Articles
PCR DETECTION OF POME AND STONE FRUIT PHYTOPLASMAS FROM ACTIVE OR DORMANT TISSUE
Article number
472_97
Pages
731 – 736
Language
Abstract
Various oligonucleotide primers, derived from the 16S rRNA, were used to detect phytoplasmas associated with apple proliferation (AP), plum leptonecrosis (PLN), pear decline (PD), apricot chlorotic leaf roll (ACLR), and X-disease.
Primers AP3-AP5 were successfully utilized for amplification of AP-, PLN,- and PD-DNA fragments from fruit trees and of AP and PLN DNA fragments from infected periwinkle.
AP and PLN PCR products were cloned into a plasmid vector and then sequenced.
An attempt was made to design PLN specific primers based on a few nucleotides that are different between PLN and AP. These primers in PCR assays gave a higher yield of products from PLN infected tissue than from AP and PD infected tissue.
The primers were more specific if used in nested PCR with PCR products obtained with AP-group specific AP3-AP5 primers.
The universal primers R16F2- R16R2 also were used for PCR detection of several phytoplasmas, including AP, ACLR and X-Disease.
Therefore, the primers can be used in PCR assays for the reliable and sensitive diagnosis of various phytoplasmas affecting pome and stone fruits.
PCR assays also were sensitive enough to amplify phytoplasma DNA fragments from dormant infected Prunus material.
Thus, PCR assays may be used for phytoplasma detection in Prunus quarantine and certification programs.
Primers AP3-AP5 were successfully utilized for amplification of AP-, PLN,- and PD-DNA fragments from fruit trees and of AP and PLN DNA fragments from infected periwinkle.
AP and PLN PCR products were cloned into a plasmid vector and then sequenced.
An attempt was made to design PLN specific primers based on a few nucleotides that are different between PLN and AP. These primers in PCR assays gave a higher yield of products from PLN infected tissue than from AP and PD infected tissue.
The primers were more specific if used in nested PCR with PCR products obtained with AP-group specific AP3-AP5 primers.
The universal primers R16F2- R16R2 also were used for PCR detection of several phytoplasmas, including AP, ACLR and X-Disease.
Therefore, the primers can be used in PCR assays for the reliable and sensitive diagnosis of various phytoplasmas affecting pome and stone fruits.
PCR assays also were sensitive enough to amplify phytoplasma DNA fragments from dormant infected Prunus material.
Thus, PCR assays may be used for phytoplasma detection in Prunus quarantine and certification programs.
Authors
L. Carraro, L. Nemchinov, A. Hadidi
Keywords
Online Articles (101)