The cultivated strawberry (Fragaria × ananassa Duch.) is a species of temperate origin, and the extension of this crop into tropical regions is dependent on genetic breeding. Breeders in the UNICENTRO’s Strawberry Breeding Program (USBP) selected, from among 2,000 hybrids, 40 hybrids that were adapted to photoperiod and temperature conditions of tropical regions. In this study, we used inter-simple sequence repeat to characterize 40 three-way hybrids developed by USBP and evaluated the genetic relationship of these hybrids with commercial cultivars, heirloom cultivars, and single hybrids. We used nine inter-simple sequence repeat primers to genotype 14 commercial cultivars, five heirloom cultivars, five single hybrids (SH), 20 three-way hybrids with short-day behavior (SDH), and 20 three-way hybrids with photoperiod-insensitive behavior (PIH). The percentage of polymorphism (100%) and both, the Nei genetic diversity (h = 0.34) and the Shannon index (I = 0.51), showed high variability and diversity, respectively, in the evaluated strawberry genotypes. Commercial cultivars showed the highest diversity indices (h = 0.30, I = 0.46), followed by PIH hybrids (h = 0.27, I = 0.41). In the dendrogram, the genotypes were distributed into three groups (commercial cultivars, heirloom cultivars, and single hybrids; SDH and PIH). This clustering was confirmed by principal coordinate analysis and Bayesian inference analysis. The overall analysis of the data revealed the efficacy of USBP in three-way hybrid development with different genetic characteristics compared with those of available commercially cultivars. These hybrids have substantial potential in becoming new strawberry cultivars.

栽培草莓（草莓 × 香蕉）Duch。）是一种温带起源物种，这种作物向热带地区的延伸取决于遗传育种。UNICENTRO草莓育种计划（USBP）的育种者从2,000个杂种中选择了40个适应热带地区的光周期和温度条件的杂种。在这项研究中，我们使用简单序列间重复来表征USBP开发的40种三向杂种，并评估了这些杂种与商业品种，传家宝品种和单一杂种的遗传关系。我们使用9个简单的序列重复引物对14个商业品种，5个传家宝品种，5个单杂种（SH），20个短期行为（SDH）的三向杂种和20个对光周期不敏感的三向杂种进行基因型分析行为（PIH）。多态性的百分比（100％）和两者h  = 0.34）和香农指数（I  = 0.51），在所评估的草莓基因型中分别表现出高变异性和多样性。商业品种表现出最高的多样性指数（h  = 0.30，I  = 0.46），其次是PIH杂种（h  = 0.27，I = 0.41）。在树状图中，基因型分为三组（商业品种，传家宝品种和单一杂种； SDH和PIH）。通过主坐标分析和贝叶斯推断分析证实了这种聚类。数据的整体分析显示，与可购得的商业品种相比，USBP在具有不同遗传特征的三向杂交发展中的功效。这些杂种具有成为新草莓品种的巨大潜力。