Abstract

In addition to its role as one of the largest staple food crops in the world, rice (Oryza sativa L.) currently serves as an important bioreactor in molecular pharming. Although some studies have been conducted on the effects of foreign protein expression on rice, most of them use mature seeds and focus on proteomic changes. In this study, the effects of fibroblast growth factor 21 (FGF21) transformation on immature rice seeds were studied by means of physiological and transcriptome sequencing method. Starch and sucrose content and their associated enzyme activities of immature seeds were determined. After extracting RNA from immature seeds, Illumina HiSeq^TM 2500 was used for RNA sequencing. The content of starch and sucrose and activities of related enzymes in transgenic FGF21 immature rice seeds were greater than those of wild type. In total, 1762 differentially expressed genes (DEGs) were identified among non-transgenic wild type (CK) and transgenic lines (T1, T2, and T3). Meanwhile, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that the top two enriched processes were starch and sucrose metabolism and plant–pathogen interactions. Most DEGs of the plant–pathogen interactions pathway were downregulated, while expression patterns were more complex for DEGs of starch and sucrose metabolic pathways. Ultimately, these results could be used to reveal mechanisms underlying transgenic seed quality changes induced by FGF21 transformation and could also serve as an exploratory study into the use of transcriptome technique for safety evaluation of rice bioreactors.

中文翻译：

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转基因FGF21未成熟水稻种子的生理和转录组分析

摘要

除了作为世界上最大的主食作物之一，稻米（Oryza sativa L.）目前还作为重要的生物反应器在分子药物领域中发挥作用。尽管已经进行了一些关于外源蛋白表达对水稻影响的研究，但大多数研究都使用成熟的种子并关注蛋白质组学变化。在这项研究中，通过生理和转录组测序方法研究了成纤维细胞生长因子21（FGF21）转化对未成熟水稻种子的影响。测定未成熟种子的淀粉​​和蔗糖含量及其相关的酶活性。从未成熟的种子中提取RNA后，将Illumina HiSeq ^TM 2500用于RNA测序。转基因淀粉和蔗糖含量及相关酶活性FGF21未成熟的水稻种子大于野生型。总共在非转基因野生型（CK）和转基因品系（T1，T2和T3）中鉴定了1762个差异表达基因（DEG）。同时，《京都基因与基因组百科全书》（KEGG）途径分析表明，富集的前两个过程是淀粉和蔗糖代谢以及植物与病原体的相互作用。植物-病原体相互作用途径中的大多数DEG均被下调，而淀粉和蔗糖代谢途径的DEG的表达模式则更为复杂。最终，这些结果可用于揭示FGF21诱导转基因种子质量变化的潜在机制 转化，也可作为转录组技术用于水稻生物反应器安全性评估的探索性研究。