Abstract
Abstract Polymerase chain reaction (PCR) primers, specific for the aggressive pathogenicity group of the rape seed pathogen Leptosphaeria maculans , were obtained according to the following experimental outline: DNA bands that differentiate between aggressive (A) and nonaggressive (NA) isolates were generated by the random amplified polymorphic DNA approach (RAPD). Differential RAPD bands were checked for pathogenicity group‐specificity and sequenced by a combination of chemical and dideoxynucleotide chain termination methods. Based on the DNA sequence longer and highly specific primer pairs for conventional PCR under stringent conditions were designed, which were tested on 97 isolates of L. maculans belonging to different pathogenicity groups. The primer pair specific for the aggressive pathogenicity group unequivocally identifies A‐isolates. The primer pair designed for the N A‐isolates. cross‐reacts with A‐isolates showing positive signals in 28 out of 63 cases. The combined application of A‐ and NA‐specific primers identifies all isolates unambiguously. The method does not require pure fungal cultures. L. macuians can be detected directly in infected plant material within a single working day. The experimental outhne presented in this communication is proposed as a general method for obtaining taxon‐specific PCR primer pairs.
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Claims 6
Claim · model-written summary
The aggressive-group primer pair unequivocally identifies A-isolates of Leptosphaeria maculans among 97 isolates tested.
- Taxon
- rape seed
_Brassica_napus_-_Habit.jpg?width=60)
Brassica napuse0 - Interactor
- Leptosphaeria maculans
Plenodomus lingame0pathogen - Outcome
- identifies A‐isolatese1
- Value
- 97 isolatese2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
_Brassica_napus_-_Habit.jpg?width=500)
Verbatim
e0“Polymerase chain reaction (PCR) primers, specific for the aggressive pathogenicity group of the rape seed pathogen Leptosphaeria maculans”
e1“The primer pair specific for the aggressive pathogenicity group unequivocally identifies A‐isolates”
e2“tested on 97 isolates of L. maculans belonging to different pathogenicity groups”
Claim · model-written summary
The NA-specific primer pair cross-reacts with A-isolates, giving positive signals in 28 of 63 cases.
- Taxon
- rape seedBrassica napuse0
- Interactor
- Leptosphaeria maculansPlenodomus lingame0pathogen
- Outcome
- cross‐reacts with A‐isolatese3
- Value
- 28 out of 63 casese3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Polymerase chain reaction (PCR) primers, specific for the aggressive pathogenicity group of the rape seed pathogen Leptosphaeria maculans”
e3“cross‐reacts with A‐isolates showing positive signals in 28 out of 63 cases”
Claim · model-written summary
The combined A- and NA-specific primers identify all isolates unambiguously.
- Taxon
- rape seedBrassica napuse0
- Interactor
- Leptosphaeria maculansPlenodomus lingame0pathogen
- Outcome
- identifies all isolates unambiguouslye4
- Value
- all isolatese4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Polymerase chain reaction (PCR) primers, specific for the aggressive pathogenicity group of the rape seed pathogen Leptosphaeria maculans”
e4“The combined application of A‐ and NA‐specific primers identifies all isolates unambiguously”
Claim · model-written summary
Leptosphaeria maculans can be detected directly in infected rape seed plant material within a single working day.
- Taxon
- rape seedBrassica napuse0
- Interactor
- Leptosphaeria maculansPlenodomus lingame0pathogen
- Outcome
- detected directly in infected plant materiale5
- Value
- within a single working daye5
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Polymerase chain reaction (PCR) primers, specific for the aggressive pathogenicity group of the rape seed pathogen Leptosphaeria maculans”
e5“can be detected directly in infected plant material within a single working day”
Claim · model-written summary
The method does not require pure fungal cultures.
- Taxon
- rape seedBrassica napuse0
- Interactor
- Leptosphaeria maculansPlenodomus lingame0pathogen
- Outcome
- does not require pure fungal culturese6
- Comparator
- pure fungal culturese6
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Polymerase chain reaction (PCR) primers, specific for the aggressive pathogenicity group of the rape seed pathogen Leptosphaeria maculans”
e6“The method does not require pure fungal cultures”
Claim · model-written summary
The authors propose this experimental outline as a general method for obtaining taxon-specific PCR primer pairs.
- Taxon
- rape seedBrassica napuse0
- Outcome
- a general method for obtaining taxon‐specific PCR primer pairse7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Polymerase chain reaction (PCR) primers, specific for the aggressive pathogenicity group of the rape seed pathogen Leptosphaeria maculans”
e7“is proposed as a general method for obtaining taxon‐specific PCR primer pairs”