Biology Reference
In-Depth Information
Materials and Methods
Greenhouse spring cultivation
Trials were carried out in central Italy (Monsampolo del Tronto-AP) and in south-
ern Italy (Pontecagnano-SA) (approval of the Italian Ministry of Health N° B/
IT/97-29). The greenhouses were rather similar and made of galvanized steel and
covered with plastic polyethylene (0.12 mm thick). An apparatus for drip-irriga-
tion was used and the soil was completely mulched. A complete randomized block
design with three replicate hybrid genotypes was adopted. Each experimental plot
measured 3.12 m2 and contained eight plants in a double row. Transplanting
was performed on March 3rd in southern Italy and on March 27th in central
Italy. The P1, P2, P5, C1, C2 and the commercial Talina hybrids were employed.
Transgenic parthenocarpic hybrids P1, P2 and P5 were obtained by crossing (as
male parent) the primary transgenic plant DR2 iaaM #34-1 with the line Tal 1/1
(P1), the primary transgenic plant DR2 iaaM #28-1 with Tal 1/1 (P2) and the
transgenic plant Tal 1/1 iaaM #1-1 with the line Tina (P5). The hybrids P1 and
P2 are homologous to C1 (DR2 × Tal1/1), except for the presence of the DefH9-
iaaM gene integrated in their genome. The transgenic hybrid P5 is homologous
to its untransformed control C2 (Tal1/1 × Tina). DR2 and Tina are parental
lines obtained through classical breeding, Tal1/1 is a double haploid line derived
from anther culture of the F1 commercial cultivar Talina. The segregation of the
marker gene nptII was checked by spraying the plants with kanamycin [14] and
allowed for the conclusion that the transgenes segregate as a single locus in the
backcrossed progenies of the three independent events analyzed (Tal iaaM 1-1: x
2
= 0.01065, P = 0.917; DR2 iaaM 34-1: x
2
= 0.0496 P = 0.824; DR2 iaaM 28-1
x
2
= 0.06467 P = 0.799). Southern blot analysis showed that DR2 iaaM 28-1 and
34-1 had a single copy of the transgene, while Tal iaaM 1-1 had three copies of the
transgene (Fig 4). Since the interaction genotype/location was not significant for
the yield, the data were computed as average of the two locations and subjected to
analysis of variance according to a randomized complete block design. Duncan's
Multiple Range Test (P = 0.05) was used for means separations.
Figure 4.
Southern blot analysis of transgenic eggplants. Numbers above the lanes indicate the independent transgenic
plant DR2iaaM#28-1 (28), DR2iaaM#34-1 (34) and Tal1/1iaaM#1-1 (Tal1/1-1). Cont indicates untransformed
plants, i.e. DR2 and Tal1/1, respectively. The probe used corresponds to the DefH9 regulatory region.
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