Agrobacterium-mediated genetic transformation of Fraxinus americana hypocotyls

Palla, Kaitlin J.; Pijut, Paula M.

doi:10.1007/s11240-014-0630-1

Cited by 27 publications

(19 citation statements)

References 21 publications

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“…We found that the appropriate rooting medium was MS medium supplemented with 0.5 mg/L IAA and 0.5 mg/L IBA, in which rooting efficiency was significantly higher than that of hormone-free MS medium (Figure 2b). Similar results have been found for other plants, such as white ash (Fraxinus americana) and grass pea (Lathyrus sativus), where IAA and IBA applications promoted rooting efficiency (Barpete et al 2014;Palla and Pijut 2015). Taken together, the results indicate that stable plantlet regeneration was optimized in cv.…”

Section: Discussionsupporting

confidence: 83%

“…In this study, the highest transformation efficiency (67%) was obtained under the conditions of an Agrobacterium concentration of OD600 = 0.6, combined with an Agrobacterium infection time of 15 min and 3 days of co-cultivation based on GUS staining analysis (Table 2). This finding is similar to the Agrobacterium concentration of OD600 = 0.4-0.6, an infection time of 10 min and a co-cultivation duration of 2-3 days found to be optimal conditions for transformation of baby bamboo (Pogonatherum paniceum) (Li et al 2015) and white ash (Fraxinus americana) (Palla and Pijut 2015). Furthermore, Agrobacterium infection for 2-10 min with 2-4 days of co-cultivation has been shown beneficial for increasing transformation efficiency in other stylo species, e.g.…”

Section: Discussionsupporting

confidence: 80%

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Improvement of plant regeneration and Agrobacterium-mediated genetic transformation of Stylosanthes guianensis

Peng-fei¹,

Liu²,

Lei³

et al. 2019

Trop. grassl.-Forrajes trop.

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Improvement of plant regeneration and Agrobacterium-mediated genetic transformation of Stylosanthes guianensisMejoramiento de la regeneración de plantas y transformación genética mediada por Agrobacterium en Stylosanthes guianensis Abstract As a pioneer tropical pasture legume, stylo (Stylosanthes guianensis) is well adapted to growth-limiting factors in acid soils. Considering the importance of stylo, there is a need to improve Agrobacterium-mediated genetic transformation to enable development of elite cultivars. In this study, S. guianensis cv. RY5 was used to systematically optimize Agrobacteriummediated transformation based on its plant regeneration. Results showed that Murashige and Skoog (MS) medium containing 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 2 mg/L 6-benzylaminopurine (6-BA) was the optimal callus induction medium. MS medium supplemented with 2 mg/L 6-BA was suitable for shoot regeneration from cotyledonderived calluses, and 0.5 mg/L indole-3-acetic acid (IAA) and 0.5 mg/L indole-3-butyric acid (IBA) applications were beneficial for rooting. The highest transformation efficiency (67%) was obtained at an Agrobacterium concentration of optical density = 0.6 combined with an infection time of 15 min and 3 days of co-cultivation. Furthermore, 200 mg/mL carbenicillin (Carb) and 0.6 mg/L Basta ® supplements were effective in eliminating excess bacterial growth and selecting transgenic plants, respectively. Subsequent polymerase chain reaction (PCR) analysis confirmed that the β-glucuronidase (GUS) and BAR genes were successfully integrated into the stylo genome. Wider testing of this improved protocol as a means of enhancing genetic improvement and gene function analysis of stylo seems warranted. ResumenComo leguminosa pionera para pasturas tropicales, stylo (Stylosanthes guianensis) es una especie bien adaptada a los factores limitantes del crecimiento en suelos ácidos. Considerando la importancia de stylo, existe la necesidad de mejorar la transformación genética mediada por Agrobacterium para permitir el desarrollo de cultivares élite. En este estudio, conducido en China tropical, se utilizó S. guianensis cv. RY5 para optimizar en forma sistemática la transformación mediada por Agrobacterium con base en la regeneración de plantas. Los resultados mostraron que el medio Murashige & Skoog (MS) que contenía 0.2 mg/L de ácido 2,4-diclorofenoxiacético (2,4-D) y 2 mg/L de 6-bencilaminopurina (6-BA) fue el medio óptimo para la inducción de tejido calloso. El medio MS suplementado con 2 mg/L de 6-BA fue adecuado para la regeneración de brotes a partir de callos derivados de cotiledones, y aplicaciones de 0.5 mg/L de ácido indol-3-acético (IAA) y 0.5 mg/L de ácido indol-3-butírico (IBA) fueron beneficiosas para la formación de raices. La mayor eficiencia de transformación (67%) se obtuvo a una concentración de Agrobacterium con densidad óptica de 0.6, Genetic transformation of Stylosanthes guianensis 481Tropical Grasslands-Forrajes Tropicales (ISSN: 2346-3775) combinada con un tiempo de infección de...

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Section: Discussionsupporting

confidence: 83%

Section: Discussionsupporting

confidence: 80%

Improvement of plant regeneration and Agrobacterium-mediated genetic transformation of Stylosanthes guianensis

Peng-fei¹,

Liu²,

Lei³

et al. 2019

Trop. grassl.-Forrajes trop.

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“…In the present study, 100 mg/l kanamycin was effective for the selection of transgenic somatic embryos. It has been reported that high concentrations of selection agent were effective for the selection of transformed callus and organs in other plant system (Li et al, 1992;Cervera et al, 1998;Saini and Jaiwal, 2007;Shrawat andGood, 2011, Junaid et al, 2012a;Junaid et al, 2012b;Liu et al, 2014;Palla and Pijut, 2015).…”

Section: Discussionmentioning

confidence: 99%

Agrobacterium-mediated genetic transformation of date palm (Phoenix dactylifera L.) cultivar "Khalasah" via somatic embryogenesis

Aslam¹,

Khan²,

Azad³

2015

Plant Sci Today

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Plant Science Today (2015) 4 days and later it was (somatic embryos) inoculated on germinating and plantlet conversion MS medium supplemented with BAP (0.75 mg/l) + kanamycin (100 mg/l). Prolific shoots developed from putatively transformed matured embryos showed 47.5 % transformation efficiency. A large number of transgenic plants were obtained and later established in black hard plastic bags. A strong GUS activity was detected in the putatively transformed plant leaves by histochemical assay and, the integration of uidA (GUS) and npt II genes into transgenic plants was confirmed by polymerase chain reaction (PCR) and Southern hybridization analysis. The established transformation protocol allows effective and quick regeneration via somatic embryogenesis, and it would be highly valuable for existing date palm orchards and improving their productivity.

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“…Many studies, including those on Dieffenbachia (Shen et al 2007), Atropa (Song and Walworth 2013), Cicer (Tripathi et al 2013) and Fraxinus (Palla and Pijut 2015) have shown that shoot organogenesis is induced in calli by the synergistic action of auxins with cytokinins. In our study, shoots could not be produced from callus derived from petiole or leaf explants from any of the cultivars.…”

Section: Discussionmentioning

confidence: 99%

Agrobacterium tumefaciens-mediated transformation of axillary bud callus of Hibiscus rosa-sinensis L. ‘Ruby’ and regeneration of transgenic plants

Trivellini

Ferrante

Hunter

et al. 2015

Plant Cell Tiss Organ Cult

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Hibiscus rosa-sinensis L. is a popular ornamental species valued for its large brightly coloured ephemeral flowers and has a range of health-promoting properties. The value of H. rosa-sinensis could be improved even further if there were ways to prolong the display life of its short-lived flowers, and to improve its frost tolerance. Development of an efficient plant transformation and regeneration procedure that allows introduction of genes into the plant will greatly facilitate this. Here we outline a transformation and regeneration procedure that is the first to produce transformed H. rosa-sinensis plants successfully. We first optimised callus induction and shoot regeneration efficiency. The highest shoot regeneration frequency of 66.7 % was achieved in the cultivar 'Ruby' when callus induced from axillary buds using a basal medium supplemented with 2.22 lM benzylaminopurine and 2.47 lM b-naphthoxyacetic acid was cultured on shoot regeneration medium. The frequency of shoot regeneration from callus was lower in 'Ben James' and absent in 'Bright Light', indicating genotypic differences. When axillary bud-derived callus of 'Ruby' was cocultured with Agrobacterium tumefaciens harbouring a bglucuronidase (GUS) reporter plasmid, 49 % of calli produced shoots on selection media. All tested plantlets were confirmed as transformed based on the presence of the GUS transgene in the genomic DNA and GUS activity measurements. Roots were induced on transgenic plantlets using half-strength basal medium supplemented with 2.85 lM indole-3-acetic acid. This simple protocol can be used to improve the ornamental, agronomic and healthpromoting traits of H. rosa-sinensis hitherto recalcitrant to A. tumefaciens-mediated transformation.

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Agrobacterium-mediated genetic transformation of Fraxinus americana hypocotyls

Cited by 27 publications

References 21 publications

Improvement of plant regeneration and <i>Agrobacterium</i>-mediated genetic transformation of <i>Stylosanthes guianensis</i>

Improvement of plant regeneration and <i>Agrobacterium</i>-mediated genetic transformation of <i>Stylosanthes guianensis</i>

Agrobacterium-mediated genetic transformation of date palm (Phoenix dactylifera L.) cultivar "Khalasah" via somatic embryogenesis

Agrobacterium tumefaciens-mediated transformation of axillary bud callus of Hibiscus rosa-sinensis L. ‘Ruby’ and regeneration of transgenic plants

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