Efficient transformation of rice (<i>Oryza sativa</i> L.) mediated by <i>Agrobacterium</i> and sequence analysis of the boundaries of the T‐DNA

Hiei, Yukoh; Ohta, Shozo; Komari, Toshihiko; Kumashiro, Takashi

doi:10.1046/j.1365-313x.1994.6020271.x

Cited by 3,221 publications

(2,176 citation statements)

References 18 publications

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“…For determination of sub-cellular localization of SNAC2 protein, the coding region of SNAC2 was fused to green fluorescence protein (GFP) reporter gene in the vector p1381-EGFP Hu et al 2006) under the control of its native promoter. All the constructs were transformed into rice Zhonghua11 (O. sativa L. ssp japonica) according to the rice genetic transformation method (Hiei et al 1994). …”

Section: Constructs and Genetic Transformationmentioning

confidence: 99%

Characterization of transcription factor gene SNAC2 conferring cold and salt tolerance in rice

et al. 2008

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Plants respond to adverse environment by initiating a series of signaling processes including activation of transcription factors that can regulate expression of arrays of genes for stress response and adaptation. NAC (NAM, ATAF, and CUC) is a plant specific transcription factor family with diverse roles in development and stress regulation. In this report, a stress-responsive NAC gene (SNAC2) isolated from upland rice IRA109 (Oryza sativa L. ssp japonica) was characterized for its role in stress tolerance. SNAC2 was proven to have transactivation and DNA-binding activities in yeast and the SNAC2-GFP fusion protein was localized in the rice nuclei. Northern blot and SNAC2 promoter activity analyses suggest that SNAC2 gene was induced by drought, salinity, cold, wounding, and abscisic acid (ABA) treatment. The SNAC2 gene was over-expressed in japonica rice Zhonghua 11 to test the effect on improving stress tolerance. More than 50% of the transgenic plants remained vigorous when all WT plants died after severe cold stress (4-8°C for 5 days). The transgenic plants had higher cell membrane stability than wild type during the cold stress. The transgenic rice had significantly higher germination and growth rate than WT under high salinity conditions. Over-expression of SNAC2 can also improve the tolerance to PEG treatment. In addition, the SNAC2-overexpressing plants showed significantly increased sensitivity to ABA. DNA chip profiling analysis of transgenic plants revealed many up-regulated genes related to stress response and adaptation such as peroxidase, ornithine aminotransferase, heavy metal-associated protein, sodium/hydrogen exchanger, heat shock protein, GDSL-like lipase, and phenylalanine ammonia lyase. Interestingly, none of the up-regulated genes in the SNAC2-overexpressing plants matched the genes up-regulated in the transgenic plants over-expressing other stress responsive NAC genes reported previously. These data suggest SNAC2 is a novel stress responsive NAC transcription factor that possesses potential utility in improving stress tolerance of rice.

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Section: Constructs and Genetic Transformationmentioning

confidence: 99%

Characterization of transcription factor gene SNAC2 conferring cold and salt tolerance in rice

et al. 2008

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“…A number of plant transformation experiments have reported plants with a mixture of non-Mendelian and Mendelian inheritance of the transgene (Christou et al 1989;Tomes et al 1990;Spencer et al 1992;Walters et al 1992;Hiei et al 1994;Register et al 1994;Peng et al 1995). In our experiments, none of the four events we studied exhibited efficient pollen transmission.…”

Section: Discussionmentioning

confidence: 67%

Expression and inheritance of the wheat Glu-1DX5 gene in transgenic maize

et al. 2002

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We have produced transgenic maize plants containing a wheat Glu-1Dx5 gene encoding the high-molecularweight glutenin subunit 1Dx5. Analysis by SDS-PAGE showed that a protein similar in size to the wheat 1Dx5 subunit accumulates in the endosperm of transgenic maize from four independent transformation events. This protein reacts with a monoclonal antibody specific to the wheat 1Dx5 subunit and was not detected in nontransgenic controls or in pollen, anthers, leaves or embryos of plants grown from seeds expressing this protein in endosperm. Genomic Southern-blot analysis is consistent with results from SDS-PAGE and indicates that the transgene integration sites are complex and are different in the four events studied. Using the presence of this protein as a phenotypic marker, we studied the inheritance of this gene through three sexual generations. Reciprocal crosses with nontransgenic plants and self-pollinations were performed, and the resulting kernels were analyzed for the presence of the 1Dx5 subunit. These data, together with PCR analysis for the transgene, suggest that the transgene is inefficiently transmitted through pollen in all four events. Abstract We have produced transgenic maize plants containing a wheat Glu-1Dx5 gene encoding the highmolecular-weight glutenin subunit 1Dx5. Analysis by SDS-PAGE showed that a protein similar in size to the wheat 1Dx5 subunit accumulates in the endosperm of transgenic maize from four independent transformation events. This protein reacts with a monoclonal antibody specific to the wheat 1Dx5 subunit and was not detected in nontransgenic controls or in pollen, anthers, leaves or embryos of plants grown from seeds expressing this protein in endosperm. Genomic Southern-blot analysis is consistent with results from SDS-PAGE and indicates that the transgene integration sites are complex and are different in the four events studied. Using the presence of this protein as a phenotypic marker, we studied the inheritance of this gene through three sexual generations. Reciprocal crosses with nontransgenic plants and selfpollinations were performed, and the resulting kernels were analyzed for the presence of the 1Dx5 subunit. These data, together with PCR analysis for the transgene, suggest that the transgene is inefficiently transmitted through pollen in all four events.

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“…Use of transgenic technologies for developing resistance to various biotic and abiotic stresses in the background of hybrid parental lines is important. Although, Agrobacterium-mediated transformation system for Japonica subspecies (Hiei et al 1994) and a few Indica rice is well established efficiency of transformation protocol remains genotype dependent (Datta et al 1999(Datta et al , 2001Khanna and Raina 1999;Tu et al 1998;Zhang et al 1998;Lin and Zhang 2005;Park et al 1996).…”

Section: Introductionmentioning

confidence: 99%

Agrobacterium-mediated genetic transformation of commercially elite rice restorer line using nptII gene as a plant selection marker

Chakraborty

Reddy

Narasu

et al. 2016

Physiol Mol Biol Plants

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Transformation of commercially important indica cultivars remains challenging for the scientific community even though Agrobacterium-mediated transformation protocols for a few indica rice lines have been well established. We report successful transformation of a commercially important restorer line JK1044R of indica rice hybrid JKRH 401. While following existing protocol, we optimized several parameters for callusing, regeneration and genetic transformation of JK1044R. Calli generated from the rice scutellum tissue were used for transformation by Agrobacterium harboring pCAMBIA2201. A novel two tire selection scheme comprising of Geneticin (G418) and Paramomycin were deployed for selection of transgenic calli as well as regenerated plantlets that expressed neomycin phosphotransferase-II gene encoded by the vector. One specific combination of G418 (30 mg l −1 ) and Paramomycin (70 mg l −1 ) was very effective for calli selection. Transformed and selected calli were detected by monitoring the expression of the reporter gene uidA (GUS). Regenerated plantlets were confirmed through PCR analysis of nptII and gus genes specific primers as well as dot blot using gus gene specific as probe.

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Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T‐DNA

Cited by 3,221 publications

References 18 publications

Characterization of transcription factor gene SNAC2 conferring cold and salt tolerance in rice

Characterization of transcription factor gene SNAC2 conferring cold and salt tolerance in rice

Expression and inheritance of the wheat Glu-1DX5 gene in transgenic maize

Agrobacterium-mediated genetic transformation of commercially elite rice restorer line using nptII gene as a plant selection marker

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