Agrobacterium-mediated Genetic Transformation for Local Cultivars of Potato (Solanum tuberosum L.) Using Marker Genes

Abstract

Genetic transformation using nodal and internodal segments from three economically important potato (Solanum tuberosum L.) varieties namely, Diamant, Cardinal and Granola was conducted using an Agrobacterium tumefaciens strain LBA4404 harbouring binary plasmid pBI12 containing the GUS and nptII genes. Node and internodal segments were used for direct regeneration as well as regeneration with the intervention of callus. Best responses were obtained for direct regeneration of shoots when the explants were cultured on MS supplemented with 4.0 mg/l BAP +1.0 mg/l IAA, 1.5 mg/l BAP + 0.5 mg/l IAA and 5.0 mg/l BAP +1.0 mg/l IAA in Diamant, Cardinal and Granola, respectively. In Diamant spontaneous in vitro microtuberization was obtained from these proliferated shoots. Further culturing of these in vitro grown green microtubers regenerated a large number of shoots on MS containing 4.0 mg/l BAP +1.0 mg/l IAA. By combining the best treatments, this protocol yielded an average transformation rate of 87% of treared explants. Stable expression of GUS gene was visualized in the various parts of transformed shoots through histochemical assay. Genomic DNA was isolated from transformed shoots and stable integration of the GUS and nptII genes was confirmed by PCR analysis.

Micropropagation of Environmental Stress Tolerant Local Potato (Solanum tuberosum L.) Varieties of Bangladesh

“…reported for nodal and internal explants (Borna et al 2010, Hussain et al 2005, Sarker and Mustafa 2002, Philip and Hampson 1995, while for leaf and internodes, NAA and BAP were reported to be the best (Khatun et al 2012). Both studies are different from our observation.…”

“…This variation may be due to the different genotypes. Borna et al (2010) reported direct regeneration from node and internode explants. Hussain et al (2005) reported direct regeneration from node and shoot apex, while indirect regeneration occurred from leaf and internode.…”

In vitro Regeneration and Agrobacterium-mediated Transformation of Potato (Solanum tuberosum L.) Cultivars Grown in Mexico

“…Establishment of an efficient in vitro regeneration protocol is a prerequisite to facilitate successful plant genetic transformation (Soto et al 2007). In this context numerous in vitro regeneration protocols have been developed for potato (Borna et al 2010, Chakravarty and Wang-Pruski 2010, Shin et al 2011, Lopéz and Chaparro 2007. Reports are also available regarding the influence of genotype, type of explants and culture conditions on the efficiency of in vitro regeneration in potato cultivars (Sarker and Mustafa 2002).…”

Protocol Optimization for In Vitro Regeneration System in Frost Sensitive Potato Genotypes: As a Basis for Genetic Transformation Studies in Solanum tuberosum L.

“…our results have no similarity with the results obtained by Hussaini et al [31] because they used combination of different growth hormones viz, BAP, GA 3 , NAA and TDZ. They found best regeneration on media supplemented with combinations of BAP, GA 3 and TDZ.…”

“…Potato is a staple food crop of forty countries around the world [2]. Potato is a tuberous crop plant which belongs to Solanacea family [3]. Potato is a source of complex carbohydrates and antioxidants that provide energy and immunity to our body.…”