原创力文档the hrpz gene of pseudomonas syringae pv. phaseolicola enhances resistance to rhizomania disease in transgenic nicotiana benthamiana and sugar beet丁香假单胞菌的hrpz基因。.pdfVIP
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the hrpz gene of pseudomonas syringae pv. phaseolicola enhances resistance to rhizomania disease in transgenic nicotiana benthamiana and sugar beet丁香假单胞菌的hrpz基因。
The hrpZ Gene of Pseudomonas syringae pv.
phaseolicola Enhances Resistance to Rhizomania Disease
in Transgenic Nicotiana benthamiana and Sugar Beet
1 1 2 1
Ourania I. Pavli , Georgia I. Kelaidi , Anastasia P. Tampakaki *, George N. Skaracis
1 Department of Crop Sciences, Agricultural University of Athens, Athens, Greece, 2 Department of Agricultural Biotechnology, Agricultural University of Athens, Athens,
Greece
Abstract
To explore possible sources of transgenic resistance to the rhizomania-causing Beet necrotic yellow vein virus (BNYVV),
Nicotiana benthamiana plants were constructed to express the harpin of Pseudomonas syringae pv. phaseolicola (HrpZPsph).
The HrpZ protein was expressed as an N-terminal fusion to the PR1 signal peptide (SP/HrpZ) to direct harpin accumulation
to the plant apoplast. Transgene integration was verified by mPCR in all primary transformants (T0), while immunoblot
analysis confirmed that the protein HrpZPsph was produced and the signal peptide was properly processed. Neither T0
plants nor selfed progeny (T1) showed macroscopically visible necrosis or any other macroscopic phenotypes. However,
plants expressing the SP/HrpZPsph showed increased vigor and grew faster in comparison with non-transgenic control
plants. Transgenic resistance was assessed after challenge inoculation with BNYVV on T1 progeny by scoring of disease
symptoms and by DAS-ELISA at 20 and 30 dpi. Transgenic and control lines showed significant differences in terms of the
number of plants that became infected, the timing of infection and the disease symptoms displayed. Plants expressing the
SP/HrpZPsph developed localized leaf necrosis in the infection
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