An easy and efficient protocol in the production of pflp transgenic banana against Fusarium wilt

Abstract

This study describes an efficient protocol for Agrobacterium tumefaciens-mediated transformation of two subgroups of genotype AAA bananas (Musa acuminata cv. Pei Chiao and Musa acuminata cv. Gros Michel). Instead of using suspension cells, cauliflower-like bud clumps, also known as multiple bud clumps (MBC), were induced from sucker buds on MS medium containing N 6 -Benzylaminopurine (BA), Thidiazuron (TDZ), and Paclobutrazol (PP333). Bud slices were co-cultivated with A. tumefaciens C58C1 or EHA105 that carry a plasmid containing Arabidopsis root-type ferredoxin gene (Atfd3) and a plant ferredoxin-like protein (pflp) gene, respectively. These two strains showed differences in transformation efficiency. The EHA105 strain was more sensitive in Pei Chiao, 51.3% bud slices were pflp-transformed, and 12.6% slices were Atfd3-transformed. Gros Michel was susceptible to C58C1 and the transformation efficiency is 4.4% for pflp and 13.1% for Atfd3. Additionally, gene integration of the putative pflp was confirmed by Southern blot. Resulting from the pathogen inoculation assay, we found that the pflp transgenic banana exhibited resistance to Fusarium oxysporum f. sp. cubense tropical race 4. This protocol is highly advantageous to banana cultivars that have difficulties in setting up suspension cultures for the purpose of quality improvement through genetic transformation. In addition, this protocol would save at least 6 months in obtaining explants for transformation and reduce labor for weekly subculture in embryogenic cell suspension culture systems.

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).…”

Control of Fusarium wilt in banana with Chinese leek

“…Scientists around the world have investigated resistance breeding (Hwang and Ko 2004 ; Smith et al2006), chemical control (Nel et al 2007), biocontrol (Saravanan et al 2003; Cao et al2005; Lian et al2009) and molecular methods (Paul et al 2011; Yip et al 2011),in search of methods to control the disease. Some of these studies were found to effectively suppress Foc growth in the laboratory and in greenhouses, but were unable to efficiently control Fusarium wilt in the field.…”

The application of the ‘Gene-deletor’ technology in banana

“…However, the performance of the banana industry is affected by biotic and abiotic stresses such as Fusarium wilt (Czislowski et al 2018;Dale et al 2017), cold stress (Dou et al 2016), typhoon (Yu et al 2014). Most banana cultivars are triploid, with long generation times, and sterility, hence, it is difficult to cultivate new resistant banana germplasm by means of cross breeding (Ghag et al 2014;Tripathi et al 2008;Yip et al 2011). In addition, the selection and breeding of new varieties by screening mutants has certain randomness and requires longer periods of time (Hasan et al 2013).…”