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Taxonomy and species diversity of the mustard family (Cruciferae or Brassicaceae) in Kazakhstan is updated. Draba bajtenovii described and hitherto known from Kungei Alatau is synonymized with D. fladnizensis for which it is a southern limit of distribution in the Eurasia, while Trans-Ili Alatau endemic D. microcarpella is found conspecific with a montane Asian D.lanceolata. Noccaea thlaspidioides (N. cochleariformis) is confirmed to occur in the country based on collections from Altai. Finally, Isatis trachycarpa and Smelowskia annua are shown to be absent in the flora of Kazakhstan due to the lack of gatherings confirming their occurrence in the country., Уточнён видовой состав крестоцветных (Cruciferae, или Brassicaceae) Казахстана. Два вида рода Draba, считавшихся эндемиками страны (D. bajtenovii и D. microcarpella, известные по единичным сборам из Кунгей и Заилийского Алатау), синонимизированы, соответственно, с арктомонтанным D. fladnizensis, для которого данное местонахождение является самым южным в Евразии, и нагорноазиатским D. lanceolata. Noccaeathlaspidioides (N.cochleariformis), факт произрастания которого в Казахстане ранее не подтверждался, вновь включён в спискок видов природной флоры РК благодаря наличию образцов из казахстанской части Алтая. Нахождение во флоре страны Isatis trachycarpa и Smelowskia annua, напротив, не получает подтверждения по причине отсутствия сборов с территории Казахстана.

Based on the study of collections of LE, RV and RWBG, information on occurrence (or absence) of certain Cruciferae (Brassicaceae) species in Russia and some neighbouring states is updated. Camelina hispida is first recorded from Russia, Camelina sativa from Turkmenistan and Goldbachia torulosa from Ukraine as alien species apparently not naturalized in relevant countries. In contrast, Isatis armena, a novelty for the flora of both Russia and Europe, along with Plagioloba persica for the first time recorded from Kyrgyzstan, appears to be aboriginal elements in the newly revealed localities. At the same time, occurrence of Isatis cardiocarpa in Europe, as well as an introduction ofGoldbachia laevigata in Ukraine does not get support. In addition, the combination Plagioloba perfoliata is validated., По материалам гербариев LE, RV и RWBG приводятся новые данные о нахождении (или ненахождении) некоторых видов крестоцветных (Cruciferae, или Brassicaceae) в России и ряде соседних стран. В качестве заносных и, по-видимому, не удержавшихся во флоре соответствующих регионов, указанных в скобках, впервые приводятся Camelina hispida (Россия), Camelina sativa (Туркменистан) и Goldbachia torulosa(Украина). Isatis armena является новинкой для России и Европы, а Plagioloba persica – для Киргизии; оба вида относятся, по всей видимости, к элементам естественной фракции флоры. Не получает подтверждения произрастание Isatis cardiocarpa в Европе, а также факт заноса Goldbachia laevigata на Украину. Кроме того, обнародована комбинация Plagioloba perfoliata.

Saudi journal of biological sciences 29(1), 190-196 (2022). doi:10.1016/j.sjbs.2021.08.072, Published by Elsevier, Amsterdam [u.a.]

Based on the transcriptome data of Isatis indigotica, a total of 110 putative glycosytransferases were identified. Through prokaryotic expression and enzymic activity assay in vitro, a novel lignan glycosyltransferase gene was screened out and named IiUGT349, which catalyzed lariciresinol into lariciresinol-4-O-β-D-glucoside and lariciresinol-4'-O-β-D-glucoside. Bioinformatics analysis suggested that IiUGT349 contained an open reading frame(ORF) of 1 401 bp encoding a protein of 467 amino acids. A protein analysis indicated that IiUGT349 have a predecited molecular weight of 52.77 kDa and pI of 5.96. Phylogenetic analysis showed that IiUGT349 belonging to UGT90 family shared low amino acid sequence identity with the reported lignan glycosyltransferases, which may represent a novel type of lignan glycosyltransferases. Quantitative real-time PCR(qRT-PCR) analysis showed that IiUGT349 was expressed in roots, stems, young leaves and leaves, with the highest expression level in stems. Further biochemical analysis showed that the optimal reaction time of IiUGT349 recombinant protein was 12 h and the optimal temperature was 45 ℃. Subcellular localization demonstrated that IiUGT349 was located in the cytoplasm and nucleus of plants. In this study, a new glucosyltransferase gene IiUGT349 from I. indigotica belonging to the UGT90 family was cloned, which laid a foundation to further investigate itsapos; function and elucidate the lignan glycosides biosynthesis pathway and plays an important role for great significance for the synthetic biology of active lignan glycosides.

Effects of perlite, vermiculite, and grass charcoal on root growth of Isatis and soil nutrients migration were studied, and the plant growth indicators and some soil properties of upper (0~5cm) and lower layer (5~15cm) were analyzed. The experiment treatments were loess (CK), loess : perlite = 3 : 1(A), loess : grass charcoal = 3 : 1(B), loess : vermiculite = 3 : 1(C), loess: perlite: grass charcoal = 6 : 1 : 1(A1), loess: perlite: vermiculite = 6 : 1 : 1(B1), loess: grass charcoal: vermiculite = 6 : 1 : 1(C1). The results showed that soil pH of vermiculite-containing treatments in the upper layer and grass charcoal treatment in the lower layer decreased significantly. Before planting, the available potassium and phosphorus of the upper layer were significantly higher than those of the lower layer, and soil organic matter (SOM) was slightly higher than that of the lower layer. After planting, the decrease of available potassium in the upper layer was less than that of the lower layer, and the decrease of available phosphorus and increase of SOM in the upper layer were slightly higher than that of the lower layer. In sum, it is preferred to choose B (loess : grass charcoal = 3 : 1) and A1 (loess: perlite: grass charcoal = 6 : 1 : 1) to improve the soil nutrient and utilization efficiency. Bangladesh J. Bot. 50(3): 947-954, 2021 (September) Special

Arsenic (As) and cadmium (Cd) belong to the group of major pollutants extremely toxic to plants. Metal hyperaccumulating plants play an important role in phytoextraction of heavy metals. Silicon (Si) plays an important role in the amelioration of heavy metal stress through physio-biochemical mechanisms, which remain poorly understood in hyperaccumulators. The main purpose of this study was to determine the impact of Si on growth and performance of As hyperaccumulator Isatis cappadocica Desv., exposed to As and Cd. Results showed that Si (especially at 1 mM level) alleviated the harmful impact of As/Cd and significantly increased the root and shoot biomass, root and shoot length and chlorophyll contents of I. cappadocica by enhancing the plant defense mechanisms. Between the two investigated harmful elements, As was accumulated in plant parts significantly more than Cd, however with considerably lower toxic growth effects. The As/Cd concentration, bioaccumulation and translocation factor and total As content both in roots and shoots of Si-supplied plant were significantly reduced as a protective mechanism, especially in Cd exposed plant. In comparison with single As/Cd treatment, Si supply reduced H2O2 content, increased total soluble protein content and enhanced glutathione S-transferase activity in shoots. The results of this study clearly showed that Si minimized As/Cd uptake and root to shoot translocation, and therefore Si cannot enhance the phytoextraction potential of this plant species. Additionally, Si-improved growth and reduced oxidative damages caused by excess of As and Cd suggested that the similar mechanisms of metal(loid) alleviation are adopted in hyperaccumulators as well as non-hyperaccumulating plants.

The lignan glycosyltransferase UGT236(belonging to the UGT71 B family) from Isatis indigotica can catalyze the production of phloridzin from phloretin in vitro. UGT236 shares high identity with P2'GT from apple. In this study, the recombinant plasmid pET28 a-MBP-UGT236 was transferred into Escherichia coli Rosetta(DE3) cells and induced by isopropyl-β-D-thiogalactoside(IPTG). The purified UGT236 protein was used for enzymatic characterization with phloretin as substrate. The results showed that UGT236 had the optimal reaction temperature of 40 ℃ and the optimal pH 8(Na_2HPO_4-NaH_2PO_4 system). The UGT236 activity was inhibited by Ni~(2+) and Al~(3+), enhanced by Fe~(2+), Co~(2+), and Mn~(2+), and did not affected by Mg~(2+), Ca~(2+), Li~+, Na~+, or K~+. The K_m, K_(cat), and K_(cat)/K_m of phloretin were 61.03 μmol·L~(-1), 0.01 s~(-1), and 157.11 mol~(-1)·s~(-1)·L, and those of UDPG were 183.6 μmol·L~(-1), 0.01 s~(-1), and 51.91 mol~(-1)·s~(-1)·L, respectively. The possible active sites were predicted by homologous modeling and molecular docking. By mutagenisis and catalytic activity detection, three key active sites, Glu391, His15, and Thr141, were identified, while Phe146 was related to product diversity. In summary, we found that the lignan glycosyltransferase UGT236 from I.indigotica could catalyze the reaction of phloretin into phloridzin. Several key amino acid residues were identified by structure prediction, molecular docking, and site-mutagenesis, which provided a basis for studying the specificity and diversity of phloretin glycoside products. This study can provide a reference for artificially producing glycosyltransferase elements with high efficiency and specific catalysis.

Jiuling Deng,1,* Ying Ma,2,* Yuqiong He,1,* Hong Yang,2 Yanhong Chen,2 Liang Wang,1 Doudou Huang,1 Shi Qiu,1,* Xia Tao,2 Wansheng Chen1 1Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, People’s Republic of China; 2Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai, 200003, People’s Republic of China*These authors contributed equally to this workCorrespondence: Wansheng ChenInstitute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, People’s Republic of ChinaTel +64 021 81886182Email chenwansheng@shutcm.edu.cnXia TaoDepartment of Pharmacy, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai, 200003, People’s Republic of ChinaTel +64 021 81886182Email taoxia2003@126.comPurpose: Isatis indigotica (Ii) is a cruciferous herb that is widely distributed in China, and its roots and leaves have been used in two renowned antipyretic detoxicate crude drugs in Chinese Pharmacopoeia, Radix (R) and Folium (F) Isatidis. However, the pharmacodynamic material basis and underlying mechanisms of the herbal efficacy remained to be elucidated.Methods: Ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was adopted for the chemical profiling of R and F Isatidis. The active ingredients were screened out through the prediction of gastrointestinal absorption and druglikeness analysis using SwissADME. A herb-ingredient-target network was constructed through target prediction of the herbal active ingredients and anti-inflammation or anti-viral properties, followed by protein–protein interaction analysis. Then, the potential relevant signaling pathways were predicted by pathway enrichment. Finally, for verification, RAW 264.7 cell line was adopted to examine the anti-inflammatory and anti-viral activities of 6 representative ingredients in Ii.Results: Seventy-three compounds have been identified from Ii through UPLC-Q-TOF-MS. A total of 17 potential active ingredients were screened through pharmacokinetics and drug-likeness evaluation using SwissADME. It was shown that key targets might include TNF, AKT1, SRC, IL2, CASP9, and CASP3 in our herb-ingredient-target network, and isovitexin, a flavonoid, tended to participate in the inflammatory response, indoles were more likely to affect the cell proliferation processes, and lignans might have a broader affinity to key targets than the other active ingredients, such as regulating immune system (targeting IL-2) and PI3K-Akt signaling pathway. In vitro, indigo and secoisolariciresinol diglucoside markedly reduced TNF-α expression in Poly (I: C)-incubated cells. Isovitexin significantly inhibited TNF-α expression, and isatin treatment markedly reduced IL-1β expression in LPS-incubated cells.Conclusion: As the pharmacodynamics material basis of Ii, indoles, lignans, and flavonoids are believed to confer beneficial properties through various cellular aspects with multiple signaling pathways involved.Keywords: Isatis indigotica, UPLC-Q-TOF-MS, network analysis, bioassay analysis

The active components, mainly derived from secondary metabolites of medicinal plants, are the material basis for the efficacy of medicinal plants. Lignans, the secondary metabolites in plants with high bioactivity, are widely distributed in a variety of plant species, and their antiviral, antitumor, antibacterial, and antioxidant activities have been proved in clinical practice. Generally, lignans are diverse in structures with many chiral centers, and most of them are optically active. The biosynthesis of lignans depends on the oxidative coupling reaction through site selection and stereo selection, which impedes synthesized lignans to form racemates, but makes them in a three-dimensional configuration. Dirigent protein(DIR) plays an important role in guiding location selection and stereo selection of lignans in biosynthesis. In vitro studies on lignan biosynthesis have shown that racemic end products are obtained in the absence of DIR proteins, while the products in a three-dimensional configuration can be yielded in the presence of DIR proteins, indicating that DIR proteins play an asymmetric role in the biosynthesis of plant secondary metabolites. The present study reviewed the biolo-gical significance of DIR protein, the cloning of DIR gene, gene structure, catalytic mechanism, and the research progress in Isatis indigotica, Eucommia ulmoides, Forsythia suspensa, Salvia miltiorrhiza, Panax pseudoginseng var. notoginseng, and Schisandra chinensis, which provides a reference for the follow-up research of DIR gene.

Pinoresinol–lariciresinol reductases (PLRs) are enzymes involved in the lignan biosynthesis after the initial dimerization of two monolignols, and this represents the entry point for the synthesis of 8-8′ lignans and contributes greatly to their structural diversity. Of particular interest has been the determination of how differing substrate specificities are achieved with these enzymes. Here, we present crystal structures of IiPLR1 from Isatis indigotica and pinoresinol reductases (PrRs) AtPrR1 and AtPrR2 from Arabidopsis thaliana, in the apo, substrate-bound and product-bound states. Each structure contains a head-to-tail homodimer, and the catalytic pocket comprises structural elements from both monomers. β4 loop covers the top of the pocket, and residue 98 from the loop governs catalytic specificity. The substrate specificities of IiPLR1 and AtPrR2 can be switched via structure-guided mutagenesis. Our study provides insight into the molecular mechanism underlying the substrate specificity of PLRs/PrRs and suggests an efficient strategy for the large-scale commercial production of the pharmaceutically valuable compound lariciresinol., Pinoresinol–lariciresinol reductases (PLRs) are enzymes involved in the lignan biosynthesis. Here, crystal structures of three PLRs in the apo, substrate-bound and product-bound states, and accompanying mutagenesis provide insight into PLRs catalytic mechanism and suggest a strategy for PLR engineering.

The complete chloroplast genome of Isatis minima, a typical ephemeral plant of Brassicaceae in the Central Asia desert, was sequenced and characterized in this study. The genome 153,642 bp in size, contains a typical quadripartite genome organization including LSC and SSC regions of 83,423 bp and 17,709 bp, and two copies of the IR regions of 26,255 bp. It has 113 unique genes, including 79 protein-coding, 30 tRNA, and four rRNA genes. Phylogenetic analysis fully resolved I. minima in a monophyletic clade with I. tinctoria. This bioinformatic data contributes to the phylogenetics systematics and evolutionary history of Brassicaceae.

The pandemic disease COVID-19, caused by SARS CoV-2, has created a global crisis. Presently, researchers across the globe are in a quest to identify/develop drugs or vaccines by targeting different non-structural proteins (Nsps) of SARS CoV-2. One such important drug target is Nsp5/main protease (Mpro) which plays a critical role in the viral replication. This cysteine protease/Mpro of SARS CoV-2 has high sequence similarity with the same protease from SARS CoV-1. Previously, it has been shown experimentally that eight polyphenols derived from the root of Isatis indigotica show inhibitory effect on the cleavage/catalytic activity of the SARS CoV-1 Mpro. But whether these polyphenols exhibit any inhibitory effect on SARS CoV-2 Mpro is unclear. To explore this possibility, here, we have adopted various computational approaches. Polyphenols that qualified the pharmacological parameters (indigo, sinigrin, hesperetin and daidzein) and two well-known Mpro inhibitors (N3 and lopinavir) were subjected to molecular docking studies. Two of them (sinigrin and hesperetin) were selected by comparing their binding affinities with N3 and lopinavir. Sinigrin and hesperetin interacted with the two most important catalytic residues of Mpro (His41 and Cys145). Molecular dynamics studies further revealed that these two Mpro-polyphenol complexes are more stable and experience less conformational fluctuations than Mpro-N3/lopinavir complex. The Mpro-hesperetin complex was more compact and less expanded than Mpro-sinigrin complex. These findings were additionally validated by MM-GBSA analysis. As a whole, our study revealed that these two polyphenols may be potent SARS CoV-2 Mpro inhibitors and may possibly be considered for COVID-19 treatment., Graphical Abstract Communicated by Ramaswamy H. Sarma

APETALA3 (AP3) and PISTILLATA (PI) are B-class MADS-box floral homeotic genes of Arabidopsis and are involved in specifying the identity of petals and stamens. In the present work, IiAP3 and IiPI, the respective orthologous genes of AP3 and PI, were cloned from Isatis indigotica. By expressing in ap3-6 and pi-1 homozygous mutant and in wild-type Arabidopsis under the control of AP3 promoter or CaMV 35S promoter, we demonstrated that IiAP3 and IiPI were functionally equivalent to AP3 and PI of Arabidopsis. Referring to previous reports and the research results in the present work, expression patterns of AP3 and PI homologs are not the same in different angiosperms possessing diverse floral structures. It suggests that the alterations in expression may contribute to the changing morphology of flowers. To further determine the relationship between IiAP3 and IiPI, the coding sequences of the different structural regions in these two proteins were swapped with each other, and the data collected from transgenic Arabidopsis plants of the chimeric constructs suggested that MADS domain was irreplaceable for the function of IiAP3, K domain of IiAP3 was involved in specifying the identity of stamens, K domain of IiPI was mainly related to the formation of petals, and C-terminal region of IiPI was involved in characterization of stamens. In addition, a complete KC region of these two proteins was more effective in phenotypic complementation of the mutants.

Eleven new sulfonated alkaloids (

Chronic hepatitis induced by hepatitis B virus (HBV) infection is a serious public health problem, leading to hepatic cirrhosis and liver cancer. Although the currently approved medications can reliably decrease the virus load and prevent the development of hepatic diseases, they fail to induce durable off-drug control of HBV replication in the majority of patients. The roots of

E-class MADS-box genes, SEPALLATA (SEP), participate in various aspects of plant development together with B-, C- and D-class MADS-box genes. IiSEP4, a homologous gene of SEP4, was cloned from Isatis indigotica. IiSEP4 was highly expressed in sepals, and its mRNA was mildly detected in leaves, inflorescences, flowers, stamens and young silicles. Constitutive expression of IiSEP4 in Arabidopsis thaliana caused early flowering, accompanied by the reduction of flowers and floral organs. Moreover, the sepals in some flowers were transformed into carpelloid structures with stigmatic papillae, and obviously accompanied by ovule formation. Yeast two-hybrid assays demonstrated that IiSEP4 interacts with other woad MADS proteins to determine the identity of floral organs. These findings reveal the important roles of IiSEP4 in floral development of I. indigotica. The results of this study can lay a foundation for further study on biological functions of MADS transcriptional factors in I. indigotica.

In this study, five different glucosinolates including progoitrin, epiprogoitrin, gluconapine, glucoerucine, glucobrassicin were determined in different plant species by using HPLC. The main constituents of I. tinctoria and I. tinctoria subsp. corymbosa roots were progoitrin and glucobrassicin, while the other three species contained gluconapine and glucobrassicin. A higher amount of glucosinolate, phenolic, flavonoid substance and protein were detected in the plant roots than in the stems. The antioxidant activity of root extracts was found higher than the stem. The root and stem extracts of plants showed significant inhibition on gram-positive bacteria, but the only root extracts showed inhibition on gram negative bacteria. The oil content of I. aucherii, I. buschiana, I. candolleana, I. tinctoria and I. tinctoria subsp. Corymbosa seeds were 30.41%, 37.55%, 38.43%, 28.79% and 36.45%, respectively. Major fatty acid component of the all plant species studied were the unsaturated fats like oleic, linoleic, alpha-linolenic, cis-11 eicosenoic and erucic acid., Bu çalışmada, HPLC analizleri sonucunda bitkilerde doğal glukozinolatlardan progoitrin, epiprogoitrin, glukonapin, glukoerusin, glukobrassisin olmak üzere beş farklı glukozinolat farklı miktarlarda belirlenmiştir. I. tinctoria ve I. tinctoria subsp. corymbosa’da köklerinde ana bileşenler progoitrin ve glukobrassisin iken, diğer üç türde glukonapin ve glukobrassisin olarak bulunmuştur. Gövdeye oranla kökte daha yüksek oranda glukozinolat, fenolik, flavonoid madde ve protein miktarı tespit edilmiştir. Ayrıca kök ekstraktlarının antioksidan aktivitesinin gövdeye oranla daha yüksek olduğu bulunmuştur. Bitkilerin kök ve gövde ekstraktlarının her ikisi de gram pozitif bakterilerin büyümesini inhibe ederken, gram negatif bakterilerin büyümesini yalnızca kök ekstraktlarının inhibe ettiği görülmüştür. I. aucherii, I. buschiana, I. candolleana, I. tinctoria ve I. tinctoria subsp. corymbosa tohumlarının yağ içerikleri sırasıyla % 30.41, 37.55, 38.43, 28.79 ve 36.45 olarak elde edilmiştir. Çalışılan tüm türlerde yağın önemli bir kısmının doymamış yağ asitlerinden (oleik, linoleik, alfa-linolenik, cis-11 eikosenoik ve erusik asit) oluştuğu belirlenmiştir.

In this study, The crude extraction of Isatis spp. (I.tinctoria, I.busichaina and I. lusitanica) were investigated for their antifungal activity. Each of methanol, ethanol, and H2O was used as a solvent. Due to find the most effective part of each species, the plant parts were used separately. The aim of this research was to determine the effect of the natural product on two common pathogenic fungi Aspergillus niger & Candida albicans. The results show that most of the used plants have a significant effect on both used fungal species. Extracted each of flower, I. tinctoria was shown the best results comparing to the other used species. Extracted leaf and flower of I. tinctoria by methanol was show the best result on Aspergillus niger & Candida albicans. The growth zone was around 90mm for control and 61mm at 75% concentration of methanolic and ethanolic extraction. The flower was for followed by the stem. Depend on the results, the methanol was shown the highest number, then ethanol and the lowest inhibition zone area was for H2O extraction.

Silver nanoparticles (AgNPs) are widely used in many applications of biotechnology, including medicine and agriculture. They are released to the nature as waste materials, which can cause physiological and biochemical effects on plants. Indigo, indirubin and tryptanthrin are valuable indole alkaloid compounds in Isatis constricta due to both medicinal effects and dye properties. This research was conducted to determine the effects of different concentrations (0, 0.25, 0.5, 1, 1.5 and 2 mg L-1) of AgNPs on the production of indigo, indirubin and tryptanthrin compounds in leaves of in vitro grown shoots of I. constricta Davis. Indigo production was 1.15-fold of control (869 +/- 8.33 mu g g(-1)) in the leaves of shoots regenerated in Murashige and Skoog supplemented with 2 mg L-1 of AgNPs (1003 +/- 11.42 mu g g(-1)) on 5 days post-treatment. Tryptanthrin production showed an increase in all applications of AgNPs, but the highest increase was observed at a concentration of 2 mg L-1 (4.59 +/- 0.046 mu g g(-1)) and this increase was 1.71-fold of control (2.68 +/- 0.031 mu g g(-1)) on 5 days post-treatment. The production of indigo and tryptanthrin decreased on 10 and 15 days post-treatment with AgNPs. The contents of indirubin decreased during day 5-10-15 and at all concentrations of AgNPs compared to the control.

Isatis tinctoria L. (woad) has been used in medicine for centuries and has demonstrated anti-inflammatory effects. However, to date, no well-defined extracts with precise analysis of active substances have been developed. The aim of this study was to develop novel extracts of Isatis tinctoria L., and to characterize their active ingredients and anti-inflammatory properties. Various extracts of Isatis tinctoria L. were analysed for their active ingredients, and screened for anti-inflammatory effects using cyclooxygenase-2 activity assays. A petroleum ether extract was found to have the best effects, and was tested in a mouse model of acute allergic contact dermatitis. In the mouse model the petroleum ether extract resulted in significantly reduced ear swelling, oedema and inflammatory cell density. In mouse skin and human keratinocyte cultures, petroleum ether extract inhibited pro-inflammatory cytokine expression. Furthermore, human mast cell degranulation was significantly inhibited in LAD2 cell cultures. In conclusion, novel woad extracts were developed and shown to have anti-inflammatory properties in a contact hypersensitivity animal model and human keratinocytes. The production of such extracts and further characterization of their specific properties will enable determination of their potential dermatological effects in the treatment of inflamed and irritated skin.

(1) Background: In recent years, the porcine reproductive and respiratory syndrome virus (PRRSV) has become a virulent pathogen that has caused devastating diseases and economic losses worldwide in the swine industry.

Background Glucosinolates (GSLs) play important roles in defending against exogenous damage and regulating physiological activities in plants. However, GSL accumulation patterns and molecular regulation mechanisms are largely unknown in Isatis indigotica Fort. Results Ten GSLs were identified in I. indigotica, and the dominant GSLs were epiprogoitrin (EPI) and indole-3-methyl GSL (I3M), followed by progoitrin (PRO) and gluconapin (GNA). The total GSL content was highest (over 20 μmol/g) in reproductive organs, lowest (less than 1.0 μmol/g) in mature organs, and medium in fresh leaves (2.6 μmol/g) and stems (1.5 μmol/g). In the seed germination process, the total GSL content decreased from 27.2 μmol/g (of seeds) to 2.7 μmol/g (on the 120th day) and then increased to 4.0 μmol/g (180th day). However, the content of indole GSL increased rapidly in the first week after germination and fluctuated between 1.13 μmol/g (28th day) and 2.82 μmol/g (150th day). Under the different elicitor treatments, the total GSL content increased significantly, ranging from 2.9-fold (mechanical damage, 3 h) to 10.7-fold (MeJA, 6 h). Moreover, 132 genes were involved in GSL metabolic pathways. Among them, no homologs of AtCYP79F2 and AtMAM3 were identified, leading to a distinctive GSL profile in I. indigotica. Furthermore, most genes involved in the GSL metabolic pathway were derived from tandem duplication, followed by dispersed duplication and segmental duplication. Purifying selection was observed, although some genes underwent relaxed selection. In addition, three tandem-arrayed GSL-OH genes showed different expression patterns, suggesting possible subfunctionalization during evolution. Conclusions Ten different GSLs with their accumulation patterns and 132 genes involved in the GSL metabolic pathway were explored, which laid a foundation for the study of GSL metabolism and regulatory mechanisms in I. indigotica.

The screening of real features among thousands of ions remains a great challenge in the study of metabolomics. In this research, a workflow designed based on the MetaboFR tool and "feature-rating" rule was developed to screen the real features in large-scale data analyses. Seventy-four reference standards were used to test the feasibility, with 83.21% of real features being obtained after MetaboFR processing. Moreover, the full workflow was applied for systematic characterization of 14 species of the genus

Stress that can occur at different levels of a person's life can cause and exacerbate various diseases. Oxidative stress and inflammation underlie this process at the cellular level. There is an urgent need to identify new and more effective therapeutic targets for the treatment of stress-induced behavioral disorders and specific drugs that affect these targets.

The current study was conducted to investigate the role of sulfur (S) and reduced glutathione (GSH) in mitigating arsenic (As) toxicity in Isatis cappadocica and Erysimum allionii. These plants were exposed for 3 weeks to different concentrations (0, 400 and 800 μM) of As to measure fresh weight, total chlorophyll, proline and hydrogen peroxide (H

Background Isatidis Radix, the root of Isatis indigotica Fort. (Chinese woad) can produce a variety of efficacious compound with medicinal properties. The tetraploid I. indigotica plants exhibit superior phenotypic traits, such as greater yield, higher bioactive compounds accumulation and enhanced stress tolerance. In this study, a comparative transcriptomic and metabolomic study on Isatidis Radix autotetraploid and its progenitor was performed. Results Through the targeted metabolic profiling, 283 metabolites were identified in Isatidis Radix, and 70 polyploidization-altered metabolites were obtained. Moreover, the production of lignans was significantly increased post polyploidization, which implied that polyploidization-modulated changes in lignan biosynthesis. Regarding the transcriptomic shift, 2065 differentially expressed genes (DEGs) were identified as being polyploidy-responsive genes, and the polyploidization-altered DEGs were enriched in phenylpropanoid biosynthesis and plant hormone signal transduction. The further integrative analysis of polyploidy-responsive metabolome and transcriptome showed that 1584 DEGs were highly correlated with the 70 polyploidization-altered metabolites, and the transcriptional factors TFs-lignans network highlighted 10 polyploidy-altered TFs and 17 fluctuated phenylpropanoid pathway compounds. Conclusions These results collectively indicated that polyploidization contributed to the high content of active compounds in autotetraploid roots, and the gene–lignan pathway network analysis highlighted polyploidy–responsive key functional genes and regulators.

Isatis indigotica is a commercial medicinal crop that is widely cultivated with high water and nutrient application, in the arid areas of northwest China. Rational irrigation and nitrogen application are key factors for successful crop management. The objective of this study was to determine the effect of water and nitrogen coupling on the photosynthetic characteristics, yield, and quality of Isatis indigotica produced in northwestern China. Field trials were conducted for 2 consecutive years at an irrigation test station. Data on photosynthetic parameters, yield, and quality were collected from individual Isatis indigotica for each treatment during 2018–2019. The application of nitrogen significantly increased photosynthetic rates and yield under the same irrigation conditions. However, the yields were reduced in the excess water treatments (W3N1 and W3N2) and in the excess nitrogen treatments (W1N3, W2N3, and W3N3) in contrast to the optimum W2N2 treatment. Moreover, the quality indicators of the W2N2 treatment decreased compared with CK, which was due to water stress and more photoassimilates being available to the roots, but the effective quality index value could be effectively improved by greatly increasing the yield.

The research was aimed to identify fruits morphological characteristics of 33 taxa belonging to the Lepidieae tribe from Brassicaceae. The characteristics of the fruit's general shape, apex shape, dimensions, colors, surface ornamentation, and beak were determined, the results indicated the importance of each of those characters in isolated species, In general, all the tribe's fruits were open capsule and short silicula. According to the dimensions of fruit, three species can be distinguished as a group with dimensions greater than 20mm, as in Brossardia, Coluteocarpus, and Didymophysa, and depending on the shape, it was possible to differentiate the species within this group, while the other species were less than 20 mm. in dimensions. They were distinguished by the general shapes, as the fruits of the Aethionema species were distinguished by their winged shaped, whereas the fruits of the genus Biscutella are bi-lobed, and the fruits have an inverted semi-triangular shape in the Capsella and peltate with prominent veins in the genus Horwoodia, or maybe peltate and longitudinally splintered with dark veins in Iberis, while the genus Sameraria fruit is circular to semi-circular characterized by intermediate appendages, while the individuals of the genus Isatis have linear-oblong fruits, and the species of both genus Lipidium and Hymenolobus were distinguished by their small and elliptic fruits and could be differentiated according to other fruits features. So could be conclude the importance of the characteristic of the general shape of the fruit firstly in species isolation then size, ornamentation, and some other traits.

Adjuvants have been used in vaccines for a long time to promote the body's immune response, reducing vaccine dosage and production costs. Although many vaccine adjuvants are developed, the use in human vaccines is limited because of either limited action or side effects. Therefore, the development of new vaccine adjuvants is required. Many studies have found that natural polysaccharides derived from Traditional Chinese medicine (TCM) possess good immune promoting effects and simultaneously improve humoral, cellular and mucosal immunity. Recently polysaccharide adjuvants have attracted much attention in vaccine preparation because of their intrinsic characteristics: immunomodulation, biocompatibility, biodegradability, low toxicity and safety. This review article systematically analysed the literature on polysaccharides possessing vaccine adjuvant activity from TCM plants, such as Astragalus polysaccharide (APS), Rehmannia glutinosa polysaccharide (RGP), Isatis indigotica root polysaccharides (IRPS), etc. and their derivatives. We believe that polysaccharide adjuvants can be used to prepare the vaccines for clinical use provided their mechanisms of action are studied in detail.

Water-soluble polysaccharides were isolated from the leaves and roots of Isatis indigotica Fort., and their structural features were studied and compared. One neutral polysaccharide fraction (WFIP-N) and three pectin fractions (WFIP-A-A, WFIP-A-B and WFIP-A-C) were obtained from the leaves, and one neutral polysaccharide fraction (WRIP-N) and two pectin fractions (WRIP-A-A and WRIP-A-B) were obtained from the roots. WFIP-A-B (Mw = 34.6 kDa) and WRIP-A-B (Mw = 29.9 kDa) were the major pectic polysaccharides. Monosaccharide composition, FT-IR, enzymatic hydrolysis, NMR and methylation analysis indicated that both WFIP-A-B and WRIP-A-B are composed of rhamnogalacturonan I (RG-I), rhamnogalacturonan II (RG-II) and homogalacturonan (HG) domains with mass ratios of 1.5:1.0:0.4 and 0.3:1.0:1.7, respectively. WFIP-A-B and WRIP-A-B were found to be rich in RG-I and HG domains, respectively, and mainly contained type II arabinogalactan (AG-II) and α-L-1,5-arabinan side chains, but those in WRIP-A-B were more numerous and longer. Our results provide structural features and differences between these polysaccharides which will help to elucidate their functional differences.

Seven alkaloids including five undescribed ones (1a/1b, 2, 3 and 5) were obtained from the leaves of Isatis indigotica Fortune. Their structures were established by extensive spectroscopic analyses. The absolute configurations of compounds 1a, 1b, 3 and 5 were determined by comparison of the experimental and calculated electronic circular dichroism (ECD) spectra. Subsequently, the neuroprotective effects of all the isolates against H2O2-induced injury in SH-SY5Y cells were evaluated in vitro by MTT assay. Moreover, Annexin V-FITC/PI double staining was performed, while the activities of antioxidant enzymes (SOD, CAT and GSH-Px) for compounds 1a and 1b were measured.

Six pairs of alkaloid enantiomers including 11 new alkaloids (1a/1b–5a/5b, 6a) were isolated from the leaves of Isatis tinctoria. Their structures were established by extensive spectroscopic analyses. Enantiomers were separated successfully by chiral chromatographic column and the absolute configurations of all isolates were determined by comparison of experimental and calculated electronic circular dichroism spectra. The isolated alkaloids were evaluated for their neuroprotective activities against H2O2-induced cell injury in human neuroblastoma SH-SY5Y cells. The results showed that 5a/5b and 6a/6b exhibited potent neuroprotective activities at 50 µM compared with the H2O2-treated group.

International audience; The pharmacologically active dichloromethane extracts of dried woad leaves (Isatis tinctoria L.), and the methanol extracts of comparable fresh leaves of the same plants, were analyzed by LC-MSn. The fresh leaf metabolite profile revealed a complex pattern of indolic compounds. Besides the known indigo precursors, isatan A, isatan B and indican, seven previously unreported indole derivatives were characterized: acetylindican, malonylindican, two dioxindole glucosides, dioxindole malonylglucoside, 6-hydroxyindole-3-carboxylic acid 6-O-glucoside and 6-hydroxyindole-3-carboxylic acid glucose ester. The integration of 122 compounds in fresh leaves and of five selected compounds (indoxyl, isatin, indigo, indirubin, and tryptanthrin) in dried leaves, formed the input data for a stepwise modelling procedure generating five predictive linear models. The structure of the predictive models and a cross validation provide evidence that the models could predict well or moderately well the accumulation of the selected lipophilic compounds, and were simple enough to be used in a woad cultivation program. PLS regression models relating each of the five selected dry leaf indolics to the fresh leaf metabolome were then fitted in order to deduct potential precursors and mechanisms leading to the formation of these lipophilic indolics in drying woad leaves. The models suggested glucobrassicin, isatan A and isatan B as the main candidate precursors of these compounds, besides a minor contribution of other fresh leaf indolics, including malonylindican, actylindican and dioxindole malonylglucoside. Dioxindole malonylglucoside was identified here as isatan C. The models further suggested that the accumulation of phenylpropanoid antioxidants in woad leaves has a negative impact on the formation of indoxyl, isatin, indigo, indirubin and tryptanthrin.

A hydroponic experiment was conducted to explore the interactive effects of selenium (Se) supplementation (0, 5, and 10 μM) and arsenic (As) toxicity (0, 200, and 400 μM) on the growth, accumulation, and oxidative damage along with defense mechanisms of metallicolous (MP) and non-metallicolous population (NMP) of Isatis cappadocica, an As-hyperaccumulator, and Brassica oleracea as reference brassica. The results revealed that As stress significantly hampered plant growth particularly in B. oleracea. It reduced plant growth due to enhanced oxidative load of As-stressed plants. Between the two Isatis populations, metallicolous plants accumulated significantly higher As, however with considerably low growth defects. Furthermore, Se supplementation counteracted the adverse effects of stress on growth and physiological performance of all studied plants. Addition of Se, particularly at higher dose (10 μM), significantly suppressed root As uptake and slightly its accumulation in shoots of B. oleracea plants treated with 400 μM As, and thus improved growth characteristics of stressed plants. Under As stress, Se supplementation increased the activities of enzymatic (peroxidase (POD) and glutathione reductase (GR)) and non-enzymatic (anthocyanins and total flavonoids) antioxidants, thereby suggesting relieved As stress by reduced oxidative damage. Taken together, these results support the beneficial role of Se in the regulation of As stress by improving growth, physiology, and antioxidant capacity, and highlight its significance for plants grown on such metal-contaminated soils.

WOS: 000464004500002 Isatis species, which are endemic across most of Turkey, exhibit antibacterial, anticancer and antiviral effects. the aim of this study was to determine of antioxidant, antimicrobial, tyrosinase inhibitor, cytotoxic activities, and phenolic profile of polar extracts of Isatis cappadocica Desv. We analyzed the antioxidant properties of extracts using total phenolic content (TPC), ferric reducing antioxidant power (FRAP), cupric reducing antioxidant capacity (CUPRAC), and 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging activity. the phenolic composition of methanolic extract of I. cappadocica was analyzed by reverse phase high performance liquid chromatography (RP-HPLC). Extracts' antimicrobial properties were evaluated based on the agar well diffusion technique. Tyrosinase inhibitory activity was measured colorimetrically. Methanolic extract yielded better FRAP and CUPRAC results and aqueous extract yielded better DPPH activity. Benzoic acid, and sinapic acid were detected as major phenolic compounds. Methanolic extract was particularly effective against all the bacteria investigated, apart from Yersinia pseudotuberculosis. Methanol extract was exhibited tyrosinase inhibitory activity. the methanol extract has caused to death of cells by dosage in the high concentrations cytotoxic activity on the PC-3 and 3T3 cell lines. the results showed that I. cappadocica could be used as a natural source in the food, cosmetic, and drug industries due to their rich antioxidant, antimicrobial, cytotoxic and tyrosinase inhibitor activities.

Isatidis Radix is the dried root of the Isatis indigotica, with pharmacological effects such as heat-clearing and detoxification, cooling blood and pharyngeal relief, antibacterial and anti-inflammatory effects. It is often used clinically to prevent and treat influenza and other diseases. In this paper, relevant domestic and foreign literatures in recent years were summarized, and it was found that Isatidis Radix lignans, indole alkaloids, polysaccharides, etc. were the main active components against influenza virus. Then its pharmacological effects and the mechanism of action were reviewed, providing a basis for in-depth research on the antiviral effect of Isatidis Radix.

The root of Isatis indigotica is used as a traditional Chinese medicine (termed Isatidis Radix) due to its antiviral effects. We examined compounds isolated from Isatidis Radix and elucidated the structures of three new natural alkaloids, and we examined the possible mechanisms or active targets of indole alkaloids occurring in blood of rats treated by gavage. Three new natural products were isolated from Radix Isatidis for the first time, including 1-methoxy-2-indoleacetonitrile, 1-hydroxy-3-indoleacetonitrile, 8-Methoxy-1, 2-dihydroquinoline, and 4 compounds isolated from this medicinal material for the first time. Their structures were elucidated using nuclear magnetic resonance. The components of Isatidis Radix were analysed using liquid chromatography tandem mass spectrometry, and 33 compounds were detected in plasma of treated rats; 24 of these compounds were indole alkaloids, and they included the newly identified compounds. Molecular docking and in vitro antithrombin activity tests showed HA inhibition activity of indoleacetonitriles.

Objectives Isatis indigotica Fort. (I. indigotica) is an herbaceous plant belonging to Cruciferae family. Its leaf (IIL) and root (IIR) are commonly used in traditional Chinese medicines (TCMs) with good clinical efficacies such as clearing away heat and detoxification, cooling blood and reducing swelling. This review aimed to provide a systematic summary on the phytochemistry, pharmacology and clinical applications of I. indigotica. Key findings This plant contains alkaloids, organic acids, flavonoids, lignans, nucleosides, amino acids, and steroids. Previous pharmacological researches indicated that I. indigotica possesses promising antivirus, antibacterial, immunoregulatory, anti-inflammation, and cholagogic effects. Importantly, it can inhibit various viruses, such as influenza, hepatitis B, mumps, herpes simplex, cytomegalovirus, and coxsachievirus. Clinically, it is frequently used to treat various viral diseases like viral influenza, parotitis and viral hepatitis. Consequently, I. indigotica may be beneficial for the prevention and treatment of coronavirus disease 2019 (COVID-19). Summary This paper reviewed the chemical constituents, pharmacological effects and clinical applications of I. indigotica which may guide further research and application of this plant.

In this paper, through the collection and collation of ancient herbs, medical books and prescriptions, combined with modern literature, the historical changes of the name, origin, position, medicinal parts, collection, processing and processing of bluegrass were systematically combed and verified.It can be seen from the research that bluegrass was first used as medicine by the fruit, namely blueberry, which was originally Polygonum tinctorium. Since the Ming and Qing Dynasties, blueberry was rarely used, and it has been no longer used medicinally. In the Wei and Jin Dynasties, the medicinal parts extended to the stems and leaves, and most of them used juice as medicine.Since the Tang Dynasty, origin has been extended to Isatis indigotica, Baphicacanthus cusia, Indigofera tinctoria, Compositae plant Wulan, etc. In the Song Dynasty, the medicinal parts extended to the roots, and thequot;Banlangenquot; began to appear, and gradually became the main medicinal parts of blue medicinal materials, the main base of which was B. cusia. Since the Qing Dynasty, I. indigotica, a Cruciferae, has gradually become a genuine indigo root, while B. cusia has become a southern indigo root. It was the first mineral dye imported from abroad for thrush, and then used as medicine, also known as clam powder. Because it was found that it had the same effect with the extract of bluegrass, it was also named indigo naturalis in China, which has lasted till now. The main stream of Isatidis Folium in the past dynasties is the dry stem and leaf of Clerodendrum cyrtophylum. Since the Qing Dynasty, the stem and leaf of Isatis indigotica, P. tinctorium and other blue grasses have been gradually mixed as substitutes and gradually become the mainstream.

The present work was conducted to assess the effects of arsenic (As, 1000 µM), diphenyleneiodonium (DPI, 10 µM) and reduced glutathione (GSH, 500 µM) on

Ionic liquids (ILs1) which are called “green solvents”, are used widely in the textile industry as adjuvants due to their many advantages. However, their persistent residues may cause ecotoxicity. The aim of the study is to explore the toxicity of different anions on imidazole ILs and their toxicological mechanism. For the experiments 1-butyl-3-methylimidazole tetrafloroborate ([C4mim]BF4) and 1- butyl -3-methylimidazolium chloride ([C4mim]Cl) were selected to study their toxic effects on Isatis tinctoria. ILs may affect the germination rate. Fresh weight, dry weight and Hill reaction activity decreased continuously with increasing of IL concentrations, showing an effect-dose relationship. Transmission electron microscopy (TEM) revealed that cell walls were fuzzy, starch granules had accumulated and the chloroplast structure was damaged. These changes will affected the function and electron transport efficiency of photosystemⅡ. Superoxide anion accumulation stimulated the activity of antioxidant enzymes (SOD, POD, CAT) and caused lipid peroxidation as well as an increased malondialdehyde content. ILs also reduced indirubin and total flavonoids contents, which reduced the pharmacological efficacy of Isatis tinctoria. This is demonstrated by three-dimensional fluorescence chromatogram. [C4mim]Cl was more toxic than [C4mim]BF4. ILs caused toxic effects to Isatis tinctoria. The ecological toxicity of ILs should be considered when using them as additives in the textile industry.

Traditional Chinese Medicine (TCM) belongs to the most elaborate and extensive systems of plant-based healing. The herb Northern Ban Lan (Isatis tinctoria) is famous for its antiviral and anti-inflammatory activity. Although numerous components isolated from I. tinctoria have been characterised so far, their modes of action have remained unclear. Here, we show that extracts from I. tinctoria exert anti-microtubular activity. Using time-lapse microscopy in living tobacco BY-2 (Nicotiana tabacum L. cv Bright Yellow 2) cells expressing GFP-tubulin, we use activity-guided fractionation to screen out the biologically active compounds of I. tinctoria. Among 54 fractions obtained from either leaves or roots of I. tinctoria by methanol (MeOH/H2 O 8:2), or ethylacetate extraction, one specific methanolic root fraction was selected, because it eliminated microtubules efficiently and rapidly. By combination of further purification with ultra-high performance liquid chromatography (UHPLC) and high-resolution (tandem) mass spectrometry (HRMS (/MS)) most of the bio-activity could be assigned to the glucosinolate compound glucobrassicin. Glucobrassicin can also affect microtubules and induce apoptosis in HeLa cells. In the light of these findings, the anti-viral activity of Northern Ban Lan is discussed in the context of microtubules being hijacked by many viral pathogens for cell-to-cell spread. This article is protected by copyright. All rights reserved.

Isatis cappadocica is a well-known arsenic-hyperaccumulator, but there are no reports of its responses to cadmium (Cd). Nitric oxide (NO) is a signaling molecule, which induces cross-stress tolerance and mediates several physio-biochemical processes related to heavy metal toxicity. In this study, the effects of Cd and sodium nitroprusside (SNP as NO donor) on the growth, defense responses and Cd accumulation in I. cappadocica were investigated. When I. cappadocica was treated with 100 and 200 μM Cd, there was an insignificant inhibition of shoot growth. However, Cd stress at Cd400 treatment decreased significantly the dry weight of root and shoot by 73 and 38%, respectively, as compared to control. The application of SNP significantly improved the growth parameters and mitigated Cd toxicity. In addition, SNP decreased reactive oxygen species (ROS) production induced by Cd. The increased total thiol and glutathione (GSH) concentrations after SNP application may play a decisive role in maintaining cellular redox homeostasis, thereby protecting plants against oxidative damage under Cd stress. Bovine hemoglobin (Hb as NO scavenger) reduced the protective role of SNP, suggesting a major role of NO in the defensive effect of SNP. Furthermore, the reduction in shoot growth and the increase of oxidative damage were more severe after the addition of Hb, which confirms the protective role of NO against Cd-induced oxidative stress. The protective role of SNP in decreasing Cd-induced oxidative stress may be related to NO production, which can lead to stimulation of the thiols synthesis and improve defense system.

Background Pseudorabies virus (PRV) is an animal virus that is globally responsible for the high economic losses in the swine industry. Isatis root is a traditional Chinese medicinal herb that possesses immune-enhancing and antiviral properties. However, the molecular mechanisms underlying the effects of the active component of the isatis root polysaccharide (IRPS) extract on immature dendritic cells remain elusive. Methods In this study, we investigated the molecular changes in primary porcine peripheral blood monocyte-derived dendritic cells (MoDCs) during PRV infection, using enzyme-linked immunosorbent assay (ELISA) and quantitative reverse transcription-polymerase chain reaction. Additionally, we studied the effect of IRPS on PRV-infected DCs. Results The results showed that IRPS stimulated the maturation of MoDCs, induced IL-12 secretion, and downregulated IL-6 expression. Conclusions Collectively, these results suggest that IRPS is a promising candidate for promoting maturation of DCs and enhancing their secretory potential after PRV infection.

Isatis indigotica Fort. (family Cruciferae), is an herb widely used in traditional herbal medicine and its dried leave was named "ISATIDIS FOLIUM". Baphicacanthus cusia (Ness) Bremek. and Polygonum tinctorium Ait. are commonly misused as ISATIDIS FOLIUM in Chinese Medicine pharmacy. For the purpose of being not misused, specific primers based on the sequence difference of chloroplast trnH-psbA intergenic spacer were designed and multiplex polymerase chain reaction method (multiplex PCR) was developed. In this study, 29 original herbal materials were analyzed and our results show that DNA size after multiplex PCR was able to distinguish variations between three herbs. DNA fragments of 464, 297, 170 base pairs (bps) were represented for I. indigotica and B. cusia and P. tinctorium, respectively. In conclusion, our investigations demonstrate that molecular identification method provides more accurate results for medicinal plants detection and good quality control of ISATIDIS FOLIUM.

Widespread arsenic (As) contamination is a severe environmental and public health concern. Isatis cappadocica, an arsenic hyperaccumulator, holds great potential to clean up As-contaminated soil and groundwater. Iron oxide is one of the most common metal oxides in the natural environment and its nanoparticulate form has been previously utilized for the removal of heavy metals/metalloids from wastewater. However, there is a paucity of information on the impact of iron oxide nanoparticles on the growth and physiological properties of I. cappadocica and its effectiveness on As removal. Current study reports for the first time the impact of superparamagnetic iron oxide nanoparticles and glutathione (GSH) modified superparamagnetic iron oxide nanoparticles (nFe3O4 and nFe3O4@GSH) on the physiological characteristic of I. cappadocica and its accumulation of As under hydroponic condition. nFe3O4@GSH alleviated the harmful impact of As and significantly increased the shoot biomass of I. cappadocica by enhancing the plant defense mechanisms. The application of GSH, nFe3O4 and nFe3O4@GSH all lowered the As concentration in plant shoots as a protective mechanism. However, the substantial shoot biomass increase due to nFe3O4@GSH resulted in a 56% higher As accumulation in plant shoots than in plants exposed to As alone, indicating the strong effectiveness of nFe3O4@GSH as a novel enhancer of the As phytoremediation by I. cappadocica. Our data further showed that the beneficial effect of nFe3O4@GSH on As phytoremediation is due to the enhancement of activities of several enzymatic and nonenzymatic antioxidants.

Glucosinolates (1-5) are important secondary metabolites found in Isatis indigotica roots. Due to their high hydrophilic and ionic nature, purified glucosinolates often contain salt impurities and moisture. Accurate assessment of their purities is important for glucosinolates being utilised as chemical markers.To develop and validate quantitative proton (Several NMR parameters such as pulse program, relaxation time, and delay time were optimised. Three qHNMR methods were developed using gluconapin (3), neoglucobrassicin (4), and sinigrin (5) for method validation and with maleic acid as internal standard.The quantification was based on the integrated area ratios of an olefinic proton (H-4 for 1-3; H-6 for 4; and H-3 for 5) of the side chain from glucosinolates relative to the olefinic proton from the internal standard using deuterated water (DThe purity of glucosinolates isolated from I. indigotica and commercial sinigrin was accurately assessed using the developed qHNMR method. The qHNMR provides a reliable and superior means to determine the purity of glucosinolates.

Chemical fractionation of the ethanolic extract of the roots of