Analysis of Varietal Differences in Pre-harvest Sprouting of Rice using RNA-Sequencing

MyoungGoo Choi, Hyen-Seok Lee, Woon-Ha Hwang, Seo-Yeong Yang, Yun-Ho Lee, Chung-gun Lee, S. Yun, Jae-Hyeok Jeong
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Abstract

Seed dormancy is an adaptive trait in which seeds do not germinate under unfavorable environmental conditions. Low dormancy seeds are easily germinated under optimal environmental conditions, and these characteristics greatly reduce the yield and quality of crops. In the present study, we compared the pre-harvest sprouting (PHS) rate of two cultivars, Joun and Jopyeong, using the Winkler scale after heading day and temperature of the test. The PHS rate increased as the Winkler scale after heading day increased from 700°C to 1100°C and the temperature of the test increased. In all conditions, the PHS rate of Jopyeong was higher than that of Joun. RNA-sequencing was used to analyze the cause of the high PHS rate. We analyzed the biological metabolic processes related to the abscisic acid (ABA) metabolite pathway using the KEGG mapper with selected differentially expressed genes in PHS seeds. We found that the expression of ABA biosynthesis genes (OsNCEDs) was down-regulated and that ABA catabolic genes (OsCYP707As) was up-regulated in PHS seeds. However, the quantitative real-time PCR results showed that Joun had a higher expression of OsNCEDs than that of Jopyeong, but OsCYP707As did not yield a significant result. Joun displayed higher ABA content than that of Jopyeong not only during ripeness time but also during PHS treatment. Taken together, we provided evidence that the ABA content remaining in the seed is important to the PHS rate, which is determined by the expression level of the ABA biosynthesis gene OsNCEDs.
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利用rna测序分析水稻收获前发芽的品种差异
种子休眠是种子在不利环境条件下不发芽的一种适应性性状。低休眠种子在最佳的环境条件下容易发芽,这些特点大大降低了作物的产量和品质。本研究采用Winkler比较法,比较了抽穗日和试验温度后两个品种“琼”和“佐平”的采前发芽率。抽穗日后Winkler标度从700℃升高到1100℃,试验温度升高,小PHS率随之升高。在所有条件下,佐平的小灵通率都高于琼。采用rna测序分析小灵通率高的原因。我们利用KEGG图谱分析了小灵通种子中与脱落酸(ABA)代谢途径相关的生物代谢过程。结果表明,小灵通种子中ABA生物合成基因OsNCEDs表达下调,ABA分解代谢基因OsCYP707As表达上调。然而,实时荧光定量PCR结果显示,Joun的osnced表达量高于Jopyeong,而OsCYP707As的表达量不显著。无论是成熟期还是小灵通处理期间,“琼”的ABA含量均高于“佐平”。综上所述,我们提供的证据表明,种子中残留的ABA含量对小灵通率很重要,而小灵通率是由ABA生物合成基因osnced的表达水平决定的。
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