A preliminary amplified fragment length polymorphism (AFLP) analysis of Spirula spirula (Cephalopoda: Spirulida)

Author(s):  
Kerstin M. Warnke
1999 ◽  
Vol 181 (15) ◽  
pp. 4469-4475 ◽  
Author(s):  
Adam Meijer ◽  
Servaas A. Morré ◽  
Adriaan J. C. Van Den Brule ◽  
Paul H. M. Savelkoul ◽  
Jacobus M. Ossewaarde

ABSTRACT The genomic relatedness of 19 Chlamydia pneumoniaeisolates (17 from respiratory origin and 2 from atherosclerotic origin), 21 Chlamydia trachomatis isolates (all serovars from the human biovar, an isolate from the mouse biovar, and a porcine isolate), 6 Chlamydia psittaci isolates (5 avian isolates and 1 feline isolate), and 1 Chlamydia pecorum isolate was studied by analyzing genomic amplified fragment length polymorphism (AFLP) fingerprints. The AFLP procedure was adapted from a previously developed method for characterization of clinical C. trachomatis isolates. The fingerprints of all C. pneumoniae isolates were nearly identical, clustering together at a Dice similarity of 92.6% (± 1.6% standard deviation). The fingerprints of the C. trachomatis isolates of human, mouse, and swine origin were clearly distinct from each other. The fingerprints of the isolates from the human biovar could be divided into at least 12 different types when the presence or absence of specific bands was taken into account. The C. psittacifingerprints could be divided into a parakeet, a pigeon, and a feline type. The fingerprint of C. pecorum was clearly distinct from all others. Cluster analysis of selected isolates from all species revealed groups other than those based on sequence data from single genes (in particular, omp1 and rRNA genes) but was in agreement with available DNA-DNA hybridization data. In conclusion, cluster analysis of AFLP fingerprints of representatives of all species provided suggestions for a grouping of chlamydiae based on the analysis of the whole genome. Furthermore, genomic AFLP analysis showed that the genome of C. pneumoniae is highly conserved and that no differences exist between isolates of respiratory and atherosclerotic origins.


2000 ◽  
Vol 38 (9) ◽  
pp. 3379-3387 ◽  
Author(s):  
Bjørn-Arne Lindstedt ◽  
Even Heir ◽  
Traute Vardund ◽  
Kjetil K. Melby ◽  
Georg Kapperud

Amplified-fragment length polymorphism (AFLP) analysis with the endonucleases BglII and MfeI was used to genotype 91 Campylobacter jejuni subsp. jejunistrains from outbreaks and sporadic cases. AFLP-generated fragments were labeled with fluorescent dye and separated by capillary electrophoresis. The software packages GeneScan and GelCompar II were used to calculate AFLP pattern similarities and to investigate phylogenetic relationships among the genotyped strains. The AFLP method was compared with two additional DNA-based typing methods, pulsed-field gel electrophoresis (PFGE) using SmaI and restriction fragment length polymorphism analysis on PCR products (PCR-RFLP) of theflaA and flaB genes. We found that AFLP analysis of C. jejuni strains is a rapid method that offers better discriminatory power than do both PFGE and PCR-RFLP. AFLP and, to a lesser extent, PCR-RFLP could differentiate strains within the same PFGE profiles, which also makes PCR-RFLP an alternative to PFGE. We were able to clearly distinguish 9 of 10 recognized outbreaks by AFLP and to identify similarities among outbreak and sporadic strains. Therefore, AFLP is suitable for epidemiological surveillance ofC. jejuni and will be an excellent tool for source identification in outbreak situations.


2006 ◽  
Vol 96 (10) ◽  
pp. 1097-1107 ◽  
Author(s):  
Larry J. Heilmann ◽  
Nadav Nitzan ◽  
Dennis A. Johnson ◽  
Julie S. Pasche ◽  
Curt Doetkott ◽  
...  

Amplified fragment length polymorphism (AFLP) using three primer sets was used to characterize 211 Colletotrichum coccodes isolates from North America, 112 of which were assigned to six vegetative compatibility groups (VCGs) using nitrate nonutilizing (nit) mutants. These isolates clustered into five corresponding groups by unweighted pairgroup method with arithmetic means-based cluster analysis of AFLP banding patterns. Isolates of C. coccodes belonging to NA-VCG1 and NA-VCG3 were closely related, as were isolates belonging to NA-VCG2 and NA-VCG5. Based on bootstrap analysis of AFLP data, the two isolates originally assigned to NA-VCG4 clustered with isolates belonging to NA-VCG2 and NA-VCG5. C. coccodes isolates that clustered with two isolates belonging to NA-VCG6 were the most diverged from other groups, including seven isolates collected from hosts other than potato. As opposed to the bootstrap analysis, a quadratic discriminant analysis (QDA) of AFLP data correctly categorized the two isolates of NA-VCG4. Furthermore, in isolates where VCG determinations had been made, this model correctly classified isolates of all VCGs. QDA classifications were identical to those made by the bootstrap analysis, with the exception of VCG4. Overall, classifications made by the QDA model were strongly correlated (r = 0.970, P < 0.001) to the VCGs assigned by traditional methods. All 99 C. coccodes isolates evaluated only by AFLP also were subjected to QDA, leading to the assignment of a presumptive VCG for each isolate. No isolates of VCG4 or VCG6 were identified by QDA within this population. Symptoms of black dot developed in plants inoculated with isolates collected from both potato and non-potato hosts. However, total yield was not significantly reduced by infection with non-potato isolates. The lack of any additional groups identified by AFLP analysis may be an indicator of a limited level of genetic variation among North American C. coccodes isolates. AFLP is a much more efficient technique for subspecific characterization in C. coccodes than VCG analysis utilizing nit mutants and will provide an effective means by which the population biology of this pathogen can be further investigated worldwide.


2007 ◽  
Vol 56 (9) ◽  
pp. 1152-1160 ◽  
Author(s):  
Stacy L. Pfaller ◽  
Timothy W. Aronson ◽  
Alan E. Holtzman ◽  
Terry C. Covert

Fine-scale genotyping methods are necessary in order to identify possible sources of human exposure to opportunistic pathogens belonging to the Mycobacterium avium complex (MAC). In this study, amplified fragment length polymorphism (AFLP) analysis was evaluated for fingerprinting 159 patient and environmental MAC isolates from southern California. AFLP analysis accurately identified strains belonging to M. avium and Mycobacterium intracellulare and differentiated between strains within each species. The method was also able to differentiate strains that were presumed to be genetically identical in two previous studies using large RFLP analysis with PFGE, or PCR-amplification of DNA segments located between insertion sequences IS1245 and IS1311. For M. avium, drinking-water isolates clustered more closely with each other than with patient or food isolates. Patient isolates were more genetically diverse. None of the environmental isolates shared identical AFLP patterns with patient isolates for either species. There were, however, environmental isolates that shared identical patterns, and patient isolates that shared identical patterns. A subset of the isolates, which are referred to as MX isolates due to their ambiguous identification with the Gen-Probe system, produced AFLP patterns similar to those obtained from M. intracellulare isolates. Sequence analysis of 16S rDNA obtained from the MX isolates suggests that they are strains of M. intracellulare that were not correctly identified by the M. intracellulare AccuProbe from Gen-Probe.


2009 ◽  
Vol 134 (4) ◽  
pp. 428-434 ◽  
Author(s):  
Salih Kafkas ◽  
Sezai Ercişli ◽  
Yıldız Doğan ◽  
Yaşar Ertürk ◽  
Ayhan Haznedar ◽  
...  

Individuals in most countries around the world drink tea (Camellia sinensis). Tea drinking has attained ceremonial status in many places as a social and medicinal beverage. Although tea is of great importance in Turkey's economy, little is known about the pattern of genetic variation among the various tea genotypes grown in Turkey. A total of 32 tea genotypes found at the Ataturk Tea and Horticulture Research Institute in the eastern Black Sea region of Turkey were sampled. Fluorescent dye amplified fragment length polymorphism (AFLP) markers and capillary electrophoresis were applied for molecular characterization. The AFLP analysis with six primer combinations generated 835 fragments of which 567 were polymorphic, corresponding to 69.8% polymorphism. Resolving powers of the AFLP primers ranged from 62.6 to 81.9, yielding a total of 437.8; the polymorphic information content (PIC) ranged from 0.76 to 0.83, with an average of 0.79. Genetic similarity values ranged from 0.68 to 0.92, with an average of 0.76. The dendrogram derived by unweighted pair group method with arithmetic mean algorithm (UPGMA) and principal coordinate analysis (PCoA) revealed that all tea genotypes could be clearly divided into four distinct clusters. The results of this study will provide valuable information to the tea cultivar breeding program for the purpose of parental selection.


2016 ◽  
Vol 73 (1) ◽  
Author(s):  
Nurita TORUAN-MATHIUS ◽  
. ENDANG-YUNIASTUTI ◽  
Ridwan SETIAMIHARJA ◽  
Murdaningsih H. KARMANA

SummaryTissue culture-derived plants of oil palmmay develop abnormal flowers in whichprimordial stamens are converted into carpel-liketissue or mantled fruits, and sterile male flowers.This abnormality can be heritable, individualpalm may show variation in mantling andreversion to the normal phenotype over time hasbeen observed. The aim of these experiments wasto analyze the differences between normal andabnormal genotypes by DNA-AFLP. DNA wasisolated from young fruits of three clones,MK152, MK209, and MK 212 each of themconsisted of normal fruits, abnormal fruits andsterile male flowers. The research consisted of (i)selection of AFLP primer which can producepolymorphic bands, (ii) genetic similaritiesanalysis, UPGMA, principal component analysisand specific DNA bands between normal orabnormal genotypes. For primers selection, 20AFLP primers with DNA from MK 152 normaland abnormal genotypes were used. The selectedprimers were then used to amplify DNA of ninegenotypes. The results show that 10 primer com-binations EcoRI/MseI produced polymorphicbands. Each primer from 10 primer producedonly one or two DNA bands indicates that thedifferences between normal and abnormalgenotypes in the same clone. However, nopolymorphism was consistently found betweennormal and abnormal clones in all the sets.Genetic similarity analysis shows that betweengenotype had high genetic similarities, around92-99%. The results of UPGMA found thedifferent clustering between normal fruit,abnormal male and abnormal fruits. The resultsshow same as clustering based on first, secondand third component. This suggest that, whilstAFLP method is an effective way of detectingvariation in tissue culture-derived plants,different approaches are required to identify thecasual basis of the mantled fruit abnormality.RingkasanTanaman kelapa sawit yang dihasilkan darikultur jaringan, umumnya dalam perkembangan-nya akan memiliki organ reproduktif yangabnormal. Abnormalitas berupa primordialstamen berkembang menjadi bentuk jaringanseperti karpel, buah mantel, atau bunga jantanmandul. Penelitian ini bertujuan untukmendapatkan pembeda DNA-AFLP antaragenotip normal dan abnormal pada klon-klonkelapa sawit. DNA diisolasi dari buah muda klonMK 152, MK 209, dan MK 212 yang masing-masing terdiri atas genotip normal, berbuahabnormal, dan berbunga jantan steril. Percobaanmencakup (i) seleksi primer AFLP yang mampumenghasilkan pita yang polimorfis, (ii) analisiskemiripan genetik, UPGMA, komponen utamadan pita pembeda antar genotip normal danabnormal. Seleksi primer dilakukan terhadap 20primer AFLP menggunakan DNA dari genotipMK 152 yang normal dan abnormal. Selanjutnyaprimer terpilih digunakan untuk mengamplifikasiDNA dari kesembilan genotip yang diuji. Hasilyang diperoleh menunjukkan bahwa 10 kombi-nasi primer EcoRI/MseI mampu menghasilkanpita yang polimorfis. Dari 10 primer yang diuji,masing-masing hanya menghasilkan satu ataudua pita DNA yang mampu membedakan genotipnormal dan abnormal dalam klon yang sama.Namun, tidak ada pita DNA spesifik yangmampu membedakan genotip normal denganabnormal untuk seluruh klon yang diuji. Analisiskemiripan genetik menunjukkan bahwa antargenotip memiliki kemiripan genetik yang sangattinggi, yaitu 92-99%. Dari hasil UPGMAdiperoleh pengelompokan yang terpisah antargenotip normal, abnormal jantan dan buahabnormal. Hasil tersebut didukung olehpengelompokan berdasarkan komponen utamasatu, dua dan tiga. Dapat disimpulkan bahwa,teknik AFLP tidak efektif untuk mendeteksipembeda antar genotip tanaman yang diperolehdari kultur jaringan, pendekatan lainnyadiperlukan untuk mengidentifikasi abnormalitas.


2002 ◽  
Vol 20 (2) ◽  
pp. 110-113
Author(s):  
Suping Zhou ◽  
Roger J. Sauve

Abstract The species identity of ‘Roy Davidson’, a popular commercial cultivar in the genus Pulmonaria, is uncertain. To determine to which species ‘Roy Davidson’ is related to, we used fifteen primer combinations for Fluorescent-Amplified Fragment Length Polymorphism (F-AFLP) analysis. UPGMA dendrograms generated from AFLP markers obtained from the primer combinations showed that ‘Roy Davidson’ shares more genetic information with P. longifolia than with P. saccharata. Several factors that can affect F-AFLP profiles (DNA isolation from leaf tissue, optimization of F-AFLP procedures, and the use of this method for genotyping pulmonarias) were also evaluated; the results are discussed in this paper.


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