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1.
ABSTRACT Spot blotch, caused by Cochliobolus sativus, is an important disease of barley in many production areas and is best controlled through the deployment of resistant cultivars. Information on the genetics of resistance in various sources can be useful in developing effective breeding strategies. Parents of the doubled haploid mapping population Calicuchima-sib/ Bowman-BC (C/B) exhibit a differential reaction to pathotypes 1 and 2 of C. sativus. To elucidate the genetics of spot blotch resistance in this population, C/B progeny were evaluated with both pathotypes at the seedling stage in the greenhouse and at the adult plant stage in the field. At the seedling stage, progeny segregated 84 resistant to 26 susceptible based on the qualitative analysis of infection response (IR) data to pathotype 1. This fit best to a 3:1 ratio, indicating that two genes were involved in conferring resistance. Quantitative analysis of the raw IR data to pathotype 1 revealed a single quantitative trait locus (QTL) on chromosome 4(4H) explaining 14% of the phenotypic variance. Adult plant resistance to pathotype 1 was conferred by QTL on chromosome 2(2H) and chromosome 3(3H), explaining 21 and 32% of the phenotypic variation, respectively. Bowman contributed the resistance alleles on chromosome 3(3H) and chromosome 4(4H), whereas Calicuchima-sib contributed the resistance allele on chromosome 2(2H). Resistance to pathotype 2 was conferred by a single gene (designated Rcs6) on chromosome 5(1H) based on qualitative analysis of data. Rcs6 was effective at both the seedling and adult plant stages and was contributed by Calicuchima-sib. This result was corroborated in the quantitative analysis of raw IR (seedling stage) and disease severity (adult plant stage) data as a single major effect (r(2) = 0.93 and 0.88, respectively) QTL was identified on chromosome 5(1H). Progeny with resistance to both pathotypes were identified in the C/B population and may be useful in programs breeding for spot blotch resistance.  相似文献   

2.
Disease observations and amplified fragment length polymorphism (AFLP) markers were used to study recent developments in the Puccinia striiformis f.sp. tritici population in Denmark. The fungus appeared spontaneously at 10 locations in Denmark in 1997 after it was not observed under natural conditions in 1996. The pattern of disease development and prevailing winds suggested that the fungus reappeared by airborne spores from the south or west. In 1998, disease incidence was more evenly distributed throughout the country. Forty-eight single lesion isolates were collected from most crops where the disease was observed in these years; all except one from 1997 belonged to two pathotypes that were not previously detected in the country, and both possessed the newly discovered Yr17 virulence. The isolates were characterized with AFLP markers together with 28 isolates representing eight of 13 pathotypes observed prior to 1996. Initial screening of 240 Pst I/ Mse I AFLP primer combinations on four isolates showed that a primer combination, on average, revealed 0·4 polymorphisms between any isolate pair. A selection of 21 primer combinations resulted in 28 AFLP markers, which revealed 16 AFLP phenotypes among all 76 isolates. The two Yr17- virulent pathotypes consisted of three AFLP phenotypes, which were observed in both 1997 and 1998; the two most frequent AFLP phenotypes occurred at most sampling locations and often within the same crop. AFLP diversity was larger among samples collected prior to 1996, and also in this period most AFLP phenotypes were observed at different sampling locations. These results are consistent with the features of an entirely asexually reproducing pathogen dispersed by aerial spores across large areas.  相似文献   

3.
Amplified fragment length polymorphism (AFLP) based genetic diversity was analyzed for 232 Colletotrichum sublineolum isolates collected between 2002 and 2004 from three geographically distinct regions of Texas, and from Arkansas, Georgia, and Puerto Rico. Results revealed significant levels of polymorphism (59%) among the isolates. Even so, genetic similarity between isolates was high, ranging from 0.78 to 1.00. Clustering of similar isolates did not correlate with either geographic origin or year of collection. Pathotypes of 20 of the isolates were determined using 14 sorghum lines previously used in Brazil and the United States and 4 from Sudan. Seventeen new pathotypes were established from the 18 isolates that gave uniform and consistent reactions on all host differentials over 2?years of greenhouse testing. Differentials BTx378 and QL3 were resistant to all isolates while BTx623 and TAM428 were universally susceptible both years. Each of these lines had shown differential responses in prior studies indicating that the pathogen population has sufficient diversity to adapt rapidly to changes in resistant host lines deployed. When the 2-step pathotype classification scheme was used, the 18 isolates examined in this study were placed in four pathotype groups (A, C, D and G), which would further then be separated into ten distinct pathotypes. Common sets of differentials and a standardized nomenclature will allow for comparison to be made among pathotypes of C. sublineolum detected from different regions and also could help direct planting of appropriate sorghum lines and aid in the development of more durable forms of resistance.  相似文献   

4.
Since 2006, verticillium wilt of olive induced by Verticillium dahliae has caused considerable economic losses in olive orchards in Tunisia. The genetic structure of V. dahliae isolates collected from different olive growing regions was investigated using virulence tests, vegetative compatibility grouping (VCG) and amplified fragment length polymorphism (AFLP) analyses. In total, 42 isolates of V. dahliae from diseased olive trees were tested. Cluster analysis and principal coordinate analysis revealed that geographic origin was the main factor determining the genetic structure of V. dahliae populations and both methods indicated a genetic separation between the central and coastal isolates. Isolates were divided into two major groups: the AFLP‐I group included all isolates from Sidi Bouzid, Kairouan, Kasserine and Sfax (centre of the country) and the AFLP‐II group included isolates from Monastir, Zaghouane, Sousse, Mahdia (coastal region), and two isolates from Sfax. Analysis of the molecular variance (amova ) indicated a significant level of genetic differentiation among (76%) and within (23%) the two populations. Analyses of both the defoliating (D) and non‐defoliating (ND) pathotypes and VCG markers indicated that most of the isolates belong to VCG 2A and 4B/ND pathotype. The disease severity was highly variable among the isolates tested (< 0·05) with no evidence of association between aggressiveness and geographical origin of the isolates. Overall, results of this study revealed a clear association between the genetic diversity of the isolates and their geographic origin, but not between genetic diversity and virulence patterns.  相似文献   

5.
Clubroot disease caused by Plasmodiophora brassicae is one of the most serious diseases in cruciferous crops. To classify isolates, we developed simple sequence repeat (SSR) markers for P. brassicae. Twenty-four Japanese isolates were used in this study: 12 isolates of an unknown pathotype from the Kyoto Prefecture, as well as 12 isolates of known pathotypes, including three single-spore lines. From the 12 isolates from Kyoto Prefecture, 11 were classified into either pathotype 2 (three isolates) or 4 (eight isolates). We designed 23 SSR markers based on the P. brassicae genome, of which 11 markers from intergenic regions showed polymorphisms in the 24 isolates. Many haploid isolates belonging to pathotypes 2 and 4 were monomorphic, and typical alleles were detected in some isolates not belonging to pathotype 4. Two bands were detected for eight SSR loci in five isolates, indicating that different genotypes were mixed in these isolates. We constructed a phylogram based on the 11 polymorphic SSRs. Pathotypes 2 and 4 formed a cluster, from which pathotypes 3 and 1 were successively placed. These results strongly suggest a close genetic relationship between isolates in pathotypes 2 and 4, consistent with our finding that isolates in these two pathotypes were found at one collection site. In combination with pathotype classification and other marker systems, the SSR markers can be used for more detailed analyses to improve the control of clubroot disease.  相似文献   

6.
ABSTRACT Spot blotch, caused by Cochliobolus sativus, is one of the most common foliar diseases of barley in the upper midwest region of the United States. To examine the genetics of host-specific virulence in C. sativus, a cross was made between isolate ND90Pr (which exhibits high virulence on barley genotype Bowman and low virulence on genotype ND 5883) and ND93-1 (which exhibits low virulence on both genotypes). Ascospore progeny segregated 48:55 for low virulence/high virulence on Bowman, indicating the presence of a single virulence gene in isolate ND90Pr. To complement the study of host-specific virulence in the pathogen, an experiment also was conducted on the genetics of specific resistance in the host. Progeny from a Bowman/ND 5883 cross were evaluated for their infection responses (IRs) to isolate ND90Pr at the seedling stage. The F(2) population segregated 1:3 for low IRs (resistant)/high IRs (susceptible), indicating the presence of a single resistance gene in genotype ND 5883. This result was confirmed in the F(3) generation, as a 1:2:1 ratio was found for homozygous resistant, segregating, and homozygous susceptible families, respectively. The data from this study demonstrate that both virulence in the pathogen and resistance in the host are under monogenic control in this specific host genotype/fungal isolate combination.  相似文献   

7.
Verticillium dahliae isolates from potato on the island of Hokkaido (potato isolates) and those belonging to pathotypes A (eggplant pathotype), B (tomato pathotype) and C (sweet pepper pathotype) were divided into three distinct groups by RAPD and REP-PCR. The three DNA groups I, II, III consisted of pathotypes A and C, pathotype B and potato isolates, respectively. The potato isolates were assigned to pathotype A on the basis of pathogenicity. Another set of potato isolates was further collected from eight potato cropping regions on Hokkaido to further examine the relationships among them in detail. Only one of these isolates was identified as DNA group II, but all the others were classified as DNA group III. Isolates from daikon, eggplant, and melon on Hokkaido also belonged to DNA group III. These results suggest that V. dahliae isolates from Hokkaido are unique at the DNA level and different from other pathotype A isolates in Japan. Received 28 February 2000/ Accepted in revised form 6 November 2000  相似文献   

8.
Tsuchiya D  Taga M 《Phytopathology》2001,91(4):354-360
ABSTRACT Cytological karyotypes with mitotic metaphase chromosomes were analyzed for Cochliobolus heterostrophus, C. carbonum, and C. sativus by the germ tube burst method (GTBM). Prior to karyotyping, procedures of GTBM suitable to Cochliobolus were established by examining several crucial conditions such as incubation period of conidia. The estimated chromosome numbers of C. heterostrophus and C. carbonum were n = 15 or 16 and n = 13 or 15 depending on the strains, respectively. In C. sativus, n = 15 was estimated. Morphological information of chromosomes including chromosome size and a threadlike-specific structure representing the nucleolar organizing region was also obtained. Our results for some standard strains are in agreement with previous estimates by pulsed field gel electrophoresis (PFGE) or PFGE coupled with restriction fragment length polymorphism genetic linkage analysis, but inconsistent with the previous estimates for other strains by conventional light microscopic cytology. Additionally, PFGE analysis of C. heterostro-phus strains indicated that chromosome number was not determinable solely by PFGE, which is hampered by comigration and clumping of DNA bands.  相似文献   

9.
Genotypic diversity in barley powdery mildew populations in northern France   总被引:1,自引:0,他引:1  
Caffier  Brändle  & Wolfe 《Plant pathology》1999,48(5):582-587
Diversity in populations of Erysiphe graminis ( Blumeria graminis ) f.sp. hordei was studied with virulence and molecular markers. Isolates were sampled in two locations in northern France from a winter barley cultivar (Plaisant) and a spring cultivar (Caruso). Only a few pathotypes (determined by virulence markers) were common. The rest of the population was diverse. Diversity within common pathotypes, estimated by five RAPD and two SCAR markers, was generally high, except for one pathotype, which was frequent on Plaisant. This pathotype carried only one virulence, Va22, out of the 11 virulences tested. It appeared as a clonal lineage, which had occurred previously, at least in 1992, in northern France, demonstrating survival of asexual lineages in populations that often reproduce sexually.  相似文献   

10.
Plasmodiophora brassicae is an obligate biotroph that causes clubroot, one of the most damaging diseases of crucifers. Differential cultivars and random amplified polymorphic DNA markers were used to assess the extent of genetic diversity among nine single-gall populations of P. brassicae and 37 single-spore isolates (SSI) derived from four of those field samples. Isolates were classified into eight pathotypes, and each isolate was associated with a unique molecular genotype. Virulence and DNA polymorphisms were detected within and between field isolates, and among SSIs from different pathotypes, hosts and geographical origins. The relatively high level of genetic diversity among field isolates was similar to that among SSIs derived from a single-club field isolate. Molecular and pathogenicity-based classifications were not clearly correlated, but isolates belonging to pathotype P1 were clustered. Two RAPD markers were specific to pathotype P1. The finding that genetic differences can occur in P. brassicae field isolates will be an important consideration in resistance genetic studies and in choosing breeding strategies to develop durable clubroot resistance.  相似文献   

11.
Roses produced or grown in the field, as well as pot‐grown and cut roses, are attacked by different fungal pathogens causing leaf spot diseases. The incorrect identification and scoring of these pathogens and the lack of information about their genetic and pathotype diversity hamper resistance breeding. This is especially true for the hemibiotrophic ascomycete Sphaceloma rosarum, which is often confused with other fungi. Here for the first time, the genetic variability between isolates at both the molecular and morphological level is analysed. Eighty leaf spot samples were collected from different rose genotypes at five different locations, and 15 single conidial isolates established. All of the samples showed high morphological similarities to the reference isolate CBS 213.33 that was obtained from a public repository. By sequencing a part of the large subunit (LSU) of the 28S ribosomal RNA and phylogenetic analysis, high sequence similarities were shown to other Sphaceloma species for 13 of the isolates and the CBS reference. One of the isolates clustered with Septoria species and another clustered with Seimatosporium species. UPGMA clustering with 145 polymorphic AFLP markers resulted in five distinct groups in the majority rule consensus tree for the 14 S. rosarum isolates, including the CBS reference. Jaccard similarities ranged from 0·31 to 0·91. A detached leaf assay using a differential set of five rose genotypes led to the classification of the five tested isolates as five distinct pathotypes. Therefore, grouping depending on the avirulence gene diversity was clearly different from clustering using selectively neutral AFLP markers that were evenly distributed throughout the genome.  相似文献   

12.
Since its initial detection in Australia in 1979, wheat yellow (stripe) rust ( Puccinia striiformis f.sp. tritici ) has evolved in Australia and New Zealand into more than 20 pathotypes with assorted virulence characteristics. This evolution is believed to have occurred in a stepwise fashion from an original single pathotype, with no subsequent new introductions. A combination of random amplified polymorphic DNA (RAPDs) and amplified fragment length polymorphisms (AFLPs) was used to examine the level of molecular variation in Australian and New Zealand isolates, and to compare this with variation amongst other isolates of P. striiformis . Using 60 RAPD primers on seven Australian isolates representing seven different pathotypes collected between 1979 and 1991, more than 300 potentially polymorphic loci were analysed and no polymorphisms were detected. Using the same primers on two UK isolates, 3% of loci showed a polymorphism. A similar level of polymorphism was found between UK isolates using AFLP primers, and between 5 and 15% of fragments were polymorphic between an isolate from the UK, an isolate from Denmark, and one from Colombia. However, no AFLP polymorphisms were found amongst 14 Australian and New Zealand isolates tested, at over 100 potentially polymorphic loci. The lack of molecular variation in the Australian and New Zealand collection is consistent with the stepwise mutation theory of pathotype evolution from a single introduction.  相似文献   

13.
Clay RP  Bergmann CW  Fuller MS 《Phytopathology》1997,87(11):1148-1159
ABSTRACT Endopolygalacturonase (EPG) of Cochliobolus sativus was produced in shake culture and purified by high-performance liquid chromatography. The enzyme had a molecular mass of 34,000 Da, an isoelectric point in the range of 9.0 to 9.5, exhibited endo activity, was nongly-cosylated, and was inhibited by polygalacturonase-inhibiting proteins from bean, pear, and tomato. The amino terminus contained a 14 amino acid region homologous to a region at the N terminus of an EPG of C. carbonum. C. sativus EPG-specific monoclonal antibodies (MAbs) were generated. Western blot analysis confirmed the specificity of the antibodies for the EPG and detected the enzyme in an extract from Hordeum vulgare (cv. Golden Promise) leaf segments infected with C. sativus. Using conventional immunogold and enzyme-gold cytochemical methods, homogalacturonan, esterified pectin, and cellulose were localized in healthy and infected barley leaf epidermis at the electron microscope level. Additionally, the leaf cell wall polysaccharides recognized by purified C. sativus EPG were localized at the electron microscope level, using the purified enzyme as a primary cytochemical reagent, followed by a gold-labeled MAb specific for the enzyme. Loss of polygalacturonic acid in the vicinity of the invading pathogen was visualized cytochemically at the electron microscope level. These observations suggest the involvement of EPG during host penetration by the fungus.  相似文献   

14.
A large collection of German rye leaf rust isolates was analysed to characterize the diversity, spatial variation and temporal dynamics of virulences. Virulence-avirulence phenotypes (=pathotypes) were determined on 23 host differentials. We found 93 pathotypes among 177 single-uredinial isolates in 2000, 201 pathotypes among 437 isolates in 2001, and 125 pathotypes among 213 isolates in 2002. In total, the 827 analyzed isolates represented 317 pathotypes. Frequency of virulences on the individual differentials varied from 2% to 97%. Eight of the differentials showed a high resistance level with virulence frequencies <10%. Virulence complexity of the isolates ranged from 3 to 21 with a mean of nine. The percentages of highly virulent isolates (>14 virulences) increased from 4 to 15% during the sampling period. A high level of virulence diversity was observed within and between individual sampling sites with Simpson indices around 0.9. Evenness indices ranged from 0.88 to 0.92. Four of the five most frequent pathotypes were found in each year but their frequency never exceeded 10%. Isolates with unusual virulence combinations could be clearly separated by principal component analysis. Location-specific pathotype frequencies were revealed in each year, but the frequency patterns varied across years. On four fields a considerable increase of highly virulent pathotypes occurred within 6 weeks during the epidemic. The high diversity of pathotypes as well as the fast accumulation of highly virulent pathotypes favour the adaptation of the pathogen to race-specific host resistances. More durable resistance might be achievable by combining new effective race-specific resistances with adult-plant and/or race-non-specific quantitative resistances.  相似文献   

15.
ABSTRACT A degree of genetic diversity may exist among Verticillium dahliae isolates within vegetative compatibility groups (VCGs) that bears phytopathological significance and is worth investigating using molecular tools of a higher resolution than VCG characterization. The molecular variability within and among V. dahliae VCGs was studied using 53 artichoke isolates from eastern-central Spain, 96 isolates from cotton, 7 from cotton soil, and 45 from olive trees in countries of the Mediterranean Basin. Isolates were selected to represent the widest available diversity in cotton- and olive-defoliating (D) and -nondefoliating (ND) pathotypes, as well as for VCG. The VCG of 96 cotton and olive isolates was determined in this present study. Molecular variability among V. dahliae isolates was assessed by fluorescent amplified fragment length polymorphism (AFLP) analysis and by polymerase chain reaction (PCR) assays for DNA fragments associated with the D (462 bp) and ND (824 bp) pathotypes, as well as a 334-bp amplicon associated with D pathotype isolates but also present in some VCG2B isolates. Isolates from cotton were in VCG1A, VCG1B, VCG2A, VCG2B, and VCG4B and those from olive trees were in VCG1A, VCG2A, and VCG4B. Artichoke isolates included representatives of VCG1A, VCG2A, VCG2B (including a newly identified VCG2Ba), and VCG4B. AFLP data were used to generate matrixes of genetic distance among isolates for cluster analysis using the neighbor-joining method and for analysis of molecular variance. Results demonstrated that V. dahliae isolates within a VCG subgroup are molecularly similar, to the extent that clustering of isolates correlated with VCG subgroups regardless of the host source and geographic origin. VCGs differed in molecular variability, with the variability being highest in VCG2B and VCG2A. For some AFLP/VCG subgroup clusterings, V. dahliae isolates from artichoke grouped in subclusters clearly distinct from those comprising isolates from cotton and olive trees. In addition, VCG2B isolates from artichoke formed two distinct clusters that correlated with PCR markers of 334 bp (VCG2B(334)) or 824 bp (VCG2B(824)). Artichoke isolates in the VCG2B(334)/2beta(334) cluster were molecularly similar to isolates of VCG1A. The molecular difference found among artichoke isolates in VCG2B correlates with virulence of isolates to artichoke and cotton cultivars demonstrated in a previous study.  相似文献   

16.
Forty-two Brassica rapa and Brassica napus lines were tested for resistance to Turnip mosaic virus (TuMV) isolates representing the three major pathotypes in Europe. Of these lines, 11 were susceptible to all pathotypes; nine were resistant to one pathotype; eight were resistant to two pathotypes; and 14 were resistant to all three pathotypes. Of the lines tested, 23 were either able to, or had the potential to, discriminate between two different pathotype-3 isolates. Genetic models for inheritance of resistance were proposed for four B. rapa lines: Jong Bai No. 2 had dominant resistance to pathotype 1 conferred by a single allele; PI418957C and Jin G 55 had recessive resistance to pathotype 4 where a single allele was required; PI418957C also had recessive resistance to pathotype 3 where a model with one of two epistatic, unlinked loci was proposed. Jong Bai No. 1 also had recessive resistance to pathotype 3, apparently conferred by alleles at three loci, where any two of the three loci were epistatic and required for resistance.  相似文献   

17.
A collection of 124 isolates of turnip mosaic virus was gathered from around the world, principally from European countries, and characterized by inoculation to four differential lines of Brassica napus (oilseed rape and swede). Three symptom phenotypes were induced—apparent immunity, local infection only, or systemic infection. Twelve distinct patterns, i.e. pathotypes, were observed. Three pathotypes were predominant in the collection; pathotype 1 isolates, which were the most common, did not overcome any of the most extreme sources of resistance in the differential lines. Of the other two, pathotype 3 isolates overcame one of the major sources of resistance and pathotype 4 isolates overcame all sources of resistance. The distribution of pathotypes within Europe was examined. No pathotype was confined to any geographical area, although pathotype 4 isolates were not found in southern Europe or Asia. Most isolates (90) originated from Brassica hosts, while others were from other cruciferae genera (19) or non-crucifers (5). The species of plant that the isolates originated from was not clearly related to the pathotype of the isolates. Resistance to pathotype 1 isolates is controlled by a dominant allele in one of the differential lines, and resistance sources are being examined in the other lines. Isolates belonging to pathotype 1 appeared to be able to mutate readily to overcome the resistance in one of the rape differential lines, but no isolates appeared to mutate to overcome the other major source of resistance in the differentials. The implications of the results for disease control strategies are discussed.  相似文献   

18.
The genetic diversity of several local populations of the fungal pathogen Cochliobolus sativus, collected over 3 years from different regions of the Czech Republic, was examined using amplified fragment length polymorphism (AFLP). A high level of variability was found even among isolates from one lesion. Measures of multilocus linkage disequilibrium suggested that recombination has a minor impact on the genetic structure of populations. Cochliobolus sativus forms genetically divergent populations (FST = 0·33), indicating a low level of geneflow between populations. This was supported by a significant correlation between genetic diversity and geographical distances up to 80–100 km. The most likely explanation for the genetic variability is that the fungus forms conidia with highly variable chromosomal rearrangements. The differentiation observed among local populations implies that genetic drift, including a founder effect, combined with restricted migration generates the structure of C. sativus populations.  相似文献   

19.
利用来自国际马铃薯中心含有单显性抗性基因的11个鉴别寄主,采用离体叶片测定方法对2007—2008年采自甘肃和内蒙古地区的共91个致病疫霉马铃薯分离物的致病型进行了测定。结果显示:来自甘肃的65株菌中共有28个致病型,有3株可以克服11个抗性基因;总共有超过一半的菌株对抗性基因R7、R9、R11有毒性,而能克服抗性基因R1、R2、R8的菌株数最少。在来自内蒙古的26株菌中,共检测到24个致病型,有2株菌能克服11个抗性基因,共有84.6%的菌株可以对含有抗性基因R7的植株表现出毒性,而R1、R2、R8被克服比例最低。因此,研究结果表明,在这两个地区可优先考虑在菌株采集地种植和选育含有R1,R2和R8这3个抗性基因的马铃薯品种。  相似文献   

20.
Puccinia triticina reproduces asexually in France and thus individual genotype is the unit of selection. A strong link has been observed between genotype identities (as assessed by microsatellite markers) and pathotypes (pools of individuals with the same combination of qualitative virulence factors). Here, we tested whether differences in quantitative traits of aggressiveness could be detected within those clonal lineages by comparing isolates of identical pathotype and microsatellite profile. Pairs of isolates belonging to different pathotypes were compared for their latent period, lesion size and spore production capacity on adult plants under greenhouse conditions, with a high number of replicates. Isolates of the same pathotype showed remarkably similar values for the measured traits, except in three situations: differences were obtained within two pathotypes for latent period and within one pathotype for sporulation capacity. One of these differences was tested again and confirmed. This indicates that the average aggressiveness level of a leaf rust pathotype may increase without any change in its virulence factors or microsatellite profile.  相似文献   

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