title: Co-expression of chimeric chitinase and a polygalacturonase-inhibiting protein in transgenic canola (Brassica napus) confers enhanced resistance to Sclerotinia sclerotiorum.
abstract: OBJECTIVE
  Sclerotinia stem rot (SSR) caused by Sclerotinia sclerotiorum is one of the major fungal diseases of canola. To develop resistance against this fungal disease, the chit42 from Trichoderma atroviride with chitin-binding domain and polygalacturonase-inhibiting protein 2 (PG1P2) of Phaseolus vulgaris were co-expressed in canola via Agrobacterium-mediated transformation.
  
  
  RESULTS
  Stable integration and expression of transgenes in T0 and T2 plants was confirmed by PCR, Southern blot and RT-PCR analyses. Chitinase activity and PGIP2 inhibition were detected by colorimetric and agarose diffusion assay in transgenic lines but not in untransformed plants. The crude proteins from single copy transformant leaves having high chitinase and PGIP2 activity (T16, T8 and T3), showed up to 44 % inhibition of S. sclerotiorum hyphal growth. The homozygous T2 plants, showing inheritance in Mendelian fashion (3:1), were further evaluated under greenhouse conditions for resistance to S. sclerotiorum. Intact plants contaminated with mycelia showed resistance through delayed onset of the disease and restricted size and expansion of lesions as compared to wild type plants.
  
  
  CONCLUSIONS
  Combined expression of chimeric chit42 and pgip2 in Brassica napus L. provide subsequent protection against SSR disease and can be helpful in increasing the canola production in Iran.
authors.affiliation: National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
authors.forename: Mahboobeh
authors.initials: M
authors.lastname: Ziaei
authors.affiliation: National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran. motalebi@nigeb.ac.ir.
authors.forename: Mostafa
authors.initials: M
authors.lastname: Motallebi
authors.affiliation: National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Zamani
authors.affiliation: National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
authors.forename: Nasim Zarin
authors.initials: NZ
authors.lastname: Panjeh
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology; D001203:Ascomycota / Q000187:drug effects / Q000472:pathogenicity*; D029688:Brassica napus / Q000235:genetics* / Q000382:microbiology*; D002688:Chitinases / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D060467:Disease Resistance; D018506:Gene Expression Regulation, Plant; D027805:Phaseolus / Q000235:genetics; D010935:Plant Diseases / Q000382:microbiology; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D030821:Plants, Genetically Modified / Q000382:microbiology; D014242:Trichoderma / Q000235:genetics
chemical_list: D000935:Antifungal Agents; C081479:PGIP protein, plant; D010940:Plant Proteins; D002688:Chitinases
keywords: Antifungal activity; Brassica napus L.; Chimeric chitinase; Polygalacturonase-inhibiting protein; Sclerotinia sclerotiorum; Sclerotinia stem rot; Transgenic plants