Soo-Yun Park

Unintended polar metabolite profiling of carotenoid-biofortified transgenic rice reveals substantial equivalence to its non-transgenic counterpart

Substantial equivalence is a critical concept for biosafety assessment of genetically modified (GM) crops. To investigate substantial equivalence among carotenoid-biofortified GM rice and five conventional rice cultivars having common white (three) and red (two) grain colors, profiles of 52 polar metabolites were analyzed using gas chromatography time-of-flight mass spectrometry. The results were compared to evaluate the differences among GM and non-GM rice cultivars using principal components analysis. The GM rice is more comparable to its non-transgenic counterpart rice variety according to the closer co-separation than for other cultivars tested. This suggests that profiling of unintended polar metabolites could be a useful tool to reveal substantial equivalence of GM rice.

Metabolite profiling approach reveals the interface of primary and secondary metabolism in colored cauliflowers (Brassica oleracea L. ssp. botrytis).

In the present study, carotenoids, anthocyanins, and phenolic acids of cauliflowers ( Brassica oleracea L. ssp. botrytis) with various colored florets (white, yellow, green, and purple) were characterized to determine their phytochemical diversity. Additionally, 48 metabolites comprising amino acids, organic acids, sugars, and sugar alcohols were identified using gas chromatography-time-of-flight mass spectrometry (GC-TOFMS). Carotenoid content was considerably higher in green cauliflower; anthocyanins were detected only in purple cauliflower. Phenolic acids were higher in both green and purple cauliflower. Results of partial least-squares discriminant, Pearson correlation, and hierarchical clustering analyses showed that green cauliflower is distinct on the basis of the high levels of amino acids and clusters derived from common or closely related biochemical pathways. These results suggest that GC-TOFMS-based metabolite profiling, combined with chemometrics, is a useful tool for determining phenotypic variation and identifying metabolic networks connecting primary and secondary metabolism.

Compositional comparative analysis between insect-resistant rice (Oryza sativa L.) with a synthetic cry1Ac gene and its non-transgenic counterpart

Composition analysis of genetically modified crops is an important consideration in the assessment of food safety. Agb0101 (Oryza sativa L. cv. Nakdongbyeo) developed in Korea is a form of insect-resistant rice that contains a synthetic truncated cry1Ac gene isolated from Bacillus thuringiensis (Bt), and demonstrates high resistance to rice leaf folder under field conditions. Nutrients, anti-nutritive components, and secondary metabolites of Agb0101 were analyzed and compared with those of its non-transgenic counterpart. The amounts of proximates, amino acids, fatty acids, minerals, vitamins, trypsin inhibitors, phytic acid, and ferulic acid in brown rice from Agb0101 were comparable to those of its non-transgenic counterpart. Statistical comparisons to test for equivalence showed that all the analyzed components in the insect-resistant rice plants were substantially equivalent to those of its non-transgenic counterpart. Furthermore, most of the measured values from Agb0101 were within the range of values reported for other commercial rice varieties.

Metabolite Profiling Based on Lipophilic Compounds for Quality Assessment of Perilla (Perilla frutescens) Cultivars

Lipophilic compounds from Korean perilla ( Perilla frutescens ) seeds were characterized to determine the diversity among their phytochemicals and to analyze relationships between their contents. Twenty-four metabolites consisting of policosanol, phytosterol, tocopherol, and fatty acids were identified. The metabolite profiles were subjected to data mining processes, including principal component analysis (PCA), partial least-squares discriminate analysis (PLS-DA), and Pearsons correlation analysis. PLS-DA could distinguish between all cultivars except between Daesil and Daeyeup cultivars. Linolenic acid contents were positively correlated with β-sitosterol (r = 0.8367, P < 0.0001) and γ-tocopherol contents (r = 0. 7201, P < 0.001) among all perilla grains. The Daesil and Daeyeup cultivars appear to be good candidates for future breeding programs because they have simultaneously high linolenic acid, phytosterol, and tocopherol levels. These results demonstrate the use of metabolite profiling as a tool for assessing the quality of food.

Metabolomics for the Quality Assessment of Lycium chinense Fruits

Lycium chinense has been used as a traditional medicine for centuries in Asia because of its positive effects on health. However, its functional components have not been elucidated. This study determines the levels of health-promoting lipophilic compounds, including carotenoids, tocopherols, and phytosterol, and those of 42 hydrophilic metabolites, including sugars, organic acids, alcohols, amines, and amino acids, in L. chinense fruit from 11 cultivars. The metabolite profiles were subjected to a principal component analysis (PCA), Pearson correlation analysis, and hierarchical clustering analysis (HCA). PCA showed the Cheongdang (LM-3) cultivar to be distinct from the others. The correlation results for a total of 55 compounds revealed strong correlations between the metabolites that participated on closely related pathways. The Cheongdang cultivar appears to be most suited for functional food production because of its high carotenoid, tocopherol, and phytosterol levels. These results indicate the usefulness of metabolite profiling as a tool for assessing the quality of food.

Metabolic Differentiation of Diamondback Moth (Plutella xylostella (L.)) Resistance in Cabbage (Brassica oleracea L. ssp. capitata)

The diamondback moth, Plutella xylostella (L.), is a major pest responsible for destroying cabbage and other Brassica vegetable crops. A diamondback moth-resistant cabbage line was studied by comparing its metabolite profiles with those of a susceptible cabbage. Fourier transform infrared spectroscopy analysis revealed that carbohydrates, aromatic compounds, and amides were the major factors that distinguished the resistant and susceptible genotypes. Gas chromatography-time-of-flight mass spectrometry profiled 46 metabolites, including 19 amino acids, 15 organic acids, 8 sugars, 3 sugar alcohols, and 1 amine in two genotypes and F1 hybrid cabbages. The levels of glycolic acid, quinic acid, inositol, fumaric acid, glyceric acid, trehalose, shikimic acid, and aspartic acid were found to be very significantly different between the resistant and susceptible genotypes with a P value of <0.0001. These results will provide a foundation for further studies on diamondback moth resistance in cabbage breeding and for the development of other herbivore-resistant crops.

Variation and correlation analysis of phenolic compounds in mungbean (Vigna radiata L.) varieties

Phenolic compounds from a wide collection of mungbean [Vigna radiata (L.) Wilczek] germplasm (56 varieties) were characterised to determine the diversity among these phytochemicals and to analyse the relationships among their contents. The profiles of 25 phenolic compounds identified from the grains were subjected to data-mining processes, including principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), Pearsons correlation analysis, and hierarchical clustering analysis (HCA). The IT212105 and IT104818 varieties separated from the others in the first two principal components of PCA. PLS-DA showed significant separation between extracts of mungbean originating from three countries: China, Japan, and Korea. HCA of these phytochemicals resulted in clusters derived from common or closely related biochemical pathways. Significant positive relationships were observed between coumaric acid and resveratrol (r=0.7195, p<0.0001). Catechin content was positively correlated with rutin (r=0.6291, p<0.0001). The IT104818 variety appears to be a good candidate for future breeding programs, as it contains high levels of phenolic compounds. These results demonstrate the use of metabolic profiling combined with chemometrics as a tool for assessing the quality of food.

Determination of phenolic acids in Korean rice (Oryza sativa L.) cultivars using gas chromatography-time-of-flight mass spectrometry

Total soluble phenolic acids (free and esterified forms) from 15 Korean rice cultivars (Oryza sativa L.) were characterized to determine the total soluble phenolic acid contents and the diversity among the phenolic acids. The individual phenolic acids were identified by GC coupled to time-of-flight MS (TOFMS) including tertbutyldimethylsilyl (TBDMS) derivatization. This method requires only 13 min for the GC-TOFMS run. Total soluble phenolic acid contents were higher in pigmented rice (168–368 μg/g) than nonpigmented rice (146–172 μg/g). Freeform phenolic acids accounted for less than 10% of the total soluble phenolic acids. The dominant compounds were ferulic and sinapic acid, which were detected as cisand trans-isomers. The content of salicylic acid in rice grain varied, ranging from 7.95 to 29.61 μg/g. The quantitative results of this study indicate that the black rice ‘Josengheugchalbyeo’ could facilitate development of phenolic-rich rice or food.

Identification and quantification of carotenoids in paprika fruits and cabbage, kale, and lettuce leaves

Twelve carotenoids were identified in Korean leafy vegetables and paprikas. by high-performance liquid chromatography, Carotenoid contents varied greatly, with red paprika having a higher antheraxanthin and capsanthin contents than other paprikas. Orange paprika had higher levels of zeaxanthin, β-cryptoxanthin, lutein, and α-carotene compared to those of other paprikas. The results of Pearson’s correlation analysis using quantitative data of carotenoids revealed that significant positive relationships were apparent between capsanthin and antheraxanthin (r=0.9870, p <0.0001), zeaxanthin and α-cryptoxanthin (r=0.9951, p <0.0001), as well as lutein and α-carotene (r=0.9612, p <0.0001). Because the correlations between carotenoids levels have provided valuable information regarding metabolic associations, this technique will contribute to identifying metabolic links for carotenoid biosynthesis.

Metabolomic Analysis and Differential Expression of Anthocyanin Biosynthetic Genes in White- and Red-Flowered Buckwheat Cultivars (Fagopyrum esculentum)

Red-flowered buckwheat ( Fagopyrum esculentum ) is used in the production of tea, juice, and alcohols after the detoxification of fagopyrin. In order to investigate the metabolomics and regulatory of anthocyanin production in red-flowered (Gan-Chao) and white-flowered (Tanno) buckwheat cultivars, quantitative real-time RT-PCR (qRT-PCR), gas chromatography time-of-flight mass spectrometry (GC-TOFMS), and high performance liquid chromatography (HPLC) were conducted. The transcriptions of FePAL, FeC4H, Fe4CL1, FeF3H, FeANS, and FeDFR increased gradually from flowering stage 1 and reached their highest peaks at flowering stage 3 in Gan-Chao flower. In total 44 metabolites, 18 amino acids, 15 organic acids, 7 sugars, 3 sugar alcohols, and 1 amine were detected in Gan-Chao flowers. Two anthocyanins, cyanidin 3-O-glucoside and cyanidin 3-O-rutinoside, were identified in Gan-Chao cultivar. The first component of the partial least-squares to latent structures-discriminate analysis (PLS-DA) indicated that high amounts of phenolic, shikimic, and pyruvic acids were present in Gan-Chao. We suggest that transcriptions of genes involved in anthocyanin biosynthesis, anthocyanin contents, and metabolites have correlation in the red-flowered buckwheat Gan-Chao flowers. Our results may be helpful to understand anthocyanin biosynthesis in red-flowered buckwheat.