Development of strain specific molecular markers for the Sclerotinia minor bioherbicide strain IMI 344141

Abstract

The plant pathogenic fungus, Sclerotinia minor IMI 344141, has been developed as a bioherbicide for broadleaf weed control in turfgrass and a means to differentiate this biocontrol agent from like organisms is required. A strain specific molecular marker was developed to detect and monitor the Sclerotinia minor IMI 344141 bioherbicide strain. The method was based on polymerase chain reaction (PCR) amplification of two sequence-characterized amplified regions (SCAR) primer pairs for a first round PCR, and another two sets of nested primers was used for a second round PCR if higher sensitivity was needed. Sclerotinia minor IMI 344141 was successfully traced from both pure cultures and environmental samples originating from bioherbicide-released field trials. DNA of the S. minor bioherbicide isolate IMI 344141 was detected in the soil 2 months after application, but was not detected in the 3- and 9-month samples after application. When applied as a bioherbicide, S. minor (IMI 344141) did not persist into the following spring season in turf environments. This molecular detection method provides a mechanism to distinguish this isolate from related organisms and a tool to monitor behavior of the biocontrol agent S. minor IMI 344141 in nature, particularly in soil.

Keywords:

• biocontrol
• bioherbicide
• detection
• mycoherbicide
• PCR
• RAPD
• SCAR
• Sclerotinia minor
• turfgrass

Acknowledgements

Financial support from the Natural Sciences and Engineering Research Council of Canada (NSERC) Idea to Innovation (I2I) grant and 4260864 Canada Inc (Sarritor Inc.) are gratefully acknowledged. The following graciously loaned S. minor isolates: G. Abawi, Cornell University, Geneva, USA; G. Boland, University of Guelph; J. Whipps, Warwick Horticulture Research International, UK; H. Huang, Agriculture and Agri-food Canada, Brandon; P. Phipps, Virginia Tech; S. O'Neill, Queensland Department Plant Industry, Brisbane; Tamrika Hind-Lanoiselet, Wagga Wagga Agricultural Institute, NSW; S. Morley, Herbarium VPRI, Victorian Department of Primary Industries, Knoxfield, Victoria, Australia; K. Elena, Benaki Phytopathological Institute, Athens, Greece; Alison Stewart, Lincoln University, Christchurch, New Zealand; Levente Kiss, Hungarian Plant Protection Institute, Budapest; P. Charue, Belgian Coordinated Collections of Microorganisms, Louvain; J. Tatnell and M. Fuhlbohm, Australian Department of Plant Industry, Kingaroy, QLD; W.G. Kim, National Institute of Agricultural Science and Technology, Suwon, Republic of Korea; F. Snippe-Claus, Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands.

Additional information

Notes on contributors

A.K. Watson

When originally published online the author name was incorrect. The online version has now been amended.