Genetic Mapping and Identification of the Gibberellin 3-Oxidase Gene GA3ox Leading to a GA-Deficient Dwarf Phenotype in Pumpkin (Cucurbita moschata D.)

Abstract

Plant height is an important indicator in the ideal plant model and contributes to optimizing yield and lodging resistance. The emergence of a dwarfing phenotype provides an opportunity for plant height improvement. In a previous study, we identified a dwarf mutant Si1 in pumpkin (Cucurbita moschata D.) obtained by ethyl methane sulfonate (EMS) mutagenesis of the inbred line N87. Phenotype identification for Si1 revealed a decrease in cell size and shorter internodes than those of wild type. Genetic analysis revealed that the dwarf mutant trait was controlled by a single recessive gene, CmaSI1. By bulked segregant analysis (BSA) and subsequent fine mapping, we mapped the CmaSI1 locus to a 463 kb region on chromosome 8 that contained 28 annotated genes in the F2 population. Only one nonsynonymous single nucleotide polymorphism (SNP) in CmoCh08G006170 was obtained according to whole-genome resequencing of the two parents. CmoCh08G006170, a homolog of Arabidopsis gibberellin 3-beta hydroxylase (GA3ox), is a key enzyme in the regulation of bioactive gibberellins (GAs). RNA-seq analysis and qRT-PCR showed that the expression level of CmoCh08G006170 in stems of Si1 was changed compared with that of wild type. The dwarf phenotype could be restored by exogenous GA3 treatment, suggesting that Si1 is a GA-deficient mutant. The above results demonstrated that CmoCh08G006170 may be the candidate gene controlling the dwarf phenotype. This study provides an important theoretical basis for the genetic regulation of vine length and crop breeding in pumpkin.