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This study aimed to analyze the effects of salt stress on the growth physiology and plant-cell ultrastructure of Isatis indigotica Fort. (I. indigotica) to evaluate its adaptability under salt stress. The effects of different concentrations of salt (NaCl; 0, 25, and 300 mmol·L−1) on the agronomic traits, activities of related enzymes, ion balance, and mesophyll-cell ultrastructure of I. indigotica were studied in a controlled pot experiment. Results showed that compared with those of the control group, the aerial-part fresh weight, underground fresh weight, tiller number, root length, root diameter, plant height, and leaf area of salt-stressed I. indigotica increased at 25 mmol·L−1 and then decreased at 300 mmol·L−1. The changes in levels of superoxide dismutase, peroxidase, ascorbate peroxidase, and catalase showed a similar trend, with significant differences compared with control group. Salt stress altered the ion balance of I. indigotica, resulting in a significant increase in Na+ content and a significant decrease in K+ content. The contents of Ca2+ and Mg2+ changed to varying degrees. The analysis of the microstructure of the root showed that under salt treatment, the epidermal cells of the root significantly thickened and the diameter of the xylem decreased. The results of ultrastructural analysis of mesophylls showed that salt stress can cause cell-membrane contraction, cell-gap enlargement, disorder in the structures of chloroplasts and mitochondria, and an increase in the number of osmiophilic particles. These changes were aggravated by the increase in NaCl concentration. This study reveals the response of I. indigotica to salt stress and provides a basis for further study on the salt-tolerance mechanism of I. indigotica. [ABSTRACT FROM AUTHOR]

Polysaccharides have been assessed as a potential natural active component in Chinese herbal medicine with anti-inflammatory properties. However, the complex and indefinite structures of polysaccharides limit their applications. This study explains the structures and anti-inflammatory potentials of three neutral polysaccharides, RIP-A1 (Mw 1.8 × 104 Da), RIP-B1 (Mw 7.4 × 104 Da) and RIP-B2 (Mw 9.3 × 104 Da), which were isolated from the roots of Isatis indigotica Fort. with sequenced ultrafiltration membrane columns, DEAE-52 and Sephadex G-100. The planar structures and microstructures of RIP-A1, RIP-B1 and RIP-B2 were further determined by HPGPC, GC–MS, methylation analysis, FT-IR, SEM and AFM, in which the structure of RIP-A1 was elucidated in detail using 1D/2D NMR. The Raw 264.7 cells were used for the anti-inflammatory activity in vitro. The results showed that RIP-A1, RIP-B1 and RIP-B2 are all neutral polysaccharides, with RIP-A1 having the smallest Mw and the simplest monosaccharide composition of the three. RIP-A1 is mainly composed of Ara and Gal, except for a small quantity of Rha. Its main structure is covered with glycosidic linkages of T-α-Araf, 1,2-α-Rhap, 1,5-α-Araf, T-β-Galp, 1,2,4-α-Rhap, 1,3,5-α-Araf and 1,6-β-Galp with 0.33:0.12:1.02:0.09:0.45:11.41:10.23. RIP-A1 significantly inhibited pro-inflammatory cytokines (NO, TNF-α, IL-6 and IL-1β) and increased anti-inflammatory cytokines (IL-4) in LPS-stimulated RAW 264.7 cells. Moreover, RIP-A1 could significantly inhibit the mRNA expression of TNF-α, IL-6 and L-1β. It could also activate IKK, p65 and IκBα (the components of the NF-κB signaling pathway). In conclusion, the above results show the structural characterization and anti-inflammatory potentials of RIP-A1 as an effective natural anti-inflammatory drug. [ABSTRACT FROM AUTHOR]

This study was to investigate the effects of three exogenous substances on chemical constituents of Isatis indigotica leaves and their efficacy in alleviating drought stress, and explore the methods of applying exogenous substances to efficient cultivation of Isatis indigotica. Polyethylene glycol (PEG) was used to simulate drought stress to deal with seeds of Isatis indigotica at the germination stage (concentration: 0, 10%, 15%, and 20%). Simultaneous operation of exogenous growth regulators [microbial inoculum (MI), γ-aminobutyric acid (GABA) and salicylic acid (SA)] and PEG were implemented in seeds of Isatis indigotica. The effects of drought stress and the mitigation of exogenous substances were observed by statistics of seed germination potential, germination rate, hypocotyl length, and radicle length of each treatment. The effects of exogenous substances on the content of alkaloids, crude protein and free amino acids in the leaves of Isatis indigotica grown in a greenhouse were determined after spraying exogenous substances on the plants. The differences of germination potential, germination rate, hypocotyl length, and radicle length among 15% PEG stress treatment, 10% PEG stress treatment and the control were significant (P<0.05). According to the predesigned germination standard, the seeds did not germinate under 20% PEG stress treatment. When the PEG concentration was 15%, the results of seed germination potential and germination rate after adding MI were significantly different from those under stress alone (P<0.05). When exposed to 10% PEG stress, the supplementation of GABA led to a notable increase in radicle length of Isatis indigotica seeds, showing significant differences compared to other three treatments. The application of MI and GABA under 15% PEG stress resulted in a significant increase in the radicle and hypocotyl length of Isatis indigotica seeds compared to other two treatments. The content of the total alkaloids in leaves of Isatis indigotica was significantly increased after spraying GABA. Meanwhile, the contents of crude protein and the total free amino acids were kept constant after spraying exogenous substances. Application of MI and GABA could alleviate drought stress of Isatis indigotica. The content of the total alkaloids in leaves of Isatis indigotica could significantly increase after spraying GABA. [ABSTRACT FROM AUTHOR]

Introduction: This study evaluated the effects of Isatis Leaf (ISL) on the growth performance, gastrointestinal tissue morphology, rumen and intestinal microbiota, rumen, serum and urine metabolites, and rumen epithelial tissue transcriptome of fattening sheep. Methods: Twelve 3.5-month-old healthy fattening sheep were randomly divided into two groups, each with 6 replicates, and fed with basal diet (CON) and basal diet supplemented with 80 g/kg ISL for 2.5 months. Gastrointestinal tract was collected for histological analysis, rumen fluid and feces were subjected to metagenomic analysis, rumen fluid, serum, and urine for metabolomics analysis, and rumen epithelial tissue for transcriptomics analysis. Results: The results showed that in the ISL group, the average daily gain and average daily feed intake of fattening sheep were significantly lower than those of the CON group (P < 0.05), and the rumen ammonia nitrogen level was significantly higher than that of the CON group (P < 0.01). The thickness of the reticulum and abomasum muscle layer was significantly increased (P < 0.05). At the genus level, the addition of ISL modified the composition of rumen and fecal microorganisms, and the relative abundance of Methanobrevibacter and Centipeda was significantly upregulated in rumen microorganisms, The relative abundance of Butyrivibrio, Saccharofermentans, Mogibacterium, and Pirellula was significantly downregulated (P < 0.05). In fecal microorganisms, the relative abundance of Papillibacter, Pseudoflavonifractor, Butyricicoccus, Anaerovorax, and Methanocorpusculum was significantly upregulated, while the relative abundance of Roseburia, Coprococcus, Clostridium XVIII, Butyrivibrio, Parasutterella, Macellibacteroides, and Porphyromonas was significantly downregulated (P < 0.05). There were 164, 107, and 77 different metabolites in the rumen, serum, and urine between the ISL and CON groups (P < 0.05). The differential metabolic pathways mainly included thiamine metabolism, niacin and nicotinamide metabolism, vitamin B6 metabolism, taurine and taurine metabolism, beta-Alanine metabolism and riboflavin metabolism. These metabolic pathways were mainly involved in the regulation of energy metabolismand immune function in fattening sheep. Transcriptome sequencing showed that differentially expressed genes were mainly enriched in cellular physiological processes, development, and immune regulation. Conclusion: In summary, the addition of ISL to the diet had the effect of increasing rumen ammonia nitrogen levels, regulating gastrointestinal microbiota, promoting body fat metabolism, and enhancing immunity in fattening sheep. [ABSTRACT FROM AUTHOR]

Lignans are a powerful weapon for plants to resist stresses and have diverse bioactive functions to protect human health. Elucidating the mechanisms of stereoselective biosynthesis and response to stresses of lignans is important for the guidance of plant improvement. Here, we identified the complete pathway to stereoselectively synthesize antiviral (−)-lariciresinol glucosides in Isatis indigotica roots, which consists of three-step sequential stereoselective enzymes DIR1/2, PLR, and UGT71B2. DIR1 was further identified as the key gene in respoJanuary 2024nse to stresses and was able to trigger stress defenses by mediating the elevation in lignan content. Mechanistically, the phytohormone-responsive ERF transcription factor LTF1 colocalized with DIR1 in the cell periphery of the vascular regions in mature roots and helped resist biotic and abiotic stresses by directly regulating the expression of DIR1. These systematic results suggest that DIR1 as the first common step of the lignan pathway cooperates with PLR and UGT71B2 to stereoselectively synthesize (−)-lariciresinol derived antiviral lignans in I. indigotica roots and is also a part of the LTF1-mediated regulatory network to resist stresses. In conclusion, the LTF1-DIR1 module is an ideal engineering target to improve plant Defenses while increasing the content of valuable lignans in plants. Proposed working model depicting the roles of LTF1-DIR1 module in lignan stereoselective biosynthesis and stress defense. [Display omitted] [ABSTRACT FROM AUTHOR]

Arsenic (As) toxicity hinders plant growth and productivity and poses human health concerns. However, silicon (Si) nutrition can help improve plant tolerance to abiotic stresses. This study evaluated the influence of Si application on plant growth and As accumulation in an As hyperaccumulator plant, Isatis (Isatis cappadocica Desv.) under As stress. Isatis seedlings were exposed to 0, 5, 25, 125, and 625 μM Na2HAsO4.7H2O (AsV) and NaAsO2 (AsIII) and then Si was applied as Na2SiO3 (1 and 2 mM). Plant exposure to As(V: 650 μM) and As(III: 125 µM) caused significant reduction in plant fresh weight by 31 and 42%, and photosynthetic pigments by 47 and 62%, respectively. The same treatments caused 37- and 4.2-fold increase in levels of reactive oxygen species. However, exogenous application of Si effectively mitigated As stress, and caused a decrease in shoot As concentration up to 15% and 21% in As(V) and As(III) treated plants, respectively. In this regard, Si application at 1 mM level was more effective. The Si-induced tolerance against As in Isatis was attributed to the activation of antioxidant defense system (increase in the activities of gluthatione-S-transferase and guaiacol peroxidase, carotenoids, proline and anthocyanin contents). In conclusion, Si application improved the As tolerance in Isatis plants by modulating plant growth, decreasing As accumulation and activating the antioxidant defense system. [ABSTRACT FROM AUTHOR]

Pre‐release host range assessments of weed biocontrol agent (BCA) candidates typically rely on no‐choice and choice feeding, oviposition, and development tests. However, these tests may exclude potentially environmentally safe BCA candidates from consideration if they develop on nontarget plant species in no‐choice tests that they would not colonize in the field because of behavioral barriers during host selection. Here, we examined the behavioral responses of Ceutorhynchus rusticus Gyllenhal (Coleoptera: Curculionidae) to olfactory and visual plant cues of nine native North American and two Eurasian confamilial plant species of invasive Eurasian mustard dyer's woad, Isatis tinctoria L. (Brassicaceae). In behavioral bioassays with olfactory, visual, and combined plant cues of one plant species vs. purified air and/or empty arms (control treatments), weevils responded with attraction to all offered I. tinctoria plant cues but showed no preference for most nontarget plant species relative to control treatments. The weevil also responded faster in these bioassays to I. tinctoria cues when compared to most nontarget species. In bioassays with olfactory, visual, or combined cues of nontarget species vs. those of I. tinctoria, C. rusticus preferred I. tinctoria over most nontarget plant species (olfactory cues: eight of 11, visual cues: two of 11, and combined cues: 11 out of 11 nontarget plant species). The results also suggest that C. rusticus mainly uses olfactory or combined olfactory and visual plant cues rather than visual cues alone in the early stages of host finding. Additionally, the findings suggest that during host finding, C. rusticus prefers cues of its Eurasian field host over those of nontarget species, reducing risks of post‐release nontarget attack in the field. Assessing behavioral responses to multimodal cues can advance our understanding of BCA candidates' host discrimination and facilitate the accurate interpretation of conventional no‐choice and choice feeding and development data during pre‐release environmental safety assessments. [ABSTRACT FROM AUTHOR]

Background: Isatis tinctoria Linnaeus and Isatis indigotica Fortune are very inconsistent in their morphological characteristics, but the Flora of China treats them as the same species. In this work, a new technology that differs from conventional barcodes is developed to prove that they are different species and to clarify their classification. Results and methods: I. indigotica was indistinguishable from I. tinctoria when using ITS2. CPGAVAS2 was used to construct the chloroplast genomes. MAFFT and DnaSP were used to calculate nucleotide polymorphism, the chloroplast genomes of the two have high diversity in the rpl32 ~ trnL-UAG short region. When using this region as a mini barcode, it was found that there are obvious differences in the base numbers of I. tinctoria and different ploidy I. indigotica were found, but diploid and tetraploid I. indigotica had the same number of bases. Moreover, the reconstruction of the maximum likelihood (ML) tree, utilizing the mini-barcode, demonstrated that I. tinctoria and both diploid and tetraploid I. indigotica are located on distinct branches. The genome size of tetraploid I. indigotica was approximately 643.773 MB, the heterozygosity rate was approximately 0.98%, and the repeat sequence content was approximately 90.43%. This species has a highly heterozygous, extremely repetitive genome. Conclusion: A new method was established to differentiate between I. indigotica and I. tinctoria. Furthermore, this approach provides a reference and basis for the directional breeding of Isatis. [ABSTRACT FROM AUTHOR]

This study aimed to establish the in vitro shoot culture of Isatis tinctoria L. and its ability to produce antioxidant bioactive compounds. The Murashige and Skoog (MS) medium variants, containing different concentrations (0.1–2.0 mg/L) of benzylaminopurine (BAP) and 1-naphthaleneacetic acid (NAA) were tested. Their influence on the growth of biomass, accumulation of phenolic compounds, and antioxidant potential was evaluated. To improve the phenolic content, agitated cultures (MS 1.0/1.0 mg/L BAP/NAA) were treated with different elicitors, including the following: Methyl Jasmonate, CaCl2, AgNO3, and yeast, as well as with L-Phenylalanine and L-Tyrosine—precursors of phenolic metabolites. The total phenolic content (TPC) of hydroalcoholic extracts (MeOH 70%) obtained from the biomass grown in vitro was determined spectrophotometrically; phenolic acids and flavonoids were quantified by RP-HPLC. Moreover, the antioxidant potential of extracts was examined through the DPPH test, the reducing power, and the Fe2+ chelating assays. The biomass extracts obtained after 72 h of supplementation with Tyr (2 g/L), as well as after 120 and 168 h with Tyr (1 g/L), were found to be the richest in TPC (49.37 ± 0.93, 58.65 ± 0.91, and 60.36 ± 4.97 mg GAE/g extract, respectively). Whereas among the elicitors, the highest TPC achieved was with CaCl2 (20 and 50 mM 24 h), followed by MeJa (50 and 100 µM, 120 h). The HPLC of the extracts led to the identification of six flavonoids and nine phenolic acids, with vicenin-2, isovitexin, syringic, and caffeic acids being the most abundant compounds. Notably, the amount of all flavonoids and phenolic acids detected in the elicited/precursor feeding biomass was higher than that of the leaves of the parental plant. The best chelating activity was found with the extract of biomass fed with Tyrosine 2 g/L, 72 h (IC50 0.27 ± 0.01 mg/mL), the strongest radical scavenging (DPPH test) for the extract obtained from biomass elicited with CaCl2 50 mM, after 24 h of incubation (25.14 ± 0.35 mg Trolox equivalents (TE)/g extract). In conclusion, the in vitro shoot culture of I. tinctoria supplemented with Tyrosine, as well as MeJa and/or CaCl2, could represent a biotechnological source of compounds with antioxidant properties. [ABSTRACT FROM AUTHOR]

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.

Key message: IiSVP of Isatis indigotica was cloned and its expression pattern was analyzed. Ectopic expression of IiSVP in Arabidopsis could delay the flowering time and reduce the size of the floral organs. SVP (SHORT VEGETATIVE PHASE) can negatively regulate the flowering time of Arabidopsis. In the present work, the cDNA of IiSVP, an orthologous gene of AtSVP in I. indigotica, was cloned. IiSVP was highly expressed in rosette leaves, inflorescences and petals, but weakly expressed in sepals, pistils and young silicles. The results of subcellular localization showed that IiSVP was localized in nucleus. Bioinformatics analysis indicated that this protein was a MADS-box transcription factor. Constitutive expression of IiSVP in Arabidopsis thaliana resulted in decrease of the number of petals and stamens, and curly sepals were formed. In IiSVP transgenic Arabidopsis plants, obvious phenotypic variations in flowers could be observed, especially the size of the floral organs. In comparison with the wild-type plants, the size of petals, stamens and pistil in IiSVP transgenic Arabidopsis plants was decreased significantly. In some transgenic plants, the petals were wrapped by the sepals. Yeast two-hybrid experiments showed that IiSVP could form higher-order complexes with other MADS proteins, including IiSEP1, IiSEP3, IiAP1 and IiSEP4, but could not interact with IiSEP2. In this work, it was proved that the flowering process and the floral development in Arabidopsis could be affected by IiSVP from I. indigotica Fortune. [ABSTRACT FROM AUTHOR]

Water shortage and wastage are critical challenges to sustainable agricultural development, especially in arid and semiarid regions worldwide. Isatis indigotica (woad), as a traditional Chinese herb, was planted in a large area in a cold and arid environment of Hexi. Regulated deficit irrigation can reduce the growth of some vegetative organs by changing the accumulation and distribution of photosynthetic products in crops, thus increasing the economic yield of crops. In agricultural production, crop productivity may be improved by mulched drip irrigation and deficit irrigation. Hence, a field experiment was conducted to investigate the responses of photosynthesis, malondialdehyde, osmotic regulators, antioxidant enzyme activities, and the yield of woad to water deficit at different growth stages. The growth stage of woad was divided in four stages: seedling, vegetative growth, fleshy root growth, and fleshy root maturity. During vegetative growth, fleshy root growth, and fleshy root maturity, three water gradients were set for plants with mild (65-75% in field water capacity, FC), moderate (55-65% in FC), and severe (45-55% in FC) deficits, respectively. In contrast, an adequate water supply (75-85% in FC) during the growth period was designed as the control (CK). The net photosynthetic rate (Pn), transpiration rate, and stomatal conductance of woad significantly decreased (P< 0.05) by moderate and severe water deficits. Still, rehydration after the water deficit could produce a noticeable compensation effect. In contrast, malondialdehyde and proline accumulation significantly increased under moderate and severe water deficits. At the same time, the superoxide dismutase, peroxidase, and catalase all had high activities (increased significantly by 19.87-39.28%, 19.91-34.26%, and 10.63-16.13% compared with CK, respectively), but yields were substantially lower, compared to CK. Additionally, the net photosynthetic rate was negatively correlated with antioxidant enzyme activity. The economic yield of plants subjected to continuous mild water deficit during both vegetative and fleshy root growth was not significantly different from that in CK. Still, the water use efficiency improved significantly. Therefore, the continuous mild water deficit during vegetative and fleshy root growth could improve the physiological and biochemical mechanisms of the plant, representing an optimal irrigation strategy for woad in cold and arid areas. [ABSTRACT FROM AUTHOR]

Natural products and their derivatives historically represent alternatives to conventional synthetic molecules for pharmacotherapy, ranging from cancer chemotherapeutics to cosmetic ingredients that exert anti-aging activities. Cellular senescence is considered a main driver of skin aging, yet natural products that target skin senescence in a specific manner are not thoroughly explored. Here, we performed a focused compound screen to identify natural products that exert anti-senescence effects. We found that Isatis tinctoria, woad extracts, displayed a senolytic effect on senescent human skin fibroblasts. Furthermore, treatment with woad extracts attenuated the expression of pro-inflammatory senescence-associated secretory phenotype (SASP), showing a senostatic activity. Intriguingly, woad extracts displayed only a marginal cytotoxic effect toward senescent human lung fibroblasts. Thus, our results reveal the potential activities of woad extracts for targeting skin senescence and suggest that woad extracts could be an attractive ingredient for cosmetics to prevent skin aging. [ABSTRACT FROM AUTHOR]

Cofactors are crucial for the biosynthesis of natural compounds, and cofactor engineering is a useful strategy for enzyme optimization due to its potential to enhance enzyme efficiency. Secoisolariciresinol dehydrogenase (SIRD) was reported to convert secoisolariciresinol into matairesinol in an NAD+-dependent reaction. Here, a SIRD designated as IiSIRD2 identified from Isatis indigotica was found to utilize NADP+ as the cofactor. To explore the structural basis for this unique cofactor preference, model-based structural analysis was carried out, and it was postulated that a variation at the GXGGXG glycine-rich motif of IiSIRD2 alters its cofactor preference. This study paves way for future investigations on SIRD cofactor specificity and cofactor engineering to improve SIRD's catalytic efficiency. [ABSTRACT FROM AUTHOR]

The bZIP transcription factor family plays important roles in plant growth and development, response to stress, and regulation of secondary metabolite biosynthesis. The identification and molecular function of bZIP gene have been deeply studied in the model plant Arabidopsis thaliana, but it has not been reported in the medicinal plant Isatis indigotica. In this study, 65 IibZIP genes were identified in the genome of I. indigotica, which were distributed on seven chromosomes, were highly conserved, could be classified into 11 subgroups. Transcriptomic and metabolomic data for leaves of I. indigotica exposed to salt stress were analyzed to construct an IibZIP gene co-expression network and metabolite correlation network. Seventeen IibZIP genes were co-expressed with 79 transcription factors, and GO and KEGG enrichment analysis showed that most of these genes were associated with abiotic stress and hormone responses of plants. 17 IibZIP genes regulated 110 metabolites through 92 transcription factor associations. In addition, IibZIP23, IibZIP38 and IibZIP51 were associated with six metabolites including three alkaloids (quinoline alkaloid stylopine, indole alkaloids tabersonine and indole-3-acetic acid), flavonoid myricetin 3-O-galactoside, and two primary metabolites 2- hydroxy-6-aminopurine, 3-dehydroshikimic acid were strongly correlated. This study provides data for identification of the IibZIP gene family and their regulation of metabolites in response to salt stress. [ABSTRACT FROM AUTHOR]

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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.

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 (H2O2) content, As and S accumulation, and guaiacol peroxidase (POD) and glutathione S-transferase (GST) along with the supplementation of 20 mg L−1 of S and 500 μM of GSH. Results revealed the significant reduction of fresh weight (especially in E. allionii), activities of POD and GST enzymes and proline content as compare to control. However, the application of S and GSH enhanced the fresh weight. Inhibition in H2O2 accumulation and improvement in antioxidant responses were measured with the application of S and GSH. Hence, the supplementation of S and GSH enhanced fresh weight and total chlorophyll in both I. cappadocica and E. allionii by alleviating the adverse effects of As stress via decreased H2O2 content and restricted As uptake. [ABSTRACT FROM AUTHOR]

The geostatistical method is one of the most advanced techniques to assess and survey groundwater quality. Using unsuitable water results in some problems such as soil salinity, reduction of soil permeability, decrease in water absorption by plant roots, diminution of crop productivity, or even menace agricultural production. Accordingly, in this study, the groundwater quality in the study's area for agricultural uses as well as industrial applications by using common diagrams and components related to the classification of water quality was investigated. ISATIS and Surfur software were used to illustrate changes in quality characteristics in the study's area graphically and Consecutive Gaussian simulations to analyze spatial connection among variables and estimate some of the quality indexes such as PH, RSC, Na%, SAR, and Ec. After data normalization, the variogram was calculated, and a sufficient model was selected to fit the experimental variogram based on the lowest SSR Error. Afterward, Consecutive Gaussian simulation in 100 samples in the study's area was examined using the fitted model based on the experimental model, and the maps resulting through simulation in different thresholds were provided. The results showed that the majority of samples have fairly suitable quality in terms of SAR, and in general, they are appropriate for agricultural uses. Percentage risk of Sodium in agricultural water in the study's area was also classified into the appropriate class. Except for four samples whose spatial connection of the forgoing cases have been interpolated and the interpolated probability maps have been provided, the water quality for industrial uses is susceptible to sedimentation in all statutes. [ABSTRACT FROM AUTHOR]

The Brassicaceae family is one of the world's top ten economically significant plant families. The genus Isatis belongs to the family Brassicaceae. Isatis species have antibacterial, anticancer and anti-viral properties. Flavonoids and phenolics are bioactive secondary metabolites commonly found in medicinal plants. The present review summarises the flavonoid and phenolic compounds of the genus Isatis and aims to highlight the recent advances in current knowledge of Isatis species as a source of flavonoids and phenolics. Many known flavonoids and phenolic compounds from Isatis species, such as quercetin, rutin, caffeic acid, kaempferol, ferulic acid, gallic acid, p-coumaric acid, p-hydroxybenzoic acid, rosmarinic acid, sinapic acid, neohesperidin, buddleoside, liquiritigenin, isorhamnetin have been isolated. [ABSTRACT FROM AUTHOR]

Isatis indigotica is a popular herbal medicine with its noticeable antiviral properties, which are primarily due to its lignan glycosides such as lariciresinol-4- O -β-D-glucoside and lariciresinol-4,4′-bis- O -β-D-glucosides (also called clemastanin B). UDP-glucose-dependent glycosyltransferases are the key enzymes involved in the biosynthesis of these antiviral metabolites. In this study, we systematically characterized the UGT72 family gene IiUGT1 and two UGT71B family genes, IiUGT4 and IiUGT71B5a, with similar enzymatic functions. Kinetic analysis showed that IiUGT4 was more efficient than IiUGT1 or IiUGT71B5a for the glycosylation of lariciresinol. Further knock-down and overexpression of these IiUGTs in I. indigotica's hairy roots indicates that they play different roles in planta : IiUGT71B5a primarily participates in the biosynthesis of coniferin not pinoresinol diglucoside, and IiUGT1 primarily participates in the biosynthesis of pinoresinol diglucoside, while IiUGT4 is responsible for the glycosylation of lariciresinol and plays a dominant role in the biosynthesis of lariciresinol glycosides in I. indigotica. Analysis of the molecular docking and site-mutagenesis of IiUGT4 have found that key residues for its catalytic activity are H373, W376, E397, and that F151 could be associated with substrate preference. This study elucidates the biosynthetic route of anti-viral lignan glycosides in I. indigotica , and provides the foundation for the production of anti-viral lignan glycosides via synthetic biology under the heterologous model. [ABSTRACT FROM AUTHOR]

Although regulated deficit irrigation may improve crop yields, little research has been conducted on the effects of water deficits on Isatis indigotica, a popular herbal medicine. Field experiments were conducted in 2016 and 2017 to study the effects of regulated deficit irrigation on the net photosynthetic rate, yield, water use efficiency (WUE), and quality of I. indigotica in northwest China. Plants at the vegetative and fleshy root growth stages were subjected to mild, moderate, and severe water deficits, and their photosynthetic physiological indexes, yield, and WUE were measured. Moderate and severe deficits, but not a mild deficit, significantly decreased the net photosynthetic rate and dry matter accumulation. The yield and WUE under mild deficit were markedly higher, reaching 8239.56 kg·ha−2 and 8390.80 kg·ha−2, respectively, in the vegetative stage and 24.11 kg·ha−2·mm−1 and 23.62 kg·ha−2·mm−1, respectively, in the fleshy root growth stage, while severe deficits significantly reduced yield and WUE. Mild and moderate deficits increased the content of (R,S)-goitrin, indirubin, and indigo, improving root quality, but severe deficits decreased these compounds. Therefore, a mild water deficit in the vegetative and fleshy root growth stages is optimal and can reduce water consumption and improve I. indigotica quality and WUE without reducing yield. [ABSTRACT FROM AUTHOR]

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 Isatis indigotica Fortune ex Lindl., a traditional Chinese medicine, were frequently used for the prevention of viral disease in China. In the present study, (−)-lariciresinol ((−)-LRSL), isolated from the roots of Isatis indigotica Fortune ex Lindl., was found to inhibit HBV DNA replication of both wild-type and nucleos(t)ide analogues (NUCs)-resistant strains in vitro. Mechanism studies revealed that (−)-LRSL could block RNA production after treatment, followed by viral proteins, and then viral particles and DNA. Promoter reporter assays and RNA decaying dynamic experiments indicated that (−)-LRSL mediated HBV RNA reduction was mainly due to transcriptional inhibition rather than degradation. Moreover, (−)-LRSL in a dose-dependent manner also inhibited other animal hepadnaviruses, including woodchuck hepatitis virus (WHV) and duck hepatitis B virus (DHBV). Combining the analysis of RNA-seq, we further found that the decrease in HBV transcriptional activity by (−)-LRSL may be related to hepatocyte nuclear factor 1α (HNF1α). Taken together, (−)-LRSL represents a novel chemical entity that inhibits HBV replication by regulating HNF1α mediated HBV transcription, which may provide a new perspective for HBV therapeutics. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

(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. IRPS has attracted extensive attention in the field of virology. However, it is not clear that IRPS has an antiviral effect on PRRSV at gene and protein levels. (2) Methods: We used transcriptomic and proteomic analysis to investigate the antiviral effect of IRPS against PRRSV. Additionally, a microbiome was used to explore the effects of IRPS on gut microbes. (3) Results: IRPS significantly extenuated the pulmonary pathological lesions and inflammatory response. We used transcriptomic and proteomic analysis to investigate the antiviral effect of IRPS against PRRSV. In the porcine model, 1669 differentially expressed genes (DEGs) and 370 differentially expressed proteins (DEPs) were identified. Analysis of the DEG/DEP-related pathways indicated immune-system and infectious-disease (viral) pathways, such as the NOD-like receptor (NLR) signaling pathway, toll-like receptor (TLR) signaling pathway, and Influenza A-associated signaling pathways. It is noteworthy that IRPS can inhibit NLR-dependent gene expression, then reduce the inflammatory damage. IRPS could exert beneficial effects on the host by regulating the structure of intestinal flora. (4) Conclusions: The antiviral effect of IRPS on PRRSV can be directly achieved by omics techniques. Specifically, the antiviral mechanism of IPRS can be better elucidated by screening target genes and proteins using transcriptome and proteome sequencing, and then performing enrichment and classification according to DEGs and DEPs. [ABSTRACT FROM AUTHOR]

Primordial black holes (PBHs) are convenient candidates to explain the elusive dark matter (DM). However, years of constraints from various astronomical observations have constrained their abundance over a wide range of masses, leaving only a narrow window open at 10 17 g ≲ M ≲ 10 22 g for all DM in the form of PBHs. We reexamine this disputed window with a critical eye, interrogating the general hypotheses underlying the direct photon constraints. We review 4 levels of assumptions: (i) instrument characteristics, (ii) prediction of the (extra)galactic photon flux, (iii) statistical method of signal-to-data comparison and (iv) computation of the Hawking radiation rate. Thanks to Isatis, a new tool designed for the public Hawking radiation code BlackHawk, we first revisit the existing and prospective constraints on the PBH abundance and investigate the impact of assumptions (i)–(iv). We show that the constraints can vary by several orders of magnitude, advocating the necessity of a reduction of the theoretical sources of uncertainties. Second, we consider an "ideal" instrument and we demonstrate that the PBH DM scenario can only be constrained by the direct photon Hawking radiation phenomenon below M max ∼ 10 20 g. The upper part of the mass window should therefore be closed by other means. [ABSTRACT FROM AUTHOR]

An experimental study of artificial weathering of two outdoor coatings pigmented with natural indigo from Isatis tinctoria was carried out. The two coatings, natural indigo‐pigmented boat varnish and outdoor paint, were exposed to cycles of ultraviolet light, water condensation and water spray for 40 days in a QUV test chamber. Synthetic indigo and commercial ultramarine pigment were employed as reference materials. The effects of artificial weathering on the coatings were evaluated with CIELab parameters, as well as visual inspection. Coating and pigment stability during artificial weathering were studied with reflectance spectrometry, hyperspectral imaging technology and Fourier Transform–infrared spectroscopy. According to the results, coatings with indigo proved to be more resistant to visual and spectral changes than unpigmented or commercially pigmented coatings. [ABSTRACT FROM AUTHOR]

Isatis indigotica Fort., belongs to the Brassicaceae family and its leaf is an important Chinese traditional folklore medicine. After many years of research, phytochemical investigations on it have led to the identification of 197 alkaloids, which could serve as valuable chemotaxonomic markers for the identification of I. indigotica. We summarize the chemotaxonomic value of alkaloids , and the results showed that I. indigotica is a typical species in the Brassicaceae family. It has some chemotaxonomic relationships with genera of Brassica L., Arabidopsis (DC.) Heynh., Thellungiella O. E. Schulz, Raphanus L., Lepidium L., together with families of Acanthaceae, Apocynaceae, Nitrariaceae, Orchidaceae and Orchidoideae. • 197 alkaloids (1 – 197) were summarized from the Isatis indigotica. • The structural characteristics of various alkaloids were described. • The chemotaxonomic significance of the compounds isolated was presumed in this paper by network analyses. [ABSTRACT FROM AUTHOR]

Sheep gastrointestinal nematode (GIN) infestation represents a limiting factor for sheep farming and milk production in Italy. The development of anthelmintic resistance to conventionally used drugs suggests the path towards the use of natural remedies as a possible alternative. The purpose of this study is to evaluate the in vitro anthelmintic efficacy of the hydroalcoholic extracts of basal leaves (It-BL), cauline leaves (It-CL) and flowers (It-F) of Isatis tinctoria (Brassicaceae), a spontaneous Sicilian species renowned as an important source of bioactive compounds. The dry extracts of the different parts of the plant were tested using the egg hatch test (EHT) in vitro to verify the efficacy against ovine GIN at different concentrations (1.00, 0.5, 0.25, 0.125 mg/mL). Thiabendazole and deionized water were used as positive and negative controls, respectively. The results obtained from EHT indicated that all the I. tinctoria extracts were highly effective (p < 0.0001) in inhibiting egg hatching within 48 h of exposure. The in vitro inhibitory effect was never less than 84% in all doses tested, and it was only slightly lower than the standard drug thiabendazole (95.6%). The current study documents the anthelmintic activity of I. tinctoria against sheep's GIN, suggesting its application as alternative natural method to limit the use of antiparasitic drugs. [ABSTRACT FROM AUTHOR]

Background: The architecture of inflorescence and the development of floral organs can influence the yield of seeds and have a significant impact on plant propagation. E-class floral homeotic MADS-box genes exhibit important roles in regulation of floral transition and differentiation of floral organs. Woad (Isatis indigotica) possesses unique inflorescence, floral organs and fruit. However, very little research has been carried out to determine the function of MADS-box genes in this medicinal cruciferous plant species. Results: SEPALLATA orthologs in I. indigotica were cloned by degenerate PCR. The sequence possessing the highest identity with SEP2 and SEP4 of Arabidopsis were named as IiSEP2 and IiSEP4, respectively. Constitutive expression of IiSEP2 in Columbia (Col-0) ecotype of Arabidopsis led to early flowering, and the number of the flowers and the lateral branches was reduced, indicating an alteration in architecture of the inflorescences. Moreover, the number of the floral organs was declined, the sepals were turned into carpelloid tissues bearing stigmatic papillae and ovules, and secondary flower could be produced in apetalous terminal flowers. In 35S::IiSEP4-GFP transgenic Arabidopsis plants in Landsberg erecta (Ler) genetic background, the number of the floral organs was decreased, sepals were converted into curly carpelloid structures, accompanied by generation of ovules. Simultaneously, the size of petals, stamens and siliques was diminished. In 35S:: IiSEP4-GFP transgenic plants of apetalous ap1 cal double mutant in Ler genetic background, the cauliflower phenotype was attenuated significantly, and the petal formation could be rescued. Occasionally, chimeric organs composed of petaloid and sepaloid tissues, or petaloid and stamineous tissues, were produced in IiSEP4 transgenic plants of apl cal double mutant. It suggested that overexpression of IiSEP4 could restore the capacity in petal differentiation. Silencing of IiSEP4 by Virus-Induced Gene Silencing (VIGS) can delay the flowering time, and reduce the number and size of the floral organs in woad flowers. Conclusion: All the results showed that SEPALLATA-like genes could influence the architecture of the inflorescence and the determinacy of the floral meristems, and was also related to development of the floral organs. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Purpose: The rate of gastrointestinal adenomatous polyps, often regarded as precancerous lesions, developing into cancer is 40 - 70 %. Endoscopic resection has been the preferred method for treating gastric polyps since the late 1960s. Surgical removal of polyps continues to play an important role in the treatment of polyps; however, the efficacy of such treatment cannot be guaranteed, and polyps may recur. Case presentation: Here, we report a 44-year-old man suffering from gastrointestinal polyps who refused surgical treatment and instead was treated with traditional Chinese herbal medicine (TCHM) for approximately 1 year. The patient was diagnosed with "qi deficiency and dampness syndrome" based on traditional Chinese medicine theory and was treated with the TCHM "strengthen qi and remove dampness formula", referred to as the shen-ling-bai-zhu powder/decoction. Conclusion: This case suggests that TCHM may play an important role in the treatment of gastrointestinal polyps. Relevant data are, however, limited and a randomized controlled trial is needed to confirm TCHM efficacy in a larger population. [ABSTRACT FROM AUTHOR]

Influenza A virus (IAV) is one of the major causes of seasonal endemic diseases and unpredictable periodic pandemics. Due to the high mutation rate and drug resistance, it poses a persistent threat and challenge to public health. Isatis tinctoria L. (Banlangen, BLG), a traditional herbal medicine widely used in Asian countries, has been reported to possess strong efficacy on respiratory viruses, including IAV. However, its effective antiIAV components and the mechanism of actions (MOAs) are not yet fully elucidated. In this study, we first summarized the chemical components and corresponding contents in BLG according to current available chemical analysis literature. We then presented a network based in silico framework for identifying potential drug candidates against IAV from BLG. A total of 269 components in BLG were initially screened by drug-likeness and ADME (absorption, distribution, metabolism, and excretion) evaluation. Thereafter, network predictive models were built via the integration of compound–target networks and influenza virus–host proteins. We highlighted 23 compounds that possessed high potential as anti-influenza virus agents. Through experimental evaluation, six compounds, namely, eupatorin, dinatin, linarin, tryptanthrin, indirubin, and acacetin, exhibited good inhibitory activity against wild-type H1N1 and H3N2. Particularly, they also exerted significant effects on drug-resistant strains. Finally, we explored the anti-IAV MOAs of BLG and showcased the potential biological pathways by systems pharmacology analysis. In conclusion, this work provides important information on BLG regarding its use in the development of anti-IAV drugs, and the network-based prediction framework proposed here also offers a powerfulful strategy for the in silico identification of novel drug candidates from complex components of herbal medicine. [ABSTRACT FROM AUTHOR]

In this study, detailed morphological and anatomical features of Isatis quadrialata were examined. With anatomical analysis, transverse sections of root, stem and leaf were examined. Epidermis is a protective tissue in root transverse sections. Transverse sections of stem contain one layered epidermis, cortex parenchyma, vascular bundles and pith cells. The cross sections of leaves have two layered palisade parenchyma and spongy parenchyma with bifacial mesophyll. Transverse sections of fruit have one layered exocarp, multi layered mesocarp and endocarp. In addition, fruit and seed micromorphologies were studied. The fruit surface ornamentation is reticulate while the seed surface ornamentation is blister. The anatomical characteristics of Isatis quadrialata were determined of taxonomic importance. [ABSTRACT FROM AUTHOR]

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 H 2 O 2 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. • Si application alleviated As/Cd toxicity in Isatis cappadocica. • As was accumulated significantly more than Cd, however with considerably low toxic growth effects. • Exogenous Si alleviated oxidative stress and enhanced antioxidant enzyme activity. • Si decreased the As/Cd accumulation in Isatis plants. • Si could be a candidate for As/Cd detoxification and phytoremediation potential of I. cappadocica. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Isatis L. genus is highly endemic in Turkey and it is traditionally suggested for the wound management. In this study, we have reported the effect of the I. glauca subsp. sivasica extract on the diabetic wound healing process. A diabetic model was generated in Wistar rats using streptozotocin injection. The rats were divided into two main groups: the control group and the Isatis group. Full thickness excisional skin wounds were created by using a biopsy punch. The Isatis group was treated with single daily dose I. glauca subsp. sivasica extract (50 mg/kg). The rats were sacrificed on day 3 or day 7 after wounding. The dominant phenolic compounds identified with RP-HPLC-DAD from the Isatis extract were the benzoic acid and the vanillic acid. The Isatis extract significantly accelerated the wound healing process considering the wound closure rates (WCR). Moreover, the levels of collagen and nitric oxide were elevated on day 3 and day 7 by Isatis administration in the diabetic wound tissue. This data suggests that, for the first time, I. glauca subsp. sivasica extract may have the potential to promote the impaired wound healing in patients with diabetes. [ABSTRACT FROM AUTHOR]

• Strong light promoted the energy metabolism of Isatis indigotica Fortune. • Light intensity affected the expression of 89 miRNAs and 16,485 mRNAs. • 12 miRNA-mRNA pairs play important roles in mitochondrial biological processes. • Light intensity affects 4 mitochondria-associated miRNA-mRNA pairs. Light is the main source of energy for plant growth. Studies have shown that I. indigotica is a light-demanding plant and its yield and various active components are positively correlated with light intensity, but no studies of light intensity affecting energy metabolism in I. indigotica have been reported. Mitochondria are the main site of energy metabolism, and miRNAs are important factors in regulating gene expression, this experiment attempts to study the effects of different light intensities on energy metabolism from the perspective of mitochondria and miRNAs. The results show that the biomass、mitochondrial structural integrity and energy metabolism in I. indigotica were found to be positively correlated with light intensity. Small RNA and transcriptome sequencing identified 241 miRNAs and 36,372 mRNAs, and degradomic technology identified 72 miRNAs targeting 106 mRNAs, among which 12 pairs of miRNA-mRNAs were annotated on mitochondria. Combined with RT-qPCR validation, it was concluded that miR167a-5p positively regulates LETM1 and affects mitochondrial structure, miR400-5p and mIR169m-p3_1ss15CT negatively regulate GRXS15 and CMC4 , respectively, affecting SDH and CCO activities, and miR395a- APS4 may affect the utilization of ATP and sulfate assimilation. In summary, the results of this study complement and enrich knowledge of light effects on mitochondria from the perspective of miRNA, while providing guidance for the cultivation of I. indigotica. [ABSTRACT FROM AUTHOR]

The AGL6 (AGMOUSE LIKE 6) gene is a member of the SEP subfamily and functions as an E-class floral homeotic gene in the development of floral organs. In this study, we cloned IiAGL6 , the orthologous gene of AGL6 in Isatis indigotica. The constitutive expression of IiAGL6 in Arabidopsis thaliana resulted in a late-flowering phenotype and the development of curly leaves during the vegetative growth period. Abnormal changes in floral organ development were observed during the reproductive stage. In woad plants, suppression of IiAGL6 using TRV-VIGS (tobacco rattle virus-mediated virus-induced gene silencing) decreased the number of stamens and led to the formation of aberrant anthers. Similar changes in stamen development were also observed in miRNA-AGL6 transgenic Arabidopsis plants. Yeast two-hybrid and BiFC tests showed that IiAGL6 can interact with other MADS-box proteins in woad; thus, playing a key role in defining the identities of floral organs, particularly during stamen formation. These findings might provide novel insights and help investigate the biological roles of MADS transcription factors in I. indigotica. • IiAGL6 was the orthologous gene of AGL6 in Isatis indigotica. • IiAGL6 transgenic Arabidopsis plants were late-flowering and produce curly leaves. • IiAGL6 overexpression resulted in abnormal changes in floral organs. • Suppression of IiAGL6 using TRV-VIGS led to aberrant stamens. • IiAGL6 play a crucial role in defining the identities of stamens. [ABSTRACT FROM AUTHOR]

Indigo naturalis (IN) is a dried powder derived from plants such as Baphicacanthus cusia (Neeks) Bremek., Polygonum tinctorium Ait. and Isatis indigotica Fork. It has a historical application as a dye in ancient India, Egypt, Africa and China. Over time, it has been introduced to China and Japan for treatment of various ailments including hemoptysis, epistaxis, chest discomfort, and aphtha. Clinical and pre-clinical studies have widely demonstrated its promising effects on autoimmune diseases like psoriasis and Ulcerative colitis (UC). Despite the documented efficacy of IN in UC patients, concerns have been raised on the development of adverse effects with long term consumption, prompting a closer examination of its safety and tolerability in these contexts. This review aims to comprehensively assess the efficacy of IN in both clinical and pre-clinical settings, with a detailed exploration of the mechanisms of action involved. Additionally, it summarizes the observed potential toxicity of IN in animal and human settings was summarized. This review will deepen our understanding on the beneficial and detrimental effects of IN in UC, providing valuable insights for its future application in patients with this condition. [ABSTRACT FROM AUTHOR]

Isatis indigotica Fort. (I. indigotica) has a long history of use in the treatment of respiratory tract infections. Here, we investigated the anti-influenza effects exerted by I. indigotica on human alveolar epithelial cells via altering virulent protein production and disrupting autophagy. I. indigotica water extract was analysed by HPLC with epigoitrin specified as the fingerprint. Cellular viability was detected via XTT assay. Hemagglutination assay, plaque assay, and quantitative reverse transcription PCR were used to evaluate the inhibitory effects on H1N1 viral infection. The initial step of the viral infection was detected by attachment and penetration assays. Western blot was applied to measure the matrix-1 (M1), matrix-2 (M2), nucleoprotein, light chain 3 beta, and Beclin-1 protein levels. The extraction rate of the I. indigotica was 12.48%. The results of the hemagglutination assay, plaque assay, and quantitative reverse transcription PCR demonstrated that I. indigotica can effectively impede viral propagation. In addition, I. indigotica inhibited the production of the M1, M2, and nucleoprotein proteins, most notably in the M2 and M1 proteins, thereby hindering viral attachment, internalisation, and penetration. After treatment with I. indigotica , light chain 3 beta II levels decreased significantly, while we noted no marked change in the production of Beclin-1. This study demonstrates that I. indigotica water extract can rescue host cells from H1N1 influenza infection by inhibiting viral attachment, penetration, and propagation. This is achieved by significantly reducing virulent protein production and disrupting the process of autophagy. Collectively, our findings indicate that I. indigotica may provide an effective alternative treatment for patients infected with the H1N1 influenza virus. • I. indigotica effectively inhibited viral propagation through the production of the M1, M2 and NP proteins. • I. indigotica induced LC3B II levels decreased significantly, while we noted no marked change in the production of Beclin-1. • I. indigotica significantly reduced virulent protein production and disrupting the process of autophagy. [ABSTRACT FROM AUTHOR]

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Isatis indigotica (2n = 14) is an important medicinal plant in China. Its dried leaves and roots (called Isatidis Folium and Isatidis Radix, respectively) are broadly used in traditional Chinese medicine for curing diseases caused by bacteria and viruses such as influenza and viral pneumonia. Various classes of compounds isolated from this species have been identified as effective ingredients. Previous studies based on transcriptomes revealed only a few candidate genes for the biosynthesis of these active compounds in this medicinal plant. Here, we report a high-quality chromosome-scale genome assembly of I. indigotica with a total size of 293.88 Mb and scaffold N50 = 36.16 Mb using single-molecule real-time long reads and high-throughput chromosome conformation capture techniques. We annotated 30,323 high-confidence protein-coding genes. Based on homolog searching and functional annotations, we identified many candidate genes involved in the biosynthesis of main active components such as indoles, terpenoids, and phenylpropanoids. In addition, we found that some key enzyme-coding gene families related to the biosynthesis of these components were expanded due to tandem duplications, which likely drove the production of these major active compounds and explained why I. indigotica has excellent antibacterial and antiviral activities. Our results highlighted the importance of genome sequencing in identifying candidate genes for metabolite synthesis in medicinal plants. [ABSTRACT FROM AUTHOR]

• Analyzed the synthase genes and compounds related to the lignan in diploid Isatis indigotica Fortune, tetraploid I.indigotica , and Isatis tinctoria Linnaeus through metabolomic and transcriptomic. • The analysis of transcription factors showed that the AP2-EREBP family (Iin24319), MYB family (Iin24843), and WRKY family (Iin08158) displayed expression patterns similar to Iin14549. • Serve as a reference for improving the quality of I. indigotica and developing germplasms with high lignan content. Lay a foundation for the functional characterization of UGTs in lignan biosynthesis pathway. Isatis indigotica Fortune is a plant species containing lignan compounds of significant economic value. Its root plays a crucial role in treating viruses and exhibits antitumor, anti-inflammatory, antibacterial, and other biological activities. Now, I. indigotica has been included in Isatis tinctoria Linnaeus. In this study, the roots of diploid I. indigotica , tetraploid I. indigotica , and Isatis tinctoria Linnaeus were analyzed using metabolome and transcriptome analysis. The metabolomic analysis detected 48 lignan metabolites, including Lirioresinol A, Vladinol A, Syringaresinol, Arctigenin, Acanthoside B, and Sesamin as characteristic compounds, without significant variations among the remaining metabolites. The transcriptomic analysis identified 41 differentially expressed phenylpropanoid synthase genes, which were further analyzed for variations in lignan transcriptome profiles across different samples. RT-qPCR analysis also revealed differential genes expression related to lignan biosynthesis pathway among the three sample groups. The analysis of transcription factors showed that the AP2-EREBP family (Iin24319), MYB family (Iin24843), and WRKY family (Iin08158) displayed expression patterns similar to Iin14549. Phylogenetic analyses also indicate that Iin14549 may play a role in lignan synthesis. These transcription factor families exhibited high expression in tetraploid I. indigotica , moderate expression in diploid I. indigotica , and low expression in I. tinctoria. The findings of this study can serve as a reference for improving the quality of I. indigotica and developing germplasms with high lignan content. Additionally, these results lay a foundation for the functional characterization of UGTs in lignan biosynthesis pathway. [ABSTRACT FROM AUTHOR]

The JAK-STAT pathway plays a crucial role in the signaling cascade associated with various cytokines that have been implicated in the pathogenesis of inflammatory diseases and myeloproliferative neoplasms (MPN). Among the isoforms of JAKs, the JAK2 subtype is primarily responsible for the function of hematopoietic system cells, making it a significant target in the treatment of MPN. However, the precise regulatory role of JAK2 in inflammatory diseases requires further investigation and confirmation. The current study employed a selective JAK2 inhibitor, ZT55, derived from Isatis indigotica roots, to examine its regulatory effects on inflammatory and immune responses in delayed-type hypersensitivity (DTH) and arthritis in mice. To evaluate the efficacy of ZT55 treatment, DNFB-induced DTH and collagen-induced arthritis (CIA) mouse models were utilized. T cells were cultured and subsequently analyzed for proliferation and activation using flow cytometry and EdU assay. Additionally, the maturation and function of dendritic cells were assessed through flow cytometry and ELISA. Our findings indicate that ZT55 significantly reduced DNFB-induced DTH and attenuated inflammation, cartilage degradation, and bone destruction in CIA mice. Moreover, ZT55 was found to inhibit the proliferation and activation of T cells and the maturation of dendritic cells by regulating the JAK2-STAT3 signaling pathway. These results suggest that selectively targeting the JAK2 isoform could have anti-inflammatory and immunosuppressive effects by regulating the adaptive and innate immune responses via the JAK2-STAT3 signaling pathway. Therefore, ZT55 has the potential to be a promising pharmaceutical candidate for the treatment of inflammatory and autoimmune diseases. [ABSTRACT FROM AUTHOR]

Drought is a major abiotic stress factor affecting plant growth and production, while utilizing beneficial endophytic fungi is one of the most promising strategies for enhancing plant growth and drought tolerance. In the present study, a pot experiment was conducted to investigate the beneficial effects of dark septate endophytes (DSE) (Acrocalymma aquatica and Alternaria alstroemeriae) combine with organic residues from sterilized or non-sterilized fermented corn straw by Trichoderma afroharzianum on the growth and drought resistance of Isatis indigotica. Drought stress (30% field water capacity) reduced the shoot biomass, root biomass, and epigoitrin content of I. indigotica by 34.30%, 80.85%, and 16.51%, respectively. The DSE inoculation alone increased shoot biomass and epigoitrin content under well-watered condition (70% field water capacity) by 13.75%− 65.98% and 52.33%− 67.59%, respectively, and shoot biomass under drought stress by 37.27%− 84.91%. The organic residues application alone increased root biomass under well-watered condition and drought stress by 71.88%− 107.06% and 463.61%− 678.69%, respectively. The mixed application of DSE and organic residues increased root biomass and epigoitrin content under well-watered condition by 66.79%− 144.66% and 89.02%− 136.63%, respectively, and under drought stress by 410.66%− 580.49% and 28.26%− 88.37%, respectively. Structural equation modeling demonstrated that DSE inoculation, organic residue application, and the mixed application improved the growth of I. indigotica under drought stress through different direct and indirect pathways. Variation partitioning showed that DSE was the primary factor improving the epigoitrin content , and organic residue was the primary factor influencing plant biomass and morphological parameters. The mixed application of DSE and organic residue had further synergistic effects on soil enzyme activities, soil organic matter, root biomass, and epigoitrin content, especially the mixed application of A. aquatica and sterilized organic residue. These results contribute to a further understanding of the ecological function of DSE in dryland soils and reveal the synergistic application value of DSE and organic residue for medicinal plant cultivation. [Display omitted] • Symbionts and organic amendment are beneficial to crop growth and drought resistance. • Dark septate endophytes was the primary factor improving the epigoitrin content. • Fermented organic residue improved soil organic matter and root development. • DSE improved the utilization of organic residues that affected DSE-plant symbiosis. • Acrocalymma aquatica and sterilized residue can be used for dryland crop cultivation. [ABSTRACT FROM AUTHOR]