Production of soft rot resistant calla lily by expressing a ferredoxin-like protein gene (pflp) in transgenic plants

• Published in: Plant cell reports 2007-04, Vol.26 (4), p.449-457
• Main Authors: Yip, Mei-Kuen, Huang, Hsiang-En, Ger, Mang-Jye, Chiu, Shih-Hua, Tsai, Yuh-Chih, Lin, Chin-I, Feng, Teng-Yung
• Format: Article
• Language: English
• Subjects: Agrobacterium; Agrobacterium tumefaciens - genetics; Biological and medical sciences; Biotechnology; Blotting, Southern; Blotting, Western; Calla; Cultivars; Disease resistance; DNA, Plant - analysis; DNA, Plant - genetics; Erwinia carotovora; Ferredoxin-like protein; Ferredoxins - genetics; Ferredoxins - metabolism; Flowers & plants; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; Methods. Procedures. Technologies; Pectobacterium carotovorum - growth & development; Plant Diseases - genetics; Plant Diseases - microbiology; Plant Proteins - genetics; Plant Proteins - metabolism; Plants, Genetically Modified; Polymerase Chain Reaction; Soft rot disease resistance; Transformation, Genetic; Transgenic; Transgenic animals and transgenic plants; Transgenic plants; Zantedeschia; Zantedeschia - genetics; Zantedeschia - metabolism; Zantedeschia - microbiology; Zantedeschia elliottiana
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s00299-006-0246-y

An efficient protocol for the Agrobacterium tumefaciens-mediated transformation of calla lily (Zantedeschia elliottiana (W. Wats.) Engl. cultivar 'Florex Gold') is described. Shoot basal discs were co-cultivated with A. tumefaciens C58C1 carrying a plasmid containing neomycin phosphotransferase (nptII) and plant ferredoxin-like protein (pflp) genes. After Agrobacterium co-cultivation, the shoot basal discs were exposed to 100 mg l-¹ kanamycin for selection. Twenty-eight out of 260 discs (10.8%) were found to have survived and produced shoot clusters. Twenty-six of these were confirmed to contain the pflp transgene by PCR, ending up in 10% transformation efficiency. The disease resistance investigation revealed that 18 transgenic plants exhibited resistance to soft rot disease caused by Erwinia carotovora subsp. carotovora. The presence of pflp gene was demonstrated by PCR, and its accumulation and activity was confirmed by Western blot and disease resistance assay. This was the first report to show the successful transformation and resistance to a bacterial pathogen in Zantedeschia. The protocol is useful for the quality improvement of calla lily through genetic transformation.