Hewett-Emmett and Tashian (Mol. Phylogenet. Evol. 1996, 5, 50 -77) mentioned that, according to DNA alignments, dioscorin from yam (D. cayenensis) was alpha-carbonic anhydrase (alpha-CA) related. In this report, we found that the purified dioscorin showed both CA dehydration activity using sodium bicarbonate as a substrate and CA activity staining after SDS-PAGE. A polyclonal antibody, which was raised against trypsin inhibitor (TI), a storage protein of sweet potato (Ipomoea batatas [L.] Lam var. Tainong 57), cross-reacted with dioscorin, which also showed TI activity determined by both activity staining after SDS-PAGE and trypsin inhibition determination.  相似文献   

3.
The potato (Solanum tuberosum L.) tuber storage protein, patatin, was purified to homogeneity with a molecular mass of 45 kDa. The purified patatin showed antioxidant or antiradical activity by a series of in vitro tests, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical (half-inhibition concentration, IC(50), was 0.582 mg/mL) scavenging activity assays, anti-human low-density lipoprotein peroxidation tests, and protections against hydroxyl radical-mediated DNA damages and peroxynitrite-mediated dihydrorhodamine 123 oxidations. Using electron paramagnetic resonance (EPR) spectrometry for hydroxyl radical detections, it was found that the intensities of the EPR signal were decreased by the increased amounts of patatin added (IC(50) was 0.775 mg/mL). Through modifications of patatin by iodoacetamide or N-bromosuccinimide, it was found that the antiradical activities of modified patatin against DPPH or hydroxyl radicals were decreased. It was suggested that cysteine and tryptophan residues in patatin might contribute to its antioxidant activities against radicals.  相似文献   

4.
Trypsin inhibitors (TIs), root storage proteins, were purified from sweet potato (Ipomoea batatas[L.] Lam cv. Tainong 57) roots by trypsin affinity column according to the methods of Hou and Lin (Plant Sci. 1997, 126, 11-19 and Plant Sci. 1997, 128, 151-158). A single band of 33 kDa TI was obtained by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels. This purified 33 kDa TI had scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. There was positive correlation between scavenging effects against DPPH (2 to 22%) and amounts of 33 kDa TI (1.92 to 46 pmol). The scavenging activities of 33 kDa TI against DPPH were calculated from linear regression to be about one-third of those of glutathione between 5 and 80 pmol. Using electron paramagnetic resonance (EPR) spectrometry for hydroxyl radical detection, it was found that 33 kDa TI could capture hydroxyl radical, and the intensities of EPR signal were significantly decreased from 1.5 to 6 pmol of 33 kDa TI compared to those of the controls. It is suggested that 33 kDa TI, one of the sweet potato root storage proteins, may play a role as an antioxidant in roots and may be beneficial to health when it is consumed.  相似文献   

5.
Fourier transform (FT)-Raman spectroscopy was employed to study the molecular structure of yam proteins isolated from three commonly consumed yam species including Dioscorea alata L., D. alata L. var. purpurea, and Dioscorea japonica. Although D. alata L. and D. alata L. var. purpurea consisted of similar amino acid residues, they still exhibited significant differences in conformational arrangement. The secondary structure of D. alata L. was mainly an alpha-helix, while D. alata L. var. purpurea was mostly in antiparallel beta-sheets. In contrast, D. japonica, which belongs to a different species, exhibited explicit differences in amino acid compositions and molecular structures of which the conformation was a mixed form of alpha-helices and antiparallel beta-sheets. FT-Raman directly proved the existence of S-S in yam proteins, implying that oligomer formation in yam proteins might be due to disulfide linking of dioscorin (32 kDa). The microenvironment of aromatic amino acids and the state of S-S in yam proteins were also discussed.  相似文献   

6.
A modified assay based on the AACC official method 86-45 (AACC, 2000) for the determination of choline in three cereals and three varieties of Dioscorea (yam) tubers was developed. When tested in wheat, rice, and oat flour, choline estimated by the modified method was 34.0-45.3% higher than that of the original AACC method. In a system with higher contents of starch and mucilage, such as Dioscorea (yam) tubers, extra procedures in sample preparation needed to be carried out to separate starch and mucilage. The choline contents of the following Dioscorea (yam) tubers using the original AACC method and the present modified AACC method through coupling an additional bubble separation procedure, respectively, were (mean +/- SD, mg/g solid) Keelung yam (D. pseudojaponicaY.) 0.92 +/- 0.09 and 2.21 +/- 0.12, Yangmingshan yam (D. alata L.) 0.77 +/- 0.09 and 1.78 +/- 0.28, and Ming-Chien yam (D. purpurea) 0.44 +/- 0.09 and 1.35 +/- 0.19. Choline was 231-306% higher than when the original AACC method was used. Dioscorea (yam) tubers were much higher in choline content than they were in cereals. Bubble separation is an appropriate procedure in the practice for the maximum assay of choline in yams. It is accurate, rapid, easy to handle, and especially good for recovering choline from a starch and polysaccharide-protein-containing system.  相似文献   

7.
The high cost and supply shortage of seed yam propagules for planting are major constraints in yam production. In the water yam (Dioscorea alata L.), aerial tubers have potential as alternative sources of planting material. In this study, we investigated morphological, molecular and ploidy variation across multiple aerial tuber producing accessions of Dioscorea alata. Initial screening of over 800 accessions from the International Institute of Tropical Agriculture germplasm collection for aerial tuber production identified a subset (15 %) of accessions, which produced aerial tubers. The aerial tuber producing accessions (along with 18 non-aerial tuber accessions) were further characterized for phenotypic and ploidy variation. In addition, using SSR markers we characterized the genetic diversity amongst all of the aerial tuber producing accessions, along with six non-aerial tuber producing accessions. Multiple Correspondence Analysis (MCA) using morphological data grouped the accessions according to their aerial tuber production. The aerial tuber production characteristics of accessions were associated with phenotypic variables and ploidy levels. The MCA analysis revealed three main groups consisting of; Group 1) all non aerial tuber producing accessions (n = 15), hastate leaf shape, less or no anthocyanin pigmentation and diploid (2n = 2x = 40), Group 2) group with some aerial tuber producing accessions, different extent of anthocyanin pigmentation, sagittate leaf shape, mainly diploid (n = 44) and three triploid (2n = 3x = 60) and 3) a group where all individuals bear aerial tuber, cordate leaf shape, intermediate anthocyanin pigmentation and majority (n = 74) tetraploid (2n = 4x = 80) and three triploid individuals. Aerial tuber production may be subject to a genome dosage effect as an increase in aerial tuber production was associated with increased ploidy level. For instance, tetraploid plants produce more aerial tubers per sprout than either triploids or diploids. Principal coordinate analysis based on SSR markers using Jaccard’s coefficient also revealed distinct groups associated with the pattern of aerial tuber formation, leaf shape and anthocyanin pigmentation. Overall our study indicates the usefulness of combining SSR markers, ploidy level and phenotypic data for identification and classification of Dioscorea alata accessions according to their extent of aerial tuber production.  相似文献   

8.
This study investigates a semicontinuous hot pressurized fluid extraction process and the scavenging activity on the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical of the extract from Taiwan yams (Dioscorea alata). Liquid-liquid extractions were preliminarily employed to generate six fractions, initially extracted by ethanol. Then, the aqueous solution of dried crude ethanol extract was sequentially fractionated by hexane, chloroform, ethyl acetate, and n-butanol. The EC50 value was defined as the UV absorption of DPPH concentrations sufficiently decreased to 50% of the original value. It was found that all peel portions have a better effect on scavenging of the DPPH free radical than meat portions, especially for the ethyl acetate partition of the peel portion of Tainung #2 yam. Its EC50 value (14.5 microg mL(-1)) was even lower than that of ascorbic acid (21.4 microg mL(-1)). Furthermore, semicontinuous hot pressurized ethanol was superior to hot pressurized water in extracting the compound scavenging the DPPH radical from the Purpurea-Roxb peel. The recovery of four unknown compounds corresponded to the scavenging ratio of DPPH free radical in the hot pressurized ethanol extract. Finally, three-level and four-factor experimental design revealed that ethanol ratio and temperature were the most effective factors in order. Conditions of 80% of aqueous ethanol, 20.0 kg/kg solid ratio, 180 psig (1.342 MPa), and 100 degrees C were preferred to extract those antioxidants from the yam peel.  相似文献   

9.
【目的】比较河北省主栽山药品种的矿质养分累积特性及营养品质的差异,以期为山药生产提质增效制定科学的养分管理措施。【方法】田间试验在河北省蠡县进行,供试山药(Dioscorea oppositifolia L.)品种包括棒药、大和白玉、紫药和小白嘴,山药4个品种的施肥量和施肥方法均一致。在成熟期,取样测定了山药地上部、根茎生物量,以及9种矿质元素(氮、磷、钾、钙、镁、铁、锰、铜和锌)含量和累积量。【结果】4个主栽山药品种根茎的鲜生物量表现为大和白玉>棒药>紫药>小白嘴,而干生物量表现为紫药>大和白玉>小白嘴>棒药。大和白玉根茎的氮、钙、镁、锰、铜、锌累积量最高,且钾、钙、镁、锰、铜、锌分配系数最高,紫药根茎的磷、钾累积量最高,且氮分配系数较高,磷分配系数最高,棒药根茎的铁累积量及分配系数均为最高。4个山药品种形成1000 kg产量对大量营养元素的需求量均表现为钾>氮>磷,对中量营养元素的需求量均表现为镁>钙,对微量营养元素的需求量均表现为铁>锌>锰>铜。以《中国食物成分表》(标准版)中山药的9种矿质营养品质指标作为参...  相似文献   

10.
11.
Oxidative stress, caused by reactive oxygen species (ROS), is responsible for modulating several pathological conditions and aging. Soluble and bound phenolic extracts of commonly consumed millets, namely, kodo, finger (Ravi), finger (local), foxtail, proso, little, and pearl, were investigated for their phenolic content and inhibition of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and ROS, namely, hydroxyl radical, peroxyl radical, hydrogen peroxide (H(2)O(2)), hypochlorous acid (HOCl), and singlet oxygen ((1)O(2)). Inhibition of DPPH and hydroxyl radicals was detrmined using electron paramagnetic resonance (EPR) spectroscopy. The peroxyl radical inhibitory activity was measured using the oxygen radical absorbance capacity (ORAC) assay. The scavenging of H(2)O(2), HOCl, and (1)O(2) was evaluated using colorimetric methods. The results were expressed as micromoles of ferulic acid equivalents (FAE) per gram of grain on a dry weight basis. In addition, major hydroxycinnamic acids were identified and quantified using high-performance liquid chromatography (HPLC) and HPLC-mass spectrometry (MS). All millet varieties displayed effective radical and ROS inhibition activities, which generally positively correlated with phenolic contents, except for hydroxyl radical. HPLC analysis revealed the presence of ferulic and p-coumaric acids as major hydroxycinnamic acids in phenolic extract and responsible for the observed effects. Bound extracts of millet contributed 38-99% to ROS scavenging, depending on the variety and the test system employed. Hence, bound phenolics must be included in the evaluation of the antioxidant activity of millets and other cereals.  相似文献   

12.
Four factorial trials were conducted with yam (Dioscorea rotundata Poir.) at Ibadan, Nigeria from 2013 to 2015, investigating effects of (1) tillage (2) fertilizer (3) intercropping (4) yam plant densities. Yam tuber yields varied between years (2013: 16.44 Mg ha?1; 2014: 10.08 Mg ha?1; 2015 26.61 Mg ha?1). In 2013 neither tillage nor fertilizer affected tuber yields. In 2014 tillage increased yields (+25.4%, P < 0.0001), fertilizer reduced yield (?10.5%; P = 0.0046). In 2015 tillage increased tuber yields by 8.1% (ns), fertilizer application increased yield (+17.5%, P = 0.0017). Across the years, tuber yields increased (P < 0.01) with increasing yam density with a constant increase in 2013 up to the highest density, yet yields leveled out above 14,815 plants ha?1 in 2014 and 2015. Intercropping with maize (66,667 plants ha?1) reduced tuber yield by 42.62% in 2013, 44.52% in 2014 and 30.68% in 2015 (P < 0.01 all years) across all yam densities. Maize grain yield was higher in sole crop in 2 years. Fertilizer increased yields in all years (P < 0.0001). Maize yield had no response to the yam densities. Ridging had a negative effect on grain yield in 2015 (?0.3 Mg ha?1, P = 0.0002). Increasing plant density appears a safe measure to increase yam yields.  相似文献   

13.
山药PAL基因全长cDNA序列的克隆、表达与分析   总被引:2,自引:0,他引:2  
为了阐明苯丙氨酸解氨酶(PAL,EC4.3.1.5)基因在山药地下块茎酚类物质积累过程中的分子机理,本研究运用普通PCR、RACE和TAIL-PCR技术从大薯(Dioscorea alataL.)’Zixiao’地下块茎中克隆到同一个PAL基因的3个片段,即长度分别为1 145bp、1 151bp和424bp的中间片段、3’端和5’端的cDNA序列;将3段cDNA序列拼接后获得大小为2 376bp的PAL基因全长cDNA序列,该序列含有一个1986bp的最大开放阅读框(ORF),一个28bp的5’端非翻译区,一个362bp含25 nt的Poly(A+)尾的3’端非翻译区,最大ORF可推测编码一个包括PAL酶活性中心的特征序列(GTITASGDLVPLSYIA)在内的661个氨基酸的多肽,分子量为71.974kDa,等电点6.310;大薯地下块茎的PAL基因分别在核苷酸与氨基酸水平上和GenBank中所选已知其他物种的同源性为73%~77%和76%~82%,说明PAL基因在系统进化上相对保守;RT-PCR分析表明该基因仅在地下块茎中表达,而且表达丰度在收获前逐渐降低。  相似文献   

14.
Tubers of six accessions of ahipa (Pachyrhizus ahipa) contained between 0.77 and 1.34% nitrogen on a dry weight basis. This corresponds to 4.8 to 8.4% crude protein based on a nitrogen to protein conversion factor of 6.25; but detailed analysis of AC230 showed that although 93% of the total N was extracted with buffer containing 1.0 M NaCl, about a third of this was lost on dialysis. It was calculated, therefore, that salt-soluble proteins comprise about 60% of the total tuber nitrogen, with low-molecular-mass nitrogenous components comprising a further 30%. Electophoretic analysis of the salt-soluble proteins showed similar patterns of components in the six accessions, with none being present in amounts sufficiently high to suggest a role as storage proteins. Furthermore, light microscopy failed to show significant deposits of protein within the tuber cells. Five "major" protein bands, which together accounted for about 19% of the total salt-soluble protein fraction were purified and subjected to N-terminal amino acid sequencing. Comparison of these with sequences in protein databases revealed similarities to alpha-amylases, chitinases and chitin binding proteins, cysteine proteinases (including major components from P. erosus tubers), a tuberization-specific protein from potato, and proteins induced in soybean and pea by stress or the plant hormone abscisic acid, respectively. It was concluded that the primary roles of these proteins are probably in aspects of tuber metabolism and development and/or conferring protection to pests and pathogens, and that true storage proteins are not present. The absence of storage proteins is consistent with the biological role of the tubers as storage organs for carbohydrates (cf cassava tuberous roots) rather than as propagules (cf yam and potato tubers).  相似文献   

15.
The influence of different fertilization practices [i.e., nitrogen (N), phosphorus (P), and potassium (K) fertilizers, crop residue, and manure] on dry matter production of white yam (Dioscorea rotundata) “kokoro” a late variety of yam, tuber biomass and dry matter distribution to the plant parts was determined in upper Oueme basin (Republic of Benin) in the years 2001, 2002 and 2003. The experimental design was a randomized complete block with four replications. Altogether there were eight plots divided into two groups consisting of four plots each, one was treated with manure (at the rate of 10 ton ha?1), second plot with mineral fertilizer (N30:P30:K60), third plot with combination of manure and mineral fertilizer, whereas the fourth plot was left as control (no application of fertilizer). In year 2002 and 2003 the same combinations were made taking crop residues (at the rate of10 ton ha?1) from external sources as a source of organic matter at the place of manure. Manure application did not have a significant effect on yam total biomass production nor on tuber yield when applied on fields after fallow. Whereas crop residue application had a positive significant effect on yam tuber and total biomass production when applied after cotton and maize and with adequate rainfall amount and distribution. Regarding partitioning pattern in yam crop, no effect of different practices of fertilization has been noticed. The crop behaves identically (i.e, partitioning rate) in both fertilized and unfertilized management practice.  相似文献   

16.
Guinea yam (Dioscorea cayenensisD. rotundata complex) is an important tuber crop that highly contributes to food security and poverty alleviation in Benin. The knowledge and understanding of the extent of genetic variation of Guinea yam germplasm is important for planning of the genetic conservation, and the utilisation of this resource. The objective of this study was to assess the genetic diversity and relationships of 64 yam landraces whose dried chips are considered as resistant to insect attacks using 41 simple sequence repeat. Among these primers, 13 were found to be polymorphic, giving 113 polymorphic alleles. The number of alleles per locus ranged from 4 (Ym50) to 13 (Ym29), with an average of 8.69. Unique allele was observed with some landraces (Singou and Tchakatchaka) and can be considered as unique gene and use in yam breeding program. The mean polymorphic information content values for all markers used was 0.76 and ranged between 0.58 and 0.91 in loci YM3 and YM32 respectively. The genetic distance of yam landraces ranged from 0.45 (Yasoubagarou) to 0.04 (Assinapeira and Alahina), indicating that the yam germplasm has a high degree of genetic diversity supported by an averagely observed heterozygosity of 0.78. Cluster analysis using unweighted pair group method with arithmetic average grouped the 64 yam landraces into two distinct clusters. This tendency was also observed in the principal coordinate analysis. The analysis of molecular variance revealed that 96 % of the variation was found within the population and only 4 % between the populations. Genetic diversity and relationship assessments among the 64 yam landraces of Benin could provide useful information for efficient use of these materials, especially for genetic improvement.  相似文献   

17.
Oca (Oxalis tuberosa Mol.) or New Zealand yam, in common with other members of this genus, contains oxalate, an antinutritive factor. Twelve South American and two New Zealand cultivars of oca were analyzed for total and soluble oxalate contents of the tubers. The range of total oxalate levels was 92-221 mg/100 g of fresh weight. Levels of soluble and total oxalate extracted from the tubers were not significantly different, suggesting that no calcium oxalate is formed in the tubers. The oxalate concentrations obtained in this study for oca suggest that previously reported values are too low and that oca is a moderately high oxalate-containing food. This is the first report of a tuber crop containing moderate to high levels of soluble oxalates in the tubers and no insoluble oxalates.  相似文献   

18.
Abstract

Fast screening methods are needed for plant breeding. The objective of this research was to evaluate the potential of near‐infrared reflectance spectroscopy (NIRS) for the simultaneous analysis of dry matter and protein contents in intact discs of fresh yam bean (Pachyrhizus spp.) tubers. Discs from 210 tubers were extracted with a punch few hours after harvesting and scanned by NIRS using a specially designed adapter. External validation revealed a close relationship between NIRS and reference methods for dry matter content (r2=0.94; standard error of performance, SEP=1.2%) and protein content (r2=0.87; SEP=1.94%). The calibration for protein content was compared with another one developed using dried‐ground tuber samples (r2=0.97; SEP=0.97%). These results suggested that NIRS can be used to determine dry matter and protein contents in fresh tuber samples of yam beans with acceptable accuracy. Further research will have to determine if additional traits can be incorporated into this scheme.  相似文献   

19.
In Nigeria, information is lacking regarding the most suitable tillage method in extensive yam production. Hence, five tillage methods were compared at two sites in 2008–2010 with reference to their effects on soil physical and chemical properties, leaf nutrient concentrations, growth and tuber yield of yam (Dioscorea rotundata Poir) on Alfisols at Owo (site A) and Akure (site B), south-west Nigeria. The tillage methods were: zero tillage (ZT), manual ridging (MR), manual mounding (MM), ploughing + harrowing (P + H) and ploughing + harrowing + ridging (P + H + R). P + H + R had lower soil bulk density than other tillage methods and resulted in higher leaf N, P, K, Ca and Mg and yam tuber yield. In ZT, bulk density, soil moisture content, soil organic C, N, P, K, Ca and Mg were significantly higher and temperature lower than other tillage methods. Results of multiple regressions revealed that bulk density significantly influenced the yield of yam rather than soil chemical properties. Compared with MR, MM, P + H and ZT, and averaged across years, P + H + R increased yam tuber yield by 12.3, 12.8, 34.9 and 50.7%, respectively, in site A and 12.9, 13.5, 25.2 and 44.5%, respectively, in site B. P + H + R was found to be most advantageous and is therefore recommended for yam cultivation.  相似文献   

20.
Time of mulching can influence the growth environment and performance of white yam (Dioscorea rotundata Poir). An on-farm trial was conducted during the 1988–1989 and 1989–1990 seasons (October–August) in Nigeria to determine the effect of time of mulching (October–February) on the hydrothermal regime and emergence, growth and tuber yield of white yam. Application of 12.5 mg ha−1 of dry Eupatorium odoratum L. (Syn. Chromolaena odorata L.) mulch on top of the mounds significantly improved soil moisture content of the 15 cm surface layer by 50–120 g kg−1 and decreased the maximum soil temperature by 2–7°C at 15 cm depth in the early growing season (March–April). The emergence and development of yam seedlings were significantly lower in unmulched plots than in mulch-treated plots. Mulching significantly increased tuber yield by about 10–15 mg ha−1 season−1. Plots mulched in October–December were more moist by 20–60 g kg−1 and cooler by 1–3°C, and had 27–44% greater emergence than those mulched in January or February. The number of leaves per plant, vine diameter and leaf area index were also significantly greater in plots mulched in October–December than plots mulched in January or February. Consequently, shoot dry weight was about 28–36% greater in yam mulched in October–December than in yam mulched in February. However, the time of mulching had no effect on soil moisture of the surface layer at the beginning of the rainy season (April), on yam emergence in May and on tuber yield and yield components. Although time of mulching did not significantly affect tuber yield, the increase (10–15%) in the tuber yield of yam mulched in December–February compared to the yam mulched in October or November was considerable. It was concluded that yam planted in October, just before the rain stops, can be mulched in January or February without detrimental effect on emergence, growth and tuber yield.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号