Amino Purine Research Articles

Different indole-3-acetic acid and 6 benzyl amino purine concentrations affect biomass, phenolic profile, and bioactivity in Mentha rotundifolia L.

Different indole-3-acetic acid and 6 benzyl amino purine concentrations affect biomass, phenolic profile, and bioactivity in Mentha rotundifolia L.

Benzyl Amino Purine (BAP), Moringa Leaf Extract and Ascorbic Acid Induced Drought Stress Tolerance in Pea (Pisum sativum L.)

Benzyl Amino Purine (BAP), Moringa Leaf Extract and Ascorbic Acid Induced Drought Stress Tolerance in Pea (Pisum sativum L.)

Effect of sterilants and plant growth regulators in regenerating commonly used cassava cultivars at the Kenyan coast

Cassava is an important root crop whose seed system is undeveloped. Micropropagation of explants in vitro has the potential of addressing the challenge of the unavailability of healthy cassava planting materials. Therefore, the study determined the effect of sterilization and plant growth regulators on cassava explants to produce certified disease-free plants of commonly used cultivars at the coast of Kenya. The apical nodes drawn from the three cultivars of cassava, Tajirika and Kibandameno and Taita, were used as explants. The sterilant sodium hypochlorite (NaOCl) at 5, 10 and 15% and 70% ethanol for 1 and 5 min and sprayed for 20 s were tested for the effect on the explant. Similarly, the effect of BAP (6-Benzyl amino purine) and NAA (1-Naphthalene acetic acid) Plant Growth Regulators (PGRs) each at 0.5, 1 and 5 mg/L under optimal conditions of sterilization was determined. Surface sterilization using 10% NaOCl followed by spraying 70% ethanol for 20 s had 85% initiation on Tajirika whereas 5% NaOCl followed by spraying 70% ethanol for 20 s had 87% and 91% initiation in Kibandameno and Taita cultivars, respectively. In Tajirika, significantly (p < 0.05) high shooting of 68% was from 5 mg/L BAP in MS media whereas approximately 50% rooting was from either 0.5 mg/L BAP or 5 mg/L NAA in MS media. Kibandameno and Taita cultivars had approximately 50% shooting from MS media without PGRs. Kibandameno had >37% rooting from 0.5 to 5 mg/L BAP or NAA in MS media whereas Taita had approximately 50% rooting from 0 to 5 mg/L NAA in MS media. This protocol showed at least 50% success rate of initiation, shooting and rooting as a rapid multiplication regeneration of Tajirika and Kibandameno and Taita cultivar plantlets with little modification of humidity and temperatures in the growth chambers. This protocol requires validation for use in large-scale production of cassava plantlets to alleviate the inadequacy of cassava planting materials among farmers.

Deciphering the Genetic Identity and Fidelity of Banana Genotype Musa acuminata through Molecular Fingerprinting

Micropropagation for extensive production of elite planting materials is an effective and superior alternative to suckers and conventional means of propagation of Musa spp. The present investigation was carried out to evaluate the genetic identity of different micropropagated banana genotypes Grand Naine, Shrimantee and Basrai. Further micropropagated banana genotypes Grand Naine comparing with mother plant during different stages of subculture using Simple Sequence Repeat (SSR) and Inter Simple Sequence Repeat (ISSR). Moreover, for micropropagation explants (suckers) were inoculated on initiation medium containing MS (Murashige and Skoog) supplemented with 5 mg/l BAP (Benzyl amino purine) and 39 mg/l AdSO4 (Adenine sulphate). The second, third and fourth subcultures were carried out to separate all multiple shoots. Furthermore, for rooting MS media fortified with growth hormones viz., 2 mg/l BAP, 1mg/l IBA (Indole-3-butyric Acid) and 200mg/l activated charcoal were used under present investigation. Findings revealed that 90 % and 75 % plantlets were successfully established during primary hardening in controlled and natural field condition. Further, 26 primers were screened from that 20 primers (ISSR and SSR) showed 146 clear, reproducible and scorable bands ranging from 100-2000 base pairs produced. The banding pattern of these randomly selected plants was monomorphic with mother plant of all selected genotype that confirmed the clonal uniformity.

Factors Affecting Callus Induction from Anther and Ovary of Okra (Abelmoschus esculentus L.)

Background: Okra [Abelmoschus esculentus (L.) Moench.] is a nutrient-rich vegetable crop widely grown in the tropics and sub-tropics mainly for its edible pods. The haploid technique has been used in plant breeding for the improvements of plants and to develop new varieties in relatively a short time. Hence, we have optimized several factors such as plant growth regulators (PGR), sucrose concentration, cold treatment, type of media and culture conditions for callus induction from the anther and ovary of okra (557 F1 hybrid). Methods: The flower buds of different sizes were collected to determine various stages of development and then subjected to cold pre-treatments. The explants were then cultured on various combinations of PGRs i.e., naphthyloxy acetic (NOA), Indole acetic acid (IAA), 2, 4-Dichlorophenoxy acetic acid (2, 4-D), Benzyl amino purine (BAP), isopentenyl adenine 2iP, Kinetin (KIN) and Thidiazuron (TDZ). Result: The optimum developmental stage of microspore for callus initiation was achieved from flower buds of okra and its size was about 11 mm long. Flower buds that emerged one week after the flowering showed significantly higher percentage of callus induction. The optimum stage for ovary and ovule culture were one or two days prior to anthesis and the flower buds stage was 35±1mm. In conclusion, our study investigated the effect of several factors that affects callus induction in okra and optimized cultured conditions for future haploid development for okra.

Changes in Java grass (Cymbopogon winterianus) physiology, essential oil production, and active constituents as influenced by catechol and some plant growth regulators applied through steaming technique.

A study was done to find out how steam-applied chemical and plant growth regulators affected plant growth and determine the most effective PGRs and chemical for maximizing java grass oil yield and quality attributes. A chemical viz. catechol and PGRs, viz. indole acetic acid (IAA), kinetin (Kn), 6-benzyl amino purine (BAP), salicylic acid (SA), and triacontanol (TRIA), ethrel, were applied to the plants at a concentration of 5 × 10−6 M at 90 days after transplantation (DAT) through an innovative technique of steaming using an electric steamer five times after a week interval. To prevent the plants from being harmed, we operated the steam jet at a certain distance from them. All the parameters were evaluated at 150 DAT. The results showed that among all these PGRs, IAA proved the best for growth, yield, and other parameters. In comparison to the control, the EO production more than doubled. Our study indicated that IAA enhanced the fresh and dry weight by 30.7% and 42.9%, total chlorophyll and carotenoids contents by 44.1% and 47.1%, nitrate reductase and carbonic anhydrase activity by 18.0% and 28.1%, respectively, over the control. Compared with control, IAA treatment increased the content and yield of essential oil by 96.8% and 155.7%, and the active constituents of EO, i.e., citronellol, citronellal, and geranial content, by 47.4%, 90.9%, and 43.9%, respectively. As a result, IAA used in this technique was very effective in increasing EO production while causing no harm to the plant.

Integrated gas chromatography-mass spectrometry and ultra-high-performance liquid chromatography-mass spectrometry renal metabolomics and lipidomics deciphered the metabolic regulation mechanism of Gushudan on kidney-yang-deficiency-syndrome rats.

Kidney-yang-deficiency-syndrome is a neuroendocrine disease caused by the dysfunction of the adrenal-pituitary-target gland axis. Gushudan is a traditional Chinese medicine prescription with the functions of tonifying the kidney and strengthening bone, and its bone-strengthening effect has been confirmed by previous anti-osteoporosis research. However, its kidney-tonifying mechanism has not been clear so far. In this study, renal metabolomics and lipidomics based on gas chromatography-mass spectrometry and ultra-high-performance liquid chromatography-high resolution mass spectrometry were integrated to find the metabolic disorders in kidney-yang-deficiency-syndrome rats. Protein precipitation and liquid-liquid extraction were used to extract metabolome and lipidome from the kidney. Gushudan regulated abnormal levels of amino acids, lipids, purines, and carbohydrates, such as L-arginine, hypoxanine, stearic acid, and phosphatidylethanolamine (P-18:1/20:4), which had effects on many metabolic pathways, such as glycerophospholipid metabolism, sphingolipid metabolism, glycine, serine and threonine metabolism and purine metabolism, and so forth. By integrating metabolomics and lipidomics, this study comprehensively revealed the abnormal metabolic activities of amino acids, lipids, and nucleotides in kidney-yang-deficiency-syndrome, and the metabolic regulation mechanism of Gushudan in preventing kidney-yang-deficiency-syndrome, as well as the improvement of Gushudan in maintaining renal cell structure, mitochondrial function, and energy supply, which also provided some new evidence and connotation for "kidney-bone" axis.

The Effect of Benzyl Amino Purine and 2,4-Dichlorophenoxyacetic Acid on in Vitro Propagation of Pranajiwa (Euchresta horsfieldii (Lesch) Benn.)

Pranajiwa (Euchresta horsfieldii (Lesch) Benn.) is an endangered medicinal plant known for its function as aphrodisiac, antidote, expectorant, and tonic. It also can be used to cure TBC. Research has found that pranajiwa is difficult to be propagated, but the tissue culture propagation method has never been applied to this species. This research is aimed to figure out the procedure to induce callogenesis of pranajiwa with the addition of Benzyl Amino Purine (BAP) and 2,4-Dichlorophenoxyacetic acid (2,4-D) and to determine the proper combination of BAP and 2,4-D to produce an optimal callus. MS-based medium with 25 combinations of BAP and 2,4-D has been used for the callus induction. Parameters observed in this research are callus appearing time, percentage of inducted calluses, and morphology observation, including the texture and color of the callus. Result shows that 90.67% calluses was successfully inducted. The optimal callus was obtained by combining BAP 1 ppm and 2,4-D 1,5 ppm with 5 days of appearing time, friable texture and green color.

Indirect organogenesis of Aceh patchouli leaf explants (Pogostemon cablin Benth) by in vitro

The Aceh patchouli plant (Pogostemon cablin. Benth) is a plantation plant that produces essential oils with great potential, known as patchouli oil. The use of in vitro culture techniques on patchouli plants needs to be developed to meet the needs of patchouli seeds. This study aimed to analyze the effect of plant growth regulators (PGRs) cytokinins 6-Benzyl Amino Purine (BAP) and Thidiazuron (TDZ) and auxin 1-Naphthaleneacetic Acid (NAA) on the organogenesis ability of Aceh patchouli leaf explants var. Lhokseumawe. The treatments analyzed were P1 = MS; P2 = BAP 0.75 mg.l−1, P3 = BAP 1.0 mg.l−1, P4 = TDZ 0.75 mg.l−1, P5 = TDZ 1.0 mg.l−1, P6 = NAA 0.5 mg.l−1, P7 = NAA 1.0 mg.l−1, P8 = BAP 0.75 mg.l−1 + NAA 0.5 mg.l−1, P9 = BAP 0.75 mg.l−1 + NAA 1.0 mg.l−1, P10 = BAP 1.0 mg.l−1 + NAA 0.5 mg.l−1, P11 = BAP 1.0 mg.l−1 + NAA 1.0 mg.l−1, P12 = TDZ 0.75 mg.l−1 + NAA 0.5 mg.l−1, P13 = TDZ 0.75 mg.l−1 + NAA 1.0 mg. l−1, P14 = TDZ 1.0 mg.l−1 + NAA 0.5 mg.l−1, P15 = TDZ 1.0 mg.l−1 + NAA 1.0 mg.l-1. The percentage of callus induction observed, percentage of shoot induction, percentage of browning, percentage of contamination and percentage of live explants at 28 DAI. The results showed that the percentage of explants forming callus was 56.4%, and the percentage of explants forming organ was 54.6%. In these two parameters the treatment of P14 and P15 gave the best results of 6.6% respectively. The percentage of explants experiencing browning was 26.1%, treatment P1 gave the highest response for this parameter, namely 6.6%. The parameter percentage of live explants was 71.3%. The best response was found in treatment P2, P3, P12 and P15, which were 6.6% each. The fastest bud formation occurred at 14 Days After Initiation (DAI) in P4 treatment. The resulting shoots look quite vigor and have formed leaves. In the combination treatment P14, showed the best results, the addition of auxin was able to increase the initiation of embryogenic callus.

Multiple bud clumps induction of banana cv. Barangan Merah (Musa acuminate Colla.) under in vitro treatment of benzyl amino purine and indole-3 acetic acid

Cultivar Barangan is one of the famous banana plants that has good taste and price. The disadvantages of this banana is susceptible to Fusarium disease. Tissue culture techniques is one ways of plants propagation for this plant. This study is aimed to determine the effective concentration of benzyl amino purines (BAP) and indole-3 acetic acid (IAA) for optimal multiple bud clumps (MBC) induction in explants of Barangan Merah banana. The Research was carried out at the Laboratory of Tissue Culture, Faculty of Agriculture, Universitas Syiah Kuala. The research design was a completely randomized design, 3x2 factorial. BAP concentration was the first factor, i.e. 5, 10 and 15 mg L−1. The second factor was the concentration of IAA, i.e. 0.1 and 0.5 mg L−1. The results of this research showed that 96,29% explants of all treatments survived. Explant contamination was 11,11% occurred by bacteria and brownish exudate that spread to the medium. MBC are significantly increased at 16 days after induction at concentration of BAP 5 mg L−1 + IAA 0.5 mg L−1. All combinations of BAP and IAA concentrations have produced MBC explants (100%) at eighth weeks after induction. The concentration of 5 mg L−1 BAP + 0.5 mg L−1 IAA resulted the highest shoots number. Analysis of variance and Least Significant Different further test showed that the longest of shoot length was founding the concentration of 5 mg L−1 BAP compared to other treatments. Increasing concentrations of BAP 10 mg L−1and 15 mg L−1 resulted in significantly lower shoot length. It can be concluded that taken together the concentration of 5 mg L−1 BAP + 0.5 mg L−1 IAA is the best to effectively multiple bud clumps induction in explants of Barangan Merah banana through tissue culture technique.

Morphological and Developmental Responses of Macro Cuttings of Vernonia Amygdalina Del. to Synthetic Growth Hormones

The morphological and developmental responses of Vernonia amygdalina macro shoot cutting to plant growth hormonal solution of 6-benzyl amino purine (BAP) and naphthalene acetic acid (NAA) were investigated in the laboratory. The concentration levels of 2.5, 5.0; 7.5 and 10.0mg/l were used alongside a control treatment (0mg/i). The growth parameters examined were shoot length, root length and leaf number. The protein and carbohydrate contents of the plants were also assessed. The result of this study showed that the root length and leaf number of the species were significantly (P=0.05) increased with increase in the concentration of plant growth regulators. Similarly, the carbohydrate and protein contents in treatment with plant growth regulators were comparatively higher than those of the control. This study suggests that plants growth regulators can be utilized to enhance growth and development activities in Vernonia amygdalina Del. and that plant growth regulators such as benzyl amino purine and naphthalene acetic acid aided to maintain an appropriate shoot to root ratio and could be used to induce rooting in stem cuttings.

Effect of dual targeting procyanidins nanoparticles on metabolomics of lipopolysaccharide-stimulated inflammatory macrophages

Inflammation plays an important role in the occurrence and development of many inflammatory diseases. The purpose of this study was to evaluate the anti-inflammatory effect and metabolic behavior of the dual targeting procyanidins (PC) nanoparticles on lipopolysaccharide (LPS)-stimulated inflammatory macrophages by metabolomics method. The double-targeting PC nanoparticles could specifically target both the CD44 receptor and mitochondria, while the single targeting PC-loaded nanoparticles that could target the CD44 receptor on the surface of macrophages. The double-targeting PC nanoparticles had better inhibitory effect than single-targeting PC nanoparticles on the leakage of lactate dehydrogenase and reactive oxygen species overexpression induced by LPS. Amino acid metabolism, energy metabolism and purine metabolism were disordered in LPS-treated group, and metabolic pathway analysis indicated that the double-targeting PC nanoparticles reversed some of LPS impacts. The changes of these potential biomarkers and their corresponding pathways are helpful to further understand the mechanism of PC nanoparticles in alleviating inflammation, and promote their application in nutrition intervention.

Kultur In Vitro Pisang Cavendish (Musa acuminata L.) Pada Media Dengan Konsentrasi Berbeda Ekstrak Jambu Batu Dan Benzyl Amino Purine

In vitro culture requires suitable media, the addition of BAP as a cytokinin is considered to accelerate the growth of Cavendish bananas. Constraints of browning in explants are also often found in the growth of banana explants. The addition of guava extract as an organic antioxidant and BAP is expected to suppress browning and improve the growth of Cavendish banana explants in vitro. The study was aimed to obtain the best concentration of guava extract (Psidium guajava) and Benzyl Amino Purine for the growth of Cavendish banana (Musa acuminata) explants in vitro. This study used a completely randomized design (CRD) with a factorial pattern of 4 x 3 with 4 replications. Treatment of guava extract added to the media (0 g/l, 1 g/l, 2 g/l, 3 g/l) and BAP (3 ppm, 6 ppm, 9 ppm). The data were analyzed using variance with the F test and continued with Duncan's Multiple Distance Test with 5% significance level. The results showed that the best result for the number of shoots were in the treatment of 1 g/l guava extract + BAP 3 ppm. In the number of leaves and high shoots the best result were in the treatment of 0 g/l guava extract + BAP 6 ppm

In vitro Propagation and Assessment of Genetic Homogeneity using RAPD and ISSR Markers in Tinospora cordifolia (Wild.) Hook. F. &amp; Thoms, An Important Medicinal Plant of Nepal

The Menispermaceae family includes the widely glabrous, succulent, climbing shrub Tinospora cordifolia (Gurjo), which has been found to have a variety of pharmacological and ethnomedicinal characteristics. T. cordifolia is also one of the most commercially exploited plants in pharmaceuticals. The nodal segment explants were initially cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of 6-benzyl amino purine (BAP), and/or Kinetin (KIN) among which MS +BAP (2.0 mg/l) induced shoot initiation after 7 days of post-inoculation. The nodal segments were then excised and treated with various concentrations of BAP, BAP with KIN, and coconut water for the proliferation among which 5 mg/l induced significant nodal segment proliferation (9.0 nodal segments/per explant) and shoot length (8.0 cm). The efficacy of coconut water in increasing the nodal segment proliferation of T. cordifolia was tested and the shoot proliferation increased significantly at 5% and 10% of coconut water, however, the maximum response of shoot number (23.0), shoot length (12 cm) was in the MS medium supplementation with BAP (5mg/l) and 10% coconut water. The genetic fidelity of these plants was also confirmed by random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) markers in wild and in vitro cultures. This protocol is an efficient way for the in vitro mass propagation of true-to-type plantlets of T. cordifolia which provides a basis for germplasm conservation and sustainable utilization.

Phenolic Compound Production Increased In Vitro Regenerated Cichorium intybus L.

Chicory (Cichorium intybus L.) is a low-height perennial or biennial herb from the family of Asteraceae. Investigation of different in vitro regeneration strategies of Cichorium intybus and increasing the number of secondary metabolites in vitro regenerated plant samples were the aims of the research. Callus and plant regenerations were achieved in basal plant growth media supplemented with plant growth regulators (PGRs). Whole plant regeneration was carried out by direct organogenesis from leaf explant in Murashige and Skoog (MS) and B5 media supplemented with naphthalene acetic (NAA) acid and indole-3-butyric acid (IBA). The highest callus quantity was produced in MS medium supplemented with indole-3-acetic acid (IAA) and benzyl amino purine (BAP). The combination and concentrations of PGRs used in MS and B5 media not only provided root and shoot formation with callus, but also caused a change in the amounts of phenolic components. In addition, some PGRs used caused an increase in the number of phenolic compounds in callus and shoots developed from the leaf explant. When plants that grow in vitro and in vivo are compared with each other, it has been determined that plants grown in vivo contain higher amounts of some phenolic compounds. In vivo and in vitro samples were extracted in ethanol/water (80:20 v/v). The analysis of phenolic compounds (caftaric, chicoric, and chlorogenic acids and esculin) were performed in high-performance liquid chromatography (HPLC) and inulin was in UV spectrophotometry. The caftaric and chlorogenic acids and inulin concentrations were higher in vivo samples than that in vitro. Contrarily, esculin, and chicoric acid concentrations were higher in the in vitro regenerated samples. The higher concentration of valuable compounds in the in vitro regenerated samples, especially in callus tissue, gives hope for large-scale production of secondary metabolites under laboratory conditions.

Optimization of bud disinfection technique and influence of growth regulators on micropropagation in ginger (Zingiber officinale Rosc.)

Ginger (Zingiber officinale Rosc.) is a spice, considered as a food medicine because of its numerous beneficial actions on health. However, its production faces several constraints, including the lack of efficient planting material. This is a limiting factor for industrial ginger production. The present study therefore aimed at developing an effective in vitro regeneration protocol for ginger. First, three disinfecting agents (sodium hypochlorite, calcium hypochlorite and mercury chloride) were tested. Then, different combinations of naphthalene acetic acid (0.5 mg/L NAA) and/or benzyl amino purine (1, 3 and 5 mg/L BAP) were evaluated on in vitro shoots regeneration. The results revealed that 3.6% sodium hypochlorite, used for 20 min, induced the best disinfection rates (100%) and healthy buds (84.66%). Furthermore, this study showed that the tested hormonal combinations significantly influenced shoots proliferation in ginger. However, the Proliferation Medium 4 (PM4) [Murashige and Skoog medium including vitamin B5 (MSB) + 0.5 mg/L of Naphthalene Acetic Acid (ANA) + 5 mg/L of Benzyl Amino Purine (BAP)] was the most effective. It induced the highest average number of shoots (22.83 shoots) with an induction rate of 80.50%. As a result of this study, 3.6% sodium hypochlorite used for 20 min and MP4 medium (MSB + 0.5 mg/L ANA + 5 mg/L BAP) were selected for in vitro ginger regeneration.

Antibacterial Mechanism of 2R,3R-Dihydromyricetin Against Staphylococcus aureus: Deciphering Inhibitory Effect on Biofilm and Virulence Based on Transcriptomic and Proteomic Analyses.

Staphylococcus aureus is a major foodborne pathogen that leads to various diseases due to its biofilm and virulence factors. This study aimed to investigate the inhibitory effect of 2R,3R-dihydromyricetin (DMY), a natural flavonoid compound, on the biofilm formation and virulence of S. aureus, and to explore the mode of action using transcriptomic and proteomic analyses. Microscopic observation revealed that DMY could remarkably inhibit the biofilm formation by S. aureus, leading to a collapse on the biofilm architecture and a decrease in viability of biofilm cell. Moreover, the hemolytic activity of S. aureus was reduced to 32.7% after treatment with subinhibitory concentration of DMY (p < 0.01). Bioinformation analysis based on RNA-sequencing and proteomic profiling revealed that DMY induced 262 differentially expressed genes and 669 differentially expressed proteins (p < 0.05). Many downregulated genes and proteins related to surface proteins were involved in biofilm formation, including clumping factor A (ClfA), iron-regulated surface determinants (IsdA, IsdB, and IsdC), fibrinogen-binding proteins (FnbA, FnbB), and serine protease. Meanwhile, DMY regulated a wide range of genes and proteins enriched in bacterial pathogenesis, cell envelope, amino acid metabolism, purine and pyrimidine metabolism, and pyruvate metabolism. These findings suggest that DMY targets S. aureus through multifarious mechanisms, and especially prompt that interference of surface proteins in cell envelope would lead to attenuation of biofilm and virulence.

Microplastics affect arsenic bioavailability by altering gut microbiota and metabolites in a mouse model

Microplastics exposure is a new human health crisis. Although progress in understanding health effects of microplastic exposure has been made, microplastic impacts on absorption of co-exposure toxic pollutants such as arsenic (As), i.e., oral bioavailability, remain unclear. Microplastic ingestion may interfere As biotransformation, gut microbiota, and/or gut metabolites, thereby affecting As oral bioavailability. Here, mice were exposed to arsenate (6 μg As g−1) alone and in combination with polyethylene particles of 30 and 200 μm (PE-30 and PE-200 having surface area of 2.17 × 103 and 3.23 × 102 cm2 g−1) in diet (2, 20, and 200 μg PE g−1) to determine the influence of microplastic co-ingestion on arsenic (As) oral bioavailability. By determining the percentage of cumulative As consumption recovered in urine of mice, As oral bioavailability increased significantly (P < 0.05) from 72.0 ± 5.41% to 89.7 ± 6.33% with PE-30 at 200 μg PE g−1 rather than with PE-200 at 2, 20, and 200 μg PE g−1 (58.5 ± 19.0%, 72.3 ± 6.28%, and 69.2 ± 17.8%). Both PE-30 and PE-200 exerted limited effects on pre- and post-absorption As biotransformation in intestinal content, intestine tissue, feces, and urine. They affected gut microbiota dose-dependently, with lower exposure concentrations having more pronounced effects. Consistent with the PE-30-specific As oral bioavailability increase, PE exposure significantly up-regulated gut metabolite expression, and PE-30 exerted greater effects than PE-200, suggesting that gut metabolite changes may contribute to As oral bioavailability increase. This was supported by 1.58–4.07-fold higher As solubility in the presence of up-regulated metabolites (e.g., amino acid derivatives, organic acids, and pyrimidines and purines) in the intestinal tract assessed by an in vitro assay. Our results suggested that microplastic exposure especially smaller particles may exacerbate the oral bioavailability of As, providing a new angle to understand health effects of microplastics.

Folate metabolism negatively regulates OAS-mediated antiviral innate immunity via ADAR3/endogenous dsRNA pathway

BackgroundFolate (FA) is an essential cofactor in the one-carbon (1C) metabolic pathway and participates in amino acid metabolism, purine and thymidylate synthesis, and DNA methylation. FA metabolism has been reported to play an important role in viral replications; however, the roles of FA metabolism in the antiviral innate immune response are unclear. ObjectiveTo evaluate the potential regulatory role of FA metabolism in antiviral innate immune response, we establish the model of FA deficiency (FAD) in vitro and in vivo. The molecular and functional effects of FAD on 2′-5′-oligoadenylate synthetases (OAS)-associated antiviral innate immunity pathways were assessed; and the potential relationship between FA metabolism and the axis of adenosine deaminases acting on RNA 3 (ADAR3)/endogenous double-stranded RNA (dsRNA)/OAS was further explored in the present study, as well as the potential translatability of these findings in vivo. MethodsFA-free RPMI 1640 medium and FA-free feed were used to establish the model of FAD in vitro and in vivo. And FA and homocysteine (Hcy) concentrations in cell culture supernatants and serum were used for FAD model evaluation. Ribonucleoprotein immunoprecipitation assay was used to enrich endogenous dsRNA, and dot-blot was further used for quantitative analysis of endogenous dsRNA. Western-blot assay, RNA isolation and quantitative real-time PCR, immunofluorescence assay, and other molecular biology techniques were used for exploring the potential mechanisms. ResultsIn this study, we observed that FA metabolism negatively regulated OAS-mediated antiviral innate immune response. Mechanistically, FAD induced ADAR3, which interacted with endogenous dsRNA, to inhibit deaminated adenosine (A) being converted into inosine (I), leading to the cytoplasmic accumulation of dsRNA. Furthermore, endogenous dsRNA accumulated in cytoplasm triggered the host immune activation, thus promoting the expression of OAS2 to suppress the replication of viruses. Additionally, injection of 8-Azaadenosine to experimental animals, an A-to-I editing inhibitor, efficiently enhanced OAS-mediated antiviral innate immune response to reduce the viral burden in vivo. ConclusionsTaken together, our present study provided a new perspective to illustrate a relationship between FA metabolism and the axis of ADAR3/endogenous dsRNA/OAS, and a new insight for the treatment of RNA viral infectious diseases by targeting the axis of ADAR3/endogenous dsRNA/OAS.

Molecular identification and high fidelity micropropagation of shell ginger (Alpinia zerumbet)

Alpinia zerumbet is a medicinal and ornamental plant of the Zingiberaceae family cultivated in many parts of the world. The plant is medicinally important and often misidentified with closely related species. The objective of this work was to establish its DNA-based identity and a protocol for high-fidelity micropropagation to support the medicinal plant industry. PCR amplification and subsequent sequencing of RuBisCO large subunit (rbcL) and Maturase K (matK) gene successfully report its DNA barcode. In vitro shoots were induced the best from rhizome buds on Murashige and Skoog (MS) medium supplemented with 6.0 mg L−1 6-benzyl amino purine and 30 g L−1 sucrose. Microshoots were rooted well in MS medium amended with 1.5 mg L−1 naphthalene acetic acid. Plantlets were satisfactorily acclimated to soils maintained in a growth chamber and subsequently in the open air for 10–20 days. Analyzing eight random amplified polymorphic DNA and eight inter-simple sequence repeat primers resulted in 53 and 60 distinct bands, respectively. Uniform banding patterns suggest the genetic uniformity of the micropropagated plants. The results offer an accurate identification of A. zerumbet from any plant tissue. The micropropagation protocol would be helpful for economically feasible and environmentally sustainable exploitation of this valuable plant.