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1.
Abstract Twenty-five isolates of dissimilatory sulfate-reducing bacteria were clustered based on similarity analysis of their phospholipid ester-linked fatty acids (PLFA). Of these, 22 showed that phylogenetic relationships based on the sequence similarity of their 16S rRNA directly paralleled the PLFA relationships. Desulfobacter latus and Desulfobacter curvatus grouped with the other Desulfobacter spp. by 16S rRNA comparison but not with the PLFA analysis as they contained significantly more monoenoic PLFA than the others. Similarly, Desulfovibrio africanus clustered with the Desulfovibrio spp. by 16S rRNA but not with them when analyzed by PLFA patterns because of higher monoenoic PLFA content. Otherwise, clustering obtained with either analysis was essentially congruent. The relationships defined by PLFA patterns appeared robust to shifts in nutrients and terminal electron acceptors. Additional analyses utilizing the lipopolysaccharide-lipid A hydroxy fatty acid patterns appeared not to shift the relationships based on PLFA significantly except when completely absent, as in Gram-positive bacteria. Phylogenetic relationships between isolates defined by 16S rRNA sequence divergence represent a selection clearly different from the multi-enzyme activities responsible for the PLFA patterns. Determination of bacterial relationships based on different selective pressures for various cellular components provides more clues to evolutionary history leading to a more rational nomenclature.  相似文献   

2.
About 20 strains of rhizobia from wild legumes were characterized based on numerical analysis of phenotypic characteristics, nodulating ability, fatty acid methyl esters (FAME) and SDS-PAGE profiles of whole cell proteins. FAME analysis revealed that palmitic (16:0), stearic (18:0) and arachidonic (20:0) were detected in most of wild-legume rhizobia, the latter being uncommon in fatty acid profiles of Rhizobium and Sinorhizobium. Numerical analysis of FAME classified strains of wild-legume rhizobia into 9 clusters and one heterogeneous group. There was both agreement and disagreement with the clustering data based on phenotypic analysis and FAME analysis. Four strains were grouped together in the same cluster based on both methods. However, 4 another strains, which were placed in one cluster of phenotypic analysis, were distributed in several clusters after FAME analysis. SDS-PAGE of whole-cell proteins revealed that the rhizobial strains exhibited protein profiles with peptide bands ranging from 5-19 band per profile and showed molar mass of 110-183 kDa. As in the case of FAME analysis, numerical analysis of protein bands was compared with clustering of phenotypic analysis. Agreement of the two methods was obvious when clustering some strains but conflicted in the classification of some other strains. However, integration of the three methods could be the basis of a polyphasic taxonomy. The twenty strains of wild-legume rhizobia were finally classified as follows: 12 strains related to Rhizobium leguminosarum, 5 strains related to Sinorhizobium meliloti and 3 strains to Rhizobium spp. Rhizobia nodulating wild herb legumes are among indigenous strains nodulating crop legumes in cultivated as well as noncultivated lands.  相似文献   

3.
Ester-linked phospholipid fatty acid (PLFA) profiles of a Pseudomonas aureofaciens strain and an Arthrobacter protophormiae strain, each isolated from a subsurface sediment, were quantified in a starvation experiment in a silica sand porous medium under moist and dry conditions. Washed cells were added to sand microcosms and maintained under saturated conditions or subjected to desiccation by slow drying over a period of 16 days to final water potentials of approximately - 7.5 MPa for the P. aureofaciens and - 15 MPa for the A. protophormiae. In a third treatment, cells were added to saturated microcosms along with organic nutrients and maintained under saturated conditions. The numbers of culturable cells of both bacterial strains declined to below detection level within 16 days in both the moist and dried nutrient-deprived conditions, while direct counts and total PLFAs remained relatively constant. Both strains of bacteria maintained culturability in the nutrient-amended microcosms. The dried P. aureofaciens cells showed changes in PLFA profiles that are typically associated with stressed gram-negative cells, i.e., increased ratios of saturated to unsaturated fatty acids, increased ratios of trans- to cis-monoenoic fatty acids, and increased ratios of cyclopropyl fatty acids to their monoenoic precursors. P. aureofaciens starved under moist conditions showed few changes in PLFA profiles during the 16-day incubation, whereas cells incubated in the presence of nutrients showed decreases in the ratios of both saturated fatty acids to unsaturated fatty acids and cyclopropyl fatty acids to their monoenoic precursors. The PLFA profiles of A. protophormiae changed very little in response to either nutrient deprivation or desiccation. Diglyceride fatty acids, which have been proposed to be indicators of dead or lysed cells, remained relatively constant throughout the experiment. Only the A. protophormiae desiccated for 16 days showed an increase in the ratio of diglyceride fatty acids to PLFAs. The results of this laboratory experiment can be useful for interpreting PLFA profiles of subsurface communities of microorganisms for the purpose of determining their physiological status.  相似文献   

4.
Because algae have become more accepted as sources of human nutrition, phylogenetic analysis can help resolve the taxonomy of taxa that have not been well studied. This can help establish algal evolutionary relationships. Here, we compare Auxenochlorella protothecoides and 23 strains of Prototheca based on their complete 16S and partial 23S plastid rDNA sequences along with nutrient utilization (auxanographic) profiles. These data demonstrate that some of the species groupings are not in agreement with the molecular phylogenetic analyses and that auxanographic profiles are poor predictors of phylogenetic relationships.  相似文献   

5.
Cultivable strains were identified from sulfate-reducing fluidized-bed reactors (FBR) treating acidic metal-containing wastewater. The FBR-communities were further characterized using culture-independent phenotypic markers, phospholipid fatty acid (PLFA) profiling. After morphological screening of 128 bacterial strains and partial sequencing of 55 strains, 17 distinct phylogenetic types were identified and characterized further. A total of 14 and 6 different bacterial strains were isolated from ethanol- and lactate-fed FBRs, respectively. Sequencing of the 16S rRNA gene showed that these strains were affiliated with members of the δ-Proteobacteria, Firmicutes and Bacteroidetes The strains were affiliated with members of the genera Desulfovibrio, Desulfotomaculum, Desulfobulbus, Desulfitobacterium, Clostridium, Caloramator, Oxobacter, and Bacteroides. Many of the strains were only distantly related to previously described species and, thus, may represent novel species or genera. A number of the strains were not detected in previously employed molecular analyses of the FBR communities, and the major component of each FBR as identified in the molecular analyses were not retrieved as cultures in this study. Most of the SRB, and two of the non-SRB utilized ethanol and lactate as a source of carbon and energy, but none of the isolates grew on acetate, an intermediate in the oxidation of ethanol and lactate. PLFA analysis revealed that the FBR community members contained large amounts of saturated fatty acids. Although the PLFA analysis showed some signatures consistent with sulfate-reducing communities, it did not show any substantial difference in the microbial communities between the reactors, an outcome that was quite contrary to the culture-independent phylogenetic analyses.  相似文献   

6.
Twenty Lactococcus lactis strains with an L. lactis subsp. lactis phenotype isolated from five traditional cheeses made of raw milk with no added starters belonging to the L. lactis subsp. lactis and L. lactis subsp. cremoris genotypes (lactis and cremoris genotypes, respectively; 10 strains each) were subjected to a series of phenotypic and genetic typing methods, with the aims of determining their phylogenetic relationships and suitability as starters. Pulsed-field gel electrophoresis (PFGE) analysis of intact genomes digested with SalI and SmaI proved that all strains were different except for three isolates of the cremoris genotype, which showed identical PFGE profiles. Multilocus sequence typing (MLST) analysis using internal sequences of seven loci (namely, atpA, rpoA, pheS, pepN, bcaT, pepX, and 16S rRNA gene) revealed considerable intergenotype nucleotide polymorphism, although deduced amino acid changes were scarce. Analysis of the MLST data for the present strains and others from other dairy and nondairy sources showed that all of them clustered into the cremoris or lactis genotype group, by using both independent and combined gene sequences. These two groups of strains also showed distinctive carbohydrate fermentation and enzyme activity profiles, with the strains in the cremoris group showing broader profiles. However, the profiles of resistance/susceptibility to 16 antibiotics were very similar, showing no atypical resistance, except for tetracycline resistance in three identical cremoris genotype isolates. The numbers and concentrations of volatile compounds produced in milk by the strains belonging to these two groups were clearly different, with the cremoris genotype strains producing higher concentrations of more branched-chain, derived compounds. Together, the present results support the idea that the lactis and cremoris genotypes of phenotypic Lactococcus lactis subsp. lactis actually represent true subspecies. Some strains of the two subspecies in this study appear to be good starter candidates.  相似文献   

7.
Although starvation survival has been characterized for many bacteria, few subsurface bacteria have been tested, and few if any have been tested in natural subsurface porous media. We hypothesized that subsurface bacteria may be uniquely adapted for long-term survival in situ. We further hypothesized that subsurface conditions (sediment type and moisture content) would influence microbial survival. We compared starvation survival capabilities of surface and subsurface strains of Pseudomonas fluorescens and a novel Arthrobacter sp. in microcosms composed of natural sediments. Bacteria were incubated for up to 64 weeks under saturated and unsaturated conditions in sterilized microcosms containing either a silty sand paleosol (buried soil) or a sandy silt nonpaleosol sediment. Direct counts, plate counts, and cell sizes were measured. Membrane phospholipid fatty acid (PLFA) profiles were quantified to determine temporal patterns of PLFA stress signatures and differences in PLFAs among strains and treatments. The Arthrobacter strains survived better than the P. fluorescens strains; however, differences in survival between surface and subsurface strains of each genus were not significant. Bacteria survived better in the paleosol than in the nonpaleosol and survived better under saturated conditions than under unsaturated conditions. Cell volumes of all strains decreased; however, sediment type and moisture did not influence rates of miniaturization. Both P. fluorescens strains showed PLFA stress signatures typical for gram-negative bacteria: increased ratios of saturated to unsaturated fatty acids, increased ratios of trans- to cis-monoenoic fatty acids, and increased ratios of cyclopropyl to monoenoic precursor fatty acids. The Arthrobacter strains showed few changes in PLFAs. Environmental conditions strongly influenced PLFA profiles.  相似文献   

8.
Shukla E  Singh SS  Singh P  Mishra AK 《Protoplasma》2012,249(3):651-661
The fatty acid methyl ester (FAME) analysis of the 12 heterocystous cyanobacterial strains showed different fatty acid profiling based on the presence/absence and the percentage of 13 different types of fatty acids. The major fatty acids viz. palmitic acid (16:0), hexadecadienoic acid (16:2), stearic acid (18:0), oleic acid (18:1), linoleic (18:2), and linolenic acid (18:3) were present among all the strains except Cylindrospermum musicola where oleic acid (18:1) was absent. All the strains showed high levels of polyunsaturated fatty acid (PUFAs; 41-68.35%) followed by saturated fatty acid (SAFAs; 1.82-40.66%) and monounsaturated fatty acid (0.85-24.98%). Highest percentage of PUFAs and essential fatty acid (linolenic acid; 18:3) was reported in Scytonema bohnerii which can be used as fatty acid supplement in medical and biotechnological purpose. The cluster analysis based on FAME profiling suggests the presence of two distinct clusters with Euclidean distance ranging from 0 to 25. S. bohnerii of cluster I was distantly related to the other strains of cluster II. The genotypes of cluster II were further divided into two subclusters, i.e., IIa with C. musicola showing great divergence with the other genotypes of IIb which was further subdivided into two groups. Subsubcluster IIb(1) was represented by a genotype, Anabaena sp. whereas subsubcluster IIb(2) was distinguished by two groups, i.e., one group having significant similarity among their three genotypes showed distant relation with the other group having closely related six genotypes. To test the validity of the fatty acid profiles as a marker, cluster analysis has also been generated on the basis of morphological attributes. Our results suggest that FAME profiling might be used as species markers in the study of polyphasic approach based taxonomy and phylogenetic relationship.  相似文献   

9.
以9株白酒酿造过程中常见的菌株为对象,研究了不同种属菌株的细胞膜特征组分磷酸脂肪酸(PLFA)的特征,以及检出量与菌株生物量之间的关系.结果表明:供试细菌、放线菌、霉菌和酵母菌的PLFA指纹图谱存在显著差异,各菌株的PLFA指纹图谱可作为区别种属的依据.不同供试菌株生物量在一定范围内与检出的总PLFA量或16:0含量呈线性关系.将不同生物量的革兰氏阳性菌G+、革兰氏阴性菌G-和真菌分别加入糟醅后,检出的PLFA相对含量与对照差异显著.基于PLFA的指纹图谱能够定量或半定量地表征糟醅微生物群落结构特征及动态变化.经对多家酿酒企业糟醅PLFA组成的检测及微生物群落结构的剖析,该方法具有普适性.  相似文献   

10.
Computed data analysis of biochemical or molecular profiles is currently used in studies of microbial taxonomy, epidemiology, and microbial diversity. We assessed the use of Partial Least Squares (PLS) regression for multivariate data analysis in bacteriology. We identified clear relationships between RAPD profiles of propionibacteria strains and their species classification, autolytic capacities, and their origins. The PLS regression also predicted species identity of some strains with RAPD profiles partially related to those of reference strains. The PLS analysis also allowed us to identify key characteristics to use to classify strains. PLS regression is particularly well adapted to i) describing a collection of bacterial isolates, ii) justifying bacterial groupings using several sets of data, and iii) predicting phenotypic characters of strains that have been classified by routine typing methods.  相似文献   

11.
A total of about 50 strains of rhizobia from two leguminous trees (Acacia andProsopis) were described and compared with 20 reference strains of rhizobia, from other tree and herb legumes on the basis of protein, fatty acid and plasmid profiles, and DNA-DNA hybridization. The rhizobia formed thirteen clusters based on protein profile analysis. These clusters were not in complete agreement with a previously published cluster analysis based on numerical taxonomy of phenotypic characteristics and lipopolysaccharide (LPS) profile analysis (Zhanget al., Int. J. Syst. Bacteriol. 41, 104, 1991; Lindström and Zahran,FEMS Microbiol. Lett. 107, 327, 1993). The fatty acid methyl esters (FAME) of representative strains of rhizobia were analyzed. The rhizobia formed fourteen different clusters based on FAME analysis but the results also conflicted with the phenotypically based methods of analysis. Strains of rhizobia classified in one cluster by any of the above methods of analysis may have shown very different fatty acid profiles. The plasmid profile analysis of the tree rhizobia., on the other hand, was more consistent with the phenotype- and LPS-based numerical analysis. Some strains of the tree rhizobia showed medium or high levels of DNA homology withRhizobium meliloti. The DNA-DNA hybridization correlated well with protein and fatty acid profiles. The described methods provide a significant taxonomic tool for discrimination between rhizobia of leguminous trees. However, further DNA-DNA hybridization studies with other recognized species of rhizobia are needed for proper identification and classification of the diverse rhizobia from leguminous trees.  相似文献   

12.
Phylogenetic relationships were inferred for six Central American and Mexican pine species by analysing RAPD marker differences among pooled DNA samples. This population level pooling strategy discounts low-frequency allelic variation within taxa, thus obtaining a 'cumulative genotype' to compare among taxa. We used the morphologically based taxonomy of pines as the basis for inference concerning molecular marker divergence. Only RAPD polymorphisms that were repeatable and intensely amplified were used. The resulting data set was found to have strong hierarchical structure. Phylogenetic analyses were carried out using both neighbour-joining and maximum parsimony. The phylogenetic relationships indicated by analysis of the pine RAPD data provide new insights on pine taxonomy. The phylogenetic analyses of the RAPD marker data placed Pinus tecunumanii and P. patula clearly into two different subgroups, strongly supporting the classification of P. tecunumanii as a distinct species, and raise a new set of issues concerning the distinctions between Pinus tecunumanii and P. caribaea . Phylogenetically informative RAPD markers will be useful tools to address ex-situ conservation issues that involve taxonomic identification and species admixture.  相似文献   

13.
我国豇豆和绿豆根瘤菌的数值分类及16S rDNA PCR-RFLP研究   总被引:1,自引:0,他引:1  
对分离自中国14个不同省(自治区)的79株豇豆和绿豆根瘤菌及12株参比菌株进行了唯一碳、氮源利用,抗生素抗性,抗逆性和酶活性等128个表型性状的测定,并用MINTS软件进行聚类分析。表型性状测定结果发现,所有菌株都有极其广泛的碳、氮源利用谱,大多数菌株可在较宽的pH(pH5·0~11·0)值范围内生长,大部分菌株能在37℃高温条件下生长,个别菌株能耐受60℃高温较长时间(20~45min)的热激。聚类分析结果表明,全部供试菌株在63·5%的相似性水平上分为两大群:一个群为慢生菌群,另一群为快生和中慢生菌群;在79%的相似性水平上分为7个亚群。在数值分类的基础上,又将参比菌株增加到22株,对79株待测菌株进行了16SrDNAPCR-RFLP分析,16SrDNAPCR产物经HaeⅢ、HinfⅠ、MspⅠ和AluⅠ4种内切酶酶切共产生34种遗传图谱类型,经GelComparⅡ软件聚类后,在79%的相似性水平上也可划分为7个亚群,与数值分类的结果有很好的一致性。  相似文献   

14.
Species evolutionary relationships have traditionally been defined by sequence similarities of phylogenetic marker molecules, recently followed by whole-genome phylogenies based on gene order, average ortholog similarity or gene content. Here, we introduce genome conservation--a novel metric of evolutionary distances between species that simultaneously takes into account, both gene content and sequence similarity at the whole-genome level. Genome conservation represents a robust distance measure, as demonstrated by accurate phylogenetic reconstructions. The genome conservation matrix for all presently sequenced organisms exhibits a remarkable ability to define evolutionary relationships across all taxonomic ranges. An assessment of taxonomic ranks with genome conservation shows that certain ranks are inadequately described and raises the possibility for a more precise and quantitative taxonomy in the future. All phylogenetic reconstructions are available at the genome phylogeny server: .  相似文献   

15.
Bacteria are very important degraders of organic substances in aquatic environments. Despite their influential role in the carbon (and many other element) cycle(s), the specific genetic identity of active bacteria is mostly unknown, although contributing phylogenetic groups had been investigated. Moreover, the degree to which phenotypic potential (i.e., utilization of environmentally relevant carbon substrates) is related to the genomic identity of bacteria or bacterial groups is unclear. The present study compared the genomic fingerprints of 27 bacterial isolates from the humic River Warnow with their ability to utilize 14 environmentally relevant substrates. Acetate was the only substrate utilized by all bacterial strains. Only 60% of the strains respired glucose, but this substrate always stimulated the highest bacterial activity (respiration and growth). Two isolates, both closely related to the same Pseudomonas sp., also had very similar substrate utilization patterns. However, similar substrate utilization profiles commonly belonged to genetically different strains (e.g., the substrate profile of Janthinobacterium lividum OW6/RT-3 and Flavobacterium sp. OW3/15-5 differed by only three substrates). Substrate consumption was sometimes totally different for genetically related isolates. Thus, the genomic profiles of bacterial strains were not congruent with their different substrate utilization profiles. Additionally, changes in pre-incubation conditions strongly influenced substrate utilization. Therefore, it is problematic to infer substrate utilization and especially microbial dissolved organic matter transformation in aquatic systems from bacterial molecular taxonomy.  相似文献   

16.
药用植物内生芽孢杆菌的多样性和系统发育研究   总被引:3,自引:0,他引:3  
[目的]了解药用植物内生芽孢杆菌的生物多样性.[方法]采用数值分类、16S rDNA PCR RFLP、BOX-PCR指纹图谱和16S rDNA序列分析技术对分离于几种药用植物的内生芽孢杆菌和已知参比菌株进行表型、遗传多样性及系统发育研究.[结果]供试菌株在数值分类聚类分析中在84%的相似水平上产生13个表观群.16S rDNAPCR-RFLP分析表明供试菌株表现出丰富的遗传多样性.BOX-PCR指纹图谱分析进一步证明药用植物的内生芽孢杆菌的基因组也具有多样性,聚群的结果与数值分类有较好一致性.用软件在Genbank中进行所得序列的同源性检索,并构建系统发育树.由16S rDNA序列分析可知,供试的代表菌株SCAU11与球形芽孢杆菌(Bacillus sphaericus)亲缘关系最近,SCAU78和SCAU25为枯草芽孢杆菌(Bacillus subtilis)的两个亚种,代表菌株SCAU39与巨大芽孢杆菌(Bacillus megaterium)的亲缘关系最近.[结论]研究结果表明药用植物内生芽孢杆菌具有明显的表型和遗传多样性.  相似文献   

17.
对分离自我国11个省24个地区49株蚕豆根瘤菌及11株参比菌株进行了唯一碳源、氮源、抗生素、耐逆性和酶活性等138个表型性状测定,并用M INTS软件进行聚类分析。结果表明,全部供试菌株在59%的相似水平上聚在一起,在80%的相似水平上可分为6个群。其中群4与参比菌株聚在一起,而其他5个群均由未知菌组成。进一步对36株菌进行了16S rDNA PCR-RFLP分析,在85%相似水平上供试菌可分为4个群和1个独立的分支,其聚群结果与数值分类结果有较好的一致性。表型及遗传型分析结果表明,我国蚕豆根瘤菌具有极大的多样性。  相似文献   

18.
对分离自我国11个省24个地区49株蚕豆根瘤菌及11株参比菌株进行了唯一碳源、氮源、抗生素、耐逆性和酶活性等138个表型性状测定,并用MINTS软件进行聚类分析。结果表明,全部供试菌株在59%的相似水平上聚在一起,在80%的相似水平上可分为6个群。其中群4与参比菌株聚在一起,而其他5个群均由未知菌组成。进一步对36株菌进行了16S rDNA PCR—RFLP分析,在85%相似水平上供试菌可分为4个群和1个独立的分支,其聚群结果与数值分类结果有较好的一致性。表型及遗传型分析结果表明,我国蚕豆根瘤菌具有极大的多样性。  相似文献   

19.
Three denitrifying strains of bacteria capable of degrading poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) were isolated from activated sludge and characterized. All of the isolates had almost identical phenotypic characteristics. They were motile gram-negative rods with single polar flagella and grew well with simple organic compounds, as well as with PHB and PHBV, as carbon and energy sources under both aerobic and anaerobic denitrifying conditions. However, none of the sugars tested supported their growth. The cellular fatty acid profiles showed the presence of C16:1omega7cis and C16:0 as the major components and of 3-OH-C10:0 as the sole component of hydroxy fatty acids. Ubiquinone-8 was detected as the major respiratory quinone. A 16S rDNA sequence-based phylogenetic analysis showed that all the isolates belonged to the family Comamonadaceae, a major group of beta-Proteobacteria, but formed no monophyletic cluster with any previously known species of this family. The closest relative to our strains was an unidentified bacterium strain LW1 (=DSM 13225) (99.9% similarity), reported previously as a 1-chloro-4-nitrobenzene degrading bacterium. DNA-DNA hybridization levels among the new isolates were more than 60%, whereas those between our isolates and strain DSM 13225 were less than 50%. The G+C content of genomic DNA of the new strains was 64 to 65 mol%. Based on these results, we concluded that the PHBV-degrading denitrifying isolates should be classified as a new genus and a new species, for which we propose the name Diaphorobacter nitroreducens. The type strain is strain NA10B (=JCM 11421=CIP 107294). We also propose to classify strain DSM 13225 as a genospecies of Diaphorobacter.  相似文献   

20.
The carbohydrate compositions of cell wall were determined in the strains of class Urediniomycetes, mainly ballistoconidium-forming yeasts and related taxa. The major component of cell wall was mannose, and glucose was included as the second component, but xylose was not detected in any strain. Out of 41 strains examined, 39 contained galactose, 14 contained arabinose and 12 contained rhamnose. As a minor component, fucose was detected in 30 strains but not in 11 strains. A phylogenetic tree based on 18S rDNA sequences indicated that the fucose-lacking strains, Erythrobasidium hasegawianum, Rhodotolura aurantiaca, R. lactosa, R. minuta, Sakaguchia dacryoidea, Sporobolomyces coprosmae, S. elongatus, S. folicola, S. gracilis, S. kluyverinielii and S. oryzicola, constituted a distinct cluster from those strains which contained fucose. This cluster corresponded to one of the five subclusters, the Erythrobasidium cluster, in the phylogenetic tree of class Urediniomycetes. The carbohydrate composition of cell wall is believed to reflect the phylogenetic relationships among basidiomycetous fungi. The presence or absence of fucose in cell wall should be regarded as an important phenotypic characteristic in the taxonomy of basidiomycetes.  相似文献   

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