Molecular Cloning and Characterization of Three Glucosinolate Transporter (GTR) Genes from Chinese Kale

Jiang, Ding; Liu, Jing; Cao, Bihao; Wu, Si-Pei; Chen, Guoju; Chen, Changming

doi:10.3390/genes10030202

Cited by 14 publications

(14 citation statements)

References 39 publications

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“…The total glucosinolate content was significantly decreased in mature leaves. 22 The concentration of sulforaphane in broccoli sprouts (1,153 mg/100 g DW) is about 10 times greater than mature broccoli (44-171 mg/100 g DW). 23 It was well-known that abiotic stress, such as supplementation of Se to plants, can lead to increased production of phenolic compounds.…”

Section: Discussionmentioning

confidence: 96%

Cytotoxicity of Selenium-Enriched Chinese Kale (Brassica oleracea var. alboglabra L.) Seedlings Against Caco-2, MCF-7 and HepG2 Cancer Cells

Luang-In¹,

Saengha²,

Buranrat³

et al. 2020

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Cultivation of Se-KS Seeds of Chinese kale (Brassica oleracea var. alboglabra L.) were obtained from Chia Tai Company Limited (Thailand). Authentication of the seeds was done by Assoc. Prof. Dr. Anut Chantiratikul from Mahasarakham University using seed morphology checking and cultivation trial in ABSTRACT Background: The Selenium-enriched Chinese kale (Brassica oleracea var. alboglabra L.) seedlings (Se-KS) have been known for its antioxidant activities, however its cytotoxic effects on various cancer cells are yet to be reported. Objective: The objective of this work was to study the cytotoxic effects of Se-KS on Caco-2, MCF-7 and HepG2 cancer cells. Materials and Methods: Freeze-dried seedlings were ground and incubated in 0.1 M citrate phosphate buffer pH 7.0 for 1 h at 37°C and extracted with dichloromethane to obtain total isothiocyanate (ITC) content which was quantified using the 1,2-benzenedithiole (BDT)-based cyclocondensation assay. The extracts from fresh seedlings were used to determine the cytotoxic effect on Caco-2, MCF-7 and HepG2 cancer cells. Results: Se-KS was found to contain total ITC content at 1.02 mmol/100 g dry weight (DW) which was significantly lower than that of 7-day old broccoli microgreens (1.60 mmol/100 g DW) as reference Cruciferous vegetables. In addition, Se-KS extract exhibited cytotoxic effects in a dose-and time-dependent manners. The lowest IC50 value of 82.83 µg/mL at 72 h was derived from HepG2 cells and the highest IC50 value of 164.00 µg/mL at 72 h was from MCF-7 cells suggesting that the Se-KS extract was most effective against HepG2 cells. Cancer cells showed signs of apoptotic bodies over 72 h and DNA fragmentations at 24 h indicating that the Se-KS extract was able to induce apoptosis in cancer cells in addition to cytotoxic effect. Conclusion: Thus, Se-KS could be a novel source of organo selenium with chemopreventive benefits for functional food development.

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Section: Discussionmentioning

confidence: 96%

Cytotoxicity of Selenium-Enriched Chinese Kale (Brassica oleracea var. alboglabra L.) Seedlings Against Caco-2, MCF-7 and HepG2 Cancer Cells

Luang-In¹,

Saengha²,

Buranrat³

et al. 2020

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“…Interestingly, the application of JA and SA to Arabidopsis and Brassica has been shown to increase the accumulation of GLSs [30][31][32] . Furthermore, recent studies showed that SA treatment stimulated the gene expression of GTR1 in Chinese kale 33 . These studies showed the possibility of interactions between SA and AtGTR1.…”

Section: Discussionmentioning

confidence: 99%

Transport Efficiency of AtGTR1 Dependents on the Hydrophobicity of Transported Glucosinolates

Chung

Cheng

Wang

et al. 2021

Preprint

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Glucosinolates (GLSs) are a group of secondary metabolites that are involved in the defense of herbivores. In Arabidopsis thaliana, Glucosinolate Transporter 1 (AtGTR1) transports GLSs with high affinity via a proton gradient-driven process. In addition to transporting GLSs, AtGTR1 also transports phytohormones, jasmonic acidisoleucine (JA-Ile), and gibberellin (GA). However, little is known about the mechanisms underlying the broad substrate specificity of AtGTR1. Here, we characterized the substrate preference of AtGTR1 by using a yeast uptake assay, and the results revealed that GLS transport rates are negatively correlated with the hydrophobicity of substrates. Interestingly, the AtGTR1 showed a higher substrate affinity for GLSs with higher hydrophobicity, suggesting a hydrophobic substrate binding pocket. In addition, a competition assay revealed that the presence of JA, salicylic acid (SA), and indole-3-acetic acid (IAA) inhibits the transport of GLSs in yeast, suggesting a potential regulatory mechanism of AtGTR1. To further characterize the functional properties of AtGTR1, mutagenesis experiments confirmed that the conserved EXXEK motif and Arg166 are essential for the GLS transport function. In addition, the purified AtGTR1 adopts a homodimeric conformation, which is possibly regulated by phosphorylation on Thr105. The phosphomimetic mutation, T105D, reduced its protein expression and completely abrogated its GLS transport function, indicating the essential role of phosphorylation on AtGTR1. In summary, this study investigated various factors associated with the GLS transport and increased our knowledge on the substrate preferences of AtGTR1. These findings contribute to understanding how the distribution of defense GLSs is regulated in plants and could be used to improve crop quality in agriculture.

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“…In B. oleracea var. chinensis Lei, two GSL transporters BocGTR1a and BocGTR1c were identified, which express predominantly in the leaves and buds ( Jiang et al, 2019 ) with a variable expression pattern that also depends on the plant developmental stages and various abiotic and biotic stress conditions. In addition to similar transmembrane topology, three-dimensional structure, and functional domains, these two transporters also showed more than 81% amino acid sequence homology with A. thaliana GTR proteins, which indicates their related functionalities.…”

Section: Plant Secondary Metabolite Transporters: Functional Relevance and Approaches For Their Discovery And Characterizationmentioning

confidence: 99%

Plant Secondary Metabolite Transporters: Diversity, Functionality, and Their Modulation

Nogia

Pati

2021

Front. Plant Sci.

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Secondary metabolites (SMs) play crucial roles in the vital functioning of plants such as growth, development, defense, and survival via their transportation and accumulation at the required site. However, unlike primary metabolites, the transport mechanisms of SMs are not yet well explored. There exists a huge gap between the abundant presence of SM transporters, their identification, and functional characterization. A better understanding of plant SM transporters will surely be a step forward to fulfill the steeply increasing demand for bioactive compounds for the formulation of herbal medicines. Thus, the engineering of transporters by modulating their expression is emerging as the most viable option to achieve the long-term goal of systemic metabolic engineering for enhanced metabolite production at minimum cost. In this review article, we are updating the understanding of recent advancements in the field of plant SM transporters, particularly those discovered in the past two decades. Herein, we provide notable insights about various types of fully or partially characterized transporters from the ABC, MATE, PUP, and NPF families including their diverse functionalities, structural information, potential approaches for their identification and characterization, several regulatory parameters, and their modulation. A novel perspective to the concept of “Transporter Engineering” has also been unveiled by highlighting its potential applications particularly in plant stress (biotic and abiotic) tolerance, SM accumulation, and removal of anti-nutritional compounds, which will be of great value for the crop improvement program. The present study creates a roadmap for easy identification and a better understanding of various transporters, which can be utilized as suitable targets for transporter engineering in future research.

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Molecular Cloning and Characterization of Three Glucosinolate Transporter (GTR) Genes from Chinese Kale

Cited by 14 publications

References 39 publications

Cytotoxicity of Selenium-Enriched Chinese Kale (Brassica oleracea var. alboglabra L.) Seedlings Against Caco-2, MCF-7 and HepG2 Cancer Cells

Cytotoxicity of Selenium-Enriched Chinese Kale (Brassica oleracea var. alboglabra L.) Seedlings Against Caco-2, MCF-7 and HepG2 Cancer Cells

Transport Efficiency of AtGTR1 Dependents on the Hydrophobicity of Transported Glucosinolates

Plant Secondary Metabolite Transporters: Diversity, Functionality, and Their Modulation

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