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Open AccessArticle

Efficient Biolistic Transformation of Immature Citrus Rootstocks Using Phosphomannose-isomerase Selection

1
Horticulture Sciences Department, Citrus Research & Education Center, University of Florida, Lake Alfred, FL 33850, USA
2
Department of Plant Pathology, Citrus Research & Education Center, University of Florida, Lake Alfred, FL 33850, USA
*
Authors to whom correspondence should be addressed.
Plants 2019, 8(10), 390; https://doi.org/10.3390/plants8100390
Received: 28 August 2019 / Revised: 26 September 2019 / Accepted: 27 September 2019 / Published: 30 September 2019
This research utilized the E. coli manA gene encoding phosphomannose isomerase (PMI) selection on sucrose/mannose medium to increase transformation efficiencies after biolistic transformation of two immature citrus rootstock cultivars. Plasmid DNA, containing the manA gene and the enhanced green fluorescent protein (egfp) reporter gene, was bombarded into epicotyl explants of immature Carrizo citrange and Swingle citrumelo. GFP positive shoots were micro-grafted onto in vitro grown immature Carrizo rootstocks. Nineteen transgenic Carrizo shoots were obtained from ten paired shots, and eight Swingle shoots from five paired shots. The mean transformation efficiency of Carrizo was 1.9 transgenics/paired shot while the transformation efficiency of Swingle was comparable at 1.6 transgenics/paired shot. The transformants were analyzed by PCR for the presence of transgenes. Southern blot analysis of eight representative Carrizo transgenic events and four Swingle transgenic events showed that all transgenics had one to three copies of the manA gene. The PMI enzyme activity in the transgenic lines was confirmed using the chlorophenol red assay. View Full-Text
Keywords: citrus; transgenic; biolistic transformation; particle bombardment; phosphomannose isomerase selection citrus; transgenic; biolistic transformation; particle bombardment; phosphomannose isomerase selection
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MDPI and ACS Style

Wu, H.; Acanda, Y.; Canton, M.; Zale, J. Efficient Biolistic Transformation of Immature Citrus Rootstocks Using Phosphomannose-isomerase Selection. Plants 2019, 8, 390.

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