Comparison of Different Sets of Primers Used in Detection and Identification of Potato Soft Rot Pectobacteriumcarotovorum subsp. carotovorum (DYE1969)

2018 ◽  
Vol 1 (1) ◽  
pp. 1-12 ◽  
Author(s):  
I. Abu-Obeid ◽  
H. Khlaif ◽  
N. Salem
2013 ◽  
Vol 163 (3) ◽  
pp. 378-393 ◽  
Author(s):  
M. Sławiak ◽  
R. van Doorn ◽  
M. Szemes ◽  
A.G.C.L. Speksnijder ◽  
M. Waleron ◽  
...  

2019 ◽  
Vol 20 (10) ◽  
Author(s):  
Tri Joko ◽  
ALAN SOFFAN ◽  
MUHAMMAD SAIFUR ROHMAN

Abstract. Joko T, Soffan A, Muhammad Saifur Rohman MS. 2019. A novel subspecies-specific primer targeting the gyrase B gene for the detection of Pectobacterium carotovorum subsp. brasiliense. Biodiversitas 20: 3042-3048. Pectobacterium carotovorum subsp. brasiliense is one of the major causative bacterial pathogens of the soft rot disease in various crops. It has a high virulence and a wide range of hosts in the tropics and the subtropics. Most often, conventional methods are not able to accurately distinguish P. carotovorum subsp. brasiliense from other subspecies. Thus, this study aimed to design a specific gyrase B gene (gyrB) -based primers for the detection and identification of soft rot pathogen. The specific primers design was based on the alignment using gyrB gene sequence data from P. carotovorum subsp. brasiliense and other data from the GenBank. The primers comprised of F-gyr-Pcb (5’-CAC AGG CAC CGC TGG CTG TT-3’) and R-gyr-Pcb (5’-CGT CGT TCC ACT GCA ATG CCA-3’) with an amplicon of 336 base pairs. The specificity of the primers pair was verified both in silico and in polymerase chain reaction (PCR) assays, where the primers could only detect P. carotovorum subsp. brasiliense. Primers’ sensitivity was determined by qualitative PCR with a detection limit of less than 0.5 ng/µL of genomic DNA. Hence, the proposed detection tool can be beneficial to advance further studies on the ecology and epidemiology of soft rot diseases.


2020 ◽  
Vol 8 (3) ◽  
pp. 358
Author(s):  
Md Niamul Kabir ◽  
Ali Taheri ◽  
C. Korsi Dumenyo

Pectobacterium and Dickeya species, usually referred to as soft rot Enterobacteriaceae, are phytopathogenic genera of bacteria that cause soft rot and blackleg diseases and are responsible for significant yield losses in many crops across the globe. Diagnosis of soft rot disease is difficult through visual disease symptoms. Pathogen detection and identification methods based on cultural and morphological identification are time-consuming and not always reliable. A polymerase chain reaction (PCR)-based detection method with the species-specific primers is fast and reliable for detecting soft rot pathogens. We have developed a specific and sensitive detection system for some species of soft rot Pectobacteriaceae pathogens in the Pectobacterium and Dickeya genera based on the use of species-specific primers to amplify unique genomic segments. The specificities of primers were verified by PCR analysis of genomic DNA from 14 strains of Pectobacterium, 8 strains of Dickeya, and 6 strains of non-soft rot bacteria. This PCR assay provides a quick, simple, powerful, and reliable method for detection of soft rot bacteria.


2014 ◽  
Vol 65 ◽  
pp. 153-167 ◽  
Author(s):  
Duy Phu Le ◽  
Mike Smith ◽  
George William Hudler ◽  
Elizabeth Aitken

Author(s):  
C.D. Humphrey ◽  
T.L. Cromeans ◽  
E.H. Cook ◽  
D.W. Bradley

There is a variety of methods available for the rapid detection and identification of viruses by electron microscopy as described in several reviews. The predominant techniques are classified as direct electron microscopy (DEM), immune electron microscopy (IEM), liquid phase immune electron microscopy (LPIEM) and solid phase immune electron microscopy (SPIEM). Each technique has inherent strengths and weaknesses. However, in recent years, the most progress for identifying viruses has been realized by the utilization of SPIEM.


2004 ◽  
Vol 171 (4S) ◽  
pp. 30-30
Author(s):  
Robert C. Eyre ◽  
Ann A. Kiessling ◽  
Thomas E. Mullen ◽  
Rachel L. Kiessling

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