Agrobacterium-mediated transformation systems of Primula vulgaris

Sadiye Hayta,Philip M Gilmartin,Jinhong Li,Mark A Smedley,Wendy A Harwood

doi:10.1186/s13007-018-0360-1

Sadiye Hayta, Philip M Gilmartin + Show 3 more

Open Access

https://doi.org/10.1186/s13007-018-0360-1

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Abstract

BackgroundGenetic transformation is a valuable tool and an important procedure in plant functional genomics contributing to gene discovery, allowing powerful insights into gene function and genetically controlled characteristics. Primulaceae species provide one of the best-known examples of heteromorphic flower development, a breeding system which has attracted considerable attention, including that of Charles Darwin. Molecular approaches, including plant transformation give the best opportunity to define and understand the role of genes involved in floral heteromorphy in the common primrose, Primula vulgaris, along with other Primula species.ResultsTwo transformation systems have been developed in P. vulgaris. The first system, Agrobacterium-mediated vacuum infiltration of seedlings, enables the rapid testing of transgenes, transiently in planta. GUS expression was observed in the cotyledons, true leaves, and roots of Primula seedlings. The second system is based on Agrobacterium tumefaciens infection of pedicel explants with an average transformation efficiency of 4.6%. This transformation system, based on regeneration and selection of transformants within in vitro culture, demonstrates stable transgene integration and transmission to the next generation.ConclusionThe two transformation systems reported here will aid fundamental research into important traits in Primula. Although, stable integration of transgenes is the ultimate goal for such analyses, transient gene expression via Agrobacterium-mediated DNA transfer, offers a simple and fast method to analyse transgene functions. The second system describes, for the first time, stable Agrobacterium-mediated transformation of Primula vulgaris, which will be key to characterising the genes responsible for the control of floral heteromorphy.

Highlights

Genetic transformation is a valuable tool and an important procedure in plant functional genomics contributing to gene discovery, allowing powerful insights into gene function and genetically controlled characteristics
One of the first steps was to assess and build a reporter binary vector to be used for the optimisation of our transformation experiments (Fig. 1)
The use of infiltration of seedlings by A. tumefaciens was described as a simple transient expression in Arabidopsis thaliana [5], and called FAST “Fast Agromediated Seedling Transformation”

Summary

Introduction

Genetic transformation is a valuable tool and an important procedure in plant functional genomics contributing to gene discovery, allowing powerful insights into gene function and genetically controlled characteristics. Molecular approaches, including plant transformation give the best opportunity to define and understand the role of genes involved in floral heteromorphy in the common primrose, Primula vulgaris, along with other Primula species. Genetic transformation is an important technique for the in vivo analysis of gene function, especially in relation to plant development through gene over-expression, gene knock-out, and gene structure function analyses. Transgenic plants are usually produced by methods that include the transformation of individual plant cells followed by regeneration of whole plants from those transformed cells. Primula species are recalcitrant to in vitro regeneration and this has limited the development of transformation systems in this genus. An efficient protocol was developed for in vitro regeneration of P. vulgaris via indirect organogenesis from adult leaf– derived explants without any seasonal limitations [14]

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