An improvedAgrobacterium-mediated transformation protocol for recalcitrant elite indica rice cultivars

Kumar, K. K.; Maruthasalam, S.; Loganathan, M.; Sudhakar, D.; Balasubramanian, P.

doi:10.1007/bf02772648

Cited by 90 publications

(80 citation statements)

References 18 publications

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“…contribute to an optimal performance in any transformation system. The vector pCAMBIA 1301 used in the present study has been proved ideal for rice transformation as reported by other groups (Hiei et al 1994;Ilag et al 2000;Chern et al 2001;Saharan et al 2004;Kumar et al 2005). Mature seed derived calli were widely considered as suitable target tissue for Agrobacterium-mediated and biolistic transformation (Kumar et al 2005).…”

Section: Discussionsupporting

confidence: 51%

“…Agrobacterium-mediated transformation has several advantages such as high transformation efficiency compared to protoplast transfer (Rao et al 2009), the ability to transfer large pieces of DNA, minimal re-arrangement of transferred DNA, and characteristic insertion into the recipient genome of a discrete segment of DNA at a low copy number (Kumar et al 2005;Tyagi et al 2007) compared to particle bombardment approach (Shrawat et al 2006;Rao et al 2009). Efficiency of Agrobacterium-mediated transformation and delivery of T-DNA into plant cells is influenced by several parameters including explant selection (Owens & Cress 1985;Hiei et al 1994;Ishida et al 1996).…”

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confidence: 99%

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The effect of different antibiotics on the elimination of Agrobacterium and high frequency Agrobacterium-mediated transformation of indica rice (Oryza sativa L.)

Priya¹,

Subramanian²,

Manikandan³

2012

Czech J. Genet. Plant Breed.

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Abstract:We report here the suitable explant with high efficiency of transformation and the positive effects of timentin over other antibiotics like carbenicillin and cefotaxime on the elimination of Agrobacterium tumefaciens during the genetic transformation of popular indica rice (Oryza sativa L.). The tissues assayed were embryogenic calli, embryos with endosperm contamination, intact seeds, leaf blades, leaf bases and coleoptiles. The frequency of transient β-glucuronidase (GUS) expression as revealed by histochemical assay was 90% for embryogenic calli, which was the highest among the explants used. On the basis of disc-diffusion assay, the maximum zone of inhibition (29 mm) at 250 mg/l was obtained for timentin. In tissue culture conditions the frequency of Agrobacterium recurrence after 20 days of infection was minimum (2.3%) at 200 mg/l of timentin. At 250 mg/l of timentin there was no Agrobacterium growth, besides, there were no negative effects on the callus growth unlike other antibiotics, hence it was selected as the optimum concentration for high frequency callus proliferation and regeneration. The effect of the parameters evaluated was determined by the callus proliferation during selection, reduction in browning, transient GUS expression and stable transformation efficiency (23.3%). The resultant plants were stable transformants as confirmed by a molecular analysis of the gus and hpt genes. The developed transformation protocol will be very helpful for the information on indica rice cultivars in general and on IR 64 in particular.

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Section: Discussionsupporting

confidence: 51%

mentioning

confidence: 99%

The effect of different antibiotics on the elimination of Agrobacterium and high frequency Agrobacterium-mediated transformation of indica rice (Oryza sativa L.)

Priya¹,

Subramanian²,

Manikandan³

2012

Czech J. Genet. Plant Breed.

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“…Based on the results, it could be concluded that significant differences existed between the cultivars in transformation, and the Hwasan 101 cultivar could be considered the most suitable cultivar for Italian ryegrass transformation. Similar to our experiment, significant genotypic variation in Agrobacterium-mediated genetic transformation potential has been reported in many monocotyledonous species (Kumar et al 2005;Wu et al 2007). …”

Section: Genotypic Variation Of Italian Ryegrass Cultivars On Transfosupporting

confidence: 89%

Genotypic variation of Agrobacterium-mediated transformation of Italian ryegrass

Lee¹,

Choi²,

Kim³

et al. 2010

Electron. J. Biotechnol.

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“…This is in agreement with a previous report, in which the use of a lower concentration (OD of 0.5) of Agrobacterium culture, as well as cocultivation media overlaid with Whatman #1 filter paper, reduced necrosis of calli. This could be because of the reduced damage to explants during Agrobacterium infection, which resulted in less phenolic production and better recovery of calli during selection (Kumar et al, 2005). The optimum cocultivation period Values represent means ± SD.…”

Section: Discussionmentioning

confidence: 99%

Ploidy stability of Oryza sativa. L cv IR64 transformed with the moth bean P5CS gene with significant tolerance against drought and salinity

Priya¹,

Krishnan²,

Ramesh³

2015

Turk J Biol

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Rice is one of the world's most important crops. Thus, it is necessary to improve it by genetic transformation. IR64 is a highyielding and disease-resistant variety, but it is sensitive to abiotic stresses, mainly salinity and drought. Therefore, an efficient transformation system is a prerequisite for transforming this indica rice with a stress-tolerant gene, the potent osmolyte proline-coding P5CS gene of Vigna aconitifolia. Successful transgene integration was confirmed by polymerase chain reaction (PCR) and Southern blot analysis. A Mendelian pattern of transgene inheritance was also observed in the progeny plants. The individual transformants expressed significant tolerance over wild-type control plants, as exhibited by NaCl tolerance, lipid peroxidation levels, and total chlorophyll content. Percentage of germination, height, and weight of the plantlet at 400 mM mannitol stress was 60%, 8.1 cm, and 92.5 mg, respectively. Total chlorophyll retention and malondialdehyde content at 200 mM NaCl stress was 65.3% and 8.2 n mol g -1 FW, respectively. These results revealed the abiotic stress tolerance conferred by Δ1-pyrroline-5-carboxylate synthetase. Additionally, flow cytometry analysis confirmed the absence of somaclonal variation. Hence, an efficient genetic transformation can be accomplished in other indica rice cultivars by using this protocol.

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An improvedAgrobacterium-mediated transformation protocol for recalcitrant elite indica rice cultivars

Cited by 90 publications

References 18 publications

The effect of different antibiotics on the elimination of Agrobacterium and high frequency Agrobacterium-mediated transformation of indica rice (Oryza sativa L.)

The effect of different antibiotics on the elimination of Agrobacterium and high frequency Agrobacterium-mediated transformation of indica rice (Oryza sativa L.)

Genotypic variation of Agrobacterium-mediated transformation of Italian ryegrass

Ploidy stability of Oryza sativa. L cv IR64 transformed with the moth bean P5CS gene with significant tolerance against drought and salinity

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