Constitutive expression of transgenes encoding derivatives of the synthetic antimicrobial peptide BP100: impact on rice host plant fitness
- Equal contributors
1 Institute of Food and Agricultural Technology (INTEA), University of Girona, Campus Montilivi, EPS-1 17071, Girona, Spain
2 Plant Genetics Department, Centre de Recerca en Agrigenòmica CSIC-IRTA-UAB-UB, Carretera de Cabrils, Km 2, 08348, Barcelona, Spain
BMC Plant Biology 2012, 12:159 doi:10.1186/1471-2229-12-159Published: 4 September 2012
Additional file 1:
Expression levels of hptII and the corresponding bp100der transgenes in three randomly chosen transgenic calluses per construct, as assessed by RT-qPCR. Transgene mRNA copy numbers were normalized with actin values (GeNorm M values below 0.5). Means and SD of the three independent events are shown. No statistical differences were found.
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Additional file 2:
Transgene DNA copy numbers of S-bp100der plants. (A) Southern blot analysis of transgenic lines. Genomic DNA was digested with the restriction enzymes EcoRI or Hind III and subjected to electrophoresis through a 0.8% agarose gel. DNAs were transferred to nylon membranes and hybridized with a thermostable alkaline phosphatase labelled probe. The migration positions and sizes of markers are indicated in base pairs on the left (MW). (B) Determination of transgene copy number by qPCR. Means of six experimental replicates are shown. RSD values were consistently below 2.5%. Transgene DNA copy numbers were normalized with actin values.
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Additional file 3:
Homozygous T2 rice lines obtained in this work and transgene mRNA expression values (relative to actin) in leaves of in vitro grown homozygous T3 plants, as assessed by RT-qPCR. Mean and SD values corresponding to each particular GM event are shown. Three biological replicates per GM event were analyzed, each with leaves of 10 plants at the two-leaf stage.
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Additional file 4:
Examples of Senia and S-bp100.2i plants at maturity.
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Additional file 5:
DNA sequences encoding the BP100 derivatives designed in this work. The sequence encoding the Pr1a signal peptide is indicated in italics. The start and stop codons are underlined.
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