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. 2023 Feb 8;12(4):758.
doi: 10.3390/plants12040758.

Metabolite Profiling to Evaluate Metabolic Changes in Genetically Modified Protopanaxadiol-Enriched Rice

Ji-Eun Sim  1 , Sung-Dug Oh  2 , Kiyoon Kang  1 , Yu-Mi Shin  1 , Doh-Won Yun  2 , So-Hyeon Baek  3 , Yong-Eui Choi  4 , Sang-Un Park  5 , Jae-Kwang Kim  1
Affiliations

Metabolite Profiling to Evaluate Metabolic Changes in Genetically Modified Protopanaxadiol-Enriched Rice

Ji-Eun Sim et al. Plants (Basel). .

Abstract

Event DS rice producing protopanaxadiol (PPD) has been previously developed by inserting Panax ginseng dammarenediol-II synthase gene (PgDDS) and PPD synthase gene (CYP716A47). We performed a gas chromatography-mass spectrometry (GC-MS)-based metabolomics of the DS rice to identify metabolic alterations as the effects of genetic engineering by measuring the contents of 65 metabolites in seeds and 63 metabolites in leaves. Multivariate analysis and one-way analysis of variance between DS and non-genetically modified (GM) rice showed that DS rice accumulated fewer tocotrienols, tocopherols, and phytosterols than non-GM rice. These results may be due to competition for the same precursors because PPDs in DS rice are synthesized from the same precursors as those of phytosterols. In addition, multivariate analysis of metabolic data from rice leaves revealed that composition differed by growth stage rather than genetic modifications. Our results demonstrate the potential of metabolomics for identifying metabolic alterations in response to genetic modifications.

Keywords: CYP716A47; PgDDS; metabolic profiling; multivariate analysis; protopanaxadiol; protopanaxadiol rice; transgenic rice.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
PCA score plot on rice seed. Abbreviations: DJ, Oryza sativa L. cv. Dongjin; DS1, line 1 of transformed rice from DJ with PgDDS and CYP716A47 genes; DS8, line 8 of transformed rice from DJ with PgDDS and CYP716A47 genes.
Figure 2
Figure 2
Score (A), loading (B) plots and variable importance in the projection (VIP) (C) of the PLS-DA models on rice seed. Abbreviations: DJ, Oryza sativa L. cv. Dongjin; DS1, line 1 of transformed rice from DJ with PgDDS and CYP716A47 genes; DS8, line 8 of transformed rice from DJ with PgDDS and CYP716A47 genes; C20-ol, eicosanol; C21-ol, heneicosanol; C22-ol, docosanol; C24-ol, tetracosanol; C26-ol, hexacosanol; C28-ol, octacosanol; C30-ol, triacontanol.
Figure 3
Figure 3
Contents of DJ, DS1, and DS8 rice metabolites with significant differences in ANOVA. Bar graphs with different letters (a, b, c) refer to the significant differences between the samples, as obtained by one-way ANOVA. Error bars represent standard deviations. Abbreviations: DJ, Oryza sativa L. cv. Dongjin; DS1, line 1 of transformed rice from DJ with PgDDS and CYP716A47 genes; DS8, line 8 of transformed rice from DJ with PgDDS and CYP716A47 genes.
Figure 4
Figure 4
Biosynthetic pathway for protopanaxadiol, phytosterols, and tocopherols. Abbreviations: IPP, isopentenyl diphosphate; FDP, farnesyl diphosphate; GGDP, geranylgeranyl diphosphate; PDP, phytyl diphosphate; MPBQ, 2-methyl-6-phytyl-1,4-benzoquinol.
Figure 5
Figure 5
Score (A) and loading (B) plots of the PCA on rice leaves. Abbreviations: DJ, Oryza sativa L. cv. Dongjin; DS1, line 1 of transformed rice from DJ with PgDDS and CYP716A47 genes; DS8, line 8 of transformed rice from DJ with PgDDS and CYP716A47 genes; C20-ol, eicosanol; C21-ol, heneicosanol; C22-ol, docosanol; C24-ol, tetracosanol; C26-ol, hexacosanol; C28-ol, octacosanol; C30-ol, triacontanol.
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