Genetic and physiological characterization of three natural allelic variations affecting the organogenic capacity in tomato (Solanum lycopersicum cv. Micro-Tom)
The study of allelic variations affecting organogenic capacity is not only relevant for manipulating plant traits but also to understand the fundamental mechanisms involved in plant development. Here, we report the characterization of three tomato (Solanum lycopersicum) loci (RG3C, RG7H and RG8F) whose alleles from its wild relative Solanum pennellii enhance in vitro shoot and root regeneration. S. pennellii alleles were introgressed into tomato cv. Micro-Tom (MT), creating near-isogenic lines. We evaluated the time taken for shoot induction and acquisition of competence by quantifying organogenesis after transferring explants, respectively, from the shoot-inducing medium (SIM) to the basal medium (BM) and from root-inducing medium (RIM) to the SIM. Concomitantly, we monitored the expression of key developmental genes. MT-Rg3C and MT-Rg7H started shoot induction, respectively, at 48 and 24 h earlier than MT and MT-Rg8F, while MT-Rg3C and MT-Rg8F acquired competence 24 h before MT. The impact of MT-Rg3C and MT-Rg8F in the acquisition of competence to assume different fates is consistent with their effect enhancing both shoot and root regeneration. MT-Rg7H seems to affect shoot induction specifically, which is in agreement with the enhanced expression of the shoot-related genes WUSCHEL and SHOOT MERISTEMLESS. Phenotypic characterization of greenhouse-grown plants showed that Rg3C has increased branching when compared to MT. Conversely, the normal branching observed in MT-Rg7H and MT-Rg8F indicates that adventitious in vitro shoot formation and ex vitro axillary bud formation/outgrowth are induced by different genetic pathways. These natural variations are thus useful for breeding highly regenerating varieties without undesirable effects on plant architecture.
KeywordsBranching Introgression lines Organogenic competence Regeneration Solanum pennellii
We thank the National Council for Scientific and Technological Development (CNPq) and the São Paulo Research Foundation (FAPESP) for fellowship and scholarship granted, respectively, to L.E.P.P. (307040/2014-3) and M.S.P. (2012/24714-0 and 2014/17553-5). We thank Gilberto B. Kerbauy, Luciano Freschi, Victor A. Vitorello and Daniel S. Moura, from the University of São Paulo, for helpful comments. Adrienne R. Washington (University of Pittsburgh) is thanked for revising the manuscript. We also thank Cassia R. F. Figueiredo and Francisco Vitti for laboratory and greenhouse assistance.
MSP was responsible for the conception and design of experiments, data analysis, drafting and editing of the manuscript. CRA contributed with RT-qPCR experiments and revised the manuscript. VAB was responsible for conception and design of RT-qPCR experiments, data analysis and manuscript editing. LEPP oversaw the study conception and design and edited the manuscript. All authors read and approved the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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