Expression of a defence-related intercellular barley peroxidase in transgenic tobacco
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Expression of a defence-related intercellular barley peroxidase in transgenic tobacco. / Kristensen, Brian K.; Brandt, Jakob; Bojsen, Kirsten; Thordal-Christensen, Hans; Kerby, Kent B.; Collinge, David B.; Mikkelsen, Jørn D.; Rasmussen, Søren K.
In: Plant Science, Vol. 122, No. 2, 28.02.1997, p. 173-182.Research output: Contribution to journal › Journal article › peer-review
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T1 - Expression of a defence-related intercellular barley peroxidase in transgenic tobacco
AU - Kristensen, Brian K.
AU - Brandt, Jakob
AU - Bojsen, Kirsten
AU - Thordal-Christensen, Hans
AU - Kerby, Kent B.
AU - Collinge, David B.
AU - Mikkelsen, Jørn D.
AU - Rasmussen, Søren K.
PY - 1997/2/28
Y1 - 1997/2/28
N2 - Tobacco plants (Nicotiana benthamiana L.) have been transformed with a T-DNA vector construct carrying the cDNA pBH6-301, encoding the major pathogen induced leaf peroxidase (Prx8) of barley, under control of an enhanced CaMV 35S promoter. Progeny from three independent transformants were analyzed genetically, phenotypically and biochemically. The T-DNA was steadily inherited through three generations. The barley peroxidase is expressed and sorted to the intercellular space in the transgenic tobacco plants. The peroxidase can be extracted from the intercellular space in two molecular forms from both barley and transgenic tobacco. The tobacco expressed forms are indistinguishable from the barley expressed forms as determined by analytical isoelectric focusing (pI 8.5) and Western-blotting. Staining for N-glycosylation showed that one form only was glycosylated. The N-terminus of purified Prx8 from transgenic tobacco was blocked by pyroglutamate, after the removal of which, N-terminal sequencing verified the transit signal-peptide cleavage site deduced from the cDNA sequence. Phenotype comparisons show that the constitutive expression of Prx8 lead to growth retardation. However, an infection assay with the tobacco powdery mildew pathogen Erysiphe cichoracearum did not indicate that the transgenic plants had achieved enhanced resistance.
AB - Tobacco plants (Nicotiana benthamiana L.) have been transformed with a T-DNA vector construct carrying the cDNA pBH6-301, encoding the major pathogen induced leaf peroxidase (Prx8) of barley, under control of an enhanced CaMV 35S promoter. Progeny from three independent transformants were analyzed genetically, phenotypically and biochemically. The T-DNA was steadily inherited through three generations. The barley peroxidase is expressed and sorted to the intercellular space in the transgenic tobacco plants. The peroxidase can be extracted from the intercellular space in two molecular forms from both barley and transgenic tobacco. The tobacco expressed forms are indistinguishable from the barley expressed forms as determined by analytical isoelectric focusing (pI 8.5) and Western-blotting. Staining for N-glycosylation showed that one form only was glycosylated. The N-terminus of purified Prx8 from transgenic tobacco was blocked by pyroglutamate, after the removal of which, N-terminal sequencing verified the transit signal-peptide cleavage site deduced from the cDNA sequence. Phenotype comparisons show that the constitutive expression of Prx8 lead to growth retardation. However, an infection assay with the tobacco powdery mildew pathogen Erysiphe cichoracearum did not indicate that the transgenic plants had achieved enhanced resistance.
KW - Erysiphe
KW - Hordeum vulgare
KW - N-glycosylation
KW - Nicotiana benthamiana
KW - Pathogen defence
KW - Signal peptide
UR - http://www.scopus.com/inward/record.url?scp=0031588809&partnerID=8YFLogxK
U2 - 10.1016/S0168-9452(96)04554-2
DO - 10.1016/S0168-9452(96)04554-2
M3 - Journal article
AN - SCOPUS:0031588809
VL - 122
SP - 173
EP - 182
JO - Plant Science
JF - Plant Science
SN - 0168-9452
IS - 2
ER -
ID: 201508699