Abstract
BackgroundPlants have the remarkable property to elaborate entire body plan from any tissue part. The conversion of lateral root primordium (LRP) to shoot is an ideal method for plant propagation and for plant researchers to understand the mechanism underlying trans-differentiation. Until now, however, a robust method that allows the efficient conversion of LRP to shoot is lacking. This has limited our ability to study the dynamic phases of reprogramming at cellular and molecular levels.ResultsHere we present an efficient protocol for the direct conversion of LRP to a complete fertile shoot system. This protocol can be readily applied to the various ecotypes of Arabidopsis. We show that, the conversion process is highly responsive to developmental stages of LRP and changes in external environmental stimuli such as temperature. The entire conversion process can be adequately analyzed by histological and imaging techniques. As a demonstration, using a battery of cell fate specific markers, we show that confocal time-lapse imaging can be employed to uncover the early molecular events, intermediate developmental phases and relative abundance of stem cell regulators during the conversion of LRP to shoot.ConclusionOur method is highly efficient, independent of genotypes tested and suitable to study the reprogramming of LRP to shoot in intact plants as well as in excised roots.Electronic supplementary materialThe online version of this article (doi:10.1186/s13007-016-0127-5) contains supplementary material, which is available to authorized users.
Highlights
Plants have the remarkable property to elaborate entire body plan from any tissue part
We examined if shoot conversion can be achieved only in excised root or if this method can be implemented to growing plants where complete root system is in the context of growing shoot
We found that our method can be used to trigger shoot formation from lateral root primordium (LRP) in intact plant (Fig. 3d)
Summary
Plants have the remarkable property to elaborate entire body plan from any tissue part. The profound capacity of somatic cells to reproduce an entire organism has been widely exploited in in vitro plant regeneration [1, 2]. This spectacular property of plant cells has great impact on both, basic plant research and biotechnological application. In vitro plant regeneration can be exploited to understand many fundamental questions pertaining to subjects such as stem cell specification, cell fate determination, cell differentiation, meristem formation and organogenesis in the absence of positional information laid down. Regeneration can be induced from the explant either directly or indirectly and are defined based on the presence or absence of an intermediate phase that leads to the formation of callus, a regenerative mass of cells. Consistent with the rapid upregulation of WUS during cytokinin-induced LRP-to-shoot conversion, inducible ectopic overexpression of WUS can convert LRP to shoot in the absence of external inductive cues and can induce shoot formation from Arabidopsis primary root meristem [6, 10]
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