caa a22 4500 46323569X CHVBK 20180405153256.0 cr unu---uuuuu 170326e20070301xx s 000 0 eng 10.1007/s11240-007-9204-9 doi (NATIONALLICENCE)springer-10.1007/s11240-007-9204-9 Expression of a polyphemusin variant in transgenic tobacco confers resistance against plant pathogenic bacteria, fungi and a virus [Elektronische Daten] [Apurva Bhargava, Milan Osusky, Benjamin Forward, Robert Hancock, William Kay, Santosh Misra] Antimicrobial cationic peptides provide a promising means of engineering plant resistance to a range of plant pathogens, including viruses. PV5 is a synthetic structural variant of polyphemusin, a cationic peptide derived from the horseshoe crab-Limulus polyphemus. PV5 has been shown to be benign toward eukaryotic membranes but with enhanced antimicrobial activity against animal pathogens. In this work, the cytotoxicity of PV5 toward tobacco protoplasts and leaf discs was assessed using TTC (2,3,5-triphenyltetrazolium chloride) and Evans blue colorimetric assays. PV5 showed no measurable cytotoxic effects even at levels as high as 60μg. As a possible approach to enhancing plant resistance, a gene encoding PV5 was fused to the signal sequence encoding the C-terminus portion of the BiP protein from Pseudotsuga menziesii, under the control of 2×35S CaMV promoter. When introduced into Nicotiana tabacum var Xanthi gene integration and expression was confirmed by both Southern and northern analyses. When transgenic plants were subsequently challenged with bacterial and fungal phytopathogens enhanced resistance was observed. Moreover, transgenic plants also displayed antiviral properties against Tobacco Mosaic Virus making PV5 an excellent candidate for conferring unusually broad spectrum resistance to plants and the first anti-plant virus antimicrobial peptide. Springer Science+Business Media B.V., 2007 Cationic peptides nationallicence Protoplast nationallicence Transgenic nationallicence Cytotoxicity nationallicence CAP : Cationic antimicrobial peptides nationallicence dH2O : Distilled water nationallicence HIV : Human immunodeficiency virus nationallicence MS : Murashige-Skoog nationallicence TMV : Tobacco mosaic virus nationallicence TTC : 2,3,5-Triphenyltetrazolium chloride nationallicence Bhargava Apurva Department of Biochemistry and Microbiology and Centre for Forest Biology, University of Victoria, V8W 3P6, Victoria, BC, Canada aut Osusky Milan Department of Biochemistry and Microbiology and Centre for Forest Biology, University of Victoria, V8W 3P6, Victoria, BC, Canada aut Forward Benjamin Department of Biochemistry and Microbiology and Centre for Forest Biology, University of Victoria, V8W 3P6, Victoria, BC, Canada aut Hancock Robert Department of Microbiology, University of British Columbia, Vancouver, BC, Canada aut Kay William Department of Biochemistry and Microbiology and Centre for Forest Biology, University of Victoria, V8W 3P6, Victoria, BC, Canada aut Misra Santosh Department of Biochemistry and Microbiology and Centre for Forest Biology, University of Victoria, V8W 3P6, Victoria, BC, Canada aut Plant Cell, Tissue and Organ Culture Kluwer Academic Publishers 88/3(2007-03-01), 301-312 0167-6857 88:3<301 2007 88 11240 https://doi.org/10.1007/s11240-007-9204-9 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11240-007-9204-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Bhargava Apurva Department of Biochemistry and Microbiology and Centre for Forest Biology, University of Victoria, V8W 3P6, Victoria, BC, Canada aut NATIONALLICENCE 700 1- Osusky Milan Department of Biochemistry and Microbiology and Centre for Forest Biology, University of Victoria, V8W 3P6, Victoria, BC, Canada aut NATIONALLICENCE 700 1- Forward Benjamin Department of Biochemistry and Microbiology and Centre for Forest Biology, University of Victoria, V8W 3P6, Victoria, BC, Canada aut NATIONALLICENCE 700 1- Hancock Robert Department of Microbiology, University of British Columbia, Vancouver, BC, Canada aut NATIONALLICENCE 700 1- Kay William Department of Biochemistry and Microbiology and Centre for Forest Biology, University of Victoria, V8W 3P6, Victoria, BC, Canada aut NATIONALLICENCE 700 1- Misra Santosh Department of Biochemistry and Microbiology and Centre for Forest Biology, University of Victoria, V8W 3P6, Victoria, BC, Canada aut NATIONALLICENCE 773 0- Plant Cell, Tissue and Organ Culture Kluwer Academic Publishers 88/3(2007-03-01), 301-312 0167-6857 88:3<301 2007 88 11240 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer