Abstract
BACKGROUND: Cereal crops are the major targets for transformation mediated crop improvement and IR36 is an early maturing, high yielding, insect and disease resistant rice variety however, it is abiotic stress sensitive. Hence, development of an efficient and reproducible micropropagation system via somatic embryogenesis and Agrobacterium tumefaciens mediated transformation is prerequisite to develop abiotic stress tolerant IR36. Further, Genetic stability of analysis of plantlets through RAPD and ISSR and Ploidy level through Flow cytometry (FCM) measurement of 2C DNA content is necessary for future application of transformed IR36. RESULTS: In this study, Mature seeds inoculated on (Murashige and Skoog) MS medium with 11.31 μM 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 0.3 μM Kinetin (Kn) had highest callus induction frequency (98%). The highest regeneration frequency (80%) was observed in MS + 13.28 μM Benzyladenine (BA) with 8.06 μM α-naphthalene acetic acid (NAA). Randomly Amplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeat (ISSR) and Flow Cytometry (FCM) analysis showed no significant variation in the 2C DNA (0.81 pg/2C) content and Ploidy level between wild type IR36 and in vitro maintained rice lines. Of the various OD bacterial culture, an optimum OD of 0.4 and inoculation duration of 10 min resulted in efficient Agrobacterium-mediated transformation. β-glucuronidase activity was maximum in callus (99.05%). CONCLUSIONS: These results described here confirm the reliability of this protocol for micropropagation and delivery of desirable gene using A. tumefaciens into indica rice.
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Claims 6
Claim · model-written summary
MS medium with 11.31 μM 2,4-D and 0.3 μM kinetin gave the highest callus induction frequency, 98%.
- Taxon
- indica rice
Oryza sativa Indica Groupt0 - Organ / stage
- Mature seedse0
- Treatment
- MS medium with 11.31 μM 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 0.3 μM Kinetin (Kn)e0
- Outcome
- callus induction frequencye0
- Direction
- higheste0
- Value
- 98%e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Mature seeds inoculated on (Murashige and Skoog) MS medium with 11.31 μM 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 0.3 μM Kinetin (Kn) had highest callus induction frequency (98%).”
t0“indica rice”
Claim · model-written summary
MS with 13.28 μM BA and 8.06 μM NAA gave the highest regeneration frequency, 80%.
- Taxon
- indica riceOryza sativa Indica Groupt0
- Treatment
- MS + 13.28 μM Benzyladenine (BA) with 8.06 μM α-naphthalene acetic acid (NAA)e0
- Outcome
- regeneration frequencye0
- Direction
- higheste0
- Value
- 80%e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The highest regeneration frequency (80%) was observed in MS + 13.28 μM Benzyladenine (BA) with 8.06 μM α-naphthalene acetic acid (NAA).”
t0“indica rice”
Claim · model-written summary
Wild type IR36 and in vitro rice lines did not differ significantly in 2C DNA content or ploidy.
- Taxon
- rice
Oryza sativae0 - Outcome
- 2C DNA (0.81 pg/2C) content and Ploidy levele0
- Direction
- no significant variatione0
- Value
- 0.81 pg/2Ce0
- Comparator
- wild type IR36e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“analysis showed no significant variation in the 2C DNA (0.81 pg/2C) content and Ploidy level between wild type IR36 and in vitro maintained rice lines.”
Claim · model-written summary
An OD of 0.4 and ten-minute inoculation yielded efficient Agrobacterium-mediated transformation.
- Taxon
- indica riceOryza sativa Indica Groupt0
- Treatment
- an optimum OD of 0.4 and inoculation duration of 10 mine0
- Interactor
- Agrobacterium tumefaciens
Agrobacterium tumefacienst1other - Outcome
- Agrobacterium-mediated transformatione0
- Direction
- efficiente0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Of the various OD bacterial culture, an optimum OD of 0.4 and inoculation duration of 10 min resulted in efficient Agrobacterium-mediated transformation.”
t0“indica rice”
t1“Agrobacterium tumefaciens”
Claim · model-written summary
β-glucuronidase activity was maximal in callus at 99.05%.
- Taxon
- indica riceOryza sativa Indica Groupt0
- Organ / stage
- calluse0
- Gene
- β-glucuronidasee0
- Outcome
- β-glucuronidase activitye0
- Direction
- maximume0
- Value
- 99.05%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“β-glucuronidase activity was maximum in callus (99.05%).”
t0“indica rice”
Claim · model-written summary
The protocol was judged reliable for micropropagation and gene delivery into indica rice.
- Taxon
- indica riceOryza sativa Indica Groupe0
- Interactor
- A. tumefaciensAgrobacterium tumefaciense0other
- Outcome
- reliability of this protocole0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“These results described here confirm the reliability of this protocol for micropropagation and delivery of desirable gene using A. tumefaciens into indica rice.”
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