Articles
TRANSFORMATION OF LILIUM LONGIFLORUM PLANTS FOR CUCUMBER MOSAIC VIRUS RESISTANCE BY PARTICLE BOMBARDMENT
Article number
568_30
Pages
209 – 214
Language
English
Abstract
Lilium species (Liliaceae) are a significant floriculture commodity and one of the three major bulb crops in the commercial market.
Lilies are monocotyledonous plants that have been generally recalcitrant to molecular genetic manipulation because of limitations that restrict utilization of transformation technologies that are routinely applied to dicotyledonous plants.
Cucumber mosaic virus (CMV) causes a serious disease of lily.
It is known that transgenic plants bearing a disabled CMV replicase gene can be resistant to that virus.
About 5000 pieces of morphogenic calli, 3-4 mm in diameter, derived from segments of sterile bulblet scales of L. longiflorum Thunb. cv. Snow Queen were microprojectile bombarded, using a Finer-type of bombardment apparatus, followed by bialaphos selection.
The plasmid p35SAc containing PAT selectable marker gene encoding phosphinothricin-N-acetyltransferase (that detoxifies the herbicide Basta) under the control of 35S promoter and the nopaline synthase (NOS) terminator; and the plasmid pSAN101 containing the CMV defective replicase gene under the control of Act promoter and NOS terminator, were used.
After several rounds of callus selection, plants were regenerated in vitro.
PCR analysis indicated that these plants contain both the PAT and the disabled CMV replicase transgenes.
Lilies are monocotyledonous plants that have been generally recalcitrant to molecular genetic manipulation because of limitations that restrict utilization of transformation technologies that are routinely applied to dicotyledonous plants.
Cucumber mosaic virus (CMV) causes a serious disease of lily.
It is known that transgenic plants bearing a disabled CMV replicase gene can be resistant to that virus.
About 5000 pieces of morphogenic calli, 3-4 mm in diameter, derived from segments of sterile bulblet scales of L. longiflorum Thunb. cv. Snow Queen were microprojectile bombarded, using a Finer-type of bombardment apparatus, followed by bialaphos selection.
The plasmid p35SAc containing PAT selectable marker gene encoding phosphinothricin-N-acetyltransferase (that detoxifies the herbicide Basta) under the control of 35S promoter and the nopaline synthase (NOS) terminator; and the plasmid pSAN101 containing the CMV defective replicase gene under the control of Act promoter and NOS terminator, were used.
After several rounds of callus selection, plants were regenerated in vitro.
PCR analysis indicated that these plants contain both the PAT and the disabled CMV replicase transgenes.
Authors
A. Lipsky, A. Cohen, V. Gaba, K. Kamo, A. Gera, A. Watad
Keywords
Lily, resistance, CMV, transformation, regeneration
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