title: Efficient transformation of Kalanchoe blossfeldiana and production of male-sterile plants by engineered anther ablation.
abstract: Engineered male sterility in ornamental plants has many applications such as facilitate hybrid seed production, eliminate pollen allergens, reduce the need for deadheading to extend the flowering period, redirect resources from seeds to vegetative growth, increase flower longevity and prevent gene flow between genetically modified and related native plants. We have developed a reliable and efficient Agrobacterium-mediated protocol for the genetic transformation of different Kalanchoe blossfeldiana commercial cultivars. Transformation efficiency for cv. 'Hillary' was 55.3% whereas that of cv. 'Tenorio' reached 75.8%. Selection was carried out with the nptII gene and increasing the kanamycin concentration from 25 to 100 mg l(-1) allowed to reduced escapes from 50 to 60% to virtually 0%. This method was used to produce male-sterile plants through engineered anther ablation. In our approach, we tested a male sterility chimaeric gene construct (PsEND1::barnase) to evaluate its effectiveness and effect on phenotype. No significant differences were found in the growth patterns between the transgenic lines and the wild-type plants. No viable pollen grains were observed in the ablated anthers of any of the lines carrying the PsEND1::barnase construct, indicating that the male sterility was complete. In addition, seed set was completely abolished in all the transgenic plants obtained. Our engineered male-sterile approach could be used, alone or in combination with a female-sterility system, to reduce the invasive potential of new ornamentals, which has become an important environmental problem in many countries.
authors.affiliation: Instituto de Biología Molecular y Celular de Plantas (CSIC.-UPV.), Ciudad Politécnica de la Innovación, Edf. 8E. C/. Ingeniero Fausto Elio s/n, Valencia, Spain.
authors.forename: Begoña
authors.initials: B
authors.lastname: García-Sogo
authors.forename: Benito
authors.initials: B
authors.lastname: Pineda
authors.forename: Lourdes
authors.initials: L
authors.lastname: Castelblanque
authors.forename: Teresa
authors.initials: T
authors.lastname: Antón
authors.forename: Mónica
authors.initials: M
authors.lastname: Medina
authors.forename: Edelín
authors.initials: E
authors.lastname: Roque
authors.forename: Claudia
authors.initials: C
authors.lastname: Torresi
authors.forename: José Pío
authors.initials: JP
authors.lastname: Beltrán
authors.forename: Vicente
authors.initials: V
authors.lastname: Moreno
authors.forename: Luis Antonio
authors.initials: LA
authors.lastname: Cañas
mesh_terms: D035264:Flowers / Q000235:genetics / Q000254:growth & development* / Q000648:ultrastructure; D018506:Gene Expression Regulation, Plant; D005818:Genetic Engineering / Q000379:methods*; D031962:Kalanchoe / Q000235:genetics* / Q000254:growth & development; D010641:Phenotype; D051479:Plant Infertility; D030821:Plants, Genetically Modified / Q000235:genetics / Q000254:growth & development; D011058:Pollen / Q000254:growth & development; D011401:Promoter Regions, Genetic; D012231:Rhizobium; D014170:Transformation, Genetic