Use of LAMP for Assessing Botrytis cinerea Colonization of Bunch Trash and Latent Infection of Berries in Grapevines

Ammour, Mélissa Si; Castaldo, Eleonora; Fedele, Giorgia; Rossi, Vittorio

doi:10.3390/plants9111538

Cited by 8 publications

(3 citation statements)

References 68 publications

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“…Mumford and Seal [26] reported that yam tissues contain some PCR-inhibitory substances that could be co-extracted with the RNA. This suggests a higher tolerance of RT-LAMP to inhibitors than RT-PCR, which has also been reported in other studies [23,44,45].…”

Section: Discussionsupporting

confidence: 87%

Improved Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for the Rapid and Sensitive Detection of Yam mosaic virus

Festus

Seal

Prempeh

et al. 2023

Viruses

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Yam (Dioscorea spp.) productivity is constrained significantly by the lack of a formal seed system. Vegetative propagation, through tuber setts as ‘seed’ yams, encourages the recycling of virus-infected planting materials, contributing to high virus incidence and yield losses. Efforts are ongoing to increase the production of high-quality seed yams in a formal seed system to reduce virus-induced yield losses and enhance the crop’s productivity and food security. Specific and sensitive diagnostic tests are imperative to prevent the multiplication of virus-infected materials contributing to a sustainable seed yam certification system. During routine indexing of yam accessions, discrepancies were observed between the results obtained from the reverse transcription loop-mediated isothermal amplification (RT-LAMP) test and those from reverse transcription polymerase chain reaction (RT-PCR); RT-LAMP failed to detect Yam mosaic virus (YMV) in some samples that tested positive by RT-PCR. This prompted the design of a new set of LAMP primers, YMV1-OPT primers. These primers detected as little as 0.1 fg/µL of purified RNA obtained from a YMV-infected plant, a sensitivity equivalent to that obtained with RT-PCR. RT-LAMP using YMV1-OPT primers is recommended for all future virus-indexing of seed yams for YMV, offering a rapid, sensitive, and cost-effective approach.

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Section: Discussionsupporting

confidence: 87%

Improved Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for the Rapid and Sensitive Detection of Yam mosaic virus

Festus

Seal

Prempeh

et al. 2023

Viruses

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“…Investigations of bunch debris at the time of thinning would determine if the fewer debris pieces in the thinned bunches were due to direct shaking of the bunch during the thinning or the removal of individual berries allowing debris to fall from the bunch over time before harvest. New methods of detecting spores in debris and potential spore production [ 85 , 86 ] from the dead plant material in the bunch could be used for a detailed study of the importance of this pathway of infection in the epidemic of vines receiving mechanical shaking or control treatments. If all debris cannot be removed by mechanical shaking and or the use of devices such as the Collard pulsed air system, then other methods to suppress inoculum within the trash such as targeted fungicides or biological control agents [ 35 ] may need to be considered if this pathway is important for harvest disease.…”

Section: Cultural Control Of Bbrmentioning

confidence: 99%

A Review of Cultural Practices for Botrytis Bunch Rot Management in New Zealand Vineyards

Mundy

Elmer

Wood

et al. 2022

Plants

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Botrytis bunch rot of grapes (BBR) causes substantial crop and wine quality issues globally. Past and present foundations for BBR control are based upon synthetic fungicides and varying forms of canopy management. Many authors regard the continued dependence on fungicides as unsustainable and have urged greater deployment of cultural, biological and nutritional strategies. However, in contrast to organic wine production, the uptake of alternative strategies in conventional vineyards has been slow based on cost and perceived reliability issues. This review summarises research from many different wine growing regions in New Zealand with the aim of demonstrating how traditional and newly developed cultural control practices have cost-effectively reduced BBR. In addition to reviewing traditional cultural practices (e.g., leaf removal), mechanical tools are described that remove floral trash and mechanically shake the vines. Multi-omics has improved our knowledge of the underlying changes to grape berries after mechanical shaking. Exogenous applications of calcium may correct calcium deficiencies in the berry skin and reduce BBR but the outcome varies between cultivar and regions. Nitrogen aids in grapevine defence against BBR but remains a complex and difficult nutrient to manage. The sustainable growth of organics and The European Green Deal will stimulate researchers to evaluate new combinations of non-chemical BBR strategies in the next decade.

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“…Various approaches have been considered for the detection of Botrytis cinerea [3], including gigahertz ultrasonic imaging of fungal spores [4], the fingerprint of the Raman spectrum of the fungal spores [5], detection from the mycelia using antibodies specific for Botrytis species [6], detection of the fungal DNA [7][8][9], or indirect detection through the measurement of indicators such as laccase activity [10], glycerol and gluconic acid [10], or plant hormones (i.e., the combined detection of salicylic, azelaic, and jasmonic acids) [11]. It is important to note that not all strains of Botrytis cinerea produce laccase.…”

Section: Introductionmentioning

confidence: 99%

Fast Electrochemical Measurement of Laccase Activity for Monitoring Grapes’ Infection with Botrytis cinerea

et al. 2022

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Grapes’ infection with the fungi Botrytis cinerea is one of the major causes of economic loss in the winemaking sector worldwide. The laccase activity of grapes is considered an appropriate indicator of this type of fungal infection, and enzymatic activity higher than 3 U/mL indicates a high risk of irreversibly damaged grape must due to enzymatic browning. This work describes a fast test for the measurement of laccase activity based on a dual optical and electrochemical detection method. A paper sensor impregnated with the enzymatic substrate dye 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) provides a semi-quantitative optical measurement. While the paper sensor can be used independently, when combined with a screen-printed electrode and amperometry measurements, it enables the quantitative detection of laccase activities down to 0.4 U/mL in only 5 min. The method was applied for monitoring the artificial infection of white, rosé, and red grapes with different strains of Botrytis cinerea. The results were confirmed by parallel analysis using the spectrophotometric method of laccase activity determination based on syringaldazine. The influence of the fungal strain and type of grape on laccase activity levels is reported. The demonstrated robustness, simplicity, and versatility of the developed method make it ideal for application on-site in the vineyard or at grape processing points.

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Use of LAMP for Assessing Botrytis cinerea Colonization of Bunch Trash and Latent Infection of Berries in Grapevines

Cited by 8 publications

References 68 publications

Improved Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for the Rapid and Sensitive Detection of Yam mosaic virus

Improved Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for the Rapid and Sensitive Detection of Yam mosaic virus

A Review of Cultural Practices for Botrytis Bunch Rot Management in New Zealand Vineyards

Fast Electrochemical Measurement of Laccase Activity for Monitoring Grapes’ Infection with Botrytis cinerea

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