Abstract
BACKGROUND: Rice genome sequencing projects have generated remarkable amount of information about genes and genome architecture having tremendous potential to be utilized in both basic and applied research. Success in transgenics is paving the way for preparing a road map of functional genomics which is expected to correlate action of a gene to a trait in cellular and organismal context. However, the lack of a simple and efficient method for transformation and regeneration is a major constraint for such studies in this important cereal crop. RESULTS: In the present study, we have developed an easy, rapid and highly efficient transformation and regeneration protocol using mature seeds as explants and found its successful applicability to a choice of elite indica rice genotypes. We have optimized various steps of transformation and standardized different components of the regeneration medium including growth hormones and the gelling agent. The modified regeneration medium triggers production of large number of shoots from smaller number of calli and promotes their faster growth, hence significantly advantageous over the existing protocols where the regeneration step requires maximum time. Using this protocol, significantly higher transformation efficiency (up to 46%) and regeneration frequency (up to 92% for the untransformed calli and 59% for the transformed calli) were achieved for the four tested cultivars. We have used this protocol to produce hundreds of independent transgenic lines of different indica rice genotypes. Upon maturity, these transgenic lines were fertile thereby indicating that faster regeneration during tissue culture did not affect their reproductive potential. CONCLUSIONS: This speedy, yet less labor-intensive, protocol overcomes major limitations associated with genetic manipulation in rice. Moreover, our protocol uses mature seeds as the explant, which can easily be obtained in quantity throughout the year and kept viable for a long time. Such an easy, efficient and generalized protocol has the potential to be a major tool for crop improvement and gene-function studies on the model monocot plant rice.
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Claims 6
Claim · model-written summary
The modified regeneration medium produced more shoots from fewer calli and promoted faster growth than existing protocols.
- Taxon
- indica rice
Oryza sativa Indica Groupt0 - Treatment
- modified regeneration mediume0
- Outcome
- production of large number of shoots from smaller number of calli and promotes their faster growthe0
- Direction
- large numbere0
- Comparator
- existing protocolse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The modified regeneration medium triggers production of large number of shoots from smaller number of calli and promotes their faster growth, hence significantly advantageous over the existing protocols”
t0“indica rice”
Claim · model-written summary
The protocol achieved transformation efficiency up to 46% in four tested cultivars.
- Taxon
- indica riceOryza sativa Indica Groupt0
- Outcome
- transformation efficiencye0
- Direction
- highere0
- Value
- 46%e0
- Qualifier
- four tested cultivarse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“significantly higher transformation efficiency (up to 46%) and regeneration frequency (up to 92% for the untransformed calli and 59% for the transformed calli) were achieved for the four tested cultivars”
t0“indica rice”
Claim · model-written summary
Regeneration frequency reached 92% for untransformed calli.
- Taxon
- indica riceOryza sativa Indica Groupt0
- Material
- untransformed callie0
- Outcome
- regeneration frequencye0
- Value
- 92%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“regeneration frequency (up to 92% for the untransformed calli and 59% for the transformed calli)”
t0“indica rice”
Claim · model-written summary
Regeneration frequency reached 59% for transformed calli.
- Taxon
- indica riceOryza sativa Indica Groupt0
- Material
- transformed callie0
- Outcome
- regeneration frequencye0
- Value
- 59%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“regeneration frequency (up to 92% for the untransformed calli and 59% for the transformed calli)”
t0“indica rice”
Claim · model-written summary
The protocol produced hundreds of independent transgenic lines of different indica rice genotypes.
- Taxon
- indica riceOryza sativa Indica Groupt0
- Outcome
- independent transgenic linese0
- Value
- hundredse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“We have used this protocol to produce hundreds of independent transgenic lines of different indica rice genotypes.”
t0“indica rice”
Claim · model-written summary
The transgenic rice lines were fertile at maturity.
- Taxon
- indica riceOryza sativa Indica Groupt0
- Organ / stage
- maturitye0
- Outcome
- transgenic lines were fertilee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Upon maturity, these transgenic lines were fertile”
t0“indica rice”
The same questions in other papers
in vitro · yield · Oryza sativa (rice)Positive and negative results
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- negative
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