Efficient transformation of Kalanchoe blossfeldiana and production of male-sterile plants by engineered anther ablation |
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Authors: | Bego?a Garc??a-Sogo Benito Pineda Lourdes Castelblanque Teresa Ant??n M??nica Medina Edel??n Roque Claudia Torresi Jos?? P??o Beltr??n Vicente Moreno Luis Antonio Ca?as |
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Institution: | 1. Instituto de Biolog??a Molecular y Celular de Plantas (CSIC.-UPV.), Ciudad Polit??cnica de la Innovaci??n, Edf. 8E. C/. Ingeniero Fausto Elio s/n, 46011, Valencia, Spain 2. Department of Applied Biology, Facolt?? di Agraria, Universit?? degli Studi di Perugia, Borgo XX Giugno 74, 06121, Perugia, Italy
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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. |
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