DETECTION OF THE CAUSAL AGENTS OF PISTACHIO GUMMOSIS BY THE POLYMERASE CHAIN REACTION

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MIRABOU ALFATHI M.; ERSHAD D.; ALI ZADEH A.; COOKE D.E.L.; DUNCAN J.M.; RAHIMIAN H.

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Journal: IRANIAN JOURNAL OF PLANT PATHOLOGY Year:2002 | Volume:38 | Issue:1-2 Page(s): 97-116

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Title

DETECTION OF THE CAUSAL AGENTS OF PISTACHIO GUMMOSIS BY THE POLYMERASE CHAIN REACTION

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Keywords

DETECTIONQ4

PISTACHIOQ4

GUMMOSISQ4

P. MEGASPERMAQ3

P. DRECHSLERIQ4

P. MELONISQ4

P. CAJANIQ4

P. SOJAEQ4

P. SINENSISQ3

ITS1Q3

ITS2Q3

RDNAQ4

PCR.Q4

Abstract

Pistachio nut (Pistacia vera) is a major nut crop in Iran and GUMMOSIS is one of the crps most serious diseases, diseased trees exuding gum from the crown and lower parts of the trunks of affected trees often die irom root and crown rot. Several species of Phytophthora have been isolated from affected trees, but the principal causes of GUMMOSIS in Kerman province, the mjor PISTACHIO-producing region in Iran, are two non-papilIate species. To date one has been described as P. MEGASPERMA and the other asP. drechsleri based on their morphological and physiologicalfactros. A technique based on the polymerase chain reaction(PCR) for the specificDETECTIONof the two main GUMMOSIS-causing Phytophthoraspecieswas developedusingthe nucleotide sequence of ITS I and ITS2 (internally transcribed spacer regions) of their genomic ribosomal RNA gene repeat (rDNA). DNA sequence from the ITS regions of the PISTACHIO P. MEGASPERMA isolates were used to develop a pair of oligonucleotide primers, pisl fwd and pisl rev. These amplified a specific fragment within ITS I and ITS2 of the all 24 isolates of P. MEGASPERMA from PISTACHIO. In further tests of specificity, the primers also generated an amplicon of the same size wiht DNA of Phytophthora sojae, P. MELONIS, P. SINENSIS, P. CAJANI, as well as P. DRECHSLERI isolates from PISTACHIO. In fact the isolates. from PISTACHIOn ascribed to P. MEGASPERMA and P. DRECHSLERI siolates, have been shown to belong to a previously undescribed species and to P. MELONIS, respectively. Together with all the aforementioned species, they form a tight clade within Phytophthora. No other Phytphthora species outside this ciade was amplified with pisl fwd and pisl rev. Amplification was highly sensitive with as little as 4 ng template DNA being detected in each case. PCR.amplification also detected both Phytophthora species, that cause GUMMOSIS in in feted roots of greenhouse-grown PISTACHIO plants. Nested PCR.with primers DC6 and ITS4, followed by the PIS 1 fwd and PIS 1 rev primers, gave clear products in every case with roots giving positive signals, in both symptomatic and asymptomatic infected plants. Digestion with Bsli of ITS amplicons (710bp) from the PISTACHIO P. MEGASPERMA isolates, created two fragments 370 and 310 bp long. These matched the two unique restriction sites within the sequence of the amplicon but were different from RFLPs generated by the same enzyme in the amplicon of P. SOJAE, P. MELONIS, P. SINENSIS, P. CAJANI. Use of the Bsl/ restriction enzyme discriminated between the two major GUMMOSIS inciting agents and distinguished them from the closely related P. SOJAE, P. MELONIS, P. SINENSIS, P. CAJANI.

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APA: Copy

MIRABOU ALFATHI, M., ERSHAD, D., ALI ZADEH, A., COOKE, D.E.L., DUNCAN, J.M., & RAHIMIAN, H.. (2002). DETECTION OF THE CAUSAL AGENTS OF PISTACHIO GUMMOSIS BY THE POLYMERASE CHAIN REACTION. IRANIAN JOURNAL OF PLANT PATHOLOGY, 38(1-2), 97-116. SID. https://sid.ir/paper/48240/en

Vancouver: Copy

MIRABOU ALFATHI M., ERSHAD D., ALI ZADEH A., COOKE D.E.L., DUNCAN J.M., RAHIMIAN H.. DETECTION OF THE CAUSAL AGENTS OF PISTACHIO GUMMOSIS BY THE POLYMERASE CHAIN REACTION. IRANIAN JOURNAL OF PLANT PATHOLOGY[Internet]. 2002;38(1-2):97-116. Available from: https://sid.ir/paper/48240/en

IEEE: Copy

M. MIRABOU ALFATHI, D. ERSHAD, A. ALI ZADEH, D.E.L. COOKE, J.M. DUNCAN, and H. RAHIMIAN, “DETECTION OF THE CAUSAL AGENTS OF PISTACHIO GUMMOSIS BY THE POLYMERASE CHAIN REACTION,” IRANIAN JOURNAL OF PLANT PATHOLOGY, vol. 38, no. 1-2, pp. 97–116, 2002, [Online]. Available: https://sid.ir/paper/48240/en

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