Molecular Techniques for Diagnosis of Bacterial Plant Pathogens

Tewari, Sakshi; Sharma, Shilpi

doi:10.1016/b978-0-12-814849-5.00027-7

Cited by 9 publications

(3 citation statements)

References 117 publications

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“…In addition, low isolation success of the bacterium from the exudates was reported earlier by McEvoy et al [4]. As shown by Tewari and Sharma [28], P. syringae pv. aesculi can be outcompeted by other fast-growing bacteria, which can lead to such a low isolation success, in particular when samples were taken from older cankers [8,13].…”

Section: Discussionmentioning

confidence: 87%

Detectability of Pseudomonassyringae pv. aesculi from European Horse Chestnut Using Quantitative PCR Compared with Traditional Isolation

Schneider

Schefer

Meyer

2021

Forests

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Bleeding cankers on horse chestnut trees (Aesculushippocastanum and Aesculus × carnea), caused by Pseudomonassyringae pv. aesculi, have been reported across Europe. In the present study, we show the successful detection of P. syringae pv. aesculi on symptomatic horse chestnut trees in Switzerland using quantitative PCR (qPCR). However, P. syringae pv. aesculi was also detected by qPCR on trees from which no isolate was obtained through cultivation. Reduced isolation success and low copy numbers of the target gene were correlated with the increasing age of symptomatic horse chestnut trees. The potential of detecting non-viable P. syringae pv. aesculi by qPCR was evaluated using an inoculation experiment with dead bacteria and detection by qPCR and cultivation. The detectability of DNA from P. syringae pv. aesculi cells dropped by 34.5% one day after inoculation and then decreased only slightly until the end of the experiment (22 days after inoculation). In contrast, no bacterial growth was observed at any time point after the inactivation of the bacteria. To protect horse chestnut trees, evaluating the viability and actual infection stage of the bacterium may play an important role.

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Section: Discussionmentioning

confidence: 87%

Detectability of Pseudomonassyringae pv. aesculi from European Horse Chestnut Using Quantitative PCR Compared with Traditional Isolation

Schneider

Schefer

Meyer

2021

Forests

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“…In addition to the above, the conventional PCR techniques also have other drawbacks such as the requirement of sophisticated instruments like thermocycler and gel documentation systems. To address these disadvantages, several isothermal amplification methods, such as loop-mediated isothermal amplification (LAMP) 13 , helicase dependant amplification (HDA) 14 , nucleic acid sequence-based amplification 15 , etc., have been developed 16 .…”

mentioning

confidence: 99%

“…The LAMP assay has been widely used to detect viruses 17 , bacteria 18,19 , and fungal pathogens 20 due to its high sensitivity and specificity compared to other conventional PCR methods 19 . The HDA assay involves in vivo DNA replication using helicase enzyme 14,16,21,22 and does not require initial heat denaturation and subsequent thermocycling steps as required by PCR. Based on the principles of the LAMP assay, several microdevices have been developed integrating foldable technology [23][24][25] .…”

mentioning

confidence: 99%

LAMP-based foldable microdevice platform for the rapid detection of Magnaporthe oryzae and Sarocladium oryzae in rice seed

Prasannakumar

Parivallal

Pramesh

et al. 2021

Sci Rep

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Rice blast (caused by Magnaporthe oryzae) and sheath rot diseases (caused by Sarocladium oryzae) are the most predominant seed-borne pathogens of rice. The detection of both pathogens in rice seed is essential to avoid production losses. In the present study, a microdevice platform was designed, which works on the principles of loop-mediated isothermal amplification (LAMP) to detect M. oryzae and S. oryzae in rice seeds. Initially, a LAMP, polymerase chain reaction (PCR), quantitative PCR (qPCR), and helicase dependent amplification (HDA) assays were developed with primers, specifically targeting M. oryzae and S. oryzae genome. The LAMP assay was highly efficient and could detect the presence of M. oryzae and S. oryzae genome at a concentration down to 100 fg within 20 min at 60 °C. Further, the sensitivity of the LAMP, HDA, PCR, and qPCR assays were compared wherein; the LAMP assay was highly sensitive up to 100 fg of template DNA. Using the optimized LAMP assay conditions, a portable foldable microdevice platform was developed to detect M. oryzae and S. oryzae in rice seeds. The foldable microdevice assay was similar to that of conventional LAMP assay with respect to its sensitivity (up to 100 fg), rapidity (30 min), and specificity. This platform could serve as a prototype for developing on-field diagnostic kits to be used at the point of care centers for the rapid diagnosis of M. oryzae and S. oryzae in rice seeds. This is the first study to report a LAMP-based foldable microdevice platform to detect any plant pathogens.

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Detection and Identification of Soil-Borne Pathogens: Classical to Recent Updates

Hubballi

Johnson

Anjali

et al. 2022

Rhizosphere Microbes

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Molecular Techniques for Diagnosis of Bacterial Plant Pathogens

Cited by 9 publications

References 117 publications

Detectability of Pseudomonassyringae pv. aesculi from European Horse Chestnut Using Quantitative PCR Compared with Traditional Isolation

Detectability of Pseudomonassyringae pv. aesculi from European Horse Chestnut Using Quantitative PCR Compared with Traditional Isolation

LAMP-based foldable microdevice platform for the rapid detection of Magnaporthe oryzae and Sarocladium oryzae in rice seed

Detection and Identification of Soil-Borne Pathogens: Classical to Recent Updates

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