Abstract
In vitro regeneration and Agrobacterium-mediated genetic transformation was studied in popular local cultivar of chilli viz., Byadagi Kaddi. Different explants (cotyledon, hypocotyls and cotyledonary nodal region) were cultured on MS medium supplemented with various levels of BAP (2–12 mg/l) and TDZ (0.1-1.0 mg/l) for adventitious shoot induction. Frequency of adventitious shoot induction was significantly more in cotyledonary nodal region explant than cotyledon and hypocotyls explants. Adventitious shoot induction response was high on medium with 0.5 mg/l TDZ. Addition of 1 mg/l of GA3 to this medium resulted in higher frequency of shoot elongation. The elongated shoots were rooted on MS medium with 0.5 mg/l IAA and 0.1 mg/l BAP. Different concentrations and combinations of growth regulators were tried to study callus induction and regeneration from cotyledon, hypocotyls and cotyledonary nodal region explants. The media combination of 0.5 mg/l 2, 4-D and 0.5 mg/l kinetin resulted in higher quantity of callus. Elimination of 2, 4-D from this media induced somatic embryos. Complete regeneration was also observed on the same media but with very low frequency (10%). Shoot apical meristems were co-cultivated with Agrobacterium strain EHA105 carrying pBinBt3 construct having cry1Ac gene. The explants pre-cultured for 48 hours, infected with agrobacterial suspension having 0.5 × 10 8 cells/ml for 10 min along with acetosyringene @ 200 μM for 48 hours yielded the transformants under in vitro conditions. Analysis of the putative transformants for the presence and expression of cry gene revealed that the transformation efficiency was 0.5 per cent. In two step RT-PCR amplicon of 1000 bp revealed the expression of transformed gene.
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Claims 10
Claim · model-written summary
Cotyledonary nodal region explants had significantly more shoot induction than cotyledon or hypocotyl explants.
Verbatim
e0“Frequency of adventitious shoot induction was significantly more in cotyledonary nodal region explant than cotyledon and hypocotyls explants”
t0“chilli”
Claim · model-written summary
Medium containing 0.5 mg/l TDZ gave high adventitious shoot induction.
- Taxon
- chillit0
- Treatment
- medium with 0.5 mg/l TDZe0
- Outcome
- Adventitious shoot induction responsee0
- Direction
- highe0
Not reportedComparator
Verbatim
e0“Adventitious shoot induction response was high on medium with 0.5 mg/l TDZ”
t0“chilli”
Claim · model-written summary
Adding 1 mg/l GA3 increased shoot elongation frequency.
Verbatim
e0“Addition of 1 mg/l of GA3 to this medium resulted in higher frequency of shoot elongation”
t0“chilli”
Claim · model-written summary
Elongated shoots rooted on MS medium with IAA and BAP.
Verbatim
e0“The elongated shoots were rooted on MS medium with 0.5 mg/l IAA and 0.1 mg/l BAP”
t0“chilli”
Claim · model-written summary
Medium with 2,4-D and kinetin produced higher callus quantity.
- Taxon
- chillit0
- Treatment
- 0.5 mg/l 2, 4-D and 0.5 mg/l kinetine0
- Outcome
- quantity of calluse0
- Direction
- highere0
Not reportedComparator
Verbatim
e0“The media combination of 0.5 mg/l 2, 4-D and 0.5 mg/l kinetin resulted in higher quantity of callus”
t0“chilli”
Claim · model-written summary
Removing 2,4-D from this medium induced somatic embryos.
Not reportedComparatorDirection
Verbatim
e0“Elimination of 2, 4-D from this media induced somatic embryos”
t0“chilli”
Claim · model-written summary
Regeneration on the same medium occurred at 10% frequency.
Verbatim
e0“Complete regeneration was also observed on the same media but with very low frequency (10%)”
t0“chilli”
Claim · model-written summary
The stated preculture and Agrobacterium infection conditions yielded transformants.
Verbatim
e0“The explants pre-cultured for 48 hours, infected with agrobacterial suspension having 0.5 × 10 8 cells/ml for 10 min along with acetosyringene @ 200 μM for 48 hours yielded the transformants under in vitro conditions”
t0“chilli”
Claim · model-written summary
Transformation efficiency was 0.5 per cent.
Verbatim
e0“the transformation efficiency was 0.5 per cent”
t0“chilli”
Claim · model-written summary
RT-PCR detected expression of the transformed gene in a 1000 bp amplicon.
Verbatim
e0“In two step RT-PCR amplicon of 1000 bp revealed the expression of transformed gene”
t0“chilli”
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