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1.
Braz. j. biol ; 83: 1-9, 2023. map, tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1468907

ABSTRACT

Cucumber mosaic virus (CMV) is a tremendous threat to vegetables across the globe, including in Pakistan. The present work was conducted to investigate the genetic variability of CMV isolates infecting pea and spinach vegetables in the Pothwar region of Pakistan. Serological-based surveys during 2016-2017 revealed 31.70% overall CMV disease incidence from pea and spinach crops. Triple-antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) revealed that all the positive isolates belong to CMV subgroup II. Two selected cDNA from ELISA-positive samples representing each pea and spinach crops were PCR-amplified (ca.1100 bp) and sequenced corresponding to the CMV CP gene which shared 93.7% nucleotide identity with each other. Both the sequences of CMV pea (AAHAP) and spinach (AARS) isolates from Pakistan were submitted to GenBank as accession nos. MH119071 and MH119073, respectively. BLAST analysis revealed 93.4% sequence identity of AAHAP isolate with SpK (KC763473) from Iran while AARS isolate shared maximum identity (94.5%) with the strain 241 (AJ585519) from Australia and clustered with some reference isolates of CMV subgroup II from UK (Z12818) and USA (AF127976) in a Neighbour joining phylogenetic reconstruction. A total of 59 polymorphic (segregating) sites (S) with nucleotide diversity (π) of 0.06218 was evident while no INDEL event was observed in Pakistani isolates. The evolutionary distance of Pakistani CMV isolates was recorded as 0.0657 with each other and 0.0574-0.2964 with other CMV isolates reported elsewhere in the world. A frequent gene flow (Fst = 0.30478 <0.33) was observed between Pakistani and earlier reported CMV isolates. In genetic differentiation analysis, the value of three permutation-based statistical tests viz; Z (84.3011), Snn (0.82456), and Ks* (4.04042) were non-significant. The statistical analysis revealed the [...].


Cucumber mosaic cucumovirus (CMV) é uma tremenda ameaça aos vegetais em todo o mundo, inclusive no Paquistão. O presente trabalho foi conduzido para investigar a variabilidade genética de isolados de CMV infectando vegetais de ervilha e espinafre na região de Pothwar, Paquistão. Pesquisas com base em sorologia durante 2016-2017 revelaram 31,70% da incidência geral da doença por CMV em safras de ervilha e espinafre. O ensaio de imunoabsorção enzimática em sanduíche de anticorpo triplo (TAS-ELISA) revelou que todos os isolados positivos pertencem ao subgrupo II do CMV. Dois cDNA selecionados de amostras positivas para ELISA representando cada safra de ervilha e espinafre foram amplificados por PCR (ca.1100 pb) e sequenciados correspondendo ao gene CMV CP que compartilhou 93,7% de identidade de nucleotídeo um com o outro. Ambas as sequências de isolados de ervilha CMV (AAHAP) e espinafre (AARS) do Paquistão foram submetidas ao GenBank como nos de acesso. MH119071 e MH119073, respectivamente. A análise BLAST revelou 93,4% de identidade de sequência do isolado AAHAP com SpK (KC763473) do Irã, enquanto o isolado AARS compartilhou a identidade máxima (94,5%) com a cepa 241 (AJ585519) da Austrália e agrupada com alguns isolados de referência do subgrupo II de CMV do Reino Unido (Z12818) e EUA (AF127976) em uma reconstrução filogenética vizinha. Um total de 59 sítios polimórficos (segregantes) (S) com diversidade de nucleotídeos (π) de 0,06218 foi evidente, enquanto nenhum evento INDEL foi observado em isolados do Paquistão. A distância evolutiva de isolados de CMV do Paquistão foi registrada como 0,0657 entre si e 0,0574-0,2964 com outros isolados de CMV relatados em outras partes do mundo. Um fluxo gênico frequente (Fst = 0,30478 < 0,33) foi observado entre os isolados de CMV do Paquistão e relatados anteriormente. Na análise de diferenciação genética, os valores de três testes estatísticos baseados em [...].


Subject(s)
Animals , Bromoviridae/genetics , Bromoviridae/pathogenicity , Pisum sativum/virology , Spinacia oleracea/virology
2.
Theor Appl Genet ; 110(2): 337-45, 2005 Jan.
Article in English | MEDLINE | ID: mdl-15551034

ABSTRACT

In melon, the Fom-1 gene confers monogenic resistance against the soil-borne fungus Fusarium oxysporum f. sp. melonis, races 0 and 2, while the closely linked Prv gene specifies resistance against the papaya ring spot virus. Markers linked to these resistance (R) genes were identified using two recombinant inbred line populations, derived from crosses between Cucumis melo Vedrantais and C. melo PI 161375, and between C. melo Vedrantais and C. melo PI 414723, respectively. Using bulked segregant analysis, as well as systematic scoring of the mapping populations, we developed two amplified fragment length polymorphism markers, two random amplified polymorphic DNA markers and five restriction fragment length polymorphism (RFLP) markers linked to this locus. Four of the RFLP sequences bear homology to nucleotide-binding site-leucine-rich repeat R genes, indicating the presence of a significant R-gene cluster in this locus. Our study provides the most closely linked markers published so far for these important traits. It also improves the resolution of the whole linkage group IX, which was difficult to order in our previous studies. Two of the markers were converted to cleaved amplified polymorphic sequence markers to facilitate their application in marker-assisted selection. Testing these two markers in several melon lines revealed different marker haplotypes in the melon germplasm and supported multiple, independent origin of the Fusarium races 0 and 2 resistance trait.


Subject(s)
Bromoviridae/pathogenicity , Carica/virology , Cucumis melo , Fusarium/pathogenicity , Genes, Plant , Genetic Markers , Immunity, Innate/genetics , Plant Diseases , Chromosome Mapping , Crosses, Genetic , Cucumis melo/genetics , Cucumis melo/microbiology , DNA, Plant/genetics , Genetic Linkage , Plant Diseases/genetics , Plant Diseases/microbiology , Polymorphism, Restriction Fragment Length , Random Amplified Polymorphic DNA Technique , Selection, Genetic
3.
Arch Virol ; 148(4): 803-12, 2003 Apr.
Article in English | MEDLINE | ID: mdl-12664302

ABSTRACT

Brome mosaic virus (BMV) requires the coat protein (CP) for cell-to-cell movement whereas Cowpea chlorotic mottle virus (CCMV), from the same genus, does not. Chimeric viruses created by exchanging the movement protein (MP) gene between the viruses can move from cell to cell. We show that interference in CP expression impaired the movement of the chimeric CCMV with the BMV MP gene but not of the chimeric BMV with the CCMV MP gene. We thus conclude that the MP gene plays a crucial role in determination of the virus-specific CP requirement in bromovirus cell-to-cell movement.


Subject(s)
Bromoviridae/physiology , Capsid Proteins/physiology , Chenopodium quinoa/virology , RNA, Viral/genetics , Viral Proteins/genetics , Amino Acid Sequence , Bromoviridae/genetics , Bromoviridae/pathogenicity , Capsid Proteins/analysis , Capsid Proteins/genetics , Genes, Viral , Green Fluorescent Proteins , Luminescent Proteins , Molecular Sequence Data , Plant Diseases/virology , Plant Viral Movement Proteins , Protoplasts/virology , RNA, Viral/analysis , Reassortant Viruses , Viral Proteins/analysis
4.
Proc Natl Acad Sci U S A ; 98(25): 14286-91, 2001 Dec 04.
Article in English | MEDLINE | ID: mdl-11717411

ABSTRACT

On entering a host cell, positive-strand RNA virus genomes have to serve as messenger for the translation of viral proteins. Efficient translation of cellular messengers requires interactions between initiation factors bound to the 5'-cap structure and the poly(A) binding protein bound to the 3'-poly(A) tail. Initiation of infection with the tripartite RNA genomes of alfalfa mosaic virus (AMV) and viruses from the genus Ilarvirus requires binding of a few molecules of coat protein (CP) to the 3' end of the nonpolyadenylated viral RNAs. Moreover, infection with the genomic RNAs can be initiated by addition of the subgenomic messenger for CP, RNA 4. We report here that extension of the AMV RNAs with a poly(A) tail of 40 to 80 A-residues permitted initiation of infection independently of CP or RNA 4 in the inoculum. Specifically, polyadenylation of RNA 1 relieved an apparent bottleneck in the translation of the viral RNAs. Translation of RNA 4 in plant protoplasts was autocatalytically stimulated by its encoded CP. Mutations that interfered with CP binding to the 3' end of viral RNAs reduced translation of RNA 4 to undetectable levels. Possibly, CP of AMV and ilarviruses stimulates translation of viral RNAs by acting as a functional analogue of poly(A) binding protein or other cellular proteins.


Subject(s)
Capsid Proteins , Capsid/metabolism , Protein Biosynthesis , RNA, Viral/genetics , RNA, Viral/metabolism , Alfalfa mosaic virus/genetics , Alfalfa mosaic virus/pathogenicity , Alfalfa mosaic virus/physiology , Bromoviridae/genetics , Bromoviridae/pathogenicity , Bromoviridae/physiology , Protein Binding , RNA, Messenger/chemistry , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Viral/chemistry , Nicotiana/virology , Virus Replication
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