The sense and antisense coat protein gene of alfalfa mosaic virus strain N20 confers protection in transgenic tobacco plants

1997 ◽  
Vol 48 (4) ◽  
pp. 503 ◽  
Author(s):  
K. W. Jayasena ◽  
B. J. Ingham ◽  
M. R. Hajimorad ◽  
J. W. Randles

The coat protein gene of a South Australian strain of alfalfa mosaic virus (AMV-N20 [NcS]) has been cloned, sequenced, and transferred into Nicotiana tabacum L. cv. Xanthi via Agrobacterium tumefaciens under the control of the CaMV 35S promoter. A number of lines (T0 generation) were selected with the coat protein gene either in sense orientation (CP+) or in antisense orientation (CP–). The T0 plants were tested for their gene expression and susceptibility to the homologous AMV strain. A significant delay in the onset of symptoms and a reduction in virus accumulation was observed in CP+ plants mechanically inoculated with AMV. CP– plants were also significantly protected but less so than the CP+ plants. Plants transformed with the expression vector only (CP0) showed a minor resistance to local infection on inoculated leaves compared with untransformed plants. The strategy of coat protein mediated protection (CPMP) using the CP gene in either messenger sense or antisense would therefore be appropriate for testing on economically important pasture legumes.

2001 ◽  
Vol 52 (1) ◽  
pp. 67 ◽  
Author(s):  
K. W. Jayasena ◽  
M. R. Hajimorad ◽  
E. G. Law ◽  
A-U. Rehman ◽  
K. E. Nolan ◽  
...  

Medicago truncatula is used as a pasture legume and a source of nitrogen for grain crops in southern Australia. Alfalfa mosaic virus (AMV) infection reduces herbage production and nodulation. The coat protein gene of a South Australian strain of AMV (AMV N20) has been transferred to Medicago truncatula cv. Jemalong 2HA using Agrobacterium-mediated transformation. The most detailed investigations were carried out with the coat protein gene in the sense orientation (CP+). Progeny (T1, T2, T3) raised from self-pollinated primary transformants (T0) containing the coat protein CP+ gene were resistant to AMV. Based on Southern analysis and segregation, the transformants contained a single gene copy. In the T3 generation, one line was immune and one line showed resistance to AMV N20. The immune line contained no detectable virus when plant sap from either inoculated or systemic leaves was bioassayed on Phaseolus vulgaris. This line was also immune to the heterologous AMV S40 isolate. A line with the coat protein gene in antisense orientation (CP–) showed delayed systemic infection but was not immune. We conclude that coat protein mediated protection (CPMP) is an effective strategy for controlling AMV infection and should be further evaluated in the field.


Virology ◽  
1991 ◽  
Vol 181 (2) ◽  
pp. 687-693 ◽  
Author(s):  
Lyda Neeleman ◽  
Antoinette C. Van Der Kuyl ◽  
John F. Bol

1987 ◽  
Vol 6 (5) ◽  
pp. 1181-1188 ◽  
Author(s):  
Nilgun E. Tumer ◽  
Keith M. O'Connell ◽  
Richard S. Nelson ◽  
Patricia R. Sanders ◽  
Roger N. Beachy ◽  
...  

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1130d-1130
Author(s):  
Guowei Fang ◽  
Rebecca Grumet

Zucchini yellow mosaic virus (ZYMV), a potyvirus, can cause major losses in cucurbit crops. With the goal of genetically engineering resistance to this disease we have engineered the ZYMV coat protein gene into a plant expression vector. The complete coat protein coding sequence, or the conserved core portion of the capsid gene, was attached to the 5' untranslated region of tobacco etch virus (TEV) in the pTL37 vector (Carrington et al., 1987, Nucl. Acid Res. 15:10066) The capsid constructs were successfully expressed by in vitro transcription and translation systems as verified by SDS-PAGE and ZYMV coat protein antibody. The constructs were then subcloned using polymerase chain reaction and attached to the CaMV 35 S transcriptional promoter on the CIBA-GEIGY pCIB710 plasmid. The constructs containing the CaMV 35S promoter, the 5' untranslated leader of TEV, and ZYMV coat protein sequences were then put between the Agrobacterium tumefaciens left and right borders in the pCIB10 vector and transferred to A. tumefaciens strain LBA4404 by triparental mating. These vectors are now being used to transform muskmelon and cucumber; resultant transgenic plants will be tested for ZYMV coat protein expression.


Virology ◽  
1994 ◽  
Vol 203 (2) ◽  
pp. 269-276 ◽  
Author(s):  
Peter E.M. Taschner ◽  
Guido Van Marle ◽  
Frans Th. Brederode ◽  
Nilgun E. Tumer ◽  
John F. Bol

2011 ◽  
Vol 26 (3) ◽  
pp. 229-243 ◽  
Author(s):  
Ivana Stankovic ◽  
Ana Vucurovic ◽  
Aleksandra Bulajic ◽  
Danijela Ristic ◽  
Janos Berenji ◽  
...  

Three-year investigation of the presence and distribution of tobacco viruses in Serbia revealed that Alfalfa mosaic virus (AMV) appeared every year with different frequency in tobacco crops. During 2008, the presence of AMV was detected in most of the tested samples (58.82%) and it was the second most common compared to all other viruses which presence was confirmed in Serbia. In 2006 and 2007, AMV was detected in a significantly lower percentage (2.80% and 13.64%, respectively). This study showed that Alfalfa mosaic virus was more commonly found in multiple infections with two, three or even four detected viruses. Single infections were detected only in 2006, in one tobacco field in the locality of Futog. During this investigation, a rapid and simple protocol was optimized and developed for molecular detection of AMV in tobacco leaves, using primers CPAMV1/CPAMV2 and commercially available kits for total RNA extraction as well as for RT-PCR (reverse transcription - polymerase chain reaction). Using RT-PCR and these primers that flank the AMV coat protein gene, a DNA fragment of 751 bp was amplified, sequenced, and compared with the sequences available in GenBank database. The sequence of isolate 196-08 (GenBank Acc. No. FJ527749) proved to be identical at the nucleotide level of 99 to 93% with those from other parts of the world. Phylogenetic analysis of 27 isolates based on 528 bp sequences of the coat protein gene did not show correlation of the isolates with their geographic origin or plant host and showed that these isolates fall into four molecular groups of strains. Serbian AMV isolate from tobacco belongs to group IV, the group that includes most of the isolates selected for phylogenetic analysis.


Crop Science ◽  
1998 ◽  
Vol 38 (6) ◽  
pp. 1661-1668 ◽  
Author(s):  
Dongmei Xu ◽  
Glenn B. Collins ◽  
Arthur G. Hunt ◽  
Mark T. Nielsen

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