基于 RNAi 的抗黑条矮缩病病毒基因工程水稻未显示出不良遗传效应:多组学分析

Chunmeng Huang, Zhi Wang, Pengyu Zhu, Wenjie Xu, Zhixin Du, Yike Qian, Wei Fu
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摘要

科学技术的进步使我们能够应对时代的挑战:饥饿、营养不良、气候变化和自然资源日益减少。粮食和农业领域最重要的创新之一就是基因工程的出现,它可以帮助我们应对上述挑战。基因工程作物在农业和粮食领域的应用可使数百万人受益。该研究比较了四个基于 RNAi 的基因工程水稻品系叶片中 siRNA、转录组和代谢组的变化程度,以评估抗黑条矮小病毒的基于 RNAi 的基因工程水稻是否发生了非基因工程水稻所没有的变化。不同长度和丰度的 siRNA 在水稻转录本中高度富集;然而,对 8 个潜在脱靶基因的相对表达分析表明,基于 RNAi 的基因工程品系中的基因表达量并没有减少,这意味着本研究中从水稻黑条矮缩病病毒反转重复序列中提取的 siRNA 并没有造成脱靶现象。本研究在基于 RNAi 的基因工程品系中发现了一些差异表达基因和差异积累代谢物,但这些基因和代谢物并不富集在对人类、动物和环境有害的途径中。通过使用双向正交偏最小二乘法模型进行综合分析,只有 30 个差异表达基因在植物激素信号转导途径中明显富集。这些基因属于水杨酸、茉莉酸、细胞分裂素、脱落酸和黄铜类固醇通路,涉及与病毒抗性或感染反应相关的通路,这些基因都发生了显著变化。
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RNAi‐based genetically engineered rice resistant to black‐streaked dwarf virus does not show adverse genetic effects: A multi‐omics analysis
Advancements in science and technology enable us to address the challenges of the times: hunger, malnutrition, climate change, and dwindling natural resources. One of the most significant innovations in food and agriculture is the advent of genetic engineering, which may help us address the aforementioned challenges. Millions of people could benefit from the application of genetically engineered crops in agriculture and food. However, it is vital that biosafety assessments of unintended effects are carried out to identify any potential risks to humans, the environment, or society. The study compared the extent of siRNA, transcriptomic and metabolomic changes in the leaves of four RNAi‐based genetically engineered rice lines to evaluate whether RNAi‐based genetically engineered rice that is resistant to black‐streaked dwarf virus experiences changes not seen in the non‐genetically engineered counterpart. siRNA, transcriptome, and metabolome profiling were conducted. siRNAs with different lengths and abundance were highly enriched in the transcript of rice; however, relative expression analysis of eight potential off‐target genes revealed that there was no decrease in gene expression in the RNAi‐based genetically engineered lines, meaning that no off‐target phenomena were caused as a result of siRNA derived from invert repeat sequences of rice black‐streaked dwarf virus in this study. This study identified some differentially expressed genes and differentially accumulated metabolites in the RNAi‐based genetically engineered lines; however, they were not enriched in pathways detrimental for humans, animals and the environment. Only 30 differentially expressed genes were significantly enriched in the plant hormone signal transduction pathway via a combined analysis using two‐way orthogonal partial least squares model. These genes belonging to salicylic acid, jasmonic acid, cytokinin, abscisic acid, and brassinosteroid pathway involved in pathways related to virus resistance or infection response were significantly changed. Taken together, there was no off‐target in RNAi‐based genetically engineered rice lines and may have a positive effect on virus resistance overall in this study.
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