scholarly journals Construction of a Ginseng Root-Meristem Sensor and a Sensing Kinetics Study on the Main Nitrogen Nutrients

Sensors ◽  
2021 ◽  
Vol 21 (3) ◽  
pp. 681
Author(s):  
Shiang Wang ◽  
Dingqiang Lu ◽  
Guangchang Pang
Keyword(s):  
Limit Of Detection ◽  
Inorganic Nitrogen ◽  
Equilibrium Analysis ◽  
Ginseng Root ◽  
Continuous Cropping ◽  
Sodium Glutamate ◽  
Sensor Sensitivity ◽  
Group A ◽  
Hardy Weinberg Equilibrium ◽  
Nitrogen Nutrients

Severe continuous cropping obstacles exist in ginseng cultivation. In order to assess these obstacles, a “sandwich” ginseng root tissue sensor was developed for the kinetic determination of five nitrogen nutrients. The results showed that the sensing parameters of the sensor reached an ultrasensitive level (limit of detection up to 5.451 × 10−24 mol/L) for the five nitrogen nutrients, and exhibited good stability and reproducibility. In the order of two-, four-, and six-year-old ginseng plants, the sensitivity to inorganic nitrogen nutrients (sodium nitrate and urea) showed an upward trend following an initial decline (the interconnected allosteric constant Ka values acted as the parameter). The fluctuations in sensor sensitivity to organic nitrogen nutrients, specifically nucleotides (disodium inosinate and disodium guanylate), were relatively small. The sensor sensitivity of two-, four-, and six-year-old ginseng plants to sodium glutamate was 9.277 × 10−19 mol/L, 6.980 × 10−21 mol/L, and 5.451 × 10−24 mol/L, respectively. Based on the survival rate of the seedlings and mortality rate of the ginseng in each age group, a Hardy–Weinberg equilibrium analysis was carried out. The results showed that the sensing ability of the root system to sodium glutamate may be an important factor affecting its survival under continuous cropping obstacles with increasing age.

scholarly journals A Panax notoginseng Root Tip Meristem Biosensor and Its Sensing Kinetics for Five Important Nitrogen Nutrients

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Zi Qing Zheng ◽  
Bo Niu ◽  
Ding Qiang Lu ◽  
Guang Chang Pang
Keyword(s):  
Sodium Nitrate ◽  
Inorganic Nitrogen ◽  
Low Cost ◽  
Panax Notoginseng ◽  
Interaction Constant ◽  
Electrode System ◽  
Plant Age ◽  
Root Tip ◽  
Sodium Glutamate ◽  
Nitrogen Nutrients

Plants absorb nitrogen mainly through their roots. Nitrogen sensing is required for the absorption and transport of different nitrogen nutrients. In this study, we constructed biosensors with immobilized Panax notoginseng root tip meristems based on a three-electrode system and successfully determined the kinetics of the interactions between the P. notoginseng root tip meristems and five important nitrogen nutrients, namely, urea, sodium nitrate, sodium glutamate, disodium inosinate, and disodium guanylate. We discovered that the biosensor’s sensing kinetics was similar to the enzyme–substrate kinetics, and the receptor–ligand interconnected allosteric interaction constant Ka (mol/L), analogous to the Michaelis constant, was calculated. The result showed that the root tip meristems of two- to four-year-old P. notoginseng plants had a higher capacity to sense inorganic nitrogen nutrients (sodium nitrate and urea) than the three organic nitrogen nutrients. The ability of the plants to sense inorganic nitrogen nutrients decreased with an increase in plant age. The sensing sensitivity of four-year-old P. notoginseng plants to disodium inosinate and disodium guanylate was 100- to 10,000-fold lower than that of the two- and three-year-old plants. Additionally, the capability to sense sodium glutamate decreased initially and then increased with an increase in plant age. The biosensors reached an ultra-sensitive level ( 1 × 10 − 22  mol/L) in sensing the five nitrogen nutrients and exhibited advantages such as good stability and reproducibility, low cost, a simple structure, and a rapid response, providing a new approach for quantitative determination of the capability of plants to sense different nitrogen nutrients.

Development of a real-time reverse transcription-PCR assay to detect and quantify group A rotavirus equine-like G3 strains

2021 ◽  
Author(s):  
Eric M. Katz ◽  
Mathew D. Esona ◽  
Rashi Gautam ◽  
Michael D. Bowen
Keyword(s):  
Viral Load ◽  
Real Time ◽  
Reverse Transcription ◽  
Dynamic Range ◽  
Limit Of Detection ◽  
Human Populations ◽  
Pcr Assay ◽  
Group A Rotavirus ◽  
Reverse Transcription Pcr ◽  
Group A

Since 2013, group A rotavirus strains characterized as novel DS-1-like inter-genogroup reassortant ‘equine-like G3’ strains have emerged and spread across five continents among human populations in at least 14 countries. Here we report a novel one-step TaqMan quantitative real-time reverse transcription-PCR assay developed to genotype and quantify the viral load for samples containing rotavirus equine-like G3 strains. Using a universal G forward primer and a newly designed reverse primer and TaqMan probe, we developed and validated an assay with a linear dynamic range of 2.3 × 10 9 – 227 copies per reaction and a limit of detection of 227 copies. The percent positive agreement, percent negative agreement, and precision of our assay were 100.00%, 99.63%, and 100.00%, respectively. This assay can simultaneously detect and quantify the viral load for samples containing DS-1-like inter-genogroup reassortant equine-like G3 strains with high sensitivity and specificity, faster turnaround time, and decreased cost and will be valuable for high-throughput screening of stool samples collected to monitor equine-like G3 strain prevalence and circulation among human populations throughout the world.

scholarly journals American Ginseng Root Rot Caused by Fusarium redolens in China

Plant Disease ◽  
2021 ◽  
Author(s):  
Sanhong Fan ◽  
Fangjie Zhao ◽  
Jiguang Zhang ◽  
Wenjing Shang ◽  
Xiaoping Hu
Keyword(s):  
Root Rot ◽  
Vascular Tissue ◽  
Morphological Characteristics ◽  
American Ginseng ◽  
Shaanxi Province ◽  
Ginseng Root ◽  
Continuous Cropping ◽  
Sterile Water ◽  
Fusarium Spp ◽  
Fusarium Redolens

American ginseng (Panax quinquefolium L.) originating from North America is one of important herbal medicine and economic crops . With the increasing market demand, China has become the third producer and the largest consumer country of American ginseng. However, continuous cropping obstacle has become the most serious problem for the production of American ginseng, and the continuous cropping of soils usually lead to accumulations of root fungal pathogens and increasing plant disease occurrence (1), root rot caused by the notorious soil-borne pathogenic fungi, Fusarium spp., results in a significant reduction of yield and quality of American ginseng. Investigation of American ginseng root rot was carried out in Liuba county, Shaanxi province, China from 2017 to 2019. About 20% of over 3-year-old American ginseng showed varied root rot symptoms in newly reclaimed fields, and more than 70% in continuous American ginseng planting fields. Among these root rot diseases, we found one kind of disease which shows symptoms of red leaves in initial stage and yellow or yellow brown lesions at the reed heads or taproots. The lesions mainly appear on the root surface; however, the vascular tissue has no discoloration. The aboveground parts become wilted and died, and the whole root appears dark brown rots. Fifteen Fusarium spp. isolates were obtained by cutting diseased rot roots into 5 × 5 mm2 pieces, disinfecting in 70% ethanol for 1 min, rinsing 2 ~ 3 times in sterile water for 1 min and isolating on PDA medium including 50 μg/mL streptomycin sulfate. All the isolates have identical morphological characteristics. The colony was white with curved and uplifted aerial hyphae in central region. The colony diameter was 48 ~51 mm after 6 days at room temperature. Microconidia were oval to cylindrical shape with 0 to 1 septa, ranged from 6.24 to10.09 μm long; the macroconidia were fusiform to conical with a hooked apical cell and a foot-shaped basal cell, usually 3 to 5 septa, ranged from 31.45 to 42.52 μm long. The chlamydospores were not found under our culture condition. Preliminary data analysis showed that the morphological characteristics of these isolates were consistent with the descriptions of Fusarium redolens (2). To clarify the fungus in the taxonomy , the rDNA internal transcribed spacer (ITS), the translation elongation factor 1 alpha (TEF1-α) and the RNA polymerase II subunit 1 (RPB1) fragments of two randomly selected isolates were amplified and sequenced. The sequences of the corresponding fragments of the two isolates were identical. The blast results in the GenBank and FUSARIUM-ID databases show the isolates belong to F. redolens (3). Previous study indicated F. redolens has an indistinguishable relative, F. hostae (4). Although the ITS sequence (MW331695) cannot provide enough information to distinguish them, the phylogenetic tree combined the sequence of TEF1-α (tempID: 2407237 ) and RPB1 (tempID: 2407229) clearly showed that the isolates are F. redolens. (Fig) The pathogenicity of a representative isolate, YP04, was tested on ginseng taproot by in vivo inoculation experiments with three replications. The taproot surface of 2-year-old healthy ginseng was washed and disinfested with 75% alcohol for 1 min and rinsed with sterile water, and dried. The surface of taproot was injured with sterilized steel needles and immersed in 1 × 106 /ml spore suspension (sterile water for control plants) for 30 min. The treatment and control plants were transplanted in 20 cm diameter flowerpots filled with sterilized humus and cultured in a greenhouse at 18-23°C. Six days after transplanting, the leaves began to turn red. The cortex of ginseng taproot showed yellow brown lesions and the vascular tissue turn to light yellow. Fifteen days after transplanting, the aboveground parts of treatment plants began to wilting and the taproots showed serious rots. no taproot rot was observed in the controls. The pathogen was re-isolated from the diseased taproots successfully. To our knowledge, this is the first report of F. redolens causing root rot of American ginseng in China.

scholarly journals Superoxide Dismutase 2 Val16Ala Polymorphism is Associated with Amiodarone-Associated Liver Injury

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Branimir Radmanovic ◽  
Jovan Jovanovic ◽  
Natasa Djordjevic ◽  
Dejan Baskic ◽  
Jelena Cukic ◽  
...  
Keyword(s):  
Liver Injury ◽  
Control Group ◽  
Single Nucleotide ◽  
Clinical Center ◽  
Group A ◽  
Hardy Weinberg Equilibrium ◽  
Pcr Method ◽  
Group B ◽  
And Control ◽  
Superoxide Dismutase 2

AbstractAssociation of SOD2 V16A single-nucleotide polymorphism (rs4880) with drug hepatotoxicity were reported but relationships with amiodarone prescriptions remained unexplored. Research was an exploratory, controlled prospective clinical trial. Patients hospitalized and treated in Clinical Center in Kragujevac, Serbia (in year 2017) were divided into experimental (using amiodarone, having liver injury, n=29, 19 males, the mean age 66.8±10.4 years), control A (neither amiodarone use nor hepatotoxicity, n=29, 19, 66.1±10.3) and control B group (using amiodarone, not having hepatotoxicity, n=29, 19, 66.8±9.8). From blood samples, among other routine biochemistry, genotyping for SOD2 polymorphism Val16Ala was conducted using real-time PCR method with TaqMan® Genotyping Master Mix and TaqMan® DME Genotyping Assay for rs4880. Patients taking amiodarone and having liver injury were mostly carriers of Val/Val (TT) genotype (13 of 24 patients, 54.2%) while Val/Ala (TC) and Ala/Ala (CC) genotypes prevailed in control group A (19 of 40, 47.5%) and control group B (9 of 23, 39.1%), respectively (2=10.409, p=0.034). Frequency of Val (T) and Ala (C) alleles were 0.51 and 0.49, respectively in the whole study sample (Hardy Weinberg equilibrium, 2=0.56, p=0.454). Carriers of TT genotype had significantly higher ALT (437.0±1158.0 vs 81.9131.5 U/L), total bilirubin (28.320.5 vs 15.313.0 mol/L) and total bile acid concentrations (10.910.2 vs 6.45.3 mol/L) compared to carriers of TC genotype (U=2.331, p=0.020, U=3.204, p=0.001 and U=2.172, p=0.030, respectively). Higher incidence of 47T allele of SOD2 was inpatients with amiodarone-associated liver injury as compared to patients on amiodarone not experiencing hepatotoxic effects.

scholarly journals Spatial-temporal dynamics of N-cycle functional genes in a temperate Atlantic estuary (Douro, Portugal)

2020 ◽  
Vol 84 ◽  
pp. 205-216
Author(s):  
P Salgado ◽  
A Machado ◽  
AA Bordalo
Keyword(s):  
Microbial Communities ◽  
Temporal Dynamics ◽  
Inorganic Nitrogen ◽  
Functional Genes ◽  
Estuarine Ecosystem ◽  
Monthly Basis ◽  
N Cycle ◽  
Low Salinity ◽  
Key Factor ◽  
Nitrogen Nutrients

Understanding the spatial and seasonal dynamics of nitrogen (N)-cycle microbial communities is pivotal for the knowledge of N biogeochemistry. The present study addressed the spatial-temporal variability of nitrification (bacterial and archaeal amoA) and denitrification (nirS, nirK, and nosZI) key genes, as well as of non-denitrifying nitrous oxide (N2O) reducers (nosZII), coupled with key environmental variables, in an estuarine ecosystem (Douro, NW Portugal). Samples were collected on a monthly basis over 1 yr, key physical-chemical parameters were measured, and specific functional gene abundances were assayed. The results revealed a clear seasonality for nirS, nosZII, and bacterial and archaeal amoA abundance, with an increase during the winter/spring seasons. This period was especially characterized by high levels of dissolved oxygen, low temperature, low salinity, and increased turbidity. Indeed, turbidity emerged as the key factor controlling the distribution of nirS, nosZII bacterial, and archaeal amoA abundance. In contrast, the abundance of nosZI increased during the summer, while nirK abundance was enhanced from the fall to late spring. Additionally, the availability of dissolved inorganic nitrogen nutrients had no commensurable effect on N-cycle functional genes. This study of the annual variation of N-cycle functional genes in a temperate Atlantic estuary provides a major contribution to the understanding of how environmental factors potentially influence the distribution and abundance of N-cycle microbial communities.

Croissance de la biomasse azotée du Phaeodactylum tricornutum (Bacillariophyceae) en cultures discontinues dialysante et non dialysante

1992 ◽  
Vol 38 (9) ◽  
pp. 945-952 ◽  
Author(s):  
Pierre Marsot ◽  
Allan D. Cembella ◽  
Khadija Mouhri
Keyword(s):  
Phaeodactylum Tricornutum ◽  
Inorganic Nitrogen ◽  
Biomass Concentration ◽  
Diffusion Limit ◽  
Active Growth ◽  
Culture Techniques ◽  
Uptake Activity ◽  
Nitrogen Nutrients ◽  
Menten Constant ◽  
Dialysis Culture

The unique conditions associated with discontinuous (batch) dialysis culture of the diatom Phaeodactylum tricornutum elicit different nutritional responses from those observed in nondialysis culture. Simultaneous determinations of the nitrogen biomass concentration and of the nitrogen nutrients (NO3− + NO2−) in the culture chamber, as well as in the seawater nutrient medium at the entrance and exit of the dialyzer, revealed that nitrogen-biomass production in dialysis cultures is achieved mainly (>90%) during postexponential growth, when the concentration of nutrients is limiting (smaller than Michaelis-Menten constant). Almost half of this biomass is produced at the diffusion limit of the apparatus, i.e., when the mass transfer of nutrient substrates, which determines the total uptake activity of the culture, reaches a maximum. In contrast, in nondialysis discontinuous cultures, in which the postexponential growth phase is short, most of the total cellular nitrogen is accumulated during active growth. Certain physiological indices relating to the active uptake and assimilation of inorganic nitrogen are consistent with the different nutritional responses elicited by these two types of culture techniques and explain the high biomass levels obtained in dialysis culture. Key words: dialysis culture, diatom, nitrogen metabolism.

scholarly journals One-step multiplex real-time RT-PCR assay for detecting and genotyping wild-type group A rotavirus strains and vaccine strains (Rotarix® and RotaTeq®) in stool samples

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e1560 ◽  
Author(s):  
Rashi Gautam ◽  
Slavica Mijatovic-Rustempasic ◽  
Mathew D. Esona ◽  
Ka Ian Tam ◽  
Osbourne Quaye ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Wild Type ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Qrt Pcr ◽  
Group A ◽  
One Step ◽  
Stool Samples ◽  
Pcr Assays ◽  
Type Strains

Background.Group A rotavirus (RVA) infection is the major cause of acute gastroenteritis (AGE) in young children worldwide. Introduction of two live-attenuated rotavirus vaccines, RotaTeq® and Rotarix®, has dramatically reduced RVA associated AGE and mortality in developed as well as in many developing countries. High-throughput methods are needed to genotype rotavirus wild-type strains and to identify vaccine strains in stool samples. Quantitative RT-PCR assays (qRT-PCR) offer several advantages including increased sensitivity, higher throughput, and faster turnaround time.Methods.In this study, a one-step multiplex qRT-PCR assay was developed to detect and genotype wild-type strains and vaccine (Rotarix® and RotaTeq®) rotavirus strains along with an internal processing control (Xeno or MS2 RNA). Real-time RT-PCR assays were designed for VP7 (G1, G2, G3, G4, G9, G12) and VP4 (P[4], P[6] and P[8]) genotypes. The multiplex qRT-PCR assay also included previously published NSP3 qRT-PCR for rotavirus detection and Rotarix® NSP2 and RotaTeq® VP6 qRT-PCRs for detection of Rotarix® and RotaTeq® vaccine strains respectively. The multiplex qRT-PCR assay was validated using 853 sequence confirmed stool samples and 24 lab cultured strains of different rotavirus genotypes. By using thermostablerTthpolymerase enzyme, dsRNA denaturation, reverse transcription (RT) and amplification (PCR) steps were performed in single tube by uninterrupted thermocycling profile to reduce chances of sample cross contamination and for rapid generation of results. For quantification, standard curves were generated using dsRNA transcripts derived from RVA gene segments.Results.The VP7 qRT-PCRs exhibited 98.8–100% sensitivity, 99.7–100% specificity, 85–95% efficiency and a limit of detection of 4–60 copies per singleplex reaction. The VP7 qRT-PCRs exhibited 81–92% efficiency and limit of detection of 150–600 copies in multiplex reactions. The VP4 qRT-PCRs exhibited 98.8–100% sensitivity, 100% specificity, 86–89% efficiency and a limit of detection of 12–400 copies per singleplex reactions. The VP4 qRT-PCRs exhibited 82–90% efficiency and limit of detection of 120–4000 copies in multiplex reaction.Discussion.The one-step multiplex qRT-PCR assay will facilitate high-throughput rotavirus genotype characterization for monitoring circulating rotavirus wild-type strains causing rotavirus infections, determining the frequency of Rotarix® and RotaTeq® vaccine strains and vaccine-derived reassortants associated with AGE, and help to identify novel rotavirus strains derived by reassortment between vaccine and wild-type strains.

scholarly journals Consistency of thresholds for eutrophication assessments, examples and recommendations

2021 ◽  
Vol 193 (10) ◽  
Author(s):  
D. Topcu ◽  
U. Brockmann
Keyword(s):  
Chlorophyll A ◽  
North Sea ◽  
Coastal Waters ◽  
Inorganic Nitrogen ◽  
Mixing Processes ◽  
The North ◽  
The North Sea ◽  
International Harmonisation ◽  
Nitrogen Nutrients ◽  
Mixing Diagrams

AbstractInternational harmonisation of management goals for eutrophication processes in coastal waters, requiring reduction of discharges and depositions of nutrients and organic matter, needs coordinated assessments and measures. This is especially necessary in open areas, connected by currents and mixing processes with trans-boundary exchanges. Management goals, defined nationally as local thresholds for nutrients and chlorophyll-a, had been applied recently (2006–2014) within international eutrophication assessments in the North Sea (OSPAR) and Baltic Sea (HELCOM). Consistency of thresholds for nitrogen nutrients and chlorophyll-a concentrations is tested by mixing diagrams and correlations between nitrogen nutrients (total and inorganic nitrogen) and chlorophyll-a. Results indicate mean consistent relations, but single deviations as in the continental coastal water of the North Sea surpassed means by a factor up to 5 for chlorophyll-a in relation to inorganic nitrogen. Thresholds differed across national borders significantly. Correlations of thresholds and assed data reflect the degree of regional deviations by comparison. Consistency of regionally applied thresholds can be achieved stepwise, by application of regionally correlated means, by adaptation to mixing and parameter relations, and finally by relations of thresholds to natural background concentrations. By this, consistency of international assessments can be improved generally, allowing coordinated management of open coastal waters.

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