RESEARCH PAPER New Biotechnology  Volume 27, Number 6  December 2010 Expression of anti-sclerotinia scFv in transgenic Brassica napus enhances tolerance against stem rot William Yajima 1 , Shiv Shankar Verma 1 , Saleh Shah 2 , Muhammad Hafizur Rahman 1 , Yue Liang 1 and Nat N.V. Kav 1, 1 Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta T6G 2P5, Canada 2 Plant Biotechnology, Alberta Innovates-Technology Futures, Vegreville, Alberta T9C 1T4, Canada Canola is an important agricultural crop imparting a significant contribution to global oilseed production. As such, optimizing yield and quality is of paramount importance and canola production can be significantly affected by sclerotinia stem rot. The utility of recombinant antibody technology in plant protection has been explored by many researchers and shows promise for the generation of new lines of agriculturally significant crops with greater resistance to diseases. The objective of the current study was to generate recombinant pathogen specific antibody (scFv)-expressing transgenic Brassica napus plants with increased tolerance to the phytopathogenic fungus, Sclerotinia sclerotiorum. Transgenic canola (B. napus) lines expressing S. sclerotiorum-specific scFv antibody showed a significant level of tolerance towards S. sclerotiorum as compared to their non-transformed counterparts. Both incidence and progression of S. sclerotiorum-induced disease symptoms were reduced in plants expressing the recombinant scFv. Introduction The contribution of canola to global oilseed production is con- siderable as approximately 46 million metric tonnes (MMT) were produced worldwide in 2005, with Canada producing about 8.4 MMT and the European Union, India, Australia and China being the other major contributors (FAOSTAT data 2006). Effective management of various abiotic and biotic stresses that this crop is exposed to is one of the factors that will need to be addressed in order to achieve any substantial increase in production. Canola diseases that can significantly reduce quality and yield include sclerotinia stem rot [caused by Sclerotinia sclerotiorum (Lib.) de Bary], alternaria blackspot [caused by Alternaria brassicae (Berk.) Sacc.] and blackleg [caused by Leptosphaeria maculans (Desmaz.) Ces. and De Not.], among others [1]. Although agronomic prac- tices and the application of effective fungicides are currently being used to control outbreaks of major canola diseases, growers must remain vigilant so as not to allow phytopathogens to interfere with optimal canola production. Furthermore, the search for newer and more efficient disease management strategies, including the gen- eration of canola with genetic resistance to phytopathogenic fungi, is warranted. Genetic modification, through traditional plant breeding or the application of biotechnology, has been used successfully to gen- erate new lines of canola with increased tolerance to fungal phytopathogens. For example, overexpression of a mitogen-acti- vated protein kinase (BnMPK4) and constitutive expression of an oxalate oxidase from Triticum aestivum led to increased tolerance to S. sclerotiorum infection in transgenic Brassica napus canola [2,3]. Additionally, higher levels of resistance to multiple fungi, includ- ing L. maculans and S. sclerotiorum, was observed in B. napus canola lines constitutively expressing an endochitinase gene, which pre- sumably encoded an enzyme that degraded the chitin found on the outer surfaces of these phytopathogenic fungi [4]. Some researchers exploring novel techniques for effective pest management in agriculturally significant crops have focused on antibody-based plant protection as a possible means to introduce Research Paper Corresponding author: Kav, Nat N.V. (nat@ualberta.ca) 816 www.elsevier.com/locate/nbt 1871-6784/$ - see front matter ß 2010 Elsevier B.V. All rights reserved. doi:10.1016/j.nbt.2010.09.010