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| Articles
by
H.M. Mazyad |
Total Records (
2 ) for
H.M. Mazyad |
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W.S. El-Araby
,
I.A. Ibrahim
,
A.A. Hemeida
,
Amal Mahmoud
,
A.M. Soliman
,
A.K. El-Attar
and
H.M. Mazyad
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Viruses are major diseases affecting potato plantations in Egypt. Different molecular and serological methods for virus detection were used to detect the Potato virus X (PVX), Potato virus Y (PVY) and Potato leaf roll virus (PLRV). ELISA, RT-PCR, immunocapture RT-PCR (IC-RT-PCR) and RT-PCR-ELISA were the detection methods used. Naturally virus-like infected potato samples were collected from different locations at El-Munofia Governorate in Egypt. PVX and PVY were isolated and identified biologically through host range and indicator plants. All detection methods mentioned above were sensitive enough to detect the three potato viruses in potato tissues. Using ELISA method, PVX, PVY and PLRV were detected in 4, 7 and 10 out of 26 samples (15.4, 26.9 and 38.5%), respectively. Mixed infection was found between PVX/PVY; PVY/PLRV and also among PVX/PVY/PLRV. According to the results, it is the use of molecular methods to confirm certain serological results i.e., the virus especially when low concentration in the infected samples. Using the advantage of IC with RT-PCR-ELISA will reduce the costs and avoid RNA degradation. Additionally, RT-PCR-ELISA could be the method of choice among the molecular methods. |
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A.M. Soliman
,
B.N. Barsoum
,
G.G. Mohamed
,
A.A. Rezk
,
A.E. Aboul-Ata
and
H.M. Mazyad
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This study, aims at determination of efficiency of micro interfering
RNA (miRNA) to develop ability of virus resistance against Egyptian PVX
isolate (PVX-Eg2) in both potato (Solanum tuberosum L. cv. Spunta)
and tobacco (Nicotiana benthamiana). RNA constructs of Sense (PVX-Eg2cpVs),
antisense (PVX-Eg2cpCs) and sense/antisense were designed, cloned and
sub- cloned for gene transfection using Agrobacterium inoculation
technique. Two to 3 leaf-stage seedlings of potato (Solanum tuberosum
L. cv. Spunta) and tobacco (Nicotiana benthamiana) were
inoculated with the three previous constructs. The construct-treated plants
were mechanically inoculated with the PVX-Eg2 isolate. Bioassay and PCR
amplification have been able to evaluate transfected-plant resistance
against PVX-Eg2 that is caused by siRNA of PVX-Eg2cp. PCR amplification
has been able to detect PVX viral genome in all challenged plants those
infiltrated with either pFGC5491 vector without insert, or with sense
construct and also with antisense construct. Bioassay has confirmed same
previous statement. Nine out of 10 sense/antisense-transfected potato
plants were negatively reacted with both bioassay and PCR amplification.
Same negative reaction has been viewed using both bioassay and PCR for
sense/antisense transfected-tobacco plants. Seven out of 10 proved they
are PVX-Eg2 resistant. |
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