Abstract
Brachiaria brizantha is a forage grass well adapted to tropical areas and cultivated in millions of hectares in Brazil. The apomictic mode of reproduction in this species, in addition to differences in ploidy between sexual and apomictic plants, impairs crossbreeding. The development of a methodology to transform apomictic cultivars will provide an option to introduce agronomic important traits to B. brizantha cv. Marandu. In addition, it will open the possibility to study in vivo the function of candidate genes involved in the apomictic reproduction. The objective of this work was to evaluate peeled seeds, isolated embryo from mature seeds, embryogenic calluses and embryogenic cell suspensions, as target explant for genetic transformation via biolistics. Plasmids bearing the marker genes gus and hptII under the control of the rice actin 1 promoter (pAct1-Os) or the maize ubiquitin 1 promoter (pUbi1Zm) were used. All the target-explants used were suitable for transient gene expression after bombardment, showing gus expression and resistance to hygromycin. Using embryogenic calluses and cell suspensions as target tissues, transgenic plants were regenerated and transgenes detected.
Highlights
IntroductionUrochloa brizantha; Shirasuna 2015) cv
Four types of explants were evaluated for genetic transformation via biolistics: (i) embryo isolated from mature seed; (ii) mature dehusked seed; (iii) embryogenic callus; (iv) cell suspension
The percentage of isolated embryos showing gus expression increased with the days in culture before bombardment reaching up to 39 % with 5 d decreasing at 7 d in culture
Summary
Urochloa brizantha; Shirasuna 2015) cv. Marandu is a forage grass well adapted to tropical areas. In Brazil Brachiaria is cultivated in millions of hectares. This cultivar is tetraploid and reproduces asexually by apomixis. Analysis of the reproductive mode, based on the embryo sac structure of 275 B. brizantha accessions from the Brazilian germplasm collection, displayed one single sexual diploid, the remaining being of Brachiaria transformation is the possibility to study the function of candidate genes involved in the apomictic reproduction (Silveira et al 2012, Guimarães et al 2013). Establishing methodology to express gene transiently may assist the investigation of promoters and genes using silencing and gain-of-function approaches in this species
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