Morphological, physiological and molecular characteristics of the seedless ‘Hongjiangcheng’ sweet orange

Seedless citrus has become one of the breeding goals due to its high edible ratio and convenience in fresh consumption and processing. In this study, the ‘Hongjiangcheng’ sweet orange (WT) and its seedless mutant (MT) after cobalt-60 radiation were selected to study the formation metabolism of citrus seedless phenotype. Compared with WT, the MT had altered primary metabolite contents, as indicated by GC-MS analysis. The mature pollen of the MT was mostly distorted and shrunken, and the orange mutant exhibited significantly lower fertility than the WT. Through pollination experiments and paraffin sectioning of the MT, we observed self-compatibility during pollen tube germination in situ, in combination with the absence of natural parthenocarpy and arrested zygotic embryo development at the fourth week after pollination. From transcriptomic analyses of ovules in the fourth week, 815 differentially expressed genes (DEGs) were identified. Furthermore, according to the annotation of gene function and qRT-PCR analysis, Cs4g10930, Cs5g21900 and orange1.1t02243 were identified as candidate genes that may govern the mechanism of seedlessness. Finally, Agrobacterium-mediated transformation verified that the overexpression of Cs4g10930 and Cs5g21900 in Newhall navel orange calli inhibited embryoid production. This study provides a better understanding of seedless formation in citrus and two key genes that may play an important role in the early selection of seedless lines in citrus breeding programs.