Plant Growth Regulators Promote Petaloidy and Modulate Related Gene Expression in Ornamental Pomegranate

Double-petal ornamental pomegranate presents for its enhanced ornamental value. Thus, cultivation techniques that promote petaloidy while modulating related gene expression are desired. To screen out the efficient treatments of plant growth regulator and key genes that enhance petaloidy, this study treated the flower buds of double- and single-petal ornamental pomegranate varieties with different concentrations of plant growth regulators naphthaleneacetic acid (NAA), methyl jasmonate (MeJA), abscisic acid (ABA), and ethephon (ETH) and quantified the number of petalized stamens (NOPSs) and the number of petals (NOPs) in both varieties. Furthermore, we investigated the expression levels of the genes flavin-containing monooxygenase (YUC), IAA-amino acid hydrolase (ILR1),indole-3-acetic acid-amido synthetase (GH3.17), auxin transporter (LAX2), auxin response factor (ARF), auxin-induced in root cultures protein (AIR12), jasmonic acid-amido synthetase (JAR1), and ABA stress ripening-induced protein (ASR) under the different treatments and analyzed their role in regulating relevant phenotypic traits. Plant growth regulator experiments demonstrated that NAA (10 mg/L) significantly increased the number of petalized stamens (NOPSs) and petals (NOPs), MeJA (100 mg/L) significantly increased the number of petalized stamens, while neither ABA nor ETH induced this morphological shift. qRT-PCR analysis confirmed that NAA upregulated ILR1, LAX2, ARF, and JAR1 in the stamens of single-petal flowers (StSi) and double-petal flowers (StDo) and petals of single-petal flowers (PeSi) and double-petal flowers (PeDo), with their expression levels strongly positively correlated with NOPS in both single- and double-petal flowers and NOP in double-petal flowers. MeJA upregulated ILR1, GH3.17, LAX2, ARF, and JAR1 in StDo and PeDo and was strongly positively correlated with NOPS and NOP in double-petal flowers. Consequently, NAA (10 mg/L) and MeJA (100 mg/L) were efficient treatments, and ILR1, GH3.17, LAX2, ARF, and JAR1 were identified as key genes in NAA- and MeJA-mediated petaloidy in ornamental pomegranates. Our results provide theoretical support for identifying the formation mechanism and improving industrial cultivation techniques for double-petal pomegranates.