A soybean (Glycine max (Linn.) Merr.) mutant displaying rolled leaves and short petioles was identified as being caused by multiple sites, named rlsp1. Compared to wild-type (WT), the phloem structure of petioles in rlsp1 was not obvious, while the thickness of leaves was significantly greater but the arrangement of leaves cells was relatively looser. Sequencing-based bulked segregant analysis (Seq-BSA) identified 10 candidate regions on chromosomes 3, 6, 8, 13, and 17 with total 6.47 Mb, containing 790 genes. 7946 and 5402 differentially expressed genes (DEGs) were also identified in leaves and petioles, respectively, by RNA-sequencing (RNA-seq). Glyma.03G128600 as orthologs of Arabidopsis BOP (BLADE ON PETIOLES) gene may be the key candidate gene causing the variation of rlsp1. The abnormal development of petiole phloem structure, inhibition of auxin signal transduction pathway, promotion of microtubule-related pathways, and repressed of carbohydrate processes were also closely associated with the formation of rlsp1. These results were further validated by the changes of phytohormones, sucrose, Ca²⁺, and chlorophyll. The candidate genes found in this study enrich the understanding of the molecular basis for the formation of rolled leaves and short petioles in soybean.

大豆（Glycine max（Linn。）Merr。）突变株显示出卷曲的叶子和短叶柄被鉴定为由多个位点引起，命名为rlsp1。与野生型（WT）相比，rlsp1中叶柄的韧皮部结构不明显，叶的厚度明显更大，但叶细胞的排列相对较松散。基于测序的大量分离子分析（Seq-BSA）在3、6、8、13和17号染色​​体上确定了10个候选区域，总区域为6.47 Mb，包含790个基因。还通过RNA测序（RNA-seq）在叶和叶柄中分别鉴定到7946和5402差异表达基因（DEG）。Glyma.03G128600作为拟南芥BOP的直系同源物（叶片上的blade基因可能是引起rlsp1变异的关键候选基因。叶柄韧皮部结构的异常发育，生长素信号转导途径的抑制，微管相关途径的促进以及糖类过程的抑制也与rlsp1的形成密切相关。这些结果通过植物激素，蔗糖，Ca ²⁺和叶绿素的变化进一步证实。这项研究中发现的候选基因丰富了对大豆卷叶和短叶柄形成的分子基础的理解。