{"title":"Effect of Transgenic Rootstock Grafting on the Omics Profiles in Tomato.","authors":"Hiroaki Kodama, Taira Miyahara, Taichi Oguchi, Takashi Tsujimoto, Yoshihiro Ozeki, Takumi Ogawa, Yube Yamaguchi, Daisaku Ohta","doi":"10.14252/foodsafetyfscj.D-20-00032","DOIUrl":null,"url":null,"abstract":"<p><p>Grafting of non-transgenic scion onto genetically modified (GM) rootstocks provides superior agronomic traits in the GM rootstock, and excellent fruits can be produced for consumption. In such grafted plants, the scion does not contain any foreign genes, but the fruit itself is likely to be influenced directly or indirectly by the foreign genes in the rootstock. Before market release of such fruit products, the effects of grafting onto GM rootstocks should be determined from the perspective of safety use. Here, we evaluated the effects of a transgene encoding β-glucuronidase (GUS) on the grafted tomato fruits as a model case. An edible tomato cultivar, Stella Mini Tomato, was grafted onto GM Micro-Tom tomato plants that had been transformed with the <i>GUS</i> gene. The grafted plants showed no difference in their fruit development rate and fresh weight regardless of the presence or absence of the <i>GUS</i> gene in the rootstock. The fruit samples were subjected to transcriptome (NGS-illumina), proteome (shotgun LC-MS/MS), metabolome (LC-ESI-MS and GC-EI-MS), and general food ingredient analyses. In addition, differentially detected items were identified between the grafted plants onto rootstocks with or without transgenes (more than two-fold). The transcriptome analysis detected approximately 18,500 expressed genes on average, and only 6 genes were identified as differentially expressed. Principal component analysis of 2,442 peaks for peptides in proteome profiles showed no significant differences. In the LC-ESI-MS and GC-EI-MS analyses, a total of 93 peak groups and 114 peak groups were identified, respectively, and only 2 peak groups showed more than two-fold differences. The general food ingredient analysis showed no significant differences in the fruits of Stella scions between GM and non-GM Micro-Tom rootstocks. These multiple omics data showed that grafting on the rootstock harboring the <i>GUS</i> transgene did not induce any genetic or metabolic variation in the scion.</p>","PeriodicalId":73044,"journal":{"name":"Food safety (Tokyo, Japan)","volume":"9 2","pages":"32-47"},"PeriodicalIF":0.0000,"publicationDate":"2021-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8254850/pdf/","citationCount":"5","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Food safety (Tokyo, Japan)","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.14252/foodsafetyfscj.D-20-00032","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2021/6/1 0:00:00","PubModel":"eCollection","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 5
Abstract
Grafting of non-transgenic scion onto genetically modified (GM) rootstocks provides superior agronomic traits in the GM rootstock, and excellent fruits can be produced for consumption. In such grafted plants, the scion does not contain any foreign genes, but the fruit itself is likely to be influenced directly or indirectly by the foreign genes in the rootstock. Before market release of such fruit products, the effects of grafting onto GM rootstocks should be determined from the perspective of safety use. Here, we evaluated the effects of a transgene encoding β-glucuronidase (GUS) on the grafted tomato fruits as a model case. An edible tomato cultivar, Stella Mini Tomato, was grafted onto GM Micro-Tom tomato plants that had been transformed with the GUS gene. The grafted plants showed no difference in their fruit development rate and fresh weight regardless of the presence or absence of the GUS gene in the rootstock. The fruit samples were subjected to transcriptome (NGS-illumina), proteome (shotgun LC-MS/MS), metabolome (LC-ESI-MS and GC-EI-MS), and general food ingredient analyses. In addition, differentially detected items were identified between the grafted plants onto rootstocks with or without transgenes (more than two-fold). The transcriptome analysis detected approximately 18,500 expressed genes on average, and only 6 genes were identified as differentially expressed. Principal component analysis of 2,442 peaks for peptides in proteome profiles showed no significant differences. In the LC-ESI-MS and GC-EI-MS analyses, a total of 93 peak groups and 114 peak groups were identified, respectively, and only 2 peak groups showed more than two-fold differences. The general food ingredient analysis showed no significant differences in the fruits of Stella scions between GM and non-GM Micro-Tom rootstocks. These multiple omics data showed that grafting on the rootstock harboring the GUS transgene did not induce any genetic or metabolic variation in the scion.
将非转基因接穗嫁接到转基因砧木上,可获得优良的农艺性状,并可生产出优质的果实供消费。在这种嫁接植物中,接穗不含任何外源基因,但果实本身可能直接或间接地受到砧木中外源基因的影响。在此类水果产品投放市场之前,应从安全使用的角度确定嫁接到转基因砧木上的效果。本文以转基因β-葡萄糖醛酸酶(GUS)基因对番茄果实嫁接的影响为研究对象。一个可食用的番茄品种,Stella Mini tomato,被嫁接到转基因的Micro-Tom番茄植株上,这些植株已经转化了GUS基因。无论砧木中是否存在GUS基因,嫁接植株的果实发育速度和鲜重均无显著差异。对水果样本进行转录组(NGS-illumina)、蛋白质组(shotgun LC-MS/MS)、代谢组(LC-ESI-MS和GC-EI-MS)和一般食品成分分析。此外,嫁接到砧木上的植株与没有转基因的植株之间存在差异检测项目(超过两倍)。转录组分析平均检测到约18500个表达基因,只有6个基因被鉴定为差异表达。对蛋白质组谱中2442个肽峰进行主成分分析,结果显示差异不显著。在LC-ESI-MS和GC-EI-MS分析中,分别鉴定出93个峰组和114个峰组,只有2个峰组的差异大于2倍。一般食品成分分析结果表明,转基因与非转基因小蓟砧木接穗果实无显著差异。这些多组学数据表明,嫁接在含有GUS转基因的砧木上,没有引起接穗的任何遗传或代谢变异。