Flower development of Lilium longiflorum - The Lilium information ...

Chapter 5
SUP PAT
1 2 3 + - 1 2 3 + -
Figure 2. PCR amplification of inserted genes. PAT gene fragment was amplified in the three recovered
A
clones, whereas the SUP gene is present only in two of the clones. Positive control (+) was carried
out with the plasmid used for transformation, and wild-type genome of Lilium longiflorum was used
as a negative control (-) of PCR reaction.
SQ 1 2 3
PAT
GAPDH
B
SQ 2
SUP
GAPDH
Figure 3. Northern analysis of gene expression in transgenic plants. (A) Total RNA isolated from leaves
derived from transgenic clones (1, 2 and 3) and a non-transgenic plant (SQ) hybridised with the PAT
probe. (B) Total RNA from floral meristems of clone 2 and from a non-transgenic plant was
hybridised with a SUP fragment. Loading control is shown by hybridisation with the constitutive
GAPDH gene visualised in the lower panels.
Discussion
Although lily (Lilium longiflorum) transformation mediated by microprojectile
bombardment is feasible, it has always shown low efficiencies. Concomitant with our
efforts of setting up an optimised protocol for lily transformation based on
bombardment, Watad et al. (1998) obtained transgenic lily plants by bombarding
morphogenic calli derived from bulblet scales. In fact, the very first transgenic lily was
produced in our lab a few years before, using a pollen-mediated transformation method
in which bombarded pollen was used to pollinate non-transgenic flowers (van der
Leede-Plegt et al., 1997). More recently, another report was published on L.
longiflorum transformation using a defective virus gene under the CaMV35S promoter
to induce resistance against the cucumber mosaic virus (CMV) (Lipsky et al., 2002).
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