Stable transformation of Theobroma cacao L. and influence of matrix attachment regions on GFP expression

Siela Maximova, C. Miller, G. Antúnez De Mayolo, S. Pishak, A. Young, Mark Guiltinan

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

We describe a protocol for Agrobacterium-mediated genetic transformation of Theobroma cacao L. using cotyledonary explants from primary somatic embryos (SEs) and A. tumefaciens strain AGL1. Transgenic plants carrying the visible marker, gene green fluorescent protein (EGFP), the selectable marker gene neomycin phosphotransferase II (NPTII), the class I chitinase gene from cacao (Chi), and tobacco nuclear matrix attachment regions (MARs) in different combinations were successfully produced via regeneration of secondary SEs. The presence of the Chi gene or MARs did not influence the number of transgenic plants produced compared to the marker genes alone. However, the inclusion of MARs contributed to increased mean GFP expression in the population of transgenics. Additionally, the presence of MARs reduced the occurrence of gene silencing and stabilized high levels of GFP expression in lines of transgenic plants multiplied via reiterative somatic embryogenesis. Ninety-four transgenic plants were acclimated in a greenhouse and grown to maturity. Detailed growth analysis indicated that there were no differences in various growth parameters between transgenic and non-transgenic SE-derived plants. Seeds produced from two genetic crosses with one of the transgenic lines were analyzed for EGFP expression - a near-perfect 1:1 segregation was observed, indicating that this line resulted from the insertion of a single locus of T-DNA.

Original languageEnglish (US)
Pages (from-to)872-883
Number of pages12
JournalPlant Cell Reports
Volume21
Issue number9
StatePublished - Jun 1 2003

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Theobroma cacao
transgenic plants
somatic embryos
genetically modified organisms
genetic markers
nuclear matrix
kanamycin kinase
gene silencing
genetic transformation
chitinase
Agrobacterium radiobacter
somatic embryogenesis
green fluorescent protein
Agrobacterium
explants
tobacco
genes
greenhouses
loci
DNA

All Science Journal Classification (ASJC) codes

  • Agronomy and Crop Science
  • Plant Science

Cite this

Maximova, Siela ; Miller, C. ; Antúnez De Mayolo, G. ; Pishak, S. ; Young, A. ; Guiltinan, Mark. / Stable transformation of Theobroma cacao L. and influence of matrix attachment regions on GFP expression. In: Plant Cell Reports. 2003 ; Vol. 21, No. 9. pp. 872-883.
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Stable transformation of Theobroma cacao L. and influence of matrix attachment regions on GFP expression. / Maximova, Siela; Miller, C.; Antúnez De Mayolo, G.; Pishak, S.; Young, A.; Guiltinan, Mark.

In: Plant Cell Reports, Vol. 21, No. 9, 01.06.2003, p. 872-883.

Research output: Contribution to journalArticle

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