Department of Grassland Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China.
Institute of Grass Research, Heilongjiang Academy of Agricultural Sciences, Harbin 150080, China.
Int J Mol Sci. 2022 Sep 23;23(19):11187. doi: 10.3390/ijms231911187.
Oat is a food and forage crop species widely cultivated worldwide, and it is also an important forage grass in plateau regions of China, where there is a high level of ultraviolet radiation and sunlight. Screening suitable reference genes for oat under UV-B and high-light stresses is a prerequisite for ensuring the accuracy of real-time quantitative PCR (qRT-PCR) data used in plant adaptation research. In this study, eight candidate reference genes (sulfite oxidase, ; victorin binding protein, ; actin-encoding, ; protein PSK SIMULATOR 1-like, ; TATA-binding protein 2-like, ; ubiquitin-conjugating enzyme E2, ; elongation factor 1-alpha, ; glyceraldehyde-3-phosphate dehydrogenase 1, ;) were selected based on previous studies and our oat transcriptome data. The expression stability of these reference genes in oat roots, stems, and leaves under UV-B and high-light stresses was first calculated using three frequently used statistical software (geNorm, NormFinder, and BestKeeper), and then the comprehensive stability of these genes was evaluated using RefFinder. The results showed that the most stably expressed reference genes in the roots, stems, and leaves of oat under UV-B stress were , and , respectively; the most stably expressed reference genes in the roots, stems, and leaves under high-light stress were , and , respectively. was the most stably expressed reference gene in all the samples. The reliability of the selected reference genes was further validated by analysis of the expression of the phenylalanine ammonia-lyase () gene. This study highlights reference genes for accurate quantitative analysis of gene expression in different tissues of oat under UV-B and high-light stresses.
燕麦是一种在全球范围内广泛种植的粮食和饲料作物，也是中国高原地区的一种重要饲料草，那里紫外线辐射和阳光强烈。筛选适合燕麦在 UV-B 和高光胁迫下的参考基因是确保植物适应研究中实时定量 PCR（qRT-PCR）数据准确性的前提。在这项研究中，根据先前的研究和我们的燕麦转录组数据，选择了 8 个候选参考基因（亚硫酸盐氧化酶、； victorin 结合蛋白、；肌动蛋白编码、；蛋白 PSK SIMULATOR 1 样、；TATA 结合蛋白 2 样、；泛素结合酶 E2、；延伸因子 1-α、；甘油醛-3-磷酸脱氢酶 1、；）。首先使用三种常用的统计软件（geNorm、NormFinder 和 BestKeeper）计算这些参考基因在燕麦根、茎和叶中在 UV-B 和高光胁迫下的表达稳定性，然后使用 RefFinder 评估这些基因的综合稳定性。结果表明，在 UV-B 胁迫下，燕麦根、茎和叶中表达最稳定的参考基因分别为、和、；在高光胁迫下，燕麦根、茎和叶中表达最稳定的参考基因分别为、和、；是所有样本中表达最稳定的参考基因。通过对苯丙氨酸解氨酶（）基因表达的分析进一步验证了所选参考基因的可靠性。这项研究突出了在 UV-B 和高光胁迫下准确定量分析燕麦不同组织中基因表达的参考基因。
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