Asian Journal of Plant Pathology1819-1541xxxx-xxxxScience International10.3923/ajppaj.2014.1.9FaragNabil S. BalabelNaglaa M.A. 1201481The possible carryover effect of three fungicides and one insecticide on results
of PCR and Multiplex-PCR for detection of bacterial pathogens was studied. The
potato brown rot caused by phylotype II, sequevar I of Ralstonia solanacearum
and the potato ring rot bacteria Clavibacter michiganensis ssp. sepedonicus
were considered in this study. Under optimized conditions for PCR activity,
the potato extract prepared from the Cara potato cultivar did not interfere
with band development of DNAs
extracted from the organisms in concern. Regardless of their mode(s) of action,
the studied pesticides exerted different effect(s) on band development according
to their chemical grouping. The dimethomorph (Acrobat), systemic fungicide,
caused complete inhibition of band development of DNA extracted from either
potato brown rot or potato ring rot bacteria which may be considered as false
negative reaction(s). The propamocarb carbamate (Previcur N), systemic fungicide,
did not inhibit band development of the organisms in concern. The dithiocarbamate
plus zinc and manganese (Diathane M45), contact fungicide however, caused a
complete band inhibition of ring rot DNA contrary to that being reported for
brown rot, indicating a preferential polymerase activity. The organophosphate
(Malathion), penetrative non-systemic insecticide, did not inhibit DNA band
development for both organisms. Multiplex-PCR of mixed DNAs
of R. solanacearum and C. michiganensis ssp. sepedonicus
caused a false negative reaction through inhibition of band development of R.
solanacearum but not for C. michiganensis ssp. sepedonicus,
under the effect of propamocarb carbamate (Previcur N) fungicide and organophosphate
(Malathion) insecticide. It could be concluded that the pesticides carryover
effect on PCR results of potato brown rot and ring rot bacteria are being variable
according to the pesticide grouping. The dimethomorph (Acrobat) fungicide caused
false negative effect with both organisms under study followed by dithiocarbamate
(Diathane) fungicide in case of ring rot bacteria. The false negative multiplex-PCR
is being evident only in case of R. solanacearum under the stress of
both propamocarb carbamate (Previcur N) fungicide and on the organophosphate
(Malthion) insecticide. The mode of action of the used pesticide, either contact
or systemic, had no effect on PCR or multiplex-PCR results. Further investigations
are needed with different groups of pesticide.]]>Anonymous,19931993Caruso, P., P. Llop, J.L. Palomo, P. Garcia, C. Morente and M.M. Lopez,1998Ralstonia Solanacearum.]]>1998pp: 128-132pp: 128-132Chaurasia, P.C.P.,2005Phytophthora infestans L.) of potato in Eastern Tarai of Nepal.]]>65761De Lomas, J.G., F.J. Sunzeri and M.P Busch,1992328385Demeke, T. and R.P. Adams,199212332334Dieffenbach, C.W., T.M.J. Lowe and G.S. Dveksler,19933S30S37Elnifro, E.M., A.M. Ashshi, R.J. Cooper and P.E. Klapper,200013559570Farag, N.S., A.A. Gomah and N.M.A. Balabel,201097378Gafar, M.O. and Y.M. Dagash,20114226230Brownie, J., S. Shawcross, J. Theaker, D. Whitcombe, R. Ferrie, C. Newton and S. Little,19972532353241Kabeil, S.S., S.M. Lashin, M.H. El-Masry, M.A. El-Saadani, M.M. Abd-Elgawad and A.M. Aboul-Einean,200844454Kelman, A.,1954Pseudomonas solanacearum to colony appearance in a tetrazolium medium.]]>44693695Kim, H., J. Kim and C. Ihm,201024175181Kudela, V.,2007433546Martin, R.R., D. James and C.A. Levesque,200038207239Mutter, G.L. and K.A. Boynton,19952314111418Powelson, M.L. and D.A. Inglis,199983265268Polz, M.F. and C.M. Cavanaugh,19986437243730Rossen, L., P. Norskov, K. Holmstrom and O.F. Rasmussen,1992173745Ruano, G., D.E. Brash and K.K. Kidd,1991714Samoucha, Y. and Y. Cohen,1990182740Schaad, N.W. and R.D. Fredric,200224250258Seal, S.E., L.A. Jackson, J.P.W. Young and M.J. Daniels,1993Pseudomonas solanacearum, Pseudomonas syzygii, Pseudomonas pickettii and the blood disease bacterium by partial 16S rRNA sequencing: Construction of oligonucleotide primers for sensitive detection by polymerase chain reaction.]]>13915871594Stephenson, F.H. and M.C. Abilock,20122012Tsai, Y.L. and B.H. Olson,19925822922295Walsh, P.S., H.A. Erlich and R. Higuchi,19921241250Wagner, A., N. Blackstone, P. Cartwright, M. Dick and B. Misof et al.,199443250261Watson, R.J. and B. Blackwell,200046633642Wilson, I.G.,19976337413751