Genetic transformation of cork oak (Quercus suber L.) for herbicide resistance

Álvarez, Rafael Jiménez-Camino; Álvarez, José Marcos Alonso; Humara, Jaime M.; Revilla, A.; Ordás, Ricardo J.

doi:10.1007/s10529-009-0033-2

Cited by 21 publications

(11 citation statements)

References 32 publications

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“…High variability in GUS activity was observed between the different transformed clones. A comparison between GUS activity and transgene copy number suggests that the different level of gene expression cannot be explained by the copy number effect [ 52 , 53 ]. Therefore, other phenomena such as the position effect in the host genome [ 54 ] or other complex configurations of the integrated T-DNA [ 7 , 55 ] should also be considered.…”

Section: Discussionmentioning

confidence: 99%

Stable Agrobacterium‐Mediated Transformation of Maritime Pine Based on Kanamycin Selection

Álvarez

Ordás

2013

The Scientific World Journal

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An efficient transformation protocol based on kanamycin selection was developed for Agrobacterium-mediated transformation of maritime pine embryonal masses. The binary vector pBINUbiGUSint, which contained neomycin phosphotransferase II (nptII) as a selectable marker gene and β-glucuronidase (uidA) as a reporter gene, was used for transformation studies. Different factors, such as embryogenic line, bacterial strain, bacterial concentration, and coculture duration, were examined and optimized. For selection of transformants, 15 mgL−1 kanamycin was used. The highest transformation efficiency (11.4 events per gram of fresh mass) was achieved when a vigorously growing embryonal mass (embryogenic line L01) was cocultivated with Agrobacterium strain AGL1 at the optical density (OD600 nm) of 0.3 for 72 h. Evidence of the stable transgene integration was obtained by polymerase chain reaction for the nptII and uidA genes and expression of the uidA gene. Maturation capacity of the transgenic lines was negatively affected by the transformation process. Induction of axillary shoots by preculturing the embryos with benzyladenine allowed overcoming the low maturation rates of some transformed lines. The transgenic embryos were germinated and the axillar shoots were rooted. Transgenic plants were transferred to potting substrate showing normal growth.

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Section: Discussionmentioning

confidence: 99%

Stable Agrobacterium‐Mediated Transformation of Maritime Pine Based on Kanamycin Selection

Álvarez

Ordás

2013

The Scientific World Journal

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“…Herbicide resistance has only been tested at laboratory level, as plantlet conversion has not been reported (Alvarez et al 2009). …”

Section: Genetic Transformationmentioning

confidence: 99%

Application of biotechnological tools to Quercus improvement

et al. 2011

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The genus Quercus, which belongs to the family Fagaceae, is native to the northern hemisphere and includes deciduous and evergreen species. The trees of the different species are very important from both economic and ecological perspectives. Application of new technological approaches (which span the fields of plant developmental biology, genetic transformation, conservation of elite germplasm and discovery of genes associated with complex multigenic traits) to these long-rotation hardwoods may be of interest for accelerating tree improvement programs. This review provides a summary of the advances made in the application of biotechnological tools to specific oak species. Significant progress has been made in the area of clonal propagation via organogenesis and somatic embryogenesis (SE). Standardized procedures have been developed for micropropagating the most important European (Q. robur, Q. petarea, Q. suber) and American (Q. alba, Q. bicolor, Q. rubra) oaks by axillary shoot growth. Although regenerated plantlets are grown in experimental trials, large-scale propagation of oak species has not been carried out. The induction of SE in oaks from juvenile explants is generally not problematic, although the use of explants other than zygotic embryos is much less efficient. During the last decade, enormous advances have been made in inducing SE from selected adult trees, mainly specimens of pedunculate oak (Q. robur) and cork oak (Q. suber). Advances in the understanding of the maturation and germination steps are required for better use of embryogenic process in clonal forestry. Quercus species are late-maturing and late-flowering, exhibit irregular seed set, and produce seeds that are recalcitrant to storage by conventional procedures. Vitrification-based cryopreservation techniques were used successfully in somatic embryos of pedunculate oak and cork oak, and an applied genbank of cork oak selected genotypes is now under development. The feasibility of genetic transformation of pedunculate oak and cork oak somatic embryos by means of co-culture techniques with several strains of Agrobacterium tumefaciens has also been demonstrated. To date, most research on the genomics of Quercus species has concerned population genetics. Approaches using functional genomics to examine the molecular and cellular mechanisms that control organogenesis and or somatic embryogenesis are still scarce, and efforts on the isolation and characterization of genes related to other specific traits should be intensified in the near future, as this would help improve the practical application of clonal forestry in recalcitrant species such as oaks.

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“…The bar gene encodes the PPT acetyltransferase enzyme which acetylates a free amino group in PPT and thereby inactivates it [ 9 ]. The bar gene was inserted into different species and hybrids of Populus [ 3 , 10 ] and Eucalyptus [ 11 , 12 ], as well as oak [ 13 ] and various coniferous [ 14 , 15 ]; however, aspen plants have not been transformed. The aim of our work was to create herbicide-resistant aspen plants by transforming Russian highly productive aspen genotypes with the bar gene and to use testing under semi-natural conditions to select lines that look promising for plantation forestry.…”

Section: Introductionmentioning

confidence: 99%

Testing Transgenic Aspen Plants with bar Gene for Herbicide Resistance under Semi-natural Conditions

et al. 2016

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Obtaining herbicide resistant plants is an important task in the genetic engineering of forest trees. Transgenic European aspen plants (Populus tremula L.) expressing the bar gene for phosphinothricin resistance have been produced using Agrobacterium tumefaciens-mediated transformation. Successful genetic transformation was confirmed by PCR analysis for thirteen lines derived from two elite genotypes. In 2014–2015, six lines were evaluated for resistance to herbicide treatment under semi-natural conditions. All selected transgenic lines were resistant to the herbicide Basta at doses equivalent to 10 l/ha (twofold normal field dosage) whereas the control plants died at 2.5 l/ha. Foliar NH4-N concentrations in transgenic plants did not change after treatment. Extremely low temperatures in the third ten-day period of October 2014 revealed differences in freeze tolerance between the lines obtained from Pt of f2 aspen genotypes. Stable expression of the bar gene after overwintering outdoors was confirmed by RT-PCR. On the basis of the tests, four transgenic aspen lines were selected. The bar gene could be used for retransformation of transgenic forest trees expressing valuable traits, such as increased productivity.

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Genetic transformation of cork oak (Quercus suber L.) for herbicide resistance

Cited by 21 publications

References 32 publications

Stable Agrobacterium‐Mediated Transformation of Maritime Pine Based on Kanamycin Selection

Stable Agrobacterium‐Mediated Transformation of Maritime Pine Based on Kanamycin Selection

Application of biotechnological tools to Quercus improvement

Testing Transgenic Aspen Plants with bar Gene for Herbicide Resistance under Semi-natural Conditions

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