Differentiation of three Bursaphelenchus species by means of RAPD- PCR
Bibliographic record
Abstract
The RAPD-PCR (Random Amplified Polymorphie DNA-Polymerase Chain Reaction) technique has been developed to differentiate the pinewood nematode, Bursaphelenchus xylophilus , from two related species. We have examined 14 isolates of B. xylophilus , 12 isolates of B. mucronatus , 7 isolates of B. fraudulentus and 2 undetermined Bursaphelenchus provenances obtained from Asia, Europe and North America. The nematodes were multiplied on Botrytis cinerea on malt agar, and tested with 13 different ten base random primers from the Operon kits 8, E, Y and Z. Informative patterns of amplified DNA fragments were obtained revealing interspecific as well as intraspecific differences. Reliable differentiation and identification of B. xylophilus and B. mucronatus was possible with primer OPY-0 1 , whereas B. xylophilus could be distinguished from B. fraudulentus using primer OPB-07. Primer OPZ-08 was most suitable for differentiation of B. mucronatus from B.fraudulentus . Our results confirmed the assignment to B. mucronatus of the C 2 isolate from Canada proposed by HARMEY and HARMEY (1993). Different RAPD patterns were obtained with B. mucronatus from Japan and China in relation to isolates from Europe, Siberia and Canada. These findings support the thesis of BECKENBACH et al. (1992) about a separate taxonomic status of the latter isolates. The RAPD patterns of two undefined provenances from the USA corresponded to patterns obtained with the European species B. fraudulentus . Further investigations using the RAPD-PCR technique will be focused on providing better intraspecific differentiation of the various provenances of B. xylophilus and B. mucronatus . Differenzierung von drei Bursaphelenchus-Arten durch RAPD-PCR Ziel der Untersuchungen war die Anwendung der RAPD-PCR-Methode (Random Amplified Polymorphie DNA-Polymerase Chain Reaction) zur taxonomischen Unterscheidung des Kiefernholznematoden, Bursaphelenchus xylophilus , von verwandten Arten. Die zu diesem Zweck verwendeten 14 Isolate von B. xylophilus , 12 Isolate von B. mucronatus , 7 Isolate von B. fraudulentus und 2 Bursaphelenchus -Isolate unbekannter Zugehorigkeit stammten aus Asien, Europa und Nordamerika und wurden an Botrytis cinerea auf Malzagar vermehrt. Sie wurden mit 13 unterschiedlichen 10-basigen Zufallsprimern der Operon-Kits B, E, Y und Z getestet. Hierbei wurden informative Bandenmuster der amplifizierten DNA-Fragmente erhalten, die inter- und intraspezifische Unterschiede widerspiegelten. Eine zuverlassige Differenzierung und Identifizierung von B. xylophilus und B. mucronatus war mit Primer OPY-01 moglich, wahrend mit Primer OPB-07 B. xylophilus gegenuber B. fraudulentus abgegrenzt werden konnte. Zur Unterscheidung von B. mucronatus und B. fraudulentus war Primer OPZ-08 am besten geeignet. Unsere Ergebnisse bestatigten die taxonomische Zuordnung des kanadischen C2-Isolates zur Art B. mucronatus durch HARMEY und HARMEY (1993). Die Unterschiede in den RAPD-Bandenmustern zwischen den Isolaten von B. mucronatus aus Japan und China einerseits und aus Europa, Sibirien und Kanada andererseits stutzen die These von BECKENBACH et al. (1992) hinsichtlich eines separaten taxonomischen Status der letztgenannten Herkunfte. Die Bandenmuster der beiden undefinierten Isolate aus den USA entsprechen denen der europaischen Art B. fraudulentus . Weitere Untersuchungen zur besseren intraspezifischen Unterscheidung innerhalb der Arten B. xylophilus und B. mucronatus sind vorgesehen.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.001 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.001 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".