HD-Zip Transcription Factor is Responsible for No-Lobed Leaf in Watermelon(Citrullus lanatus L.)

Leaf is a vital organ of plants that plays an essential role in photosynthesis and respiration.As an important agronomic trait in leaf development,leaf shape is classified into lobed,entire(no-lobed),and serrated in most crops.In this study,two-lobed leaf watermelon inbred lines WT2 and WCZ,and a no-lobed leaf watermelon inbred line WT20 were used to create two F_(2)populations.Segregation analysis suggested that lobed leaves were dominant over the no-lobed leaves,and it was controlled by a signal gene.A locus on watermelon chromosome 4 controlling watermelon lobed/no-lobed leaves was identified through BSA-seq strategy combined with linkage analysis.The candidate gene was fine-mapped to a 61.5 kb region between 21,224,481 and 21,285,957 bp on watermelon chromosome 4 using two F_(2)populations.Four functional genes were annotated in the candidate region,while sequences blast showed that there was a single-base deletion(A/-)only in the exon of Cla018360,which resulted in premature termination of translation in the no-lobed leaf lines.Function prediction showed that Cla018360 encodes an HD-Zip protein that has been reported to regulate the development of leaf shape.The single-base deletion also occurred in the HD-Zip domain.We inferred that the Cla018360 gene is the candidate gene for regulating the development of lobed/no-lobed leaves in watermelon.Gene expression analysis showed that Cla018360 was highly expressed in young leaves.Phylogenetic analysis showed that Cla018360 had a close genetic relationship with AtHB51,which had been reported to regulate the formation of leaf shape in Arabidopsis.Furthermore,transcriptome analysis showed that a total of 333 differentially expressed genes were identified between WT2 and WT20,of which 115 and 218 genes were upregulated and downregulated in no-lobed leaved watermelon WT20.This study not only provides a good entry point for studying leaf development but also provides foundational insights into breeding for special plant architecture in watermelon.