Haploid Strategies for Functional Validation of Plant Genes

Increasing knowledge of plant genome sequences requires the development of more reliable and efficient genetic approaches for genotype–phenotype validation. Functional identification of plant genes is generally achieved by a combination of creating genetic modifications and observing the according phenotype, which begins with forward-genetic methods represented by random physical and chemical mutagenesis and move towards reverse-genetic tools as targeted genome editing. A major bottleneck is time need to produce modified homozygous genotypes that can actually be used for phenotypic validation. Herein, we comprehensively address and compare available experimental approaches for functional validation of plant genes, and propose haploid strategies to reduce the time needed and cost consumed for establishing gene function.

TrendsRecent advances in plant genomics and sequencing technology revealed numerous associations between phenotypes and plant genes. However, definitive functional annotations after in vivo validation have been thoroughly established for few of these genes.Genetic methods for establishing gene functions are achieved by creating genetic modifications that cause phenotypes of interest. Diploid plants are usually heterozygous for the modified region in the first generation. More additional generations are required to obtain homozygous genotypes that can actually be used for validation of their phenotypic effect.Efforts to obtain haploid plants have been undertaken in many plant species of almost all families in the plant kingdom. We proposed the strategies based on haploid mutagenesis and haploid transformation, respectively, which contribute to accelerating functional validation of plant genes.