Abstract
The comparison of direct and indirect somatic embryogenesis (DSE and ISE) for biolistic transformation of sugarcane by minimal expression cassettes indicated the highest transformation efficiency with ISE (2.2 indepen- dent transgenic plants per shot) and the most rapid pro- duction of transgenic plants with DSE (12 weeks from explant to plants in soil). Microprojectiles of 0.3 lm diameter produced 5 times more transgenic lines than 1 lm microprojectiles when used at the same weight basis per shot. A significant reduction of the number of hybridization signals in the Southern blot was also observed with 0.3 lm microprojectiles when compared to 1.0 lm microprojec- tiles. This suggests that the lower DNA carrying capacity and greater number of the smaller microprojectiles con- tributes to more transgenic lines with less complex trans- gene integration. When geneticin sulfate, was used for selection following DSE, significantly more (4.8 times) transgenic plants were produced than with paromomycin sulfate and an equal number of non-expressing plants were produced with both selection agents. In conclusion, opti- mization of two alternative morphogenic routes for regeneration (DSE and ISE), biolistic and selection parameters generated rooted, transgenic plants of a com- mercially important sugarcane cultivar with simple trans- gene integration and within 12 or 19 weeks of culture initiation, respectively. Reducing the complexity of the transgene integration and minimizing the time in tissue culture will likely enhance the performance of the trans- genic events.
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Claims 7
Claim · model-written summary
Indirect somatic embryogenesis gave the highest transformation efficiency in sugarcane, at 2.2 independent transgenic plants per shot, compared with direct embryogenesis.
Verbatim
e0“The comparison of direct and indirect somatic embryogenesis (DSE and ISE) for biolistic transformation of sugarcane”
e1“indicated the highest transformation efficiency with ISE (2.2 indepen- dent transgenic plants per shot)”
Claim · model-written summary
Direct somatic embryogenesis gave the most rapid production of transgenic sugarcane plants, within 12 weeks from explant to plants in soil.
Not reportedComparator
Verbatim
e0“The comparison of direct and indirect somatic embryogenesis (DSE and ISE) for biolistic transformation of sugarcane”
e2“the most rapid pro- duction of transgenic plants with DSE (12 weeks from explant to plants in soil)”
Claim · model-written summary
Microprojectiles of 0.3 µm diameter produced 5 times more transgenic lines in sugarcane than 1 µm microprojectiles.
Verbatim
e0“The comparison of direct and indirect somatic embryogenesis (DSE and ISE) for biolistic transformation of sugarcane”
e3“Microprojectiles of 0.3 lm diameter produced 5 times more transgenic lines than 1 lm microprojectiles when used at the same weight basis per shot”
Claim · model-written summary
Significantly fewer hybridization signals in Southern blots were observed with 0.3 µm microprojectiles than with 1.0 µm microprojectiles in sugarcane.
Verbatim
e0“The comparison of direct and indirect somatic embryogenesis (DSE and ISE) for biolistic transformation of sugarcane”
e4“A significant reduction of the number of hybridization signals in the Southern blot was also observed with 0.3 lm microprojectiles when compared to 1.0 lm microprojec- tiles”
Claim · model-written summary
Geneticin sulfate produced 4.8 times significantly more transgenic sugarcane plants than paromomycin sulfate after direct embryogenesis.
Verbatim
e0“The comparison of direct and indirect somatic embryogenesis (DSE and ISE) for biolistic transformation of sugarcane”
e5“When geneticin sulfate, was used for selection following DSE, significantly more (4.8 times) transgenic plants were produced than with paromomycin sulfate”
Claim · model-written summary
The number of non-expressing sugarcane plants was equal under both selection agents.
Verbatim
e0“The comparison of direct and indirect somatic embryogenesis (DSE and ISE) for biolistic transformation of sugarcane”
e6“an equal number of non-expressing plants were produced with both selection agents”
Claim · model-written summary
Optimized protocols generated rooted transgenic plants of a sugarcane cultivar with simple transgene integration within 12 or 19 weeks of culture initiation.
Verbatim
e7“generated rooted, transgenic plants of a com- mercially important sugarcane cultivar with simple trans- gene integration and within 12 or 19 weeks of culture initiation”