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Plant Cell Reports

, Volume 30, Issue 3, pp 425–436 | Cite as

Highly efficient Agrobacterium-mediated transformation of banana cv. Rasthali (AAB) via sonication and vacuum infiltration

  • Kondeti Subramanyam
  • Koona Subramanyam
  • K. V. Sailaja
  • M. Srinivasulu
  • K. LakshmideviEmail author
Original Paper

Abstract

A reproducible and efficient transformation method was developed for the banana cv. Rasthali (AAB) via Agrobacterium-mediated genetic transformation of suckers. Three-month-old banana suckers were used as explant and three Agrobacterium tumefaciens strains (EHA105, EHA101, and LBA4404) harboring the binary vector pCAMBIA1301 were used in the co-cultivation. The banana suckers were sonicated and vacuum infiltered with each of the three A. tumefaciens strains and co-cultivated in the medium containing different concentrations of acetosyringone for 3 days. The transformed shoots were selected in 30 mg/l hygromycin-containing selection medium and rooted in rooting medium containing 1 mg/l IBA and 30 mg/l hygromycin. The presence and integration of the hpt II and gus genes into the banana genome were confirmed by GUS histochemical assay, polymerase chain reaction, and southern hybridization. Among the different combinations tested, high transformation efficiency (39.4 ± 0.5% GUS positive shoots) was obtained when suckers were sonicated and vacuum infiltered for 6 min with A. tumefaciens EHA105 in presence of 50 μM acetosyringone followed by co-cultivation in 50 μM acetosyringone-containing medium for 3 days. These results suggest that an efficient Agrobacterium-mediated transformation protocol for stable integration of foreign genes into banana has been developed and that this transformation system could be useful for future studies on transferring economically important genes into banana.

Keywords

Banana suckers GUS histochemical assay Sonication Vacuum infiltration Acetosyringone 

Abbreviations

CaMV 35S

Cauliflower mosaic virus 35S promoter

SAAT

Sonication-assisted Agrobacterium-mediated transformation

MS

Murashige and Skoogs medium

YEP

Yeast extract peptone medium

BA

6-Benzylaminopurine

NAA

α-Naphthaleneacetic acid

IBA

Indole-3-butyric acid

hpt

Hygromycin phospho transferase gene

gus

β-Glucuronidase gene

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Kondeti Subramanyam
    • 1
  • Koona Subramanyam
    • 2
  • K. V. Sailaja
    • 1
  • M. Srinivasulu
    • 3
  • K. Lakshmidevi
    • 1
    Email author
  1. 1.Department of BiochemistrySri Krishnadevaraya UniversityAnantapurIndia
  2. 2.Department of Biotechnology, Sreenidhi Institute of Science and Technology (Autonomous)Jawaharlal Nehru Technological UniversityHyderabadIndia
  3. 3.Department of MicrobiologySri Krishnadevaraya UniversityAnantapurIndia

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