Abstract
n this study, we report our efforts to develop an efficient Agrobacterium-mediated transformation protocol for the transformation of Indica rice IR58025B with mannose as a selection agent. Various factors, such as basal medium, culture temperature, strains for infection, air-drying duration of calli after infection, and salt concentration of coculture medium, were systematically optimized so that the highest transformation frequency (TF) reached to 74%. About 94% of mature embryos were induced to generate calli of high quality on modified NB medium. Both callus quantity and quality were improved when callus was induced from endosperm-free embryos in comparison to intact mature seeds. Higher temperature (32°C) showed improvements when compared to lower temperatures at (25°C and 28°C) for both callus induction and transformation. Agrobacterium strains of EHA105 produced the best TF in comparison with EHA101, LBA4404 and AGL1. However, LBA4404 produced the highest single copy frequency. Prior to co-cultivation, air-drying of inoculated calli was performed in a laminar flow hood for 4-7 hours and produced a higher TF. Lower salt concentration for co-cultivation medium enhanced both DNA delivery and TF.
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Claims 10
Claim · model-written summary
The optimized rice transformation protocol reached 74% transformation frequency.
- Taxon
- Indica Rice
Oryza sativa Indica Groupt0 - Material
- IR58025Bt0
- Outcome
- transformation frequency (TF)e0
- Value
- 74%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“highest transformation frequency (TF) reached to 74%.”
t0“Indica Rice Transformation of IR58025B”
Claim · model-written summary
Modified NB medium induced high-quality calli from about 94% of mature embryos.
- Taxon
- Indica RiceOryza sativa Indica Groupt0
- Material
- IR58025Bt0
- Treatment
- modified NB mediume0
- Outcome
- mature embryos were induced to generate calli of high qualitye0
- Direction
- inducede0
- Value
- 94%e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“About 94% of mature embryos were induced to generate calli of high quality on modified NB medium.”
t0“Indica Rice Transformation of IR58025B”
Claim · model-written summary
Endosperm-free embryos improved callus quantity and quality relative to intact mature seeds.
- Taxon
- Indica RiceOryza sativa Indica Groupt0
- Material
- IR58025Bt0
- Treatment
- endosperm-free embryose0
- Outcome
- callus quantity and qualitye0
- Direction
- improvede0
- Comparator
- intact mature seedse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Both callus quantity and quality were improved when callus was induced from endosperm-free embryos in comparison to intact mature seeds.”
t0“Indica Rice Transformation of IR58025B”
Claim · model-written summary
At 32°C, callus induction improved relative to 25°C and 28°C.
- Taxon
- Indica RiceOryza sativa Indica Groupt0
- Material
- IR58025Bt0
- Treatment
- Higher temperature (32°C)e0
- Outcome
- callus inductione0
- Direction
- improvementse0
- Comparator
- lower temperatures at (25°C and 28°C)e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Higher temperature (32°C) showed improvements when compared to lower temperatures at (25°C and 28°C) for both callus induction and transformation.”
t0“Indica Rice Transformation of IR58025B”
Claim · model-written summary
At 32°C, transformation improved relative to 25°C and 28°C.
- Taxon
- Indica RiceOryza sativa Indica Groupt0
- Material
- IR58025Bt0
- Treatment
- Higher temperature (32°C)e0
- Outcome
- transformatione0
- Direction
- improvementse0
- Comparator
- lower temperatures at (25°C and 28°C)e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Higher temperature (32°C) showed improvements when compared to lower temperatures at (25°C and 28°C) for both callus induction and transformation.”
t0“Indica Rice Transformation of IR58025B”
Claim · model-written summary
EHA105 yielded the best transformation frequency among the tested Agrobacterium strains.
- Taxon
- Indica RiceOryza sativa Indica Groupt0
- Material
- IR58025Bt0
- Treatment
- EHA105e0
- Interactor
- Agrobacterium strains of EHA105e0other
- Outcome
- TFe0
- Direction
- beste0
- Comparator
- EHA101, LBA4404 and AGL1e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Agrobacterium strains of EHA105 produced the best TF in comparison with EHA101, LBA4404 and AGL1.”
t0“Indica Rice Transformation of IR58025B”
Claim · model-written summary
LBA4404 produced the highest single-copy frequency.
- Taxon
- Indica RiceOryza sativa Indica Groupt0
- Material
- IR58025Bt0
- Treatment
- LBA4404e0
- Outcome
- single copy frequencye0
- Direction
- higheste0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“However, LBA4404 produced the highest single copy frequency.”
t0“Indica Rice Transformation of IR58025B”
Claim · model-written summary
Air drying inoculated calli for 4-7 hours before cocultivation raised transformation frequency.
- Taxon
- Indica RiceOryza sativa Indica Groupt0
- Material
- IR58025Bt0
- Treatment
- air-drying of inoculated callie0
- Outcome
- TFe0
- Direction
- highere0
- Qualifier
- for 4-7 hourse0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Prior to co-cultivation, air-drying of inoculated calli was performed in a laminar flow hood for 4-7 hours and produced a higher TF.”
t0“Indica Rice Transformation of IR58025B”
Claim · model-written summary
Lower cocultivation medium salt concentration enhanced DNA delivery.
- Taxon
- Indica RiceOryza sativa Indica Groupt0
- Material
- IR58025Bt0
- Treatment
- Lower salt concentration for co-cultivation mediume0
- Outcome
- DNA deliverye0
- Direction
- enhancede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Lower salt concentration for co-cultivation medium enhanced both DNA delivery and TF.”
t0“Indica Rice Transformation of IR58025B”
Claim · model-written summary
Lower cocultivation medium salt concentration enhanced transformation frequency.
- Taxon
- Indica RiceOryza sativa Indica Groupt0
- Material
- IR58025Bt0
- Treatment
- Lower salt concentration for co-cultivation mediume0
- Outcome
- TFe0
- Direction
- enhancede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Lower salt concentration for co-cultivation medium enhanced both DNA delivery and TF.”
t0“Indica Rice Transformation of IR58025B”
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