AFLP analysis and improved phytoextraction capacity of transgenic gshI-poplar clones (Populus x canescens L.) for copper in vitro.
Clone stability and in vitro phytoextraction capacity of vegetative clones of P. 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 P. 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 microM 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.
Gyulai
Gábor
G
St. Stephanus University, Department of Genetics and PB, Gödöllö, Hungary. gyulai.gabor@mkk.szie.hu
Humphreys
Mervyn
M
Bittsánszky
András
A
Skøt
Kirsten
K
Kiss
József
J
Skøtc
Leif
L
Gullner
Gábor
G
Heywood
Sue
S
Szabó
Zoltán
Z
Lovatt
Alan
A
Radimszky
László
L
Roderick
Hywel
H
Rennenberg
Heinz
H
Abberton
Michael
M
Komíves
Tamás
T
Heszky
László
L
D001483:Base Sequence; D001673:Biodegradation, Environmental; D003300:Copper / Q000493:pharmacokinetics*; D017931:DNA Primers; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification; D005784:Gene Amplification; D018515:Plant Leaves; D030821:Plants, Genetically Modified / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D032107:Populus / Q000378:metabolism*; D015183:Restriction Mapping
D017931:DNA Primers; D018744:DNA, Plant; D003300:Copper