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

被引：45

作者：

Maximova, S ^{[1
]}

Miller, C ^{[1
]}

de Mayolo, GA ^{[1
]}

Pishak, S ^{[1
]}

Young, A ^{[1
]}

Guiltinan, MJ ^{[1
]}

机构：

[1] Penn State Univ, Coll Agr Sci, Dept Hort, University Pk, PA 16802 USA

来源：

PLANT CELL REPORTS | 2003年 / 21卷 / 09期

关键词：

Theobroma cacao; Agrobacterium tumefaciens; genetic transformation; matrix attachment region; somatic embryogenesis;

D O I：

10.1007/s00299-003-0596-7

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

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 nontransgenic 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.

引用

页码：872 / 883

页数：12

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