Strategies for the Detection of Potential Beet necrotic yellow vein virus Genome Recombinations which might Arise as a Result of Growing A type Coat Protein Gene-expressing Sugarbeets in Soil Containing B Type Virus

Nicotiana benthamiana plants transformed with the coat protein gene of tomato bushy stunt virus (TBSV) failed to elicit effective virus resistance when inoculated with wild-type virus. Subsequently, R1 and R2 progeny from 13 transgenic lines were inoculated with a TBSV mutant containing a defective coat protein gene. Mild symptoms typical of those elicited in nontransformed plants infected with the TBSV mutant initially appeared. However, within 2 to 4 weeks, up to 20% of the transgenic plants sporadically began to develop the lethal syndrome characteristic of wild-type virus infections. RNA hybridization and immunoblot analyses of these plants and nontransformed N. benthamiana inoculated with virus from the transgenic lines indicated that wild-type virus had been regenerated by a double recombination event between the defective virus and the coat protein transgene. Similar results were obtained with a TBSV deletion mutant containing a nucleotide sequence marker, and with a chimeric cucumber necrosis virus (CNV) containing the defective TBSV coat protein gene. In both cases, purified virions contained wild-type TBSV RNA or CNV chimeric RNA derived by recombination with the transgenic coat protein mRNA. These results thus demonstrate that recombinant tombusviruses can arise frequently from viral genes expressed in transgenic plants.

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Construction of Plant Transformation Vectors Carrying Beet Necrotic Yellow Vein Virus Coat Protein Gene (I) - Transformation Vectors

Biotechnology & Biotechnological Equipment ◽

10.1080/13102818.2005.10817195 ◽

2005 ◽

Vol 19 (2) ◽

pp. 80-86

Author(s):

N. Nagl ◽

I. Atanassov ◽

K. Roussanov ◽

S. Paunovich ◽

A. Atanassov ◽

...

Keyword(s):

Coat Protein ◽

Coat Protein Gene ◽

Plant Transformation ◽

Protein Gene ◽

Yellow Vein ◽

Virus Coat Protein ◽

Virus Coat ◽

Transformation Vectors

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Cloning of the coat protein gene from beet necrotic yellow vein virus and its expression in sugar beet hairy roots

Theoretical and Applied Genetics ◽

10.1007/bf00224989 ◽

1991 ◽

Vol 81 (6) ◽

pp. 777-782 ◽

Cited By ~ 22

Author(s):

U. Ehlers ◽

U. Commandeur ◽

R. Frank ◽

J. Landsmann ◽

R. Koenig ◽

...

Keyword(s):

Coat Protein ◽

Sugar Beet ◽

Hairy Roots ◽

Coat Protein Gene ◽

Protein Gene ◽

Yellow Vein

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Nucleotide sequence of the coat protein gene of a strain of clover yellow vein virus from New Zealand: conservation of a stem-loop structure in the 3? region of potyviruses

Archives of Virology ◽

10.1007/bf01314631 ◽

1992 ◽

Vol 124 (1-2) ◽

pp. 133-146 ◽

Cited By ~ 16

Author(s):

G. T. Bryan ◽

R. C. Gardner ◽

R. L. S. Forster

Keyword(s):

New Zealand ◽

Nucleotide Sequence ◽

Coat Protein ◽

Coat Protein Gene ◽

Protein Gene ◽

Yellow Vein ◽

Loop Structure ◽

Stem Loop ◽

Stem Loop Structure ◽

Clover Yellow Vein Virus

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New species emergence via recombination among isolates of the Brazilian tomato infecting Begomovirus complex

Pesquisa Agropecuária Brasileira ◽

10.1590/s0100-204x2006000800018 ◽

2006 ◽

Vol 41 (8) ◽

pp. 1329-1332 ◽

Cited By ~ 7

Author(s):

Alice Kazuko Inoue-Nagata ◽

Darren Patrick Martin ◽

Leonardo Silva Boiteux ◽

Leonardo de Britto Giordano ◽

Isabel Cristina Bezerra ◽

...

Keyword(s):

New Species ◽

Coat Protein ◽

Coat Protein Gene ◽

Intergenic Region ◽

Protein Gene ◽

Nucleotide Sequences ◽

Yellow Vein ◽

Streak Virus ◽

Recombination Analysis ◽

Leaf Curl Virus

Partial nucleotide sequences of five tomato infecting Begomovirus isolates were determined from DNA-A fragments, corresponding to the 5' region of the replication associated protein gene, the intergenic region and the 5' region of the coat protein gene. Isolate DFM shared 95% identity with Tomato mottle leaf curl virus (TMoLCV), isolates 34, PA-05, and Ta4 were 88% identical to Tomato yellow vein streak virus and isolate DF-BR3 shared 77% identity with TMoLCV. Recombination analysis indicated that isolate DF-BR3 was a chimaera, and it provided evidence that there is a complex and actively recombining population of tomato infecting begomoviruses in Brazil.

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Advances in development of transgenic resistance to beet necrotic yellow vein virus (BNYVV) in sugar beet

Genetika ◽

10.2298/gensr0503181n ◽

2005 ◽

Vol 37 (3) ◽

pp. 181-189

Author(s):

Nevena Nagl ◽

Ivan Atanasov ◽

Krasimir Rusanov ◽

Svetlana Paunovic ◽

Lazar Kovacev ◽

...

Keyword(s):

Coat Protein ◽

Sugar Beet ◽

Coat Protein Gene ◽

Protein Gene ◽

Transformation Method ◽

Yellow Vein ◽

Bar Gene ◽

Axillary Shoots ◽

Viral Cdna

Fragments of viral cDNA containing the coat protein gene of beet necrotic yellow vein virus were cloned in plant transformation vector pCAMBIA3301M with the bar gene as selectable marker. Vector pC3301MCPL carrying coat protein gene with leader sequence, and pC3301MCPS with coat protein gene, were used in Agrobacterium - mediated transformation of sugar beet. The transformation method used was based on the fact that sugar beet develops axillary shoots in in vitro conditions, when placed on media with citokinins. Since this ability is not genotype or ploidy dependant it is widely used for sugar beet vegetative multiplication. Sterile seedlings, with removed cotyledons and lower half of hypocotyl, were used as starting material. After transformation ex-plants were put on micropropagation medium with cephotaxime and phosphinotricyn (ppt), where axillary shoots started to develop. Since concentration of ppt was not selective enough, after two subcultivations it was increased twofold. Only one sample, transformed with pC3301MCPS preserved morphogenetic potential for micropropagatio, and it was tested for presence of COS fragment and bar gene bz PCR with soecific primers.

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