Phytoplasma diseases of plants: molecular diagnostics and way forward

Nair, Smita; Manimekalai, R.

doi:10.1007/s11274-021-03061-y

Cited by 18 publications

(17 citation statements)

References 137 publications

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“…Taxonomic keys and descriptions have been developed and widely used for identifying potatoes. Moreover, a loop-mediated isothermal amplification (LAMP) method has been implemented for the rapid and accurate detection of pathogens in plants [25,26]. However, a significant constraint in LAMP assays lies in the design of proper primers.…”

Section: Introductionmentioning

confidence: 99%

Volatile Organic Compounds and Physiological Parameters as Markers of Potato (Solanum tuberosum L.) Infection with Phytopathogens

Steglińska

Pielech‐Przybylska²,

Janas³

et al. 2022

Molecules

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The feasibility of early disease detection in potato seeds storage monitoring of volatile organic compounds (VOCs) and plant physiological markers was evaluated using 10 fungal and bacterial pathogens of potato in laboratory-scale experiments. Data analysis of HS-SPME-GC-MS revealed 130 compounds released from infected potatoes, including sesquiterpenes, dimethyl disulfide, 1,2,4-trimethylbenzene, 2,6,11-trimethyldodecane, benzothiazole, 3-octanol, and 2-butanol, which may have been associated with the activity of Fusarium sambucinum, Alternaria tenuissima and Pectobacterium carotovorum. In turn, acetic acid was detected in all infected samples. The criteria of selection for volatiles for possible use as incipient disease indicators were discussed in terms of potato physiology. The established physiological markers proved to demonstrate a negative effect of phytopathogens infecting seed potatoes not only on the kinetics of stem and root growth and the development of the entire root system, but also on gas exchange, chlorophyll content in leaves, and yield. The negative effect of phytopathogens on plant growth was dependent on the time of planting after infection. The research also showed different usefulness of VOCs and physiological markers as the indicators of the toxic effect of inoculated phytopathogens at different stages of plant development and their individual organs.

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Section: Introductionmentioning

confidence: 99%

Volatile Organic Compounds and Physiological Parameters as Markers of Potato (Solanum tuberosum L.) Infection with Phytopathogens

Steglińska

Pielech‐Przybylska²,

Janas³

et al. 2022

Molecules

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show abstract

“…However, the inaccuracy of the nested PCR method using universal primers R16mF2/R16mR1 and R16F2n/R16R2 for detection of SPLDaP suggested the need to develop a specific method for further disease diagnosis. To date, an array of molecular methods has been developed for the detection of phytoplasmas, including PCR amplification, nested PCR, closed tube quantitative PCR assays, ddPCR, and the loop-mediated isothermal amplification (LAMP) assay [ 16 ], and more non-ribosomal genes, such as RpoB [ 23 ], SecY [ 24 ], Tuf [ 25 , 26 ], and Cpn60 [ 27 ] have been used as targets. However, the time consumption, cost, labour, laboratory equipment requirement, personal experience, and detection accuracies and efficiencies varied for these published methods [ 16 ].…”

Section: Discussionmentioning

confidence: 99%

“…To date, an array of molecular methods has been developed for the detection of phytoplasmas, including PCR amplification, nested PCR, closed tube quantitative PCR assays, ddPCR, and the loop-mediated isothermal amplification (LAMP) assay [ 16 ], and more non-ribosomal genes, such as RpoB [ 23 ], SecY [ 24 ], Tuf [ 25 , 26 ], and Cpn60 [ 27 ] have been used as targets. However, the time consumption, cost, labour, laboratory equipment requirement, personal experience, and detection accuracies and efficiencies varied for these published methods [ 16 ]. In all, nested PCR based on the 16S rRNA gene is still the most basic molecular detection method of phytoplasma [ 16 ], especially for the diagnosis of new phytoplasma disease, although it sometimes amplifies nontarget bacterial species due to the high conservation of the 16Sr RNA gene among various bacteria [ 18 , 19 , 20 , 21 ].…”

Section: Discussionmentioning

confidence: 99%

“…An accurate method for the detection of phytoplasmas in host plants and vectors is important; it would play a key role in disease diagnosis. So far, various molecular methods for phytoplasma detection have been developed [ 16 ]. In particular, nested PCR assays using universal primers (e.g., P1/P7 or R16mF2/R16mR1, followed by R16F2n/R16R2) based on the 16S rRNA gene have been widely used for phytoplasma detection in different host plants [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

“…In particular, nested PCR assays using universal primers (e.g., P1/P7 or R16mF2/R16mR1, followed by R16F2n/R16R2) based on the 16S rRNA gene have been widely used for phytoplasma detection in different host plants [ 17 ]. This assay involves two rounds of amplifications via PCR using universal or group specific primers and has higher sensitivity than direct PCR [ 16 ]. However, it sometimes amplifies nontarget bacterial species in some host plants and vectors, due to the high conservation of the 16Sr RNA gene among various species of bacteria, causing a problem of false positives [ 18 , 19 , 20 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

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Development of a Specific Nested PCR Assay for the Detection of 16SrI Group Phytoplasmas Associated with Sisal Purple Leafroll Disease in Sisal Plants and Mealybugs

Wang

Tan

et al. 2022

Plants

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Sisal purple leafroll disease (SPLD) is currently the most destructive disease affecting sisal in China, yet its aetiology remains unclear. In our previous research, it was verified to be associated with phytoplasmas, and nested PCR based on the 16S rRNA gene using universal primers R16mF2/R16mR1 followed by R16F2n/R16R2 was confirmed as the most effective molecular method for the detection of phytoplasmas associated with SPLD (SPLDaP). However, the method has a shortcoming of inaccuracy, for it could produce false positive results. To further manage the disease, accurate detection is needed. In this study, we developed a specific nested PCR assay using universal primers R16F2n/R16R2, followed by a set of primers designed on 16Sr gene sequences amplified from SPLDaP, nontarget bacteria from sisal plants, and other phytoplasma subgroups or groups. This established method is accurate, specific, and effective for detection of 16SrI group phytoplasma in sisal, and its sensitivity is up to 10 fg/μL of total DNA. It also minimized the false positive problem of nested PCR using universal primers R16mF2/R16mR1 followed by R16F2n/R16R2. This method was further used to verify the presence of phytoplasma in Dysmicoccusneobrevipes, and the results showed that D. neobrevipes could be infected by SPLDaP and thus could be a candidate for vector transmission assays.

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Detection and Identification of Plant Viruses, Viroids, and Phytoplasma Based on High-Throughput Molecular Approaches

Rithesh,

Paul,

Amanthra Keloth

et al. 2024

Interdisciplinary Biotechnological Advances

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Phytoplasma diseases of plants: molecular diagnostics and way forward

Cited by 18 publications

References 137 publications

Volatile Organic Compounds and Physiological Parameters as Markers of Potato (Solanum tuberosum L.) Infection with Phytopathogens

Volatile Organic Compounds and Physiological Parameters as Markers of Potato (Solanum tuberosum L.) Infection with Phytopathogens

Development of a Specific Nested PCR Assay for the Detection of 16SrI Group Phytoplasmas Associated with Sisal Purple Leafroll Disease in Sisal Plants and Mealybugs

Detection and Identification of Plant Viruses, Viroids, and Phytoplasma Based on High-Throughput Molecular Approaches

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