scholarly journals Identification of a New Badnavirus in the Chinaberry (Melia azedarach) Tree and Establishment of a LAMP-LFD Assay for Its Rapid and Visual Detection

Viruses ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2408
Author(s):  
Huixin Lu ◽  
Jintian Tang ◽  
Kai Sun ◽  
Xiaoping Yu
Keyword(s):  
Virus Transmission ◽  
Lamp Assay ◽  
Melia Azedarach ◽  
Small Interfering Rnas ◽  
Tree Seedling ◽  
Traditional Medicines ◽  
Tree Virus ◽  
Pathogenic Viruses ◽  
Leaf Deformation ◽  
Development And Validation

The Chinaberry tree, a member of the Meliaceae family, is cultivated in China for use in traditional medicines. In 2020, Chinaberry trees with leaf deformation symptoms were found in Hangzhou, Zhejiang province, China. In order to identify possible pathogenic viruses, a symptomatic sample was subjected to deep sequencing of small interfering RNAs. Assembly of the resulting sequences led to the identification of a novel badnavirus, provisionally designated Chinaberry tree badnavirus 1 (ChTBV1). With the recent development of China’s seedling industry and increasing online shopping platforms, the risk of tree virus transmission has increased substantially. Therefore, it is important to detect the occurrence of ChTBV1 to ensure the safety of the Chinaberry tree seedling industry. Here, we describe the development and validation of a sensitive and robust method relying on a loop-mediated isothermal amplification (LAMP) assay, targeting a 197 nt region, to detect ChTBV1 from Chinaberry tree leaves. The LAMP assay was also adapted for rapid visualization of results by a lateral flow dipstick chromatographic detection method.

Nightshade curly top virus: A possible new virus of the genus Topocuvirus infecting Solanum nigrum in China

Plant Disease ◽  
2020 ◽  
Author(s):  
Kai Sun ◽  
Yan Liang ◽  
Xueting Zhong ◽  
Xuenan Hu ◽  
Pengjun Zhang ◽  
...  
Keyword(s):  
Amino Acid ◽  
Phylogenetic Analyses ◽  
Solanum Nigrum ◽  
Open Reading Frames ◽  
Small Interfering Rnas ◽  
Protein Amino Acid ◽  
Sequence Identity ◽  
Pathogenic Viruses ◽  
Leaf Deformation ◽  
Symptomatic Sample

Virus-like symptoms, including leaf deformation and curling, were observed on nightshade (Solanum nigrum) in Zhejiang province, China. To identify possible pathogenic viruses or viroids, a symptomatic sample was subjected to deep sequencing of small interfering RNAs. Assembly of the resulting sequences led to identification of a novel geminivirus, provisionally designated nightshade curly top virus (NCTV). The complete genomic DNA sequence is 2,867 nucleotides that encodes seven open reading frames. NCTV shares 77.1 % overall nucleotide sequence identity, 86.3 % coat protein amino acid, and 78.9 % replication-associated protein amino acid sequence identity with Topocuvirus tomato pseudo-curly top virus (TPCTV). Polymerase chain reaction screening of nightshade field isolates indicated that NCTV is widely distributed in Zhejiang. Agrobacterium-mediated inoculation revealed that NCTV is highly infectious to Nicotiana benthamiana, Solanum nigrum, Solanum lycopersicum, and Solanum tuberosum. Based on pairwise comparisons and phylogenetic analyses, NCTV is proposed as a provisional member of the genus Topocuvirus.

scholarly journals Subgenomic flavivirus RNA binds the mosquito DEAD/H-box helicase ME31B and determines Zika virus transmission by Aedes aegypti

2019 ◽  
Vol 116 (38) ◽  
pp. 19136-19144 ◽  
Author(s):  
Giel P. Göertz ◽  
Joyce W. M. van Bree ◽  
Anwar Hiralal ◽  
Bas M. Fernhout ◽  
Carmen Steffens ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Zika Virus ◽  
Affinity Purification ◽  
Mechanistic Model ◽  
Virus Transmission ◽  
Protein Translation ◽  
Mass Spectrometry Analysis ◽  
Small Interfering Rnas ◽  
Spectrometry Analysis ◽  
Mosquito Infection

Zika virus (ZIKV) is an arthropod-borne flavivirus predominantly transmitted by Aedes aegypti mosquitoes and poses a global human health threat. All flaviviruses, including those that exclusively replicate in mosquitoes, produce a highly abundant, noncoding subgenomic flavivirus RNA (sfRNA) in infected cells, which implies an important function of sfRNA during mosquito infection. Currently, the role of sfRNA in flavivirus transmission by mosquitoes is not well understood. Here, we demonstrate that an sfRNA-deficient ZIKV (ZIKVΔSF1) replicates similar to wild-type ZIKV in mosquito cell culture but is severely attenuated in transmission by Ae. aegypti after an infectious blood meal, with 5% saliva-positive mosquitoes for ZIKVΔSF1 vs. 31% for ZIKV. Furthermore, viral titers in the mosquito saliva were lower for ZIKVΔSF1 as compared to ZIKV. Comparison of mosquito infection via infectious blood meals and intrathoracic injections showed that sfRNA is important for ZIKV to overcome the mosquito midgut barrier and to promote virus accumulation in the saliva. Next-generation sequencing of infected mosquitoes showed that viral small-interfering RNAs were elevated upon ZIKVΔSF1 as compared to ZIKV infection. RNA-affinity purification followed by mass spectrometry analysis uncovered that sfRNA specifically interacts with a specific set of Ae. aegypti proteins that are normally associated with RNA turnover and protein translation. The DEAD/H-box helicase ME31B showed the highest affinity for sfRNA and displayed antiviral activity against ZIKV in Ae. aegypti cells. Based on these results, we present a mechanistic model in which sfRNA sequesters ME31B to promote flavivirus replication and virion production to facilitate transmission by mosquitoes.

scholarly journals Lamalbid, Chlorogenic Acid, and Verbascoside as Tools for Standardization of Lamium album Flowers—Development and Validation of HPLC–DAD Method

Molecules ◽  
2020 ◽  
Vol 25 (7) ◽  
pp. 1721
Author(s):  
Monika E. Czerwińska ◽  
Eleonora Kalinowska ◽  
Dominik Popowski ◽  
Agnieszka Bazylko
Keyword(s):  
Chlorogenic Acid ◽  
High Performance ◽  
Dry Weight ◽  
Upper Respiratory Tract ◽  
Topical Medication ◽  
Traditional Medicines ◽  
Array Detection ◽  
Upper Respiratory ◽  
Development And Validation ◽  
Lamium Album

Preparations from the flowers or herb of the white dead nettle (Lamium album L.) are recommended for the treatment of upper respiratory tract disorders or as a topical medication for mild inflammation of the throat, mouth, and skin. Taking into consideration the significance of L. album in traditional medicines across Europe, as well as the lack of studies describing the quantities of their most abundant constituents, we aimed to design a high-performance liquid chromatography coupled with diode-array detection (HPLC–DAD) method for potential standardization procedures of extracts from flowers of L. album. The HPLC–DAD method was developed and validated for quantification of iridoids (lamalbid), phenolic acids/depsides (chlorogenic acid), phenylpropanoids (verbascoside), and flavonoids (rutin; quercetin malonylhexoside; tiliroside) in aqueous and ethanolic-aqueous extracts of Lamii albi flos. The method was specific, accurate, and precise. Lamalbid was the most abundant compound both in aqueous (39.09 ± 1.02 m/g dry weight) and ethanolic-aqueous (26.66 ± 0.64 m/g dry weight) extracts. The quantities of selected compounds, except for chlorogenic acid and tiliroside, were higher in the aqueous extract than in the ethanolic-aqueous one. In conclusion, the method developed allowed for quantitation of compounds from different classes. In particular, chlorogenic acid and verbascoside have been proposed as reference compounds for routine quantitative control of Lamii albi flos.

Development and Validation of a Rapid and Efficient Method for Simultaneous Determination of Scopolin and Scopoletin in Convolvulus Species by Ultra- high-performance Liquid Chromatography-Tandem Mass Spectrometry

2020 ◽  
Vol 16 (5) ◽  
pp. 494-503
Author(s):  
Asma’a Al-Rifai ◽  
Ahmad Aqel ◽  
Muneera Al-Saleem
Keyword(s):  
Bioactive Compounds ◽  
Plant Extracts ◽  
High Performance ◽  
Ethanolic Extract ◽  
Traditional Medicines ◽  
Medicinal Plant Extracts ◽  
Liquid Chromatography Tandem Mass ◽  
Recovery Percentage ◽  
Development And Validation ◽  
Tof Ms

Background: Convolvulus species are extensively used in traditional medicines for the treatment of peptic ulcer diseases primarily associated with the presence of many bioactive compounds, such as coumarins. Methods: A validation and improvement of a rapid, sensitive and simple technique for bioactive compounds analysis of scopolin and scopoletin in Convolvulus pilosellifolius (CP) and Convolvulus austroaegyptiacus (CA) plant extracts using UHPLC-MS/MS were performed. Both plants extracts were subjected to high-resolution DART-ToF-MS technique for fast profiling of their constituents. Results: DART-ToF-MS spectra proved the presence of experimental mass at 193.05137 and 355.10371 m/z in the extract of CA plant and at 193.05084 and 355.10365 m/z in CP plant extract for scopolin and scopoletin compounds, respectively. The total chromatographic analysis time was less than 10 min with excellent resolution and negligible matrix effect. The validated parameter showed good linearity (R2 ≥0.998) over a wide concentration range of 0.05-10.0 μg mL-1 for both scopolin and scopoletin, detection limits of scopolin and scopoletin were 0.03 and 0.01 μg mL-1, respectively. The method also showed good intraday and interday precisions (RSD<4.33%). The recovery percentage values were between 97.04 and 99.97% at three added concentration limits. The highest content of scopolin (3.485 mg g-1) and scopoletin (0.795 mg g-1) was detected in CA ethanolic extract. The studied plants were finally compared with all previous reports in terms of scopolin and scopoletin content. Conclusion: The results indicate that the suggested method can be used for regular analysis of active compounds in medicinal plant extracts.

scholarly journals Double-Stranded RNAs in Plant Protection Against Pathogenic Organisms and Viruses in Agriculture

Acta Naturae ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 13-21 ◽  
Author(s):  
S. Y. Morozov ◽  
A. G. Solovyev ◽  
N. O. Kalinina ◽  
M. E. Taliansky
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Transgenic Plants ◽  
Virulence Genes ◽  
Plant Protection ◽  
Pathogen Resistance ◽  
Pathogenic Microorganisms ◽  
Small Interfering Rnas ◽  
Pathogenic Viruses ◽  
Pathogenic Organisms

Recent studies have shown that plants are able to express the artificial genes responsible for the synthesis of double-stranded RNAs (dsRNAs) and hairpin double-stranded RNAs (hpRNAs), as well as uptake and process exogenous dsRNAs and hpRNAs to suppress the gene expression of plant pathogenic viruses, fungi, or insects. Both endogenous and exogenous dsRNAs are processed into small interfering RNAs (siRNAs) that can spread locally and systemically through the plant, enter pathogenic microorganisms, and induce RNA interference-mediated pathogen resistance in plants. There are numerous examples of the development of new biotechnological approaches to plant protection using transgenic plants and exogenous dsRNAs. This review summarizes new data on the use of transgenes and exogenous dsRNAs for the suppression of fungal and insect virulence genes, as well as viruses to increase the resistance of plants to these pathogens. We also analyzed the current ideas about the mechanisms of dsRNA processing and transport in plants.

scholarly journals Exogenous RNAs for Gene Regulation and Plant Resistance

2019 ◽  
Vol 20 (9) ◽  
pp. 2282 ◽  
Author(s):  
Alexandra S. Dubrovina ◽  
Konstantin V. Kiselev
Keyword(s):  
Crop Protection ◽  
Pathogen Resistance ◽  
Plant Viruses ◽  
Small Interfering Rnas ◽  
Down Regulation ◽  
Exogenous Rna ◽  
Pathogenic Viruses ◽  
Plant Surfaces ◽  
Uptake Mechanisms ◽  
Provided Examples

Recent investigations documented that plants can uptake and process externally applied double-stranded RNAs (dsRNAs), hairpin RNAs (hpRNAs), and small interfering RNAs (siRNAs) designed to silence important genes of plant pathogenic viruses, fungi, or insects. The exogenously applied RNAs spread locally and systemically, move into the pathogens, and induce RNA interference-mediated plant pathogen resistance. Recent findings also provided examples of plant transgene and endogene post-transcriptional down-regulation by complementary dsRNAs or siRNAs applied onto the plant surfaces. Understanding the plant perception and processing of exogenous RNAs could result in the development of novel biotechnological approaches for crop protection. This review summarizes and discusses the emerging studies reporting on exogenous RNA applications for down-regulation of essential fungal and insect genes, targeting of plant viruses, or suppression of plant transgenes and endogenes for increased resistance and changed phenotypes. We also analyze the current understanding of dsRNA uptake mechanisms and dsRNA stability in plant environments.

scholarly journals Application of Ethnobotanical Indices on the Use of Traditional Medicines against Common Diseases

2014 ◽  
Vol 2014 ◽  
pp. 1-21 ◽  
Author(s):  
Imran Khan ◽  
Naser M. AbdElsalam ◽  
Hassan Fouad ◽  
Akash Tariq ◽  
Riaz Ullah ◽  
...  
Keyword(s):  
Plant Species ◽  
Withania Somnifera ◽  
Modern Medicine ◽  
Melia Azedarach ◽  
Traditional Medicines ◽  
Alternate Fuel ◽  
Semistructured Interviews ◽  
Unexplored Area ◽  
Over 40 Years ◽  
Ethnomedicinal Knowledge

The present study was aimed at documenting the detailed ethnomedicinal knowledge of an unexplored area of Pakistan. Semistructured interviews were taken with 55 informants randomly chosen regarding detailed ethnomedicinal and sociocultural information. The study exposed 67 medicinal plant species used to prepare 110 recipes and the major modes of herbal formulation were decoction and powdering (20% each). The disease categories with the highest Fic values were gastrointestinal and dermatological (0.87 each). The study determined 3 plant species, i.e.,Acacia modestaWall.,Caralluma tuberculataR.Br., andWithania somnifera(L.) Dunal with a FL of 100%. DMR results showed thatOlea ferruginea(Sol.) Steud. ranked first,Morus albaL. ranked second, andMelia azedarachL. ranked third. Among the 55 informants, the male concentration was high (61%) and most of them were over 40 years old while a leading quantity of respondents (45%) was uneducated. There is a dire need to take necessary steps for the conservation of important medicinal plants by inhibiting overgrazing and providing alternate fuel resources. Young generations should be educated regarding the importance of ethnomedicinal knowledge and plants with high Fic and FL values should be further checked chemically and pharmacologically for future exploration of modern medicine.

scholarly journals Rapid and reliable distinguish of Morinda officinalis How by loop-mediated isothermal amplification (LAMP) assay

2020 ◽  
Author(s):  
Shuhan Wang ◽  
Aihua Sui ◽  
Yafei Han ◽  
Quan Zhou ◽  
Hang Xu ◽  
...  
Keyword(s):  
Low Cost ◽  
Lamp Assay ◽  
Its2 Sequence ◽  
Isothermal Amplification ◽  
Nucleic Acid Amplification ◽  
Nuclear Ribosomal Dna ◽  
Traditional Medicines ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Method ◽  
Human Disorders

Abstract The medicinal plant Morinda officinalis How (MO), especially the root, has been frequently used in traditional medicines around the world as an herbal drug for treating variable human disorders and diseases. Various adulterations of MO were found for economic or production limitations. However, authentication of MO from its adulterants by LAMP has not yet been established. The present study introduces a commercially available nucleic acid amplification method, loop-mediated isothermal amplification (LAMP) assay for the distinguish of MO from its adulterants. In this method, we combined DNA barcodes technology to design 2 pairs independent LAMP primers, which based on the internal transcribed spacer 2 (ITS2) sequence of MO’s nuclear ribosomal DNA. Our results showed that the LAMP could amplify the samples as expected and successfully identify target MO, and the limit for DNA template preciseness was verified as 1 × 10− 1 pg/µl. All the visual or real-time turbidity detection was performed within 60 min at approximately 63 °C. The result showed that the LAMP assay and primers we designed have high accuracy and efficiency for the differentiation of MO and its adulterants. Our results illustrated that the proposed low-cost, fast and reliable LAMP assay without the need for expensive equipment or specialized techniques could be a good way for MO rapid authentication.

scholarly journals Exploration of Antibacterial Potential of Melia Azedarach L.

2017 ◽  
Vol 1 (1) ◽  
pp. 19-26
Author(s):  
T. Munir ◽  
A Mohyuddin ◽  
Z Khan ◽  
R Haq
Keyword(s):  
Antimicrobial Agents ◽  
Herbal Medicines ◽  
Antibacterial Activities ◽  
Antimicrobial Activities ◽  
Melia Azedarach ◽  
Bacterial Strains ◽  
Traditional Medicines ◽  
Medicinal Value ◽  
Great Attraction ◽  
Causes Of Deaths

Melia azedarach L. belongs to one of the most versatile medicinal plants family meliaceae (mahogany) which has great attraction for researchers. The plant was selected for research because it was one of the least explored members. The presence of saponin, alkaloids, tannins and flavonoids in the leaves extracts of plant indicated its medicinal value. These compounds have pharmacological effects against cancer, viral and malarial infections that are one of the main causes of deaths. With passage of time most of bacterial strains develop resistance against traditional medicines so they are needed to be upgraded or replaced. There is a need to develop antimicrobial agents with more effectiveness and minimum side effects. There are some reports from last two decades that Melia azedarach is a potential source of novel antibodies. Its extracts have both antioxidant and antimicrobial activities. Powdered leaves of M. azedarach were extracted with methanol and extract was preliminary examined by phytochemical tests and thin layer chromatography. The different concentrations of extracts showed good antibacterial activities against three pathogenic bacterial strains Staphylococcus aureus, Escherichia coli and Bacillus thuringiensis. The results indicated that M. azedarach L. could be an effective source of herbal medicines against infectious diseases.

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