Adzuki bean (Vigna angularis var. angularis), an important food legume, is grown in more than 30 countries, especially in East Asia. Almost 25% of the world population is affected by iron deficiency. Iron is very important on adzuki bean yield and quality. The identification of suitable reference genes in adzuki bean is important for exploring gene expression and the molecular mechanisms of gene, yet few reference genes have been reported for adzuki bean. This research systematically evaluated the stability of 11 candidate reference genes (ACT, ACT1, CYP, CYP2, SKIP16, ELF1B, HDC, Lectin, TUA, PEPKR1, and 18SrRNA) using the geNorm, NormFinder, BestKeeper, and RefFinder protocols under iron-deficient stress. The best combined number of reference genes for all samples was two. ELF1B/Lectin was best for qRT-PCR analysis of roots and combined analysis of roots, stems, and leaves under iron deficiency, ELF1B/CYP was optimum for leaves, and SKIP16/PEPKR1 was optimum for stems. The identification of these reference genes will enable accurate quantification of gene expression under iron deficiency and enhance our comprehension of reference genes in adzuki bean.

中文翻译：

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使用定量实时 PCR 鉴定和验证缺铁条件下小豆 (Vigna angularis) 中的参考基因

红豆 (Vigna angularis var. angularis) 是一种重要的食用豆科植物，在 30 多个国家种植，尤其是在东亚。世界上近 25% 的人口受到铁缺乏症的影响。铁对红豆的产量和品质非常重要。鉴定红豆中合适的内参基因对于探索基因表达和基因的分子机制具有重要意义，但目前鲜有报道用于红豆的内参基因。本研究系统地评估了 11 个候选参考基因（ACT、ACT1、CYP、CYP2、SKIP16、ELF1B、HDC、Lectin、TUA、PEPKR1 和 18SrRNA）的稳定性，在缺铁条件下使用 geNorm、NormFinder、BestKeeper 和 RefFinder 协议压力。所有样品的最佳参考基因组合数为 2。ELF1B/Lectin最适合缺铁条件下根的qRT-PCR分析和根、茎和叶的联合分析，ELF1B/CYP最适合叶片，SKIP16/PEPKR1最适合茎。这些参考基因的鉴定将能够准确量化缺铁条件下的基因表达，并增强我们对小豆参考基因的理解。