Genome-wide identification and testing of superior reference genes for transcript normalization during analyses of flesh development in Asian pear cultivars
Genome-wide identification and testing of superior reference genes for transcript normalization during analyses of flesh development in Asian pear cultivars
复制标题
在亚洲果肉发育分析过程中，对转录本标准化的优质参考基因进行全基因组鉴定和测试
DOI：
10.1016/j.scienta.2020.109459
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发表时间：
2020-09-20
影响因子：
4.3
通讯作者：
Chen, Jianqing
中科院分区：
文献类型：
--
作者：
Chen, Changmei;Yuan, Meng;Chen, Jianqing
定量实时聚合酶链反应（qRT-PCR）是目前分析基因表达和转录组的最佳选择。在这种情况下，重要的是使用理想的参考基因（RG）来标准化待比较样品的表达数据。因此，本研究旨在鉴定和验证亚洲梨品种果实发育qRT-PCR分析的上级RGs。共有7个“常用”的RGs，7个“传统”管家基因（HKG），和4个新的基因被选为候选RGs的基础上，35个公开的转录组库，包括数据的果实发育在5个亚洲梨品种。转录组学和qRT-PCR分析一致地显示，新型RG比“常用”RG和“传统”HKG更稳定地表达。在新的RG中，BPS 1和ICDH 1是最高和最稳定表达的基因。此外，这两个基因形成了在梨果肉发育分析期间标准化基因表达数据的最佳RG组合。这些研究结果可能有助于研究人员优化他们的RGS的选择，为未来的亚洲梨品种果肉发育过程中的基因表达的调查。
Quantitative real-time polymerase chain reaction (qRT-PCR) is currently the best available option for analyzing gene expression and transcriptomes. In this context, it is important to use ideal reference genes (RGs) to normalize the expression data for the samples to be compared. Thus, the present study was established to identify and validate superior RGs for qRT-PCR analyses of fruit development in Asian pear cultivars. A total of seven "commonly used" RGs, seven "traditional" housekeeping genes (HKGs), and four novel genes were selected as candidate RGs based on 35 publicly available transcriptome libraries that include data for fruit development in five Asian pear cultivars. Transcriptomic and qRT-PCR analyses consistently revealed that the novel RGs were expressed more stably than "commonly used" RGs and "traditional" HKGs. Among the novel RGs, BPS1 and ICDH1 were the most highly and stably expressed genes. Moreover, these two genes formed the optimal RG combination for normalizing gene expression data during analyses of pear flesh development. These findings may help researchers optimize their selection of RGs for future investigations of gene expression during flesh development in Asian pear cultivars.