scholarly journals Initial infection of roots and leaves reveals different resistance phenotypes associated with coat protein gene-mediated resistance to Potato mop-top virus

2002 ◽  
Vol 83 (5) ◽  
pp. 1201-1209 ◽  
Author(s):  
Anna Germundsson ◽  
Maria Sandgren ◽  
Hugh Barker ◽  
Eugene I. Savenkov ◽  
Jari P. T. Valkonen
Keyword(s):  
Coat Protein ◽  
Nicotiana Benthamiana ◽  
Protein Gene ◽  
Resistance Mechanism ◽  
Spongospora Subterranea ◽  
Mechanical Inoculation ◽  
Systemic Movement ◽  
Cp Gene ◽  
Transgenic Potatoes ◽  
Potato Mop Top Virus

Resistance to the pomovirus Potato mop-top virus (PMTV) was studied in potato (Solanum tuberosum cv. Saturna) and Nicotiana benthamiana transformed with the coat protein (CP) gene of PMTV. The incidence of PMTV infections was reduced in tubers of the CP-transgenic potatoes grown in the field in soil infested with the viruliferous vector, Spongospora subterranea. However, in those tubers that were infected, all three virus RNAs were detected and virus titres were high. The CP-transgenic N. benthamiana plants were inoculated with PMTV using two methods. Following mechanical inoculation of leaves, no RNA 3 (the CP-encoding RNA homologous to the transgene) was detected in leaves, but in some plants low amounts of RNA 3 were detected in roots; RNA 2 was readily detected in leaves and roots of several plants. Inoculation of roots using viruliferous S. subterranea resulted in infection of roots in all plants and the three PMTV RNAs were detected. However, no systemic movement of PMTV from roots to the above-ground parts was observed, indicating a novel expression of resistance. These data indicate that the CP gene-mediated resistance to PMTV specifically restricts accumulation of PMTV RNA 3, and is more effective in leaves than roots. Furthermore, expression of resistance is different depending on whether leaves or roots are inoculated. Data do not exclude the possibility that both a protein-mediated and an RNA-mediated resistance mechanism are involved.

scholarly journals Molecular studies of Potato mop-top virus (PMTV) in transgenic N. benthamiana and S. tuberosum

2002 ◽  
Vol 38 (SI 1 - 6th Conf EFPP 2002) ◽  
pp. 183-185
Author(s):  
A. Germundsson ◽  
M. Sandgren ◽  
H. Barker ◽  
E.I. Savenkov ◽  
J.P.T. Valkonen
Keyword(s):  
Nicotiana Benthamiana ◽  
Transgenic Potato ◽  
Infested Soil ◽  
Systemic Movement ◽  
Potato Plants ◽  
Cp Gene ◽  
Molecular Studies ◽  
Infested Field ◽  
Transgenic Potato Plants ◽  
Potato Mop Top Virus

Resistance to Potato mop-top virus (PMTV) was studied in Nicotiana benthamiana and potato (Solanum tuberosum cv.<br />Saturna) transformed with the coat protein (CP) gene of PMTV. In N. benthamiana plants mechanically inoculated with<br />PMTV, RNA2 was detected in leaves and roots in several plants in the absence of RNA3. When N. benthamiana was<br />grown in infested soil, viral RNA was detected in roots, but no systemic movement of PMTV to above-ground parts was<br />observed. The incidence of PMTV infections was reduced in the CP-transgenic potato plants grown in an infested field<br />in Sweden. However, in infected tubers, all three virus RNAs were detected.

scholarly journals Transformation of Nicotiana benthamiana with the Potato Mop-Top Virus Coat Protein Gene Produces a Novel Resistance Phenotype Mediated by the Coat Protein

1998 ◽  
Vol 11 (7) ◽  
pp. 626-633 ◽  
Author(s):  
Hugh Barker ◽  
Brian Reavy ◽  
Kara D. McGeachy ◽  
Sheila Dawson
Keyword(s):  
Steady State ◽  
Coat Protein ◽  
Nicotiana Benthamiana ◽  
Infectivity Assay ◽  
Cp Gene ◽  
Transgenic Lines ◽  
Steady State Levels ◽  
Potato Mop Top Virus ◽  
Virus Coat ◽  
Rna Transcript

Transformed lines of Nicotiana benthamiana expressing a translatable transgene (CPtrans) encoding the coat protein (CP) gene of potato mop-top virus (PMTV) differed considerably in steady-state levels of transgene RNA transcript and CP that were positively correlated. A mean of 88% of transgenic T1 plants from seven transgenic lines were resistant to manual PMTV inoculation. Resistance was identified by failure to develop symptoms in plants and inability to recover infectious virus with a sensitive infectivity assay: many of the 12% of transgenic plants that gave a positive infectivity assay did not produce symptoms and virus was not detected in repeat assays on the same plants. Resistant plants did not express a “recovery” phenotype and resistance was not overcome by use of RNA inoculum. Strong resistance was expressed in all CPtrans lines irrespective of the amount of transcript RNA or CP detected, and there was no evidence that resistance was mediated by a “transgene silencing” mechanism. Lines transformed with PMTV CP gene sequence in a nontranslatable form (CPnontrans) also contained differing steady-state levels of RNA transcript, but were not resistant to PMTV, although in some lines symptoms were slightly milder and were slower to develop than those in nontransgenic control plants. These results suggest that CPtrans is unique because, although it depends on CP translation to be effective, it mediates very strong resistance in 10 independent transformed lines (tested here and in other work).

scholarly journals Nicotiana benthamiana plants transgenic for PPV-SWC coat protein are resistant to PPV infection

2017 ◽  
Vol 38 (SI 2 - 6th Conf EFPP 2002) ◽  
pp. 608-612 ◽  
Author(s):  
S. Comes ◽  
A. Fanigliulo ◽  
P. Piazzolla ◽  
A. Crescenzi
Keyword(s):  
Monoclonal Antibody ◽  
Agrobacterium Tumefaciens ◽  
Coat Protein ◽  
Transgenic Plants ◽  
Nicotiana Benthamiana ◽  
Protein Gene ◽  
Variable Degree ◽  
Cp Gene ◽  
Homologous Virus ◽  
Transgenic Lines

Two constructs containing the coat protein gene (CP) of PPV-SwC – in (+) and (–) sense – were obtained and used to transform Nicotiana benthamiana plants via Agrobacterium tumefaciens. Transformed lines were analysed for the integration of the CP gene by PCR and Southern blot. Sixty transgenic lines were selected. Expression of CP gene was investigated by immuno-western blot using a monoclonal antibody specific for PPV-SwC. Various levels of CP expression were detected in transgenic plants. R1 lines were challenged with the homologous virus and with isolates belonging to D, M and EA PPV groups. A variable degree of resistance was obtained, going from complete susceptibility to immunity. Susceptible plants showed a slight delay in symptoms expression when compared to non transformed control. No correlation was established by CP expression level and observed resistance.

scholarly journals Quantitative Analysis of Efficient Endogenous Gene Silencing in Nicotiana benthamiana Plants Using Tomato bushy stunt virus Vectors That Retain the Capsid Protein Gene

2007 ◽  
Vol 20 (6) ◽  
pp. 609-618 ◽  
Author(s):  
Daniela Pignatta ◽  
Pavan Kumar ◽  
Massimo Turina ◽  
Abhaya Dandekar ◽  
Bryce W. Falk
Keyword(s):  
Gene Silencing ◽  
Nicotiana Benthamiana ◽  
Protein Gene ◽  
Fluorescent Protein ◽  
Infectious Clone ◽  
Tomato Bushy Stunt Virus ◽  
Green Fluorescent Protein Gene ◽  
Capsid Protein Gene ◽  
Endogenous Gene ◽  
Cp Gene

Tomato bushy stunt virus (TBSV) coat protein (CP) replacement vectors have been used previously to silence transgenes (e.g., the green fluorescent protein gene) but have not been effective for silencing endogenous plant genes. New TBSV vectors which retained the CP gene were developed by engineering an XhoI restriction site in three positions (3f, CEB, and CEA) of the pTBSV-100 infectious clone. Magnesium chelatase (ChlH) and phytoene desaturase (PDS) were chosen as targets for endogenous gene silencing. Initial experiments using CP replacement vectors with a 230-bp sense or antisense ChlH insert gave a silencing phenotype prominent only in the first new leaves above those inoculated. No silencing phenotype was apparent beyond these leaves whereas, for PDS, no silencing phenotype was observed. When plants were inoculated with the XhoI insert vectors containing ChlH and PDS sequences, plants showed a silencing phenotype extensively throughout the challenged plant, indicating an improved ability for virus movement and silencing in Nicotiana benthamiana host plants. Silencing efficiencies were quantified using real-time reverse-transcription polymerase chain reaction, indicating specific silencing effects of each individual silencing vector. Only one recombinant vector (pPD-3f5), where the XhoI insert was at the 3′ end of the CP gene, failed to give effective silencing. Here, we show that our new CP-retaining TBSV vectors (CEA-CEB) form typical TBSV virions, retain silencing inserts of variable lengths (110 to 260 nucleotides), and can systemically silence endogenous genes in N. benthamiana.

scholarly journals Potato mop-top virus: the coat protein-encoding RNA and the gene for cysteine-rich protein are dispensable for systemic virus movement in Nicotiana benthamiana

2003 ◽  
Vol 84 (4) ◽  
pp. 1001-1005 ◽  
Author(s):  
Eugene I. Savenkov ◽  
Anna Germundsson ◽  
Andrey A. Zamyatnin ◽  
Maria Sandgren ◽  
Jari P. T. Valkonen
Keyword(s):  
Coat Protein ◽  
Nicotiana Benthamiana ◽  
Virus Movement ◽  
Cysteine Rich Protein ◽  
Protein Encoding ◽  
Potato Mop Top Virus

scholarly journals PVYNTN-CP coat protein gene mediated virus resistance of transgenic potato plants

2009 ◽  
Vol 7 (4) ◽  
pp. 41-50 ◽  
Author(s):  
Zenon Stasevski ◽  
Olga N Ilinskaya
Keyword(s):  
Coat Protein ◽  
Coat Protein Gene ◽  
Virus Resistance ◽  
Protein Gene ◽  
Solanum Tuberosum L ◽  
Transgenic Potato ◽  
Potato Plants ◽  
Transgenic Potato Plants ◽  
Transgenic Potatoes ◽  
Virus Strains

PVY<sup style="line-height:1.6em">NTN</sup>-CP <span style="line-height:1.6em">coat protein gene from a necrotic strain of potato virus </span>Y (pvy<sup style="line-height:1.6em">ntn</sup>) <span style="line-height:1.6em">has been transferred into two potato </span>Solanum tuberosum L. <span style="line-height:1.6em">cultivars </span>Mindenes <span style="line-height:1.6em">and </span>Somogyi kifli via Agrobacterium tumefaciens <span style="line-height:1.6em">transformation. Expression of integrated PVY</span><sup style="line-height:1.6em">NTN</sup><span style="line-height:1.6em">-CP gene were confirmed for 33 (89 %) of 37 and 3 (75 %) of 4 kanamycin-resistant regenerants of potato cultivars Mindenes and Somogyi kifli respectively. The level of virus resistance against two virus strains </span>(PVY°, PVY<sup style="line-height:1.6em">NTN</sup>) <span style="line-height:1.6em">of independent lines of transgenic potatoes varied between extreme resistance to susceptibility. The three independent lines of transgenic potatoes proved to be extreme resistant against both PVY strains.</span>

scholarly journals Coat protein gene of a PStV-Bm isolate from West Nusa Tenggara, Indonesia

2020 ◽  
Vol 21 (3) ◽  
Author(s):  
Nur Indah Julisaniah ◽  
SUHARJONO ◽  
RETNO MASTUTI ◽  
ESTRI LARAS ARUMINGTYAS
Keyword(s):  
Coat Protein ◽  
Coat Protein Gene ◽  
Genetic Similarity ◽  
Protein Gene ◽  
Rna Virus ◽  
Genetic Relationships ◽  
Cp Gene ◽  
Similar Cluster ◽  
Similar Index ◽  
The Usa

Abstract. Julisaniah NI, Suharjono, Mastuti R, Arumingtyas EL. 2020. Coat protein gene of a PStV-Bm isolate from West Nusa Tenggara, Indonesia. Biodiversitas 21: 903-909. Peanut stripe virus (PStV) is a single-stranded positive-sense RNA virus capable of infecting peanut plants. An isolate of PStV (PStV-Bm) was collected from a peanut field in the Bima District, West Nusa Tenggara Province, Indonesia and the coat protein (CP) gene of this virus (CP-PStV) was extracted from the viral RNA and analyzed using reverse transcription-polymerase chain reaction methods. The CP-PStV gene of PStV-Bm was aligned with several PStV genes deposited in the Genbank (http://www.ncbi.nml.nih.gov). Based on the nucleotide sequence of the CP gene, PStV-Bm was grouped into a similar cluster with other PStVs that originated from Indonesia with a similar index, ranging from 96.8% to 98.9%. Genetic similarity (about 96.1%) was also observed between PStV-Bm and PStV from the USA. This genetic similarity indicated that viruses from adjacent regions have high genetic relationships. Some amino acid differences were observed in PStV-Bm that may be typical of this strain.

scholarly journals Characteristic of Tobacco mosaic virus isolated from cucumber and tobacco collected from East Java, Indonesia

2019 ◽  
Vol 20 (10) ◽  
Author(s):  
LISTIHANI ◽  
SRI HENDRASTUTI HIDAYAT ◽  
SURYO WIYONO ◽  
Tri Asmira Damayanti
Keyword(s):  
Phylogenetic Analysis ◽  
Amino Acid ◽  
Nucleic Acid ◽  
Coat Protein ◽  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Protein Gene ◽  
Cp Gene ◽  
Emerging Virus ◽  
Tomato Isolate

Abstract. Listihani, Hidayat SH, Wiyono S, Damayanti TA. 2019. Characteristic of Tobacco mosaic virus isolated from cucumber and tobacco collected from East Java, Indonesia. Biodiversitas 20: 2937-2942. Tobacco mosaic virus (TMV) is a newly emerging virus infecting cucumbers in Java, Indonesia. The basic characters of the TMV isolated from cucumber need further study to investigate its differences with that from tobacco. Thus, the research aimed to study the character of both isolates based on their biological, symptomatology and nucleic acid of coat protein (CP) gene properties. The TMV isolates from both cucumber and tobacco were able to infect similar indicator plants with differing symptom expressions, especially on eggplant. Homology of nucleotide and amino acid of coat protein gene among isolates were about 90.3% and 91.0%, and homology to other isolates was about 87.6 to 93.8% and 89.3 to 96.8%, respectively. There were 17 amino acid differences in the CP gene which is presumed to differentiate those two isolates. Phylogenetic analysis CP gene sequences compared with corresponding isolates in GenBank showed the two isolates separated in different clades. The cucumber isolate from Kediri is closely related to tomato isolate from China in clade I, while tobacco isolate from Jember closely related to tobacco isolate from China in clade IIindicating the existence of two differ TMV variants. It will increase the difficulty to manage TMV in the fields.

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