Background : Light-Emitting Diodes (LEDs) have been reported to alter the composition of the secondary metabolites present in many plants. For example, light exposure has been reported to significantly affect secondary metabolite biosynthesis in plants, and irradiance levels have been reported to affect the concentration and production of both phenylpropanoids and carotenoids. Therefore, the objective of the present study was to determine the most effective LED light source in enhancing growth and secondary metabolites (polyphenols and carotenoids) and investigate the effect of LED illumination on production the of primary and secondary metabolites (polyphenols and carotenoids) in Vigna unguiculata L. Walp. sprouts Methods and Results : In order to determine the effect of light-emitting diodes (LEDs) on plant metabolism, the present study examined the primary and secondary metabolite profiles of Vigna unguiculata L. Walp. sprouts that were exposed to red, blue, white, or a combination of red and blue LEDs using high-performance liquid chromatography (HPLC), electrospray ionization-mass spectrometry (ESI-MS), gas chromatography-mass spectrometry (GC-MS), and gas chromatography time-of-flight mass spectrometry (GC-TOF-MS). A total of 39 hydrophilic compounds were identified and quantitated using GC-TOF-MS, and six phenylpropanoids and six carotenoids were quantified using HPLC. The plants grown under blue LED light contained the highest level of total carotenoids (253.72 ± 17.27 ㎍/g) and phenylpropanoids (2,600.51 ± 4.90 ㎍/g). Thus, the current study provides a new approach for enhancing the carotenoid and phenylpropanoid production of V. unguiculata. Conclusion : This study suggests that blue LED light source is the most appropriate for the sprout growth and production of phenolic compounds and carotenoids in cowpea sprouts. Furthermore, these findings confirm that HPLC, GC-MS, and GC-TOF-MS are suitable for investigating metabolic relationships and offer a tenable strategy for enhancing secondary metabolite production using LED light sources.

Background : Fagopyrum esculentum Moench (common buckwheat) is an important pseudocereal due to high agricultural and medicinal values. It contains various minerals, fiber, and flavonoids. Additionally, flavonoids in buckwheat have various health effects. Thus, this study is aim to optimize the concentration of chitosan, salicylic acid (SA), and jasmonic acid (JA), for the production of phenolics in germinated buckwheat using high-performance liquid chromatography (HPLC). Methods and Results : The treatment with 0.1% chitosan increased the accumulation of all 7 phenolic compounds compared with the control, 0.01 and 0.5% chitosan treatments (p < 0.05). Furthermore, the germinated buckwheat treated with JA at the specific concentrations of 50, 100, and 150 μM increased the accumulation of total phenolic compounds. The germinated buckwheat grown in 150 μM of JA showed the highest amount of total phenolics which was approximately 2.47 times higher than that of control. Particularly, the accumulation of gallic acid, rutin, catechin, chlorogenic acid, and (-)-epicatechin were approximately 2.00, 2.38, 1.76, 2.81, and 7.95 times higher in JA-treated buckwheat than in the control buckwheat samples. A total of seven phenolics, including gallic acid, catechin, chlorogenic acid, caffeic acid, (-)-epicatechin, benzoic acid, and rutin, were detected in germinated buckwheat. Apparently, JA and chitosan treatment enhanced the accumulation of phenolic compounds in the germinated buckwheat. Particularly, the treatments with 0.1 % chitosan and 150 μM JA were the most effective on the accumulation of phenolic compounds. According to the time-course analysis, a 72 h chitosan treatment enhanced the production of phenolics. Similarly, the germinated buckwheat treated with 48 and 72 h showed the accumulation of higher levels of phenolic compounds than the control buckwheat. Conclusion : This study aimed to optimize the concentrations and treatment period of elicitors, chitosan and JA, for the enhanced production of phenolic compounds in germinated buckwheat. Thus, these results might help build sturdy strategies to enhance the production of phenolics in germinated buckwheat as a good nutritional source for human consumption.

Background : Radish sprouts or young seedlings are important nutritional vegetables in Asian countries. In the present study, we investigated plant growth and levels of glucosinolate accumulation in radish sprouts in response to treatments with different carbon sources. Plant growth and accumulation of glucosinolates were inversely correlated in treatments with different carbon sources. Methods and Results : All the carbon sources used in this study inhibited the growth of shoot and root in radish sprouts and facilitated significantly the accumulation of glucosinolates. Seven different glucosinolate compounds were detected in radish sprouts treated with different carbon sources. The total as well as individual amounts of all the identified glucosinolates increased after treatments with different carbon sources, except for 4-methoxyglucobrassicin and glucoiberin. In particular, the supplementation of sucrose, galactose, and glucose resulted in highest glucosinolate accumulation in radish sprouts. Radish sprouts treated with sucrose showed the highest levels of total glucosinolates, which was 1.22-fold higher than that of untreated sprouts. Furthermore, sucrose treatment resulted in higher production of gluconapoleiferin, glucoerucin, glucoraphasatin, and glucobrassicin compared with that in untreated sprouts, and the treatment of galactose and glucose similarly enhanced glucosinolate production when compared with untreated sprouts. Conclusion : seven glucosinolates were identified by HPLC and great differences in glucosinolates levels were observed in radish sprouts in response to different carbon sources. The glucosinolate production was positively affected by most of the stimuli used in this study. The results presented herein provide information about optimal cultivation conditions, particularly the suitable carbon sources, that will enhance glucosinolate production in radish sprouts.

Background : Morus alba L. (M. alba L.) belongs to the family Moraceae and has been industrially used as a food source for the silkworm and as a sedative in traditional medicine due to their adaptogenic, hypotensive, anti-inflammatory, hypoglycemic, and anti-adipogenic properties. In this study, metabolic profiling of diploid and tetraploid in Morus alba L. ‘Cheongil’ Methods and Results : Carotenoids are secondary metabolites found in most of the higher plants. In the current study, the levels of carotenoids were much higher in tetraploid of M. alba L. ‘Cheongil’ than diploid of M. alba L. ‘Cheongil’. Particularly, the levels of lutein, 13z-β-carotene, α-carotene, E-β-carotene, and 9Z-β-carotene were 2.51, 5.53, 7.68, 4.43, and 4,46 times higher in tetraploid of M. alba L. ‘Cheongil’, respectively. Additionally, hierarchical cluster analysis (HCA) was carried out using Pearson’s correlation results in order to determine the relationships between metabolites of diploid and tetraploid in Morus alba L. ‘Cheongil’. The results of HCA revealed the degree of correlation among 42 metabolites, which were identified and quantitated by GC-TOFMS. Among TCA organic acids, succinic acid was positively correlated with fumaric acid (r = 0.8250, p < 0.0001) and malic acid (r = 0.9436, p < 0.0001). Among carbohydrates, fructose was positively correlated with glucose (r = 0.9398, p < 0.0001) and mannose (r = 0.9394, p < 0.0001). Conclusion : In this study, we investigated primary metabolites and secondary metabolites (carotenoids) in diploid and tetraploid in Morus alba L. ‘Cheongil’ in order to provide information on the difference between

Background : Morus alba L. (M. alba L.), belonging to the family Moraceae, is widely distributed in East Asia. Fruits of M. alba L. have been used in traditional herbal medicine due to their antioxidant, anticancer, and antidiabetic properties. Phenolics play a main role for the growth, development, and pigment accumulation of plants. In this study, metabolic profiling of white (M. alba L. ‘Turkey’) and red (M. alba L. ‘Cheongil’) fruits during maturation. Methods and Results : Phenolic compounds are secondary metabolites found in most of the higher plants. In the current study, the levels of phenolic compounds decreased during the maturation of Turkey and Cheongil fruits. Particularly, the Turkey fruits showed a dramatic decrease in the accumulation of phenolics. Principal component analysis (PCA) is one of powerful tools to identify overall patterns in the multivariat experimental data. The PCA score plots results revealed a clear classification between Cheongil and Turkey. Additioanlly, each group spread left to right in the X-axis by maturity. Two principal components of the score plot explained 71.1% of the total variance. Principal component 1 was associated with the separation of each group by maturity and isolation of Turkey 1. Conclusion : In this study, we investigated primary metabolites and secondary metabolites (phenolics) in the white fruits (M. alba L. ‘Turkey’) and red fruits (M. alba L. ‘Cheongil’) in order to provide information on change in metabolite patterns during maturation.

Background : Agastache rugosa (A. rugosa), belonging to the Lamiaceae family, is a medicinal plant mainly distributed in Korea and contains various phenolic compounds revealing anti-fungal and anti-HIV properties. This study is aim to investigate change in phenylpropanoid content of flowers at different developmental stages using high performance liquid chromatography (HPLC) and quantitative real time polymerase chain reaction (qRT-PCR). Methods and Results : The variation in the transcriptional level of phenylpropanoid biosynthetic genes and phenylpropanoid contents in the flowers of A. rugosa at different developmental stages was analyzed. The transcript levels of phenylpropanoid biosynthesis genes, including ArPAL (phenylalanine ammonia-lyase), ArC4H (cinnamate 4-hydroxylase), and ArCHS (Chalcone synthase), were high in flowers at 1st stage compared with flowers at 2nd and 3rd stages. On the other hand, the expression levels of flavonoid biosynthesis genes, including ArTAT (tyrosine amino transferase), ArHPPR (hydroxyl phenylpyruvate reductase), and ArRAS (rosmarinic acid synthase), were higher in flowers at 3rd stage than those of flowers at 1st and 2nd. These results were consistent with HPLC analysis revealing that most phenolic compounds were higher in flowers at 1st and 2nd stage but the level of rosmarinic acid was the highest in 3rd stage. Conclusion : Our findings provide the information on change in phenylpropanoid biosynthesis in A. rugosa flowers at different developmental stages.

Background : Galantamine is mainly obtained from the bulbs and flowers of Galanthus caucasicus, Galanthus woronowii, and other related genera such as Narcissus tazetta, Narcissus pseudonarcissus, Leucojum aestivum, and Lycoris radiata. Galantamine is used to treat Alzheimer’s disease (AD) and as an AD painkiller. Narcissus tazetta (N. tazetta), belonging to the Amaryllidaceae family, is a ornamental plant containing galantamine. In this study, metabolic profiling of N. tazetta different organs was performed. Methods and Results : The amount of galantamine in bulb of N. tazetta is the highest levels. About 0.61 ± 0.09 ㎎/g in bulb, 0.15 ± 0.17 ㎎/g in root, and 0.10 ± 0.0 ㎎/g in leaf. Contents of galantamine in root and leaf are not statistically significant. The total phenolic contents in leaf are the highest level. Rutin and kaempferol are identified all part of N. tazetta. On the other hands, 4-hydroxybenzoic acid is existed in leaf and caffeic acid is only existed in root. None of the bulbs except rutin and kaempferol are identified. Because plant secondary metabolism is closely related to plant primary metabolism, we used GC-TOF-MS on the levels of hydrophilic low-molecular-weight molecules in the N. tazetta. A total of 41 metabolites, including sugars, amino acids, organic compounds, and phenolic acids, were identified and measured, and the resulting quantitative data were subjected to principal components analysis (PCA). The results of PCA of metabolic profiles clearly showed the lack of marked variance among different organs of L. radiata. Two principal components of the score plot explained 86.79% of the total variance (component 1; 55.40%, component 2; 31.39%). Component 1 resolved the separation of leaves from the other plant parts. Conclusion : Narcissus tazetta belongs to amaryllidaceae family. These family has various alkaloids, in particular, galantamine is beneficial to Alzheimer patients. All parts of N. tazetta produce galantamine, in particular, the highest level is existed in the bulb. In contrast, phenolic compounds are identified

Background : Members of Amaryllidaceae family produce several alkaloids with unique structures and a variety of medicinal properties. Galantamine, in particular, is one of the alkaloids approved by the Food and Drug Administration (FDA), and the European Registration Bureau for treatment of Alzheimer’s disease. Lycoris radiata (L. radiata), belonging to the Amaryllidaceae family, is a bulbous plant containing galantamine, which exhibits selective and dominant acetylcholinesterase inhibition. In this study, metabolic profiling of L. radiata different organs was performed. Methods and Results : Galantamine in root, bulb, and leaf of L.radiata analyzed by high performance liquid chromatography (HPLC). The amount of galantamine in leaf is about 1.07 ± 0.17 ㎎/g and it is the higher than bulb (0.88 ± 0.01 ㎎/g) and root (0.75 ± 0.01 ㎎/g). These results are statistically significant. Six phenolics are identified in L. radiata through high performance liquid chromatography. Total amounts of phenolics are the highest in bulb. Because plant secondary metabolism is closely related to plant primary metabolism, we used GC-TOF-MS on the levels of hydrophilic low-molecular-weight molecules in the L. radiata. A total of 41 metabolites, including sugars, amino acids, organic compounds, and phenolic acids, were identified and measured, and the resulting quantitative data were subjected to principal components analysis (PCA). The results of PCA of metabolic profiles clearly showed the lack of marked variance among different organs of L. radiata. Two principal components of the score plot explained 89.4% of the total variance (component 1, 51.86%; component 2, 37.54%). Component 1 resolved the separation of leaves from the other plant parts. Conclusion : Amaryllidaceae family synthesize galantamine belonging to alkaloids. Particularly, in bulb of Lycoris radiata, galantamine contents are the highest level. Thus, bulb is very beneficial for Alzheimer’s disease because the galantamine is well known as treatment of dementia of Alzheimer type.

Background : Narcissus tazetta (N. tazetta), belonging to the Amaryllidaceae family, is a bulbous plant distributed in Korea, China, and Japan. Amaryllidaceae family plants contained galantamine exhibiting dominant and selective acetylcholinesterase inhibition. In this study, transcriptome analysis of N. tazetta was carried out. Methods and Results : The results of studies conducted in duplicate revealed the presence of a total of 305,228 and 370,567 unigenes, acquired from 69,605,788 and 59,770,506 raw reads, respectively, after trimming the raw reads using CutAdapt, assembly using Trinity package, and clustering using CD-Hit-EST. The resulting unigenes were annotated based on the NCBI Non-redundant protein database, as N. tazetta is genetically closer to Phoenix dactylifera and Elaeis guineensis. The unigenes of N. tazetta were clustered into three major categories: biological processes, cellular components, and molecular functions, with 51 functional sections. A large number of unigenes (11,371 and 15,535 from replicates 1 and 2, respectively) were categorized in the biological process cluster, followed by the cellular component cluster, and the molecular function cluster. With respect to the biological process category, the unigenes were assigned to 23 functional sections. The majority of unigenes were involved in cellular processes. Among the unigenes clustered as the cellular component with 14 sections, most genes were associated with the cell and cell parts. Furthermore, 156,584 and 201,353 unigenes, respectively, matched the molecular function cluster with 14 sections, of which most unigenes were related to metabolic process and cellular process. Conclusion : This study provides functional information of N. tazetta and highlights the use of the Illumina platform for transcriptome research.

Background : Lycoris radiata belongs to the Amaryllidaceae family and is a bulbous plant native to South Korea, China, and Japan. Galantamine, a representative alkaloid of Amaryllidaceae plants, including L. radiata, exhibits selective and dominant acetylcholinesterase inhibition. In this study, transcriptome analysis of L. radiata was performed. Methods and Results : Genes for galantamine biosynthesis were used to design primers for qRT-PCR. Quantitative RT-PCR was performed with LrActin as a reference gene for normalization. The RT-PCR results reveal the expression of LrPAL A and LrC4H at an early stage in the pathway. Interestingly, the expression of these genes was significantly higher in roots. However, the expression levels of LrNNR and LrN4OMT, which are closely involved in galantamine biosynthesis, were significantly higher in bulbs than leaves and roots. The expression levels of LrPAL B, LrTYDC, LrCYP98A3 and LrCYP76T were not significantly different among the different parts of the plants tested. HPLC analysis confirmed the presence of galantamine in all the organs, including the root (0.53 ± 0.07 ㎎/g dry weight), bulb (0.27 ± 0.04 ㎎/g dry weight), and leaf (0.75 ± 0.09 ㎎/g dry weight). The galantamine level in the bulb was 1.42 and 2.78 times higher than that in the root and leaf, respectively. The results of qRT-PCR for the eight galantamine genes revealed relatively high levels of genes expressed early, including LrPAL A, LrPAL B, LrC4H, and LrTYDC in the roots. However, in the bulbs, the levels of LrNNR and LrN4OMT were higher, which are crucial for galantamine biosynthesis. It also explains why bulbs contain high amounts of galantamine, which is likely due to the increased expression of LrNNR and LrN4OMT and the high levels of LrCYP96T, although the genes expressed early were expressed at high levels in the root. Conclusion : Transcript data of plants grown in a growth chamber revealed high expression levels of LrNNR and LrN4OMT genes that are closely involved in galantamine biosynthesis, and, as expected, we observed higher amounts of galantamine in the bulbs than in the root and leaves.

Background : Lycoris radiata (L. radiata), which belongs to Amaryllidaceae family, is native to Northeast Asia including Korea, Japan, and China. It is known for its high ornamental and medicinal values. Extensive research has been conducted in a several fields, including molecular biology, morphology, pharmacology, physiology, palynology, and chromosomal biology. The plant is notable for its various biological activities, including anti-cancer, anti-malarial, anti-microbial, reduction in blood pressure, anti-inflammatory, cytotoxicity, and neuroprotective effects. Methods and Results : The results of studies conducted in duplicate revealed the presence of a total of 325,609 and 404,019 unigenes, acquired from 9,913,869,968 and 10,162,653,038 raw reads, respectively, after trimming the raw reads using CutAdapt, assembly using Trinity package, and clustering using CD-Hit-EST. The resulting unigenes were annotated based on the NCBI Non-redundant protein database, as L. radiata is genetically closer to Elaeis guineensis and Phoenix dactylifera. The unigenes of L. radiata were clustered into three major categories: biological processes, cellular components, and molecular functions, with 51 functional sections. A large number of unigenes (203,157 and 224,813 from replicates 1 and 2, respectively) were categorized in the biological process cluster, followed by the cellular component cluster, and the molecular function cluster. With respect to the biological process category, the unigenes were assigned to 23 functional sections. The majority of unigenes were involved in cellular processes. Among the unigenes clustered as the cellular component with 14 sections, most genes were associated with the cell and cell parts. Furthermore, 78,017 and 88,817 unigenes, respectively, matched the molecular function cluster with 14 sections, of which most unigenes were related to binding and catalytic activity. Conclusion : This study provides functional information of L. radiata and highlights the use of the Illumina platform for transcriptome research.

Background : Nasturtium officinale, belongs to the Brassicaceae, is a perennial plant growing in and around natural watersystem. It is commonly called watercress and commercially consumed as a salad crop in many countries. Hairy root cultures (HRCs), transformed by Agrobacterium rhizogenes (A. rhizogenes), have been noted for an experimental model system in plant metabolic engineering for the synthesis of natural products since hairy roots have fascinating properties including biochemical and genetic stability, high biosynthetic capacity for secondary metabolites and rapid growth rates. Methods and Results : The dry weights (DW) of watercress hairy roots was measured after 4-days freeze dryer. The highest weight was recorded after HRCs in SH and half-strength SH medium, followed by the levels of DW in MS and half-strength medium. However, the level of DW after HRCs in half-strength medium was lowest. SH medium is the most suitable for the growth of watercress hairy roots. Glucosinolate contents in the hairy roots varied responding to the basal media. 1/2 SH basal media resulted in the lowest value of total glucosinolate. MS basal media, however, resulted in the highest value of total glucosinolate, as well as the highest accumulation of each glucosinolate. The hairy roots cultured in 1/2 MS basal media showed the second highest value of total glucosinolate, followed by B5, SH and 1/2 B5. The accumulation of 4-Methoxyglucobrassicin and gluconasturtiin was also secondly higher in the hairy roots in 1/2 MS. In contrast, the level of 4-Hydroxyglucobrassicin and glucobrassicin were the second highest after culturing in SH and B5, respectively. Conclusion : In this study, media played a main role in growth and glucosinolate accumulation in watercress hairy roots. SH and half-strength SH media enabled the rate growth of hairy roots to be highest. In contrast, the highest accumulation of glucosinolate was recorded after HRCs in MS media. The current study suggests HRCs of watercress could be one of an effective alternative approaches for the enhanced production of glucosinolates

Background : Agastache rugosa (A.rugosa), belongs to the Labiatae family, is a perennial plant distributed in Korea, Japan, Taiwan and China. It is commonly called korean mint and commercially consumed as a medicinal plant in many countries since the crop contains monoterpenes and phenylpropanoids including rosmarinic acid, tilianin and acacetin. Achievement of hairy root cultures (HRCs) through infection of A rhizogenes is a valuable alternative approach, resulting from genetic and biochemical stability, rapid growth rates and synthesis of natural products. Methods and Results : The hairy root, obtained from the explant of A.rugosa, was cultured in the basal half-strength MS (Murashige & Skoog) medium. The dry weights (DW) of hairy roots was measured after 4-days freeze dryer. The highest levels of DW were obtained at hairy roots cultured in the basal medium supplemented with glucose, galactose and sucrose. The lowest weight was recorded after HRCs in the control, meant that the medium did not contain any carbon sources. Sucrose, glucose and galactose are the most suitable for the growth of korean mint hairy roots. the rosmarinic acid contents in the hairy roots varied responding to various carbohydrates. The basal media added with sucrose resulted in the highest value of rosmarinic acid, followed by the basal media with galactose and glucose. The control showed the lowest amount of rosmarinic acid. Conclusion : In this study, carbon source are of importance for growth and accumulation of rosmarinic acid accumulation in korean mint hairy roots. Especially, the accumulation of rosmrinic acid and hairy root growth was the most appropriate carbohydrate. The current study suggests HRCs of korean mint could provide an valuable alternative approaches for the enhanced production of rosmarinic acid.

Background : Mulberry (Morus alba L.), renowned for their medicine benefits and the leave as the sole food for silkworm (Bombyx mori). To understanding the molecular mechanism of color formation and nutritive value in different mulberry fruit varieties, we use high-throughput transcriptome sequencing technique to investigated the anthocyanin and betulinic biosynthesis pathway related functional genes. In addition, the total antosyanin and betuinic acid contend were also measured. Methods and Results : The resulting cDNA library was then sequenced using an Illumina HiSeq™ 2000 system. The clean reads were assembled using Trinity software, Then perform gene family clustering to get final unigenes. The pH differential method was used to determine the total anthocyanin content (TAC) of methanol extract from the red and white mulberry, and High-performance liquid chromatography (HPLC) analysis was used to quantify the triterpenes content. In this study, total 50,149 unigenes with an average length of 1,125 nt and N50 equaling 1,861 nt were generated. Using these transcriptome sequecing, cDNAs encoding anthocyanin biosynthetic genes and triterpene biosynthetic genes were isolated. In addition, total anthocyanins and betulinic acid content were analyzed. A great amount of total anthocyanins (59.16 mg/g) were found in fully ripe fruit of Cheongil. Accumulation of betulin and betulinic acid were also detected in all stages of Cheongil and Turkey fruits with small amount. Conclusion : The results of transcriptome sequencing provide useful information at molecular lever in mulberry research, such as interesting gene discovering, marker assisted molecular breeding, and interesting metabolic pathway investigate. The gene expression results could help us understanding of the molecular mechanisms of different fruit color determining factor.