Creation of Transgenic Bananas Expressing Human Lysozyme Gene for Panama Wilt Resistance

Abstract

Human lysozyme (HL) inhibits Fusarium oxysporum (FocR4) growth in vitro. To obtain transgenic bananas (Musa spp.) that are resistant to Panama wilt (F. oxysporum), we introduced an HL gene that is driven by a constitutive cauliflower mosaic virus 35S promoter into the banana via Agrobacteriummediated transformation. PCR confirmed that 51 transgenic plants were obtained. The development of Panama wilt symptoms were examined after the plants had been grown in pots. The non-transgenic plants developed typical fusarium symptoms 60 d after FocR4 inoculation, whereas 24 of 51 transgenic plants remained healthy. The transgenic banana plants that showed resistance to FocR4 in the pots were then planted in a field that was heavily infected with FocR4 for further investigation. Eleven of 24 plants developed symptoms before bud emergence; another 11 plants showed symptoms after bud emergence and the remaining two plants, H-67 and H-144, remained healthy and were able to fruit. Northern blotting analysis demonstrated that H-67 and H-144, bearing the strongest resistance to Panama wilt, had the highest level of HL expression and that the expression of HL was well correlated with the FocR4 resistance of transgenic plants. We conclude that Agrobacterium-mediated transformation, with the assistance of particle bombardment, is a powerful approach for banana transformation and that a transgenic HL gene can cause resistance of the crop to FocR4 in the field.

Management of Fusarium wilt of banana: A review with special reference to tropical race 4

“…For example, no results are available from race 1 field trials (Subramaniam et al, 2006;Paul et al, 2011;Ghag et al, 2012), and field results for TR4 tolerance are often absent (Hu et al, 2013;Mahdavi et al, 2012;Yip et al, 2011). In a single paper published 9 years ago, two of 51 Cavendish transformants that expressed the human lysozyme (HL) gene fruited in a TR4 field trial (Pei et al, 2005). Whether subsequent work with these or similar lines has been conducted is not known.…”

Cell suspension culture and plant regeneration of a Brazilian plantain, cultivar Terra

“…through nonconventional strategies, i.e. protoplast culture and fusion, mutagenesis, somaclonal variation and genetic transformation (Ganapathi et al, 2001;Matsumoto et al, 2002;Assani et al, 2005;Houllou-Kido et al, 2005;Kumar et al, 2005;Pei et al, 2005). In addition, the incorporation of biotechnological procedures in breeding programs depends upon effi cient and reliable plant regeneration protocols.…”

Genetic engineering in Banana and Plantain

“…Ma et al [24] transformed somatic embryos of banana cv Grand Naine, through particle bombardment using of a chimeric β-glucuronidase (GUS). Somatic embryos of multiplication stage were bombarded with microprojectile coated with PBPFA5 and pDH 1001 plasmid.…”

“…To obtain transgenic bananas (Musa spp.) that are resistant to Panama wilt (F. oxysporum), Pei et al [24] introduced an HL gene that is driven by a constitutive CaMV 35S promoter into the banana via Agrobacterium-mediated transformation with the assistance of particle bombardment. In this study 51 individual transgenic plants harboring the HL gene were obtained and 24 displayed resistance to FocR4 in the early vegetative growth stage.…”