Omics Profiles of Non-transgenic Scion Grafted on Transgenic RdDM Rootstock

H. Kodama, Yukiko Umeyama, Taira Miyahara, Taichi Oguchi, Takashi Tsujimoto, Y. Ozeki, Takumi Ogawa, Y. Yamaguchi, D. Ohta
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引用次数: 1

Abstract

Abstract Grafting of commercial varieties onto transgenic stress-tolerant rootstocks is attractive approach, because fruit from the non-transgenic plant body does not contain foreign genes. RNA silencing can modulate gene expression and protect host plants from viruses and insects, and small RNAs (sRNAs), key molecules of RNA silencing, can move systemically. Here, to evaluate the safety of foods obtained from sRNA-recipient plant bodies, we investigated the effects of rootstock-derived sRNAs involved in mediating RNA-directed DNA methylation (RdDM) on non-transgenic scions. We used tobacco rootstocks showing RdDM against the cauliflower mosaic virus (CaMV) 35S promoter. When scions harboring CaMV 35S promoter sequence were grafted onto RdDM-inducing rootstocks, we found that RdDM-inducing sRNAs were only weakly transported from the rootstocks to the scion, and we observed a low level of DNA methylation of the CaMV 35S promoter in the scion. Next, wild-type (WT) tobacco scions were grafted onto RdDM-inducing rootstocks (designated NT) or WT rootstocks (designated NN), and scion leaves were subjected to multi-omics analyses. Our transcriptomic analysis detected 55 differentially expressed genes between the NT and NN samples. A principal component analysis of proteome profiles showed no significant differences. In the positive and negative modes of LC-ESI-MS and GC-EI-MS analyses, we found a large overlap between the metabolomic clusters of the NT and NN samples. In contrast, the negative mode of a LC-ESI-MS analysis showed separation of clusters of NT and NN metabolites, and we detected 6 peak groups that significantly differed. In conclusion, we found that grafting onto RdDM-inducing rootstocks caused a low-level transmission of sRNAs, resulting in limited DNA methylation in the scion. However, the causal relationships between sRNA transmission and the very slight changes in the transcriptomic and metabolomic profiles of the scions remains unclear. The safety assessment points for grafting with RdDM rootstocks are discussed.
转基因RdDM砧木嫁接非转基因接穗组学分析
摘要将商业品种嫁接到转基因耐胁迫砧木上是一种有吸引力的方法,因为来自非转基因植物体的果实不含外源基因。RNA沉默可以调节基因表达,保护宿主植物免受病毒和昆虫的侵害,而RNA沉默的关键分子小RNA(sRNA)可以系统移动。在这里,为了评估从sRNA受体植物体获得的食物的安全性,我们研究了参与介导RNA定向DNA甲基化(RdDM)的砧木衍生的sRNA对非转基因接穗的影响。我们使用表现出针对花椰菜花叶病毒(CaMV)35S启动子的RdDM的烟草砧木。当携带CaMV 35S启动子序列的接穗嫁接到RdDM诱导的砧木上时,我们发现RdDM诱发的sRNA仅从砧木弱地转运到接穗,并且我们观察到接穗中CaMV 35S启动子的DNA甲基化水平较低。接下来,将野生型(WT)烟草接穗嫁接到RdDM诱导的砧木(指定为NT)或WT砧木(指定的NN)上,并对接穗叶片进行多组学分析。我们的转录组分析在NT和NN样本之间检测到55个差异表达的基因。蛋白质组图谱的主成分分析显示没有显著差异。在LC-ESI-MS和GC-EI-MS分析的阳性和阴性模式中,我们发现NT和NN样品的代谢组簇之间有很大的重叠。相反,LC-ESI-MS分析的阴性模式显示NT和NN代谢物簇的分离,我们检测到6个显著不同的峰组。总之,我们发现嫁接到RdDM诱导的砧木上会导致sRNA的低水平传递,导致接穗中的DNA甲基化有限。然而,sRNA传播与接穗转录组和代谢组谱的微小变化之间的因果关系尚不清楚。讨论了RdDM砧木嫁接的安全性评价要点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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