AFLP analysis and improved phytoextraction capacity of transgenic gshI-poplar clones (Populus canescens L.) in vitro

Gyulai, Gábor and Humphreys, M. and Bittsánszky, András and Kiss, József and Gullner, Gábor and Szabó, Zoltán and Radimszky, László and Kőmíves, Tamás and Heszky, László (2005) AFLP analysis and improved phytoextraction capacity of transgenic gshI-poplar clones (Populus canescens L.) in vitro. ZEITSCHRIFT FÜR NATURFORSCHUNG C - A JOURNAL OF BIOSCIENCES, 60. pp. 300-306. ISSN 0939-5075

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Abstract

Clone stability and in vitro phytoextraction capacity of vegetative clones of R x canescens (2n = 4x = 38) including two transgenic clones (ggs11 and lgl6) were studied as in vitro leaf disc cultures. Presence of the gshI-transgene in the transformed clones was detected in PCR reactions using gshI-specific primers. Clone stability was determined by fAFLP (fluorescent amplified DNA fragment length polymorphism) analysis. In total, 682 AFLP fragments were identified generated by twelve selective primer pairs after EcoRI-MseI digestion. Four fragments generated by EcoAGT-MseCCC were different (99.4% genetic similarity) which proves an unexpectedly low bud mutation frequency in R x canescens. For the study of phytoextraction capacity leaf discs (8 mm) were exposed to a concentration series of ZnSO4 (10(-1) to 10(-5) m) incubated for 21 days on aseptic tissue culture media WPM containing 1 mu m Cu. Zn2+ caused phytotoxicity only at high concentrations (10(-1) to 10(- 2) m). The transgenic poplar cyt-ECS (ggs11) clone, as stimulated by the presence of Zn, showed elevated heavy metal (Cu) uptake as compared to the non-transformed clone. These results suggest that gshI-transgenic poplars may be suitable for phytoremediation of soils contaminated with zinc and copper.

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