Revised cocultivation conditions produce effective Agrobacterium-mediated genetic transformation of carnation (Dianthus caryophyllus L.)

Production and rooting behaviour of rolB-transgenic plants of grape rootstock 'Richter 110' (Vitis berlandieri 3 V. rupestris) Abstract Genetic improvement of grape rootstocks is aimed at protection against grape phylloxera and other soil-borne pests and diseases, good rooting and graft compatibility as well as adaptability to a wide range of soil and climatic conditions. Apart from the long evaluation period required, breeding is complicated by the high heterozygosity in grapes. As an alternative to traditional crossing, gene transfer permits addition of single traits, largely without affecting the genetic background of existing valuable cultivars. Here we report on the production and rooting behaviour of transgenic grape rootstock 'Richter 110' carrying the Agrobacterium rhizogenes rolB gene, which is known to promote rooting. Transformation was achieved by co-cultivation of somatic embryogenic callus with Agrobacterium tumefaciens LBA4404 harbouring plasmid pHrB. The T-DNA of pHrB contains the hpt gene, conferring hygromycin resistance, and the rolB gene under control of its own promoter. PCR using transgene-specific primers verified the presence of hpt in all 36 hygromycin resistant clones selected, while only 24 clones also possessed the rolB gene. Rooting behaviour was examined in vitro, using tip, node and internode explants, and in aeroponic culture in the greenhouse, using single-node cuttings. Compared to internodes of non-transgenic 'Richter 110', those of rolB-transgenic clones in general showed significantly higher rooting ability and, in contrast to the former, were able to root profusely even in the absence of auxin. Cuttings of three rolB-transgenic clones in aeroponic culture produced almost twice as many primary roots as those of the non-transgenic control.

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“…Such incomplete transfers or integrations have been reported previously (e.g. Dean et al 1988;Sedira et al 2001;Nontaswatsri et al 2004).…”

Section: Discussionsupporting

confidence: 76%

Production and rooting behaviour of rolB-transgenic plants of grape rootstock ‘Richter 110’ (Vitis berlandieri × V. rupestris)

Geier

Eimert

Scherer

et al. 2008

Plant Cell Tiss Organ Cult

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“…It was observed that the media with AS promoted transformation; this is in agreement with a previous report (Ozawa, 2009) stating that the omission of AS reduced transformation efficiency. The addition of AS in the inoculation step induced genetic transformation; further addition of AS in the cocultivation step produced higher transformation rates than those achieved by incorporating it in the inoculation step alone (Nontaswatsri et al, 2004). The addition of 100 μM AS resulted in a higher intensity of GUS expression than 200 μM AS (Cui et al, 2004;Svabova and Griga, 2008).…”

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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“…Genetic engineering is a valuable tool, currently used in classical breeding methods for the production of novel carnation varieties, and these technologies have been exploited to introduce new traits into commercial cultivars. In the last 20 years, Agrobacterium tumefaciens -mediated transformation systems have been developed using different plant explants like leaves, petals, and stems Van Altvorst et al 1995, 1996Yantcheva et al 1997b ;Nontaswatsri et al 2004 ). Direct gene transfer by using microprojectile bombardment or with Agrobacterium was also successfully conducted (Zuker et al 1995(Zuker et al , 1999 in creating transgenic carnations with novel traits.…”

Section: Genetic Transformation Of Carnationmentioning

confidence: 99%

“…In most cases, bacterial strain and density, period of cocultivation, selective pressure, and regeneration method were identifi ed to be very crucial for the effi ciency of transformation. Different Agrobacterium strains like AGLO, LBA 4404, and EHA 105 were used and bacterial density level varies widely (0.5-1 OD 6oo ) in tested transformation experiments (Zuker et al 1999 ;Nontaswatsri et al 2004 ;Szoke et al 2006 ;Arici and Koc 2009 ). The positive effect of acetosyringone in different concentrations was reported in different research studies (Nontaswatsri and Fukai 2006 ;Arici and Koc 2009 ).…”

Section: Genetic Transformation Of Carnationmentioning

confidence: 99%

Somatic Embryogenesis and Genetic Transformation of Carnation (Dianthus caryophyllus L.)

Iantcheva

2015

Somatic Embryogenesis in Ornamentals and Its Applications

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The chapter summarizes the progress of somatic embryogenesis and genetic transformation of carnation obtained during the last 40 years. Factors that determine the processes of indirect and direct somatic embryogenesis of this ornamental plant are described and discussed. The present chapter outlines the current results in the fi eld and focuses on the primary guidelines of genetic transformation of carnation. Future applications and intentions identifi ed by Bulgarian research group in this direction have also been highlighted.

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Revised cocultivation conditions produce effective Agrobacterium-mediated genetic transformation of carnation (Dianthus caryophyllus L.)

Cited by 19 publications

References 22 publications

Production and rooting behaviour of rolB-transgenic plants of grape rootstock ‘Richter 110’ (Vitis berlandieri × V. rupestris)

Production and rooting behaviour of rolB-transgenic plants of grape rootstock ‘Richter 110’ (Vitis berlandieri × V. rupestris)

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

Somatic Embryogenesis and Genetic Transformation of Carnation (Dianthus caryophyllus L.)

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