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A reverse genetics approach identifies novel mutants in light responses and anthocyanin metabolism in petunia

Abstract

Flower color and plant architecture are important commercially valuable features for ornamental petunias (Petunia x hybrida Vilm.). Photoperception and light signaling are the major environmental factors controlling anthocyanin and chlorophyll biosynthesis and shade-avoidance responses in higher plants. The genetic regulators of these processes were investigated in petunia by in silico analyses and the sequence information was used to devise a reverse genetics approach to probe mutant populations. Petunia orthologs of photoreceptor, light-signaling components and anthocyanin metabolism genes were identified and investigated for functional conservation by phylogenetic and protein motif analyses. The expression profiles of photoreceptor gene families and of transcription factors regulating anthocyanin biosynthesis were obtained by bioinformatic tools. Two mutant populations, generated by an alkalyting agent and by gamma irradiation, were screened using a phenotype-independent, sequence-based method by high-throughput PCR-based assay. The strategy allowed the identification of novel mutant alleles for anthocyanin biosynthesis (CHALCONE SYNTHASE) and regulation (PH4), and for light signaling (CONSTANS) genes.

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Acknowledgments

The authors would like to acknowledge Rauly Moretti for the excellent technical assistance. Petunia research is supported by a FAPESP grant (2006/06306-0) to VQ. VQ is the recipient of a CNPq productivity research fellowship (307031/2010-1). The present work is part of the MSc. dissertation of ASB in Tropical and Subtropical Agriculture at the Instituto Agronômico (IAC), Campinas, Brazil (Genetics, Plant Breeding and Biotechnology).

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Correspondence to Vera Quecini.

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Berenschot, A.S., Quecini, V. A reverse genetics approach identifies novel mutants in light responses and anthocyanin metabolism in petunia. Physiol Mol Biol Plants 20, 1–13 (2014). https://doi.org/10.1007/s12298-013-0212-4

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  • DOI: https://doi.org/10.1007/s12298-013-0212-4

Keywords

  • Data mining
  • Flavonoid
  • Light signaling
  • Mutagenesis
  • Solanaceae
  • TILLING