Novel and efficient transformation of wild passion fruit ( Passiflora cincinnata Mast.) using sonication-assisted Agroba
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Novel and efficient transformation of wild passion fruit (Passiflora cincinnata Mast.) using sonication-assisted Agrobacterium-mediated transformation Maurecilne Lemes da Silva 1 & Daniela Lopes Paim Pinto 2 & Andreia Barcelos Passos 3 & Francismar Corrêa Marcelino-Guimarães 4 & Ana Aparecida Bandini Rossi 5 & William Krause 6 & Ilio Fealho de Carvalho 1 & Diego Silva Batista 7 & Diego Ismael Rocha 8 & Wagner Campos Otoni 9 Received: 15 May 2020 / Accepted: 27 October 2020 / Editor: Harold Trick # The Society for In Vitro Biology 2020
Abstract Passiflora cincinnata stands out among Passifloraceae because of its medicinal properties and its resistance to pathogens, which promotes its use as a rootstock for other Passiflora species. A valuable strategy for obtaining disease-resistant passion fruit plants is represented by genetic transformation; however, this requires efficient regeneration. Here, we aimed to establish an efficient protocol for generating transgenic passion fruit plants using sonication-assisted Agrobacterium-mediated transformation of somatic embryos. The latter were obtained from anthers, sonicated, and exposed to an Agrobacterium suspension for 15 or 30 s. As a comparison, non-sonicated embryos were also exposed to the same bacterial treatment, whereas non-infected embryos were used as controls. Plant identity was confirmed by PCR, qPCR, and histochemical assay. Transgenic plants were obtained at higher rates in the treatments applying sonication. Overall, 171 plantlets were regenerated, 38 of which showed stable uidA reporter gene expression. Of these 38 transgenic plants, 22 (57.89%) and 13 (34.21%) were obtained by sonication-assisted Agrobacterium-mediated transformation for 30 s and 15 s, respectively, whereas the remaining 3 (7.89%) were exposed for 30 s but without prior sonication. Our results indicate that sonication-assisted Agrobacterium-mediated transformation for 30 s enhanced transformation efficiency in P. cincinnata. We believe that this system will allow for more efficient production of transgenic passion fruit plants. Keywords Passion fruit . Genetic transformation . Somatic embryogenesis . SAAT . Reporter genes
* Maurecilne Lemes da Silva [email protected] 1
Department of Biology, State University of Mato Grosso, Tangará da Serra, Mato Grosso 78300-000, Brazil
2
Department of Biological Sciences, University of Rhode Island, Kingston, RI 02881, USA
3
Department of Biology, Federal University of Espírito Santo, São Mateus, Espírito Santo 29932-540, Brazil
4
Embrapa Soybean, Warta District, Londrina, Paraná 86001-970, Brazil
5
Department of Biology, State University of Mato Grosso, Alta Floresta, Mato Grosso 78580-000, Brazil
6
Department of Agronomy, State University of Mato Grosso, Tangará da Serra, Mato Grosso 78300-000, Brazil
7
Department of Agriculture, Federal University of Paraíba, Campus III, Bananeiras, Paraíba 58220-000, Brazil
8
Institute of Biological Sciences, Federal University of Jataí, Campus Jatobá, Jataí, Goiás 75801-615, Bra
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