Chrysanthemum chlorotic mottle viroid (CChMVd) is a serious pathogen infecting chrysanthemum worldwide. To improve and enhance the detection procedure, the colorimetric loop-mediated isothermal amplifi cation (LAMP) technique was developed. Six LAMP primers were newly designed and tested the optimal conditions using a recombinant plasmid of CChMVd as a DNA template. The optimal conditions for colorimetric LAMP were incubation at 65°C for 60 min. At these conditions, the ladder-liked pattern LAMP products were detected along with the change of color from pink to yellow in the positive reactions. The sensitivity of colorimetric LAMP was up to 2.18 × 103 copies of recombinant plasmid concentration which was 104-fold greater than that of polymerase chain reaction (PCR). The developed colorimetric LAMP was not cross reacted to other viruse s and viroid. Effi ciency of colorimetric reverse transcription (RT)-LAMP for detecting CChMVd in chrysanthemum plantlets obtained from meristem tip culture was evaluated to verify and ensure CChMVd-free plantlets production. The results showed that 18 chrysanthemum plantlets were free of CChMVd contamination. Therefore, the colorimetric RT-LAMP can be used to detect CChMVd routinely due to its accuracy, rapidness and sensitivity.
In Thailand, strawberry production is confined mainly to Chiang Mai and Chiang Rai Provinces, located in the mountainous northern region, where the cool climate from December to March permits production (Pipattanawong, 2015). Strawberry plants are highly susceptible to high temperatures due to the reduction of strawberry flower formation and fruit quality (Klamkowski & Treder, 2008). Moreover, higher temperatures generate a higher demand for water, creating water stress in plants during dry seasons (Moretti et al., 2010). Therefore, strawberry is often harvested in winter and spring, but not in summer (Yamasaki, 2013). The greenhouse cultivation of strawberry, which provides protection from rain, pathogens and insects, and which ensures an optimal temperature environment, can be carried out throughout the year (Shigeoki et al., 2017).To produce strawberry all year round, greenhouse cultivation with an evaporative cooling system is often used for strawberry cultivation in Thailand (Pipattanawong, 2015). Like all commercially cultivated fruits, strawberries face a number of threats. Common diseases include leaf scorch, leaf spot, leaf blight, powdery mildew and anthracnose (Pipattanawong, 2015).However, strawberry plants are especially susceptible to fungal pathogens which can affect almost all parts of strawberry (Maas, 2012), and the occurrence of several fungi that infect strawberry crowns and roots has given rise to concerns that production
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