Embryogenic callus proliferation and regeneration conditions for genetic transformation of diverse sugarcane cultivars

Basnayake, Shiromani W. V., Moyle, Richard and Birch, Robert G. (2011) Embryogenic callus proliferation and regeneration conditions for genetic transformation of diverse sugarcane cultivars. Plant Cell Reports, 30 3: 439-448. doi:10.1007/s00299-010-0927-4


Author Basnayake, Shiromani W. V.
Moyle, Richard
Birch, Robert G.
Title Embryogenic callus proliferation and regeneration conditions for genetic transformation of diverse sugarcane cultivars
Journal name Plant Cell Reports   Check publisher's open access policy
ISSN 0721-7714
Publication date 2011-03
Year available 2010
Sub-type Article (original research)
DOI 10.1007/s00299-010-0927-4
Volume 30
Issue 3
Start page 439
End page 448
Total pages 10
Place of publication Germany
Publisher Springer
Collection year 2011
Language eng
Abstract Amenability to tissue culture stages required for gene transfer, selection and plant regeneration are the main determinants of genetic transformation efficiency via particle bombardment into sugarcane. The technique is moving from the experimental phase, where it is sufficient to work in a few amenable genotypes, to practical application in a diverse and changing set of elite cultivars. Therefore, we investigated the response to callus initiation, proliferation, regeneration and selection steps required for microprojectile-mediated transformation, in a diverse set of Australian sugarcane cultivars. 12 of 16 tested cultivars were sufficiently amenable to existing routine tissue-culture conditions for practical genetic transformation. Three cultivars required adjustments to 2,4-D levels during callus proliferation, geneticin concentration during selection, and/or light intensity during regeneration. One cultivar gave an extreme necrotic response in leaf spindle explants and produced no callus tissue under the tested culture conditions. It was helpful to obtain spindle explants for tissue culture from plants with good water supply for growth, especially for genotypes that were harder to culture. It was generally possible to obtain several independent transgenic plants per bombardment, with time in callus culture limited to 11-15 weeks. A caution with this efficient transformation system is that separate shoots arose from different primary transformed cells in more than half of tested calli after selection for geneticin resistance. The results across this diverse cultivar set are likely to be a useful guide to key variables for rapid optimisation of tissue culture conditions for efficient genetic transformation of other sugarcane cultivars. © 2010 Springer-Verlag.
Keyword Particle bombardment
Saccharum
Selection
Tissue culture
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Special Issue: Plant biotechnology in support of the Millennium Goals

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2011 Collection
School of Biological Sciences Publications
 
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Created: Wed, 23 Mar 2011, 10:15:04 EST by Dr Richard Moyle on behalf of School of Biological Sciences