Detection of the Bacterial Potato Pathogens Pectobacterium and Dickeya spp. Using Conventional and Real-Time PCR

Humphris, Sonia; Cahill, Greig; Elphinstone, J. G.; Kelly, Rachel; Parkinson, Neil; Pritchard, Leighton; Toth, Ian K.; Saddler, G. S.

doi:10.1007/978-1-4939-2620-6_1

Cited by 35 publications

(15 citation statements)

References 34 publications

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“…Subsequently, 200 μl of homogenate suspension was transferred into 1,800 μl PEB (0.32 g/l of MgSO 4 , 1.08 g/l of (NH 4 ) 2 SO 4 , 1.08 g/l of K 2 HPO 4 , and 1.7 g/l of sodium polypectate) liquid medium (Perombelon and Van Der Wolf, 2002; Czajkowski et al, 2010) and incubation was continued under the same conditions for 48 h. The bacterial culture was centrifuged to collect bacteria for total DNA extraction using the HiPure Bacterial DNA Kit (Magen, China). The presence of the Pcc pathogen was detected through polymerase chain reaction (PCR) using the following two Pcc -specific primers: EXPCCF (5′-GAA CTT CGC ACC GCC GAC CTT CTA-3′) and EXPCCR (5′-GCC GTA ATT GCC TAC CTG CTT AAG-3′) as previously described (Kang et al, 2003; Humphris et al, 2015). Data were analyzed using the Chi-squared test ( p < 0.05).…”

Section: Methodsmentioning

confidence: 99%

Biocontrol of Soft Rot of Chinese Cabbage Using an Endophytic Bacterial Strain

Cui

Munir

et al. 2019

Front. Microbiol.

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Soft rot caused by Pectobacterium carotovorum subsp. carotovorum ( Pcc ) is a major constraint in the production of Chinese cabbage. The objective of this study was to demonstrate that the causative agent Pcc may be successfully managed by Bacillus amyloliquefaciens KC-1, both in vitro and in vivo . Chinese cabbage seedlings were cultivated in organic substrate termed bio-organic substrate using a floating-seedling system with B. amyloliquefaciens KC-1. This approach was applied in a greenhouse to evaluate the management of soft rot. The results showed that the extent of soft rot, as well as the transmission of Pcc to the stem progeny and its survival in the rhizosphere, was reduced following inoculation with B. amyloliquefaciens KC-1. In contrast, the population diversity of B. amyloliquefaciens KC-1 persisted in the Chinese cabbage stems after germination. These findings revealed that B. amyloliquefaciens KC-1 was able to survive and suppress the growth of Pcc in Chinese cabbage and its rhizosphere, protecting the host from the pathogen. The use of B. amyloliquefaciens KC-1 throughout the growth period of plants may be an effective strategy for the prevention of soft rot in Chinese cabbage.

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Section: Methodsmentioning

confidence: 99%

Biocontrol of Soft Rot of Chinese Cabbage Using an Endophytic Bacterial Strain

Cui

Munir

et al. 2019

Front. Microbiol.

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“…A number of methods have been suggested for the identification and differentiation of SRP (reviewed in [12,13]). Pat has been detected by means of conventional PCR [14][15][16][17] and loop-mediated isothermal DNA amplification (LAMP) [18,19].…”

Section: Introductionmentioning

confidence: 99%

Development of qPCR Detection Assay for Potato Pathogen Pectobacterium atrosepticum Based on a Unique Target Sequence

Lukianova

Evseev

Stakheev

et al. 2021

Plants

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The recent taxonomic diversification of bacterial genera Pectobacterium and Dickeya, which cause soft rot in plants, focuses attention on the need for improvement of existing methods for the detection and differentiation of these phytopathogens. This research presents a whole genome-based approach to the selection of marker sequences unique to particular species of Pectobacterium. The quantitative real-time PCR assay developed is selective in the context of all tested Pectobacterium atrosepticum strains and is able to detect fewer than 102 copies of target DNA per reaction. The presence of plant DNA extract did not affect the sensitivity of the assay.

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“…As there are not sufficiently effective protocols for mass treatment, one of the best agro-technical approaches is to reject the infected plants. For all these reasons, timely detecting P. atrosepticum is crucial to effectively protect plants and produce potato seed tubers [ 7 , 8 ].…”

Section: Introductionmentioning

confidence: 99%

Pectobacterium atrosepticum Biosensor for Monitoring Blackleg and Soft Rot Disease of Potato

et al. 2020

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Pectobacterium atrosepticum (Pba) is a quarantine and threatening phytopathogen known as the causal agent of blackleg and soft rot disease of potatoes in many areas. Its early detection is then important to have healthy potato tubers and reduce economic losses. Today, conventional methods such as enzyme-linked immunosorbent-assay (ELISA) and polymerase chain reaction (PCR) are typically used for Pba detection, but they are expensive and time-consuming. Here we report on the optimization of an alternative approach based on an electrochemical impedance immunosensor combining a microfluidic module and a microelectrodes array, and having advantages in terms of low cost, ease of use and portability. For validation and for assessing its performance, the lab-on-chip platform has been compared with two standard methods (ELISA and PCR).

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Detection of the Bacterial Potato Pathogens Pectobacterium and Dickeya spp. Using Conventional and Real-Time PCR

Cited by 35 publications

References 34 publications

Biocontrol of Soft Rot of Chinese Cabbage Using an Endophytic Bacterial Strain

Biocontrol of Soft Rot of Chinese Cabbage Using an Endophytic Bacterial Strain

Development of qPCR Detection Assay for Potato Pathogen Pectobacterium atrosepticum Based on a Unique Target Sequence

Pectobacterium atrosepticum Biosensor for Monitoring Blackleg and Soft Rot Disease of Potato

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