Journal of Agricultural Science and Technology
Journal of Agricultural Science and Technology
JAST
Agriculture
http://jast.modares.ac.ir
1
admin
1680-7073
2345-3737
10.22034/jast
en
jalali
1391
6
1
gregorian
2012
9
1
14
5
online
1
fulltext
en
A Dual Coat Protein Construct Establishes Resistance to Passionfruit Woodiness and Cucumber Mosaic Viruses
There is a high degree (>95%) of intraspecies similarity in the coat protein (CP) amino acid sequences within <em>Passionfruit woodiness virus </em>(PWV) and <em>Cucumber mosaic virus</em> (CMV), both infecting passionfruit vine in New South Wales. On this basis, a dual transgene containing the translatable cDNAs coding for the CPs of PWV and CMV was constructed in the binary vector pBI121 and used for transformation of <em>Nicotiana benthamiana</em>, a susceptible host to both viruses. The transformation was achieved by co-cultivation of the agrobacteria with the leaf disks prepared from the surface- sterilized leaves. Five transgenic lines including 1-1, 1-5, 1-7, 1-12 and 1-24 were regenerated. Insertion and transcription of the dual construct were confirmed, however, only the CMV CP was feasibly detectable by DAS-ELISA in the lines. Low level accumulation of CMV and/or PWV was evident in the lines. In the initail challenge trial where 1:10 dilution of plant sap was used, a 5-day delay in symptom was generally shown. Inoculations with 1:100 plant sap also gave similar results as with 1:10 dilution. Lines 1-5 and 1-12, which were inoculated with 1:1000 dilution of sap, remained uninfected by CMV till 27 dpi, whereas with PWV, 1-12 became infected by 11 dpi. Four cuttings of line 1-12 reacted diffferently to the challenge inoculations i.e. three of them resisted PWV, whereas two of them were susceptible to CMV. Since PWV CP was not detectable in the transgenic lines but evidence of resistance to PWV was found in them, this was suggestive of an RNA silencing mechnaism involved in the resistance. Because the CMV CP was detectable in the transgenic lines, this suggested requirement for the CP expression in the resistance . The resistance, or apparent immunity, was manifested by an apparent delay in symptom expression and accumulation of relatively low levels of the viruses.
Coat Protein,Cucumber mosaic virus,Dual construct,Passionfruit woodiness virus
1105
1120
http://jast.modares.ac.ir/browse.php?a_code=A-23-1000-4263&slc_lang=en&sid=23
N.
Sokhandan- Bashir
N.
Sokhandan- Bashir
100319475328460046218
100319475328460046218
Yes
Department of Plant Protection, College of Agriculture, University of Tabriz, Tabriz, Islamic Republic of Iran.
Department of Plant Protection, College of Agriculture, University of Tabriz, Tabriz, Islamic Republic of Iran.
M.
Gillings
M.
Gillings
100319475328460046219
100319475328460046219
No
Department of Biological Sciences, Macquarie University, Sydney, Australia.
Department of Biological Sciences, Macquarie University, Sydney, Australia.
J.
Bowyer
J.
Bowyer
100319475328460046220
100319475328460046220
No
Department of Crop Sciences, The University of Sydney, New South Wales 2006 Australia.
Department of Crop Sciences, The University of Sydney, New South Wales 2006 Australia.