• Title/Summary/Keyword: Glucosinolate biosynthesis

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Metabolic engineering of aliphatic glucosinolates in Chinese cabbage plants expressing Arabidopsis MAM1, CYP79F1, and CYP83A1

  • Zang, Yun-Xiang;Kim, Jong-Hoon;Park, Young-Doo;Kim, Doo-Hwan;Hong, Seung-Beom
    • BMB Reports
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    • v.41 no.6
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    • pp.472-478
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    • 2008
  • Three Arabidopsis cDNAs, MAM1, CYP79F1, and CYP83A1, required for aliphatic glucosinolate biosynthesis were introduced into Chinese cabbage by Agrobacterium tumefaciens-mediated transformation. The transgenic lines overexpressing MAM1 or CYP83A1 showed wild-type phenotypes. However, all the lines overexpressing CYP79F1 displayed phenotypes different from wild type with respect to the stem thickness as well as leaf width and shape. Glucosinolate contents of the transgenic plants were compared with those of wild type. In the MAM1 line M1-1, accumulation of aliphatic glucosinolates gluconapin and glucobrassicanapin significantly increased. In the CYP83A1 line A1-1, all the aliphatic glucosinolate levels were increased, and the levels of gluconapin and glucobrassicanapin were elevated by 4.5 and 2 fold, respectively. The three CYP79F1 transgenic lines exhibited dissimilar glucosinolate profiles. The F1-1 line accumulated higher levels of gluconapoleiferin, glucobrassicin, and 4-methoxy glucobrassicin. However, F1-2 and F1-3 lines demonstrated a decrease in the levels of gluconapin and glucobrassicanapin and an increased level of 4-hydroxy glucobrassicin.

Effect of cultivars on hairy root induction and glucosinolate biosynthesis in a hairy root culture of Kimchi cabbage (Brassica rapa L. ssp. Pekinensis

  • Sang Un Park;Sook Young Lee
    • Korean Journal of Agricultural Science
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    • v.49 no.1
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    • pp.51-60
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    • 2022
  • Cruciferous vegetables are rich in biologically active compounds such as glucosinolates and have various health benefits. Among these vegetables, Kimchi cabbage (Brassica rapa L. ssp. Pekinensis) is one of the most popular leafy vegetables due to the presence of the highest amounts of numerous vital phytonutrients, minerals, vitamins, and antioxidants. This study aims to investigate the effects of six cultivars (Chundong 102, Asia No Rang Mini, Hwimori Gold, Asia Seoul, Wol Dong Chun Chae, and Asia Bbu Ri) on hairy root induction and glucosinolate biosynthesis in the hairy root cultures of Kimchi cabbage. Seven different glucosinolates, in this case sinigrin, gluconapin, glucoerucin, glucobrassicin, 4-methoxyglucobrassicin, gluconasturtiin, and neoglucobrassicin, were detected in the hairy root cultures of Kimchi cabbage. Among the different cultivars, Asia No Rang Mini was the most promising candidate for hairy root stimulation, as it achieved the highest values for the growth rate, root number, root length, transformation efficiency, and total glucosinolate content. Overall, the Asia No Rang Mini cultivar of Kimchi cabbage performed best as a promising cultivar hairy root culture for glucosinolate production.

Glucosinolate Content Varies and Transcriptome Analysis in Different Kale Cultivars (Brassica oleracea var. acephala) Grown in a Vertical Farm (수직농장에서 자란 케일(Brassica oleracea var. acephala) 품종에 따른 글루코시놀레이트 함량의 변화 및 전사체 분석)

  • Nguyen, Thi Kim Loan;Lee, Ga Oun;Jo, Jung Su;Lee, Jun Gu;Lee, Shin-Woo;Son, Ki-Ho
    • Journal of Bio-Environment Control
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    • v.31 no.4
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    • pp.332-342
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    • 2022
  • Kale (Brassica oleracea var. acephala) is one of the most frequently consumed leafy vegetables globally, as it contains numerous nutrients; essential amino acids, phenolics, vitamins, and minerals, and is particularly rich in glucosinolates. However, the differences in the biosynthesis of glucosinolates and related gene expression among kale cultivars has been poorly reported. In this study, we investigated glucosinolates profile and content in three different kale cultivars, including green ('Man-Choo' and 'Mat-Jjang') and red kale ('Red-Curled') cultivars grown in a vertical farm, using transcriptomic and metabolomic analyses. The growth and development of the green kale cultivars were higher than those of the red kale cultivar at 6 weeks after cultivation. High-performance liquid chromatography (HPLC) analysis revealed five glucosinolates in the 'Man-Choo' cultivar, and four glucosinolates in the 'Mat-Jjang' and 'Red-Curled' cultivars. Glucobrassicin was the most predominant glucosinolate followed by gluconastrutiin in all the cultivars. In contrast, other glucosinolates were highly dependent to the genotypes. The highest total glucosinolates was found in the 'Red-Curled' cultivar, which followed by 'Man-Choo' and 'Mat-Jjang'. Based on transcriptome analysis, eight genes were involved in glucosinolate biosynthesis. The overall results suggest that the glucosinolate content and accumulation patterns differ according to the kale cultivar and differential expression of glucosinolate biosynthetic genes.

Elicitation of Indole-3-ylmethyl Glucosinolate Biosynthesis in Turnip Culture Cells and Their Relationship with Plant Resistance to Botrytis cinerea (잿빛곰팡이병 추출물을 이용한 순무배양세포의 Indole-3-ylmethyl glucosinolate의 생합성유도와 병원성연구)

  • Kwon, Soon Tae;Zhang, Vivian
    • Korean Journal of Plant Resources
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    • v.30 no.5
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    • pp.542-548
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    • 2017
  • Two different races of Botryris cinerea were selected by the response of plant leaves to the pathogen infection. Based on lesion size of the pathogen on the leaves, turnip showed susceptible response to 'Grape-01' race, and resistant to 'Orange' race. Turnip leaves infected with resistant pathogen race, "Orange", showed significantly higher content of indole-3-ylmethyl glucosinolate (I3M) than those infected with susceptible race, 'Grape-01'. Contents of I3M in the leaves with resistant 'Orange' race was 2.5 times as high as that in uninfected leaves, whereas I3M in the leaves infected with susceptible 'Grape-01' race showed lower content than in untreated leaves. Growth of turnip suspension cells was significantly inhibited by the treatment of MeOH extract or water extract of 'Orange' race as compared with the treatment of susceptible race, 'Grape-01'. Treatment of MeOH or water extract from 'Orange' race to turnip suspension cells, strongly inhibited cell viability up to 22.7% or 16.5%, respectively. However, plant cells treated with MeOH or water extract from resistant race, 'Orange' showed higher I3M content than that from susceptible race, 'Grape-01'. These results suggest that accumulation and degradation of I3M glucosinolate in turnip cells closely related to the resistance and susceptibility of turnip cells to Botrytis cinerea.

Metabolic Engineering of Indole Glucosinolates in Chinese Cabbage Plants by Expression of Arabidopsis CYP79B2, CYP79B3, and CYP83B1

  • Zang, Yun-Xiang;Lim, Myung-Ho;Park, Beom-Seok;Hong, Seung-Beom;Kim, Doo Hwan
    • Molecules and Cells
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    • v.25 no.2
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    • pp.231-241
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    • 2008
  • Indole glucosinolates (IG) play important roles in plant defense, plant-insect interactions, and stress responses in plants. In an attempt to metabolically engineer the IG pathway flux in Chinese cabbage, three important Arabidopsis cDNAs, CYP79B2, CYP79B3, and CYP83B1, were introduced into Chinese cabbage by Agrobacterium-mediated transformation. Overexpression of CYP79B3 or CYP83B1 did not affect IG accumulation levels, and overexpression of CYP79B2 or CYP79B3 prevented the transformed callus from being regenerated, displaying the phenotype of indole-3-acetic acid (IAA) overproduction. However, when CYP83B1 was overexpressed together with CYP79B2 and/or CYP79B3, the transformed calli were regenerated into whole plants that accumulated higher levels of glucobrassicin, 4-hydroxy glucobrassicin, and 4-methoxy glucobrassicin than wild-type controls. This result suggests that the flux in Chinese cabbage is predominantly channeled into IAA biosynthesis so that coordinate expression of the two consecutive enzymes is needed to divert the flux into IG biosynthesis. With regard to IG accumulation, overexpression of all three cDNAs was no better than overexpression of the two cDNAs. The content of neoglucobrassicin remained unchanged in all transgenic plants. Although glucobrassicin was most directly affected by overexpression of the transgenes, elevated levels of the parent IG, glucobrassicin, were not always accompanied by increases in 4-hydroxy and 4-methoxy glucobrassicin. However, one transgenic line producing about 8-fold increased glucobrassicin also accumulated at least 2.5 fold more 4-hydroxy and 4-methoxy glucobrassicin. This implies that a large glucobrassicin pool exceeding some threshold level drives the flux into the side chain modification pathway. Aliphatic glucosinolate content was not affected in any of the transgenic plants.

Identification of glucosinolate-associated QTLs in cabbage (Brassica oleracea L. var. capitata)

  • Oh, Sang Heon;Choi, Su Ryun;Pang, Wenxing;Rameneni, Jana Jeevan;Yi, So Young;Kim, Man-Sun;Im, Su Bin;Lim, Yong Pyo
    • Korean Journal of Agricultural Science
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    • v.45 no.1
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    • pp.1-8
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    • 2018
  • Glucosinolates are one of the important plant secondary metabolites that are produced mainly in Brassicaceae plants. The compounds are primarily involved in defense responses to biotic and abiotic resistance in plants and play important biological roles during plant growth and development. In this study, the glucosinolate profiles in leaves of two different Brassica oleracea populations were compared using high-performance liquid chromatography (HPLC). The nine major glucosinolates compounds in cabbage leaves were identified as belonging to the aliphatic and indolic groups. Among them, sinigrin, which belongs to the aliphatic group, was recorded to be 41% whereas glucobrassicin and 4-methoxyglucobrassicin, which belong to the indolic group, were recorded to be 53.8%. In addition, we performed a genetic analysis to identify regions of the genome regulating glucosinolates biosynthesis in the $F_3$ population of Brassica oleracea. A total of 9 glucosinolates were used for the quantitative trait locus (QTL) analysis. Out of 9, a total of 3 QTLs were identified and they were associated with sinigrin, glucobrassicin, and 4-methoxyglucobrassicin synthesis located in Chromosome 1 and Chromosome 8, respectively. The results of this study will provide valuable information for the breeding of cabbage containing high glucosinolate content, and our next target is to develop component-specific and tightly linked markers for various glucosinolates.

Effect of methyl jasmonate on the glucosinolate contents and whole genome expression in Brassica oleracea (유묘기 양배추류에서 메틸자스모네이트에 의한 글루코시놀레이트 함량 변화 및 전사체 발현 분석)

  • Lee, Jeongyeo;Min, Sung Ran;Jung, Jaeeun;Kim, HyeRan
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.189-204
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    • 2019
  • In this study, we analyzed the changes in glucosinolate content and gene expression in TO1000DH3 and Early big seedling upon methyl jasmonate (MeJA) treatment. Analysis of glucosinolate contents after MeJA treatment at $200{\mu}M$ concentration showed that the total glucosinolate content increased by 1.3-1.5 fold in TO1000DH3 and 1.3-3.8 fold in Early big compared to those before treatment. Aliphatic glucosinolates, progoitrin and gluconapin, were detected only in TO1000DH3, and the changes in the content of neoglucobrassicin were the greatest at 48 hours after MeJA treatment in TO1000DH3 and Early big. The transcriptomic analysis showed that transcripts involved in stress or defense reactions, or those related to growth were specifically expressed in TO1000DH3, while transcripts related to nucleosides or ATP biosynthesis were specifically expressed in Early big. GO analysis on transcripts with more than two-fold change in expression upon MeJA treatment, corresponding to 12,020 transcripts in TO1000DH3 and 13,510 transcripts in Early big, showed that the expression of transcripts that react to stimulus and chemical increased in TO1000DH3 and Early big, while those related to single-organism and ribosome synthesis decreased. In particular, the expression increased for all transcripts related to indole glucosinolate biosynthesis, which is associated with increase in glucobrassicin and neoglucobrassicin contents. Upon MeJA treatment, the expression of AOP3 (Bo9g006220, Bo9g006240), TGG1 (Bo14804s010) increased only in TO1000DH3, while the expression of Dof1.1 (Bo5g008360), UGT74C1 (Bo4g177540), and GSL-OH (Bo4g173560, Bo4g173550, Bo4g173530) increased specifically in Early big.