Optimisation of Adventitious Shoot Regeneration and Agrobacterium-mediated Transformation in Canna × generalis (Canna Lily)

Rani Singh, Arvind Kumar Dubey, Indraneel Sanyal

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

Canna being ornamental plants has a significant role in agriculture, medical, economy and food industry. Canna has a limited vase life due to the rapid loss of moisture from its perianth. For improving its market value, cuticularisation of the perianth can be achieved by the expression of a heterologous cutin producing gene, using the tissue culture and transformation protocol developed in this study. Efficient, rapid and direct adventitious shoot regeneration was successfully established in Canna × generalis using recalcitrant rhizome explants. The explants were cultured on MS medium supplemented with 6-benzylaminopurine (6-BA), thidiazuron (TDZ), and kinetin. Among the four genotypes taken for tissue culture, the ‘Trinacria variegata’ was the best responding cultivar. And 2 mg · L −1 6-BA or 1.5 mg · L −1 TDZ along with 0.1 mg · L −1 IAA was optimum for their regeneration. The highest regeneration was achieved in ‘Trinacria variegata’ (36%) on 6-BA, 33% on TDZ while kinetin failed to evoke any regenerative responses. Regeneration was enhanced by supplementation of 100 mg · L −1 ascorbic acid (AsA), while, 100 mg · L −1 of L-cysteine or 100 mg · L −1 dithiothreitol (DTT), inhibited regeneration. Shoots were observed to develop 3–5 fibrous roots on MS medium supplemented with 0.5 mg · L −1 indole-3-butyric acid (IBA). The plantlets were transplanted into pots and acclimatised in glasshouse with 100% survival. For transformation of Canna, rhizome explants were co-cultivated for 60 min in Agrobacterium suspension. The explants were washed with 500 mg · L −1 cefotaxime solution, subjected to 100 mg · L −1 kanamycin selection followed by excision of the shoots and culturing them on IBA-supplemented media for root development. Transgene integration in the putative transformants was confirmed by PCR assay and copy number by Southern blot hybridisation analysis.

Original languageEnglish
Pages (from-to)39-46
Number of pages8
JournalHorticultural Plant Journal
Volume5
Issue number1
DOIs
StatePublished - 1 Jan 2019
Externally publishedYes

Keywords

  • 6-BA
  • Agrobacterium
  • Canna
  • TDZ
  • UidA
  • ascorbic acid
  • regeneration
  • rhizome
  • transformation

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Ecology
  • Renewable Energy, Sustainability and the Environment
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)
  • Plant Science

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