Integrated BSA-Seq and WGCNA Analyses Reveal Candidate Genes Associated with Winter Bud Dormancy Maintenance in Fruit Mulberry (Morus spp.)

The excessively concentrated ripening period of mulberries causes seasonal surplus. Fruit mulberry (<i>Morus</i> spp.) exhibits the unique trait of "simultaneous flowering and leaf flushing", rendering budburst timing closely correlated with fruit ripening time. Thus, deciphering the molecular mechanism underlying winter bud dormancy maintenance in fruit mulberry is urgently needed. Herein, an F₁ hybrid population comprising 337 individuals, derived from <i>Morus wittiorum</i> (♀) and '322' (♂), was utilized as research material. Through Bulked Segregant Analysis Sequencing (BSA-Seq), we successfully mapped a dormancy-associated QTL interval designated as LB (Late Burst), spanning 9,990,001-11,990,000 bp on Chromosome 13. Integrating Weighted Gene Co-expression Network Analysis (WGCNA) results, <i>MaSVP</i> was identified as a candidate gene within this interval. Virus-induced gene silencing (VIGS) of <i>MaSVP</i> in winter buds of <i>Morus wittiorum</i> significantly accelerated budburst compared to the control, demonstrating that <i>MaSVP</i> represses winter bud dormancy release and plays a crucial role in regulating dormancy maintenance in fruit mulberry. Dynamic expression profiling of dormancy-related genes revealed that the transcript levels of <i>MaSVP</i>, <i>MaSAPK3</i>, <i>MaCASL2</i>, and <i>MaPYR8</i> were significantly downregulated (Tukey's test, <i>p</i> < 0.05) as budburst approached, whereas those of <i>MaFT</i> and <i>MaGA20ox1-D</i> were significantly upregulated (Tukey's test, <i>p</i> < 0.05). These results indicate that winter bud dormancy maintenance in <i>Morus wittiorum</i> is associated with abscisic acid (ABA) and gibberellin (GA) metabolism. Collectively, this study provides critical insights into the biological basis of winter bud dormancy maintenance in fruit mulberry and offers valuable genetic resources for breeding late-maturing cultivars.