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PubMed2005

SecY gene sequence analysis for finer differentiation of diverse strains in the aster yellows phytoplasma group.

Aster yellows (AY) group (16SrI) phytoplasmas are associated with more than 100 economically important diseases worldwide and represent the most diverse and widespread phytoplasma group. Phylogenetic analysis of secY gene sequences resolved 10 genetically distinct lineages. The 10 lineages coincide with those delineated by phylogenetic analysis based on ribosomal protein (rp) gene sequences. However, greater genetic variability among the 10 lineages was revealed based on secY gene sequences. The distinct phylogenetic lineages can be readily identified through restriction fragment length polymorphism (RFLP) analysis of secY gene sequences. Ten subgroups were differentiated among the AY group phytoplasmas based on RFLP analysis of secY gene sequences. Phylogenetic analysis based on secY gene sequences in this study, and previous studies on the 16S rRNA gene, tuf gene, and rp gene sequences reinforced the notion that most subgroups identified by RFLP analysis of secY and rp gene sequences represent distinct phylogenetic lineages.

DNA, Bacterial↗
PubMed2000

Genetic variation in the endemic and endangered Rosmarinus tomentosus Huber-Morath & Maire (Labiatae) using RAPD markers. Random amplified polymorphic DNA.

Rosmarinus tomentosus Huber-Morath & Maire, an endemic species of southern Spain, is critically endangered as a consequence of habitat destruction by anthropogenic activities. Random amplified polymorphic DNA (RAPD) markers were used for initial evaluation of genetic variation in this species; among zones, among populations (within zones and independently of zones), and among individuals (within populations and zones). The eight primers used to study five populations provided 126 reliable bands, of which 109 (86%) were polymorphic. Analyses of the RAPD markers conducted using different approaches (cluster analysis, Shannon's diversity measure and analysis of molecular variance-AMOVA) showed similar results concerning genetic structure of populations of R. tomentosus. The three zones studied, and their corresponding populations, were clearly defined through UPGMA analysis. AMOVA analyses indicated that 18.0% of the total genetic variation was attributable to differences among zones. When total variance was partitioned without considering the zonal distribution of the populations, 34.1% was attributable to differences among populations, and 65.9% to differences among individuals within populations. Similar results were obtained using Shannon's information measure. Significant genetic distances (Ph(ist) values) were obtained between any two populations. This strong genetic differentiation among zones, populations and subpopulations may be explained by geographical distances and limited gene flow. This study will be of considerable help during subsequent formulations of conservation strategies for the endangered species R. tomentosus.

Asteraceae↗
PubMed1999

Identification by 16S rDNA fragment amplification and determination of genetic diversity by random amplified polymorphic DNA analysis of Pasteurella pneumotropica isolated from laboratory rodents.

Pasteurella pneumotropica is an opportunistic bacterium frequently isolated from colonies of various laboratory rodents. Identification of this species, including its differentiation into two distinct biotypes (Jawetz and Heyl), is usually based on the use of conventional bacteriologic methods. In this study, a 16S rDNA fragment amplification procedure was developed for use as an alternative method for identification and differentiation of P. pneumotropica. Polymerase chain reaction (PCR) products were two distinctive fragments of 937 and 564 bp specific for biotypes Jawetz and Heyl, respectively. Specificity of PCR products could be achieved by EcoRI cleavage, leading to 596 plus 341-bp and 346 plus 218-bp fragments for each of the amplification products. Use of this procedure confirmed identification of 34 field isolates and allowed definitive identification of some strains that could not have been done by use of bacteriologic examinations. Field isolates subjected to random amplified polymorphic DNA (RAPD) analysis had high genetic diversity among biotype Jawetz strains in contrast to biotype Heyl strains. In conclusion, RAPD could represent an additional means for identification of ambiguous strains of biotype Heyl and a valuable epidemiologic tool for identification of biotype Jawetz strains of P. pneumotropica.

Animals↗
PubMed1992

X-linked nephrogenic diabetes insipidus: from the ship Hopewell to RFLP studies.

Nephrogenic diabetes insipidus (NDI; designated 304800 in Mendelian Inheritance in Man) is an X-linked disorder with abnormal renal and extrarenal V2 vasopressin receptor responses. The mutant gene has been mapped to Xq28 by analysis of RFLPs, and tight linkage between DXS52 and NDI has been reported. In 1969, Bode and Crawford proposed, under the term "the Hopewell hypothesis," that most cases in North America could be traced to descendants of Ulster Scots who arrived in Nova Scotia in 1761 on the ship Hopewell. They also suggested a link between this family and a large Mormon pedigree. DNA samples obtained from 13 independent affected families, including 42 members of the Hopewell and Mormon pedigrees, were analyzed with probes in the Xq28 region. Genealogical reconstructions were performed. Linkage between NDI and DXS304 (probe U6:2.spl), DXS305 (St35-691), DXS52 (St14-1), DXS15 (DX13), and F8C (F814) showed no recombination in 12 families, with a maximum lod score of 13.5 for DXS52. A recombinant between NDI and DXS304, DXS305, was identified in one family. The haplotype segregating with the disease in the Hopewell pedigree was not shared by other North American families. PCR analysis of the St14 VNTR allowed the distinction of two alleles that were not distinguishable by Southern analysis. Carrier status was predicted in 24 of 26 at-risk females. The Hopewell hypothesis cannot explain the origin of NDI in many of the North American families, since they have no apparent relationship with the Hopewell early settlers, either by haplotype or by genealogical analysis. We confirm the locus homogeneity of the disease by linkage analysis in ethnically diverse families. PCR analysis of the DXS52 VNTR in NDI families is very useful for carrier testing and presymptomatic diagnosis, which can prevent the first manifestations of dehydration.

Blotting, Southern↗
PubMed1999

Quasispecies of hepatitis C virus in serum and in three different parts of the liver of patients with chronic hepatitis.

Hepatitis C virus (HCV) has been known to infect hosts as a quasispecies. Several reports have shown this using serum samples, but there is little information about quasispecies in the liver. In this study, we evaluated quasispecies in serum and in 3 different parts of the liver in 8 patients with varying severity of chronic hepatitis C by calculating nucleotide diversity, entropy, type of substitution and by phylogenetic analysis. Nucleotide diversity of HCV was different in each sample and ranged from 0.37% +/- 0.31% to 4.10% +/- 1.06%. However, the degree of HCV diversity in serum correlated with that in the liver in each patient (P <.01). Common HCV clones were found both in serum and liver samples in all 6 noncirrhotic patients, but all serum clones were different from the clones from the 2 cirrhotic livers. Phylogenetic analysis showed that the degree of genetic diversity of HCV among the 3 liver samples was significantly high in the 4 patients with fibrosis. These genetic compartmentalizations of HCV did not depend on the type of substitution or the viral load of each liver sample. HCV quasispecies within the liver may be closely related to the viral life cycle and the pathogenesis of persistent infection of HCV.

Adult↗
PubMed2002

Analysis of hantavirus genetic diversity in Argentina: S segment-derived phylogeny.

Nucleotide sequences were determined for the complete S genome segments of the six distinct hantavirus genotypes from Argentina and for two cell culture-isolated Andes virus strains from Chile. Phylogenetic analysis indicates that, although divergent from each other, all Argentinian hantavirus genotypes group together and form a novel phylogenetic clade with the Andes virus. The previously characterized South American hantaviruses Laguna Negra virus and Rio Mamore virus make up another clade that originates from the same ancestral node as the Argentinian/Chilean viruses. Within the clade of Argentinian/Chilean viruses, three subclades can be defined, although the branching order is somewhat obscure. These are made of (i) "Lechiguanas-like" virus genotypes, (ii) Maciel virus and Pergamino virus genotypes, and (iii) strains of the Andes virus. Two hantavirus genotypes from Brazil, Araraquara and Castello dos Sonhos, were found to group with Maciel virus and Andes virus, respectively. The nucleocapsid protein amino acid sequence variability among the members of the Argentinian/Chilean clade does not exceed 5.8%. It is especially low (3.5%) among oryzomyine species-associated virus genotypes, suggesting recent divergence from the common ancestor. Interestingly, the Maciel and Pergamino viruses fit well with the rest of the clade although their hosts are akodontine rodents. Taken together, these data suggest that under conditions in which potential hosts display a high level of genetic diversity and are sympatric, host switching may play a prominent role in establishing hantavirus genetic diversity. However, cospeciation still remains the dominant factor in the evolution of hantaviruses.

Animals↗
PubMed2001

A worldwide analysis of AG molecular diversity inferred from serology.

Ten population samples from different geographic origins were tested serologically for the AG polymorphism of human beta-lipoproteins. Their haplotype frequencies were used with previously published data to perform a wide analysis of AG genetic differentiations throughout the world. Coancestry coefficients were computed from weighted F(ST)s among populations by using a matrix of molecular distances among AG haplotypes, which is here determined on the basis of DNA studies. Coancestry coefficients derived from unweighted F(ST)s and more classical Prevosti distances were computed on the same data and used for a comparison. In all cases a highly significant correlation was found between genetics and geography on a worldwide scale, while the significance of the correlation with linguistics differed. A test of significance of the pairwise F(ST)s among populations also gave different results depending on whether the molecular distance matrix among AG haplotypes was included. Globally, this study shows that in spite of being highly significantly correlated to each other, different genetic distance measures can lead to different interpretations of the same data set. Moreover, the elucidation of the molecular models related to the presently known serological polymorphisms may represent an additional tool for analyzing such polymorphisms in human population genetics studies.

Amino Acid Substitution↗
PubMed2006

Genetic diversity of source germplasm of Upland cotton in China as determined by SSR marker analysis.

The genetic diversity of 43 sources of Upland cotton germplasm with different parental origins, breeding periods, and ecological growing areas in China were studied on the basis of simple sequence repeat (SSR) markers. A total of 130 gene alleles with 80% polymorphism were detected from 36 SSR primers. The number of alleles per primer ranged from two to eight with an average of 3.6. The polymorphism information content (PIC) range was 0.278-0.865, with an average of 0.62. The average genotype diversity index (H') was 1.102, the highest was 2.039 and the lowest was 0.451. The average coefficient of the genetic similarity of SSR markers among source germplasm was 0.610, ranging from 0.409 to 0.865. These indicated that the genetic diversity at the genomic level of the selected source germplasm was rich, and was representative of the diversity of the germplasms, in general. The diversity at the genome level of the base germplasm from the second and third breeding periods was decreased compared to that of the first period, indicating that the cotton genetic background in China became narrow gradually. The diversity of SSR markers among the base germplasm from early maturity cotton growing areas in the north was higher than those from the Huanghe and Yangtze growing areas. The molecular marker genetic similarity index of the domestic varieties was higher than that in the introduced varieties, which indicates that the genetic diversity in domestic cultivars was lower than that in the introduced varieties. This study gives an overview of the genetic diversity of the cotton germplasm base in China, and provides a guide for breeders to develop new cultivars efficiently.

China↗
PubMed1992

PCR-based RFLP analysis of DNA sequence diversity in the gastric pathogen Helicobacter pylori.

DNA sequence diversity among 60 independent isolates of the gastric pathogen Helicobacter pylori was assessed by testing for restriction fragment length polymorphisms (RFLPs) in several PCR-amplified gene segments. 18 Mbol and 27 HaeIII RFLPs were found in the 2.4 kb ureA-ureB (urease) segment from the 60 strains; this identified 44 separate groups, with each group containing one to four isolates. With one exception, each isolate not distinguished from the others by RFLPs in ureA-ureB was distinguished by Mbol digestion of the neighboring 1.7 kb ureC-ureD segment. The 1.5 kb flaA (flagellin) gene, which is not close to ure gene cluster, was also highly polymorphic. In contrast, isolates from initial and followup biopsies yielded identical restriction patterns in each of the three cases tested. The potential of this method for detecting population heterogeneity was tested by mixing DNAs from different strains before amplification: the arrays of restriction fragments obtained indicated co-amplification from both genomes in each of the five pairwise combinations tested. These results show that H. pylori is a very diverse species, that indicate PCR-based RFLP tests are almost as sensitive as arbitrary primer PCR (RAPD) tests, and suggest that such RFLP tests will be useful for direct analysis of H. pylori in biopsy and gastric juice specimens.

Base Sequence↗
PubMed2001

Analysis of codon usage diversity of bacterial genes with a self-organizing map (SOM): characterization of horizontally transferred genes with emphasis on the E. coli O157 genome.

With increases in the amounts of available DNA sequence data, it has become increasingly important to develop tools for comprehensive systematic analysis and comparison of species-specific characteristics of protein-coding sequences for a wide variety of genomes. In the present study, we used a novel neural-network algorithm, a self-organizing map (SOM), to efficiently and comprehensively analyze codon usage in approximately 60,000 genes from 29 bacterial species simultaneously. This SOM makes it possible to cluster and visualize genes of individual species separately at a much higher resolution than can be obtained with principal component analysis. The organization of the SOM can be explained by the genome G+C% and tRNA compositions of the individual species. We used SOM to examine codon usage heterogeneity in the E. coli O157 genome, which contains 'O157-unique segments' (O-islands), and showed that SOM is a powerful tool for characterization of horizontally transferred genes.

Algorithms↗
PubMed1988

Toward a population genetic analysis of Salmonella: genetic diversity and relationships among strains of serotypes S. choleraesuis, S. derby, S. dublin, S. enteritidis, S. heidelberg, S. infantis, S. newport, and S. typhimurium.

Variation in the chromosomal genomes of 1527 isolates of eight common serotypes (O and H antigen profiles) of Salmonella was assessed by analysis of electrophoretically demonstrable allelic polymorphism at 23 metabolic enzyme loci. Seventy-one distinctive electrophoretic types, representing multilocus genotypes, were identified. A basically clonal population structure was indicated by the presence of strong linkage disequilibrium among enzyme loci, the association of each serotype with a relatively small number of multilocus enzyme genotypes, and the global distribution of certain genotypes. For each of six of the serotypes, 83-96% of isolates were members of a single clone. The occurrence of each of four serotypes (S. derby, S. enteritidis, S. infantis, and S. newport) in isolates of clones belonging to several evolutionary lineages, some of which are distantly related, suggests that the horizontal transfer and recombination of chromosomal genes mediating expression of cell-surface antigens has been a significant process in the evolution of the salmonellae. Two divergent clone clusters of S. derby differ in the relative frequency with which they cause disease in birds versus mammals, and two major lineages of S. newport differ in the frequency with which their clones are associated with disease in humans versus animals.

Alleles↗
PubMed2004

Characterization of bacterial diversity in pulque, a traditional Mexican alcoholic fermented beverage, as determined by 16S rDNA analysis.

The bacterial diversity in pulque, a traditional Mexican alcoholic fermented beverage, was studied in 16S rDNA clone libraries from three pulque samples. Sequenced clones identified as Lactobacillus acidophilus, Lactobacillus strain ASF360, L. kefir, L. acetotolerans, L. hilgardii, L. plantarum, Leuconostoc pseudomesenteroides, Microbacterium arborescens, Flavobacterium johnsoniae, Acetobacter pomorium, Gluconobacter oxydans, and Hafnia alvei, were detected for the first time in pulque. Identity of 16S rDNA sequenced clones showed that bacterial diversity present among pulque samples is dominated by Lactobacillus species (80.97%). Seventy-eight clones exhibited less than 95% of relatedness to NCBI database sequences, which may indicate the presence of new species in pulque samples.

Agave↗
PubMed2006

[Genetic diversity of Eucommia ulmoides by RAPD analysis].

OBJECTIVE: To determine the genetic diversity of Eucommia ulmoides. METHOD: 260 samples of 20 populations were analyzed through radom amplified polymorphic DHA (RAPD). RESULT: Total polymorphic loci percentage was 96.36 and the average was 38.92. 110 bands were produced with 10 random primers and 106 were polymorphic. Nei's gene diversity (H) was 0.246 1, Shannon's Information index(I) was 0.386 8, Gst was 0.424 4, indicating that 42.44% of the genetic variation was distributed among populations and 57.65% within populations. CONCLUSION: The genetic variation was relatively high in E. ulmoides, so the genetic diversity conservation principle should mainly focus on protection of the populations.

China↗
PubMed2000

[A study of the genetic diversity of common wild rice (O. rufipogon Griff.) and cultivated rice (O. sativa L.) by RFLP analysis].

The genetic diversity of cultivated rice and common wild rice from 10 countries in Asia were evaluated by RFLP analysis using 44 probes of single copy. We used the parameters such as proportion of polymorphic loci (P), average number of alleles per locus (A), average number genotype per locus (Ng), degree of heterozygate per locus (Ho) and average gene diversity (Hs) showing genetic diversity. It is indicated that the common wild rice from China has the largest genetic diversity in the 10 countries, and the secondary is the common wild rice from India. The results also showed that the genetic diversity in Chinese common wild rice is larger than the genetic diversity in common wild rice of South and Southeast Asia. The average gene diversity (Hs) of South Asian wild rice is higher than Southeast Asian wild rice, while the parameters value of proportion of polymorphic loci (P), average number of alleles per locus (A) and average number genotype per locus (Ng), Southeast Asian wild rice is higher than South Asian wild rice. It is found that the genetic diversity in cultivated rice is obviously lower than in common wild rice. In the detected 44 loci, the number of polymorphic loci on O. sativa is as the same as 3/4 in O. rufipogon, the number of alleles in O. sativa is as the same as 60% in O. rufipogon, the number of genotype in O. sativa is as same as 1/2 in O. rufipogon. In O. sativa, the genetic diversity of indica is larger than that of japonica. O. rufipogon from China have highest value in degree of hetrozygate per locus (Ho), and Ho of wild rice is as two times as cultivated rice. It is showed that in the proceeding of wild rice evolved into cultivated rice, Ho and number of alleles was reduced, and the genetic diversity was decreased.

Genetic Variation↗
PubMed2002

[RAPD analysis for the genetic diversity of Brassica rapa in Tibet].

Tibet, a most beautiful place, locating in southwestern China. She has been called as the third pole of the earth. Unique geological history, complex land surface and climatic zones, various soil types, all different wild vegetations etc., all of these make Tibet a very typical area of vertical agricultural ecosystem. The ecosystem in Tibet may be the most complex in the world, which varies from place to place. Genetic differentiation of 107 accessions of Brassica rapa from Tibet plateau was studied by DNA PAPD analysis using 2210 bp random primers, the genetical distribution in 107 accession of Brassica rapa from Tibet plateau was found. The results are as follows: (1) Total 236 bands were produced from 107 Tibet oilseed accession of B. rapa germaplasm resource in Tibet, of 210 bands amplified from B. rapa germaplasm resource showed polymorphism, with the ratio 88.98%. The result showed that oilseed accession of B. rapa in Tibet has richer genetic diversity; (2) Dendrogram constructed from DNA RAPDs showed that 107 accessions of B. rapa from Tibet plateau were divided into 11 cluster by calculating genetic distance, the cluster analysis showed that the genetic variation among oilseed accessions of B. rapa was closely related with their eco-geographic distribution; extensive variation existed among the accessions from Tibet Province. Based on the analysis of unique geological history, complex land surface and climatic zones, various soil types, complex growing environments, long agricultural history, different cropping systems, and natural and artificial selection as well as plant geography, plant evolution theory, it concludes that Tibet is one of the oil seed gene centers in the word.

Brassica rapa↗
PubMed1997

Analysis of VH gene diversity in rainbow trout (Oncorhynchus mykiss): both nonsynonymous and synonymous nucleotide changes are more frequent in CDRs than in FRs.

Forty-six immunoglobulin VH gene sequences of rainbow trout were compiled to analyze the extent of variations and the frequency of nucleotide changes in CDRs and FRs. The results show that the frequency of nonsynonymous (amino acid replacing) changes (Ka) are on average 4.9 times higher in complementarity determining regions (CDRs) than in FRs, thus contributing more diversity in CDRs. Unexpectedly, however, the frequency of synonymous (silent) changes (Ks) show the same tendency: it was 5.3 times higher in CDRs than in framework regions (FRs). The distribution of Ks/Ka ratios of each comparison shows no segregation between CDRs and FRs. The same analysis applied to five germline VH genes of Heterodontus francisci shows the same result as was found with the rainbow trout. In contrast, the results from mouse data show that, while the CDR/FR ratio for Ka is much higher (7.4), the CDR/FR ratio for Ks is only slightly higher (1.8). The distribution of Ks/Ka ratios in mouse indicates clear segregation between CDRs and FRs. This suggests that CDR germline diversity is largely generated by gene conversion in VHs of lower vertebrates such as rainbow trout or shark. This mechanism might be advantageous to lower vertebrates in generating V gene diversity faster than other mechanisms such as point mutation and selection.

Animals↗
PubMed2005

Analysis of the genetic diversity of the Plasmodium falciparum multidrug resistance gene 5' upstream region.

Recent findings indicating a low level of polymorphism in the Plasmodium falciparum genome have led to the hypothesis that existent polymorphisms are likely to have functional significance. We tested this hypothesis by developing a map of the polymorphism in the P. falciparum multidrug resistance 1 (pfmdr1) gene 5' upstream region and assaying its correlation with drug resistance in a sample of field isolates from Dakar, Senegal. A comparison of six geographically diverse laboratory strains showed that the 1.94-kb 5'-untranslated region is highly monomorphic, with a total of four unique single nucleotide polymorphisms (SNPs) being identified. All of the mutations were localized to a 462-basepair region proximal to the transcription start point. Analysis of this region in field isolates shows the prevalence of one SNP throughout the entire population of parasites, irrespective of drug resistance status. The SNP frequency of the pfmdr1 upstream region is lower than that found in the noncoding region of other genes.

ATP-Binding Cassette Transporters↗
PubMed1999

Molecular analysis of the microbial diversity present in the colonic wall, colonic lumen, and cecal lumen of a pig.

Random clones of 16S ribosomal DNA gene sequences were isolated after PCR amplification with eubacterial primers from total genomic DNA recovered from samples of the colonic lumen, colonic wall, and cecal lumen from a pig. Sequences were also obtained for cultures isolated anaerobically from the same colonic-wall sample. Phylogenetic analysis showed that many sequences were related to those of Lactobacillus or Streptococcus spp. or fell into clusters IX, XIVa, and XI of gram-positive bacteria. In addition, 59% of randomly cloned sequences showed less than 95% similarity to database entries or sequences from cultivated organisms. Cultivation bias is also suggested by the fact that the majority of isolates (54%) recovered from the colon wall by culturing were related to Lactobacillus and Streptococcus, whereas this group accounted for only one-third of the sequence variation for the same sample from random cloning. The remaining cultured isolates were mainly Selenomonas related. A higher proportion of Lactobacillus reuteri-related sequences than of Lactobacillus acidophilus- and Lactobacillus amylovorus-related sequences were present in the colonic-wall sample. Since the majority of bacterial ribosomal sequences recovered from the colon wall are less than 95% related to known organisms, the roles of many of the predominant wall-associated bacteria remain to be defined.

Anaerobiosis↗