New Zealand Journal of Crop and Horticultural Science abstracts

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Differentiation of a specific Trichoderma biological control agent
by restriction fragment length polymorphism (RFLP) analysis

J. K. BOWEN^1*
S. C. FRANICEVIC¹
R. N. CROWHURST²
M. D. TEMPLETON²
A. STEWART¹⁺

1School of Biological Sciences
University of Auckland
Auckland, New Zealand

2The Horticulture and Food Research
Institute of New Zealand
Mt Albert Research Centre
Private Bag 92 169
Auckland, New Zealand

^*Present address: The Horticulture and Food Research Institute of New Zealand, Mt Albert Research Centre, Private Bag 92 169, Auckland, New Zealand.

⁺Present address: Department of Plant Science, Lincoln University, Canterbury, New Zealand.

Abstract  A single biological control isolate belonging to the species aggregate Trichoderma harzianum Rifai was differentiated from other closely related Trichoderma species and commonly isolated mycoflora by molecular assay. Restriction fragment length polymorphism (RFLP) analysis enabled the differentiation of the single isolate; a hybridising band of 1.1 kb being diagnostic. Cross-hybridisation occurred against the DNA from 12 Trichoderma isolates, representative of three species aggregates. The hybridisation probe used during the RFLP analysis was an amplification product produced during random amplification of polymorphic DNA (RAPD) polymerase chain reaction (PCR) analysis using DNA from T. harzianum isolate C65 as template. This amplification product was not visible following separation of amplification products generated during PCR using DNA from a test array of fungi as template. Diagnostic PCR, using primers designed from the sequence of the specific polymorphic amplification product failed to discriminate between C65 and other Trichoderma isolates, since products with equivalent size were generated. The ability to differentiate the isolate of T. harzianum from other closely related fungi will enable monitoring of the specific isolate in field trials to assess biological control ability.

Keywords  Trichoderma harzianum; Sclerotinia sclerotiorum; Actinidia deliciosa; biological control agent; RAPD PCR; RFLP; molecular marker

New Zealand Journal of Crop and Horticultural Science, 1996, Vol. 24: 207-217

0114-0671/96/2403-0207 $2.50/0 (c) The Royal Society of New Zealand 1996

PDF file of entire paper: medium quality (2224K); (scanned from paper original: notes about this process)

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