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Ensifer-mediated transformation: an efficient non-Agrobacterium protocol for the genetic modification of rice.

Zuniga-Soto E, Mullins E, Dedicova B - Springerplus (2015)

Bottom Line: Ensifer adhaerens (OV14) is a soil-related bacterium with the proven ability to genetically modify the model plant A. thaliana and the staple crop S. tuberosum (Wendt et al., Trans Res 21:567-578, 2012).The results indicated that E. adhaerens (OV14) exhibits infection efficiencies ranging between 50-70 %, 90-100 % and 90-95 % for Curinga, Nipponbare and IR64 respectively.Integration analyses conducted on 24 transgenic rice lines illustrated that T-DNA insertion occurred randomly throughout the rice genome for EMT (and AMT), with similar integration patterns in the rice genomic DNA observed for both bacterial species.

View Article: PubMed Central - PubMed

Affiliation: International Center for Tropical Agriculture (CIAT) Transformation Platform, International Center for Tropical Agriculture (CIAT), A.A. 6713 Cali, Colombia ; Department of Crop Science, Teagasc Crops Research Centre, Oak Park, Carlow, Ireland.

ABSTRACT
While Agrobacterium-mediated transformation (AMT) remains the most widely used technique for gene transfer in plants, interest exists for the use of non-Agrobacterium gene delivery systems due to freedom-to-operate issues that remain with AMT across several jurisdictions. In addition, the plant pathogenic mode of action of Agrobacterium tumefaciens significantly increases the costs to passage engineered cultivars through the regulatory process. Ensifer adhaerens (OV14) is a soil-related bacterium with the proven ability to genetically modify the model plant A. thaliana and the staple crop S. tuberosum (Wendt et al., Trans Res 21:567-578, 2012). While previous work was relevant for dicotyledonous species, in this study, the efficacy of Ensifer adhaerens (OV14)-mediated transformation (EMT) was determined on two japonica rice varieties, Curinga and Nipponbare, and the recalcitrant indica variety, IR64. The results indicated that E. adhaerens (OV14) exhibits infection efficiencies ranging between 50-70 %, 90-100 % and 90-95 % for Curinga, Nipponbare and IR64 respectively. Curinga and Nipponbare plants transformed with E. adhaerens (OV14) and A. tumefaciens (LBA4404 and EHA105) were regenerated achieving transformation efficiencies of 16 % and 26-32 % for Curinga and 7 and 4 % for Nipponbare respectively. Separately, the transformation of IR64 was only recorded via EMT (transformation efficiency ~1 %). Integration analyses conducted on 24 transgenic rice lines illustrated that T-DNA insertion occurred randomly throughout the rice genome for EMT (and AMT), with similar integration patterns in the rice genomic DNA observed for both bacterial species.

No MeSH data available.


Related in: MedlinePlus

Map positions of T-DNA insertions derived from independent transgenic lines. The 12 rice chromosomes are displayed in blue; mitochondrial and chloroplast DNA are indicated as black circles. T-DNA insertions belonging to E. adhaerens and A. tumefaciens are indicated in blue and red respectively
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Fig3: Map positions of T-DNA insertions derived from independent transgenic lines. The 12 rice chromosomes are displayed in blue; mitochondrial and chloroplast DNA are indicated as black circles. T-DNA insertions belonging to E. adhaerens and A. tumefaciens are indicated in blue and red respectively

Mentions: Mapping the T-DNA flanking sequences on the rice chromosomes revealed no apparent chromosomal position preference. T-DNA insertions appeared to be randomly distributed for both E. adhaerens and A. tumefaciens (both strains) obtained events. Moreover, the insertions appeared to be evenly distributed along the chromosomes within the rice genome (Fig. 3).Fig. 3


Ensifer-mediated transformation: an efficient non-Agrobacterium protocol for the genetic modification of rice.

Zuniga-Soto E, Mullins E, Dedicova B - Springerplus (2015)

Map positions of T-DNA insertions derived from independent transgenic lines. The 12 rice chromosomes are displayed in blue; mitochondrial and chloroplast DNA are indicated as black circles. T-DNA insertions belonging to E. adhaerens and A. tumefaciens are indicated in blue and red respectively
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4628045&req=5

Fig3: Map positions of T-DNA insertions derived from independent transgenic lines. The 12 rice chromosomes are displayed in blue; mitochondrial and chloroplast DNA are indicated as black circles. T-DNA insertions belonging to E. adhaerens and A. tumefaciens are indicated in blue and red respectively
Mentions: Mapping the T-DNA flanking sequences on the rice chromosomes revealed no apparent chromosomal position preference. T-DNA insertions appeared to be randomly distributed for both E. adhaerens and A. tumefaciens (both strains) obtained events. Moreover, the insertions appeared to be evenly distributed along the chromosomes within the rice genome (Fig. 3).Fig. 3

Bottom Line: Ensifer adhaerens (OV14) is a soil-related bacterium with the proven ability to genetically modify the model plant A. thaliana and the staple crop S. tuberosum (Wendt et al., Trans Res 21:567-578, 2012).The results indicated that E. adhaerens (OV14) exhibits infection efficiencies ranging between 50-70 %, 90-100 % and 90-95 % for Curinga, Nipponbare and IR64 respectively.Integration analyses conducted on 24 transgenic rice lines illustrated that T-DNA insertion occurred randomly throughout the rice genome for EMT (and AMT), with similar integration patterns in the rice genomic DNA observed for both bacterial species.

View Article: PubMed Central - PubMed

Affiliation: International Center for Tropical Agriculture (CIAT) Transformation Platform, International Center for Tropical Agriculture (CIAT), A.A. 6713 Cali, Colombia ; Department of Crop Science, Teagasc Crops Research Centre, Oak Park, Carlow, Ireland.

ABSTRACT
While Agrobacterium-mediated transformation (AMT) remains the most widely used technique for gene transfer in plants, interest exists for the use of non-Agrobacterium gene delivery systems due to freedom-to-operate issues that remain with AMT across several jurisdictions. In addition, the plant pathogenic mode of action of Agrobacterium tumefaciens significantly increases the costs to passage engineered cultivars through the regulatory process. Ensifer adhaerens (OV14) is a soil-related bacterium with the proven ability to genetically modify the model plant A. thaliana and the staple crop S. tuberosum (Wendt et al., Trans Res 21:567-578, 2012). While previous work was relevant for dicotyledonous species, in this study, the efficacy of Ensifer adhaerens (OV14)-mediated transformation (EMT) was determined on two japonica rice varieties, Curinga and Nipponbare, and the recalcitrant indica variety, IR64. The results indicated that E. adhaerens (OV14) exhibits infection efficiencies ranging between 50-70 %, 90-100 % and 90-95 % for Curinga, Nipponbare and IR64 respectively. Curinga and Nipponbare plants transformed with E. adhaerens (OV14) and A. tumefaciens (LBA4404 and EHA105) were regenerated achieving transformation efficiencies of 16 % and 26-32 % for Curinga and 7 and 4 % for Nipponbare respectively. Separately, the transformation of IR64 was only recorded via EMT (transformation efficiency ~1 %). Integration analyses conducted on 24 transgenic rice lines illustrated that T-DNA insertion occurred randomly throughout the rice genome for EMT (and AMT), with similar integration patterns in the rice genomic DNA observed for both bacterial species.

No MeSH data available.


Related in: MedlinePlus