scholarly journals The molecular characterization of the coat protein sequence and differentiation of CMV- subgroup I on tobacco from native flora in Turkey

2020 ◽  
Vol 48 (2) ◽  
pp. 523-534
Author(s):  
Mustafa USTA ◽  
Abdullah GÜLLER ◽  
Abidin GÜNAY
Keyword(s):  
Coat Protein ◽  
Sequence Similarity ◽  
Coat Protein Sequence ◽  
Rt Pcr ◽  
Plant Host ◽  
Native Flora ◽  
Subgroup Ii ◽  
Escherichia Coli Jm109 ◽  
First Time

Cucumber mosaic virus (CMV) has a broad plant-host range and a wide ecological zone distribution. Virus-like symptoms were observed on tobacco fields of Adiyaman province (Turkey) showing conspicuous mottling, greenish mosaic patterns and severe malformations of leaves. A total of forty tobacco samples tested positive against CMV by reverse transcription polymerase chain reaction (RT-PCR) using coat protein gene specific primers. Five randomly chosen CMV isolates were cloned into pGEM T-Easy vector and transformed into Escherichia coli JM109 strain. The recombinant bacterial clones containing insert-DNA were further purified and sequenced bidirectionally. In multiplex-RT-PCR studies carried out, it was found that all 40 CMV isolates belong to Subgroup I by resulting a 593 bp long DNA fragments. CMV subgroup IA was found to predominate in 4 out of 5 tobacco samples and CMV subgroup IB was found in 1 out of 5 CMV-positive samples by comparing the isolates with CMV reference isolates in phylogenetic tree. However, no Subgroup II sequences were found by multiplex RT-PCR using discriminating primers. The nucleic acid sequences were analyzed for the investigation of diversity of coat protein (CP) sequences of 5 CMV isolates. The sequence similarity ranged from 94.2-100% with the CMV subgroup I isolates infecting diverse plants in other regions of the world. The evolutionary tree revealed that the CMV IA Adiyaman isolates exhibited a genetic affinity with Australian and Spanish isolates. However, the CMV IB Adiyaman isolate showed a close genetic relationship with only the Australian isolates. To our knowledge, this study shows for the first time the occurrence of CMV IA and IB isolates infecting cultured tobacco plants in Adiyaman province.

scholarly journals Preliminary molecular characterization of some Citrus tristeza Closterovirus isolates infecting Croatian citrus

2017 ◽  
Vol 38 (SI 2 - 6th Conf EFPP 2002) ◽  
pp. 264-266
Author(s):  
S. Černi ◽  
D. Škorić ◽  
M. Krajačić
Keyword(s):  
Coat Protein ◽  
Protein Sequence ◽  
Coastal Region ◽  
Coat Protein Sequence ◽  
Rt Pcr ◽  
Double Stranded Rna ◽  
The World ◽  
Citrus Tristeza Closterovirus ◽  
Citrus Tristeza

Citrus tristeza Closterovirus (CTV) is widespread in major citrus-growing regions of the world often causing destructive diseases. Citrus samples were taken from orchards in the Croatian coastal region. CTV was detected in two symptomless field trees of Satsuma mandarins and one diseased lemon tree. Double-stranded RNA was isolated from the field trees and the dsRNA patterns were compared in polyacrylamide gels. The same dsRNA extracts were used as templates in RT-PCR experiments amplifying the CTV coat protein sequence. Amplicons were subjected to SSCP and RFLP analyses. The results indicate greater similarity between CTV isolates from Satsuma mandarins than between these two and the lemon isolate.

Serological Characterization of Hungarian Plum Pox Virus Isolates

1997 ◽  
Vol 52 (5-6) ◽  
pp. 391-395
Author(s):  
Juan José López-Moya ◽  
Dionisio López-Abella ◽  
José-Ramón Díaz-Rúiz ◽  
Belén Martinez-Garcia ◽  
Richard Gáborjányi
Keyword(s):  
Monoclonal Antibodies ◽  
Coat Protein ◽  
Blot Analysis ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
Dot Blot ◽  
Serological Characterization ◽  
Spanish Isolate ◽  
Virus Isolates

Abstract Three Hungarian (No.2, 4 and 9), and a Moldavian (K) plum pox virus isolates were compared with a characterized Spanish isolate (5.15) by RT-PCR, ELISA, dot-blot and West­ern blot analysis. Monoclonal antibodies prepared against the external, intermediate and internal sequences of the coat protein of the Spanish isolate were able to differentiate the four isolates. Hungarian isolate No. 2 proved to be serologically identical to the Spanish isolate, while No. 4 showed appreciable differences and No. 9 could be recognized only by the monoclonal antibodies representing the intermedial and internal parts of the coat protein. K isolate showed a more distant relationship to other isolates. Our experiment provided the first demonstration of the presence of D type isolates in Hungary.

scholarly journals First Report of Tobacco mild green mosaic virus in Capsicum chinense in Venezuela

Plant Disease ◽  
2006 ◽  
Vol 90 (8) ◽  
pp. 1108-1108 ◽  
Author(s):  
C. Córdoba ◽  
A. García-Rández ◽  
N. Montaño ◽  
C. Jordá
Keyword(s):  
Mosaic Virus ◽  
Seed Transmission ◽  
Tomato Mosaic Virus ◽  
Capsicum Chinense ◽  
Rt Pcr ◽  
Plant Host ◽  
First Report ◽  
Coat Protein Region ◽  
Pcr Product ◽  
First Time

In July 2003, noticeable deformations of leaves were observed on a local variety of Capsicum chinense, also called ‘Aji dulce’, from a pepper plantation located in Venezuela, (Monagas State). ‘Aji dulce’ is a basic ingredient of the Venezuelan gastronomy with an estimated cultivated area of 2,000 ha. The seeds of this local pepper are obtained by the growers who reproduce and multiply their own seeds every year. Seeds of affected plants were sent to our laboratory, and a group of approximately 100 seeds was sown in a controlled greenhouse that belongs to the Polytechnic University of Valencia, Spain. Three months later, obvious curling and bubbling developed on the leaves of the plants. Extracts of symptomatic plants tested negative for Tomato mosaic virus (ToMV), Tobacco mosaic virus (TMV), Pepper mild mottle virus (PMMV), and Tobacco etch virus (TEV) by double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA) with policlonal antibodies specific to each virus (Loewe Biochemica GMBH, Sauerlach, Germany; Phyto-Diagnostics, INRA, France). Total RNA was isolated from 0.5 g of original seed sent from Venezuela and from 25 samples of leaves of plants grown in the greenhouse with an RNeasy Plant Mini Kit (Qiagen Sciences, Germantown, Maryland). The RNA isolated was used in reverse transcription-polymerase chain reaction (RT-PCR) with specific primers for Tobacco mild green mosaic virus (TMGMV) (1) predicted to amplify a 530 bp of the coat protein region. From all samples, a RT-PCR product of the expected size was obtained and then sequenced. BLAST analysis of one sequence (GenBank Accession No. DQ460731) showed high levels of identity with TMGMV isolates, with more than 99% nucleotide identity with the DSMZ PV-112 isolate (GenBank Accession No. AJ429096). The symptomatology observed on pepper plants, the TMGMV RT-PCR assay, and the consensus of sequenced regions with TMGMV lead us to conclude that TMGMV was the causal agent of the diseased C. chinense plants. Although TMGMV has a wide plant host range occurring worldwide (1), to our knowledge, this is not only the first time TMGMV has been detected in Venezuela, but also the first report of TMGMV in C. chinense in Venezuela and the first reliable probe of the TMGMV seed transmission. Reference: (1) J. Cohen et al. Ann. Appl. Biol. 138:153, 2001.

scholarly journals Identification and molecular characterization of onion yellow dwarf virus (OYDV) in Allium cepa crops in Poland

2021 ◽  
Vol 61 (3) ◽  
pp. 214-220
Keyword(s):  
Phylogenetic Analysis ◽  
Molecular Characterization ◽  
Coat Protein Sequence ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Onion Yellow Dwarf Virus ◽  
Rt Pcr ◽  
Pcr Products ◽  
Yellow Dwarf Virus

Onion yellow dwarf virus is distributed worldwide significantly reducing yield of crops from the Allium genus. The aim of the study was the detection and molecular characterization of newly identified OYDV isolates infecting onions in Poland. The virus was detected by transmission electron microscopy and RT-PCR techniques using two pairs of diagnostic primers: OYDV-NibCPF1/R1 and OYDV-CPF2/R2. The specificity of obtained RT-PCR products was confirmed by Sanger sequencing and received viral coat protein sequence was used for phylogenetic analysis. The phylogenetic analysis was carried out using CP sequences of the new Polish onion isolate obtained in this study and 37 other sequences of OYDV retrieved from GenBank. The analysis revealed that the Polish OYDV isolate is the most similar to the OYDV isolates derived from onions from Argentina and Germany, which may indicate their common origin. Moreover, it was observed that the Polish onion and garlic isolates are very diverse and belong to different phylogroups.

scholarly journals Characterization of a U.S. Isolate of Beet black scorch virus

2007 ◽  
Vol 97 (10) ◽  
pp. 1245-1254 ◽  
Author(s):  
John J. Weiland ◽  
David Van Winkle ◽  
Michael C. Edwards ◽  
Rebecca L. Larson ◽  
Weilin L. Shelver ◽  
...  
Keyword(s):  
Coat Protein ◽  
Protein Gene ◽  
Sequence Similarity ◽  
Systemic Infection ◽  
Amino Acid Sequence Similarity ◽  
Adenosine Nucleotides ◽  
Tetragonia Expansa ◽  
Extension Analysis ◽  
Beet Black Scorch Virus

The first reported U.S. isolate of Beet black scorch necrovirus (BBSV) was obtained and characterized. Host range of the virus for localized and occasionally systemic infection included the Chenopodiaceae and Tetragonia expansa; Nicotiana benthamiana supported symptomless systemic infection by the virus. The complete nucleotide sequence of the genomic RNA of the virus, designated BBSV-Co, exhibits 93% similarity to the genome of the ‘Ningxia’ isolate of BBSV from China. Amino acid sequence similarity in predicted genes ranged from 95% in the p4 gene to 97% in the p82 and coat protein genes. A potential additional gene exists within the U.S. isolate of BBSV that is absent from Chinese isolates of BBSV due to nucleotide differences between these isolates within the coat protein gene. Coat protein analysis by isoelectric focusing and by mass spectroscopy indicated the presence of phosphorylated residues. Using primer extension analysis of the 5′ end of the genome and site-directed mutants of genomic clones of BBSV-Co from which infectious RNA was produced, the native 5′ end of the BBSV-Co genome was determined to be 5′-GAAACCTAACC…3′, lacking the two terminal adenosine nucleotides in the published sequences of BBSV from China.

A Human Lymphoid Progenitor Capable of Circulating and Seeding the Thymus.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2216-2216
Author(s):  
Emmanuelle M. Six ◽  
Delphine Bonhomme ◽  
Marta Monteiro ◽  
Kheira Beldjord ◽  
Alexandrine Garrigue ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bone Marrow ◽  
T Cell ◽  
T Cell Development ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Rt Pcr ◽  
Pcr Assay ◽  
First Time

Abstract Identification of a thymus-seeding progenitor originating from human bone marrow constitutes a key milestone in understanding and correcting defects in T-cell development. Here, we report the characterization of a novel human bone marrow lymphoid-restricted subpopulation which is part of the lineage-negative CD34+CD10+ progenitor population and which can be distinguished from B-cell-committed precursors by the absence of CD24 expression. We demonstrated that these Lin-CD34+CD10+CD24- progenitors lack myeloid and erythroid potential but can generate B, T and NK lymphocytes following culture on MS5 or OP9-hDelta1 stroma. The gene expression profile of this population, analyzed by a multiplex RT-PCR assay, revealed co-expression of RAG1, TdT, PAX5, CD3ε and IL-7Rα. These progenitors are not only present in the bone marrow but also in the blood throughout life, suggesting an ability to circulate. Moreover we showed that the Lin-CD34+CD10+CD24- cells also correspond to the most immature population of the thymus which gives rise to Lin-CD34+CD7+ T-cell precursors. Taken as a whole these findings unravel for the first time the existence of a postnatal lymphoid-restricted population which is capable of migrating from the bone marrow to the thymus.

scholarly journals Detection and partial molecular characterization of Grapevine fleck virus, Grapevine virus D, Grapevine leafroll-associated virus -5 and -6 infecting grapevines in Brazil

2012 ◽  
Vol 42 (12) ◽  
pp. 2127-2130 ◽  
Author(s):  
Thor Vinícius Martins Fajardo ◽  
Marcelo Eiras ◽  
Osmar Nickel ◽  
Carla Rosa Dubiela ◽  
Eliezer Rodrigues de Souto
Keyword(s):  
Coat Protein ◽  
Molecular Characterization ◽  
Rna Binding ◽  
Rna Binding Protein ◽  
Amino Acid Sequences ◽  
Viral Diseases ◽  
Grapevine Virus ◽  
Available Information ◽  
First Time

Grapevine fleck, rugose wood and leafroll are three grapevine viral diseases whose causal agents (or associated viruses) respectively are Grapevine fleck virus (GFkV), Grapevine virus D (GVD) and Grapevine leafroll-associated virus 5 and 6 (GLRaV-5 and -6). The objective of this work was to perform a partial molecular characterization of local isolates of these four viral species that infect grapevines. The nucleotide and deduced amino acid sequences of complete genes of the coat protein (CP) (of GFkV), the CP and the RNA binding protein (of GVD), the CP and the partial hHSP70 gene (of GLRaV-5) and the partial hHSP70 gene (of GLRaV-6) were aligned and compared in silico with other isolates. These data extend the available information about Brazilian isolates of GFkV, GLRaV-5 and -6, and reports for the first time the GVD occurrence in Brazil.

scholarly journals Biological and Molecular Characterization of Moroccan watermelon mosaic virus and a Potyvirus Isolate from Eastern Sudan

Plant Disease ◽  
2001 ◽  
Vol 85 (5) ◽  
pp. 547-552 ◽  
Author(s):  
Hervé Lecoq ◽  
Gasim Dafalla ◽  
Cécile Desbiez ◽  
Catherine Wipf-Scheibel ◽  
Brigitte Delécolle ◽  
...  
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Distinct Species ◽  
Coat Protein Sequence ◽  
Watermelon Mosaic Virus ◽  
Protein Sequence Analysis ◽  
Leaf Deformation ◽  
Core Part ◽  
Eastern Sudan

A potyvirus (Su-94-54) was isolated from a naturally infected snake cucumber (Cucumis melo var. flexuosus) plant with severe mosaic and leaf deformation symptoms collected in Eastern Sudan. This isolate has a host range limited to cucurbits and is serologically distantly related to Moroccan watermelon mosaic virus (MWMV) and to Papaya ringspot virus (PRSV). Coat protein sequence analysis of Su-94-54 and MWMV and comparison with other potyviruses indicate that Su-94-54 is more closely related to MWMV than to any other potyvirus. Based on the amino acid sequence identity in the core part of the coat protein with MWMV (86%), this isolate could be regarded as a distinct species. However, because of biological, cytological, and serological affinities with MWMV, we propose that this isolate be considered as a strain of MWMV, possibly an evolutionary intermediate between MWMV and PRSV, until more is known on the structure of the PRSV subgroup within the genus Potyvirus.

scholarly journals Biological and Molecular Characterization of a Novel Tobamovirus with a Unique Host Range

Plant Disease ◽  
2003 ◽  
Vol 87 (10) ◽  
pp. 1190-1196 ◽  
Author(s):  
Scott Adkins ◽  
Ivanka Kamenova ◽  
Diann Achor ◽  
Dennis J. Lewandowski
Keyword(s):  
Coat Protein ◽  
Host Range ◽  
Protein Sequence ◽  
Genome Organization ◽  
Protein Gene ◽  
Coat Protein Sequence ◽  
Plant Viruses ◽  
Experimental Host Range ◽  
Solanaceous Plants

Tobamoviruses are among the best characterized and most studied plant viruses. Three subgroups of tobamoviruses correspond to viral genome sequence and host range to include those viruses infecting (i) solanaceous plants, (ii) brassicas, or (iii) cucurbits or legumes. We isolated a virus from Florida landscape plantings of the malvaceous plant hibiscus (Hibiscus rosasinensis) that appears to be a tobamovirus based upon its virion morphology, genome organization, and coat protein sequence. The experimental host range of this virus included five malvaceous species but excluded all tested brassica, cucurbit, and legume species and 12 of the 19 solanaceous species tested. The unique host range and comparison of coat protein gene and protein sequences with those of recognized tobamoviruses indicate that this is a novel to-bamovirus. A limited survey revealed that this virus is widespread in hibiscus and related species in the Florida landscape.

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