Abstract:, Edelclaiton Daros
RESUMOO raquitismo-da-soqueira, causado por Leifsonia xyli subsp. xyli é considerado uma das mais importantes doenças bacterianas da cana-de-açúcar (Saccharum spp.). Para verificar a possível ocorrência de Leifsonia xyli subsp. xyli na região noroeste do Paraná, foi realizado um levantamento em 32 áreas de plantio de duas usinas, utilizando-se o método sorológico "Dot blot" e um protocolo baseado na reação em cadeia da polimerase. As amostras foram coletadas em áreas cultivadas com 8 varied… Show more
“…Few studies have been conducted regarding PCR feasibility directly from vascular fluid over the years Comstock and Irey [4] Lopes et al [15] Wang et al [7]. Results from this work shows that fast methods of DNA extraction, which took approximately 2 hours for a total of 100 samples, may be as effective as more time-demanding and elaborated protocols that require around 24 hours to complete [16][17][18][19][20]. Therefore, large scale diagnosis of leaf scald in sugarcane may be performed through the DNA extraction techniques M2, M4 or M5, since these methods are fast, simple and do not use phenol as one of its reagents [21][22][23].…”
Among the techniques used for Leaf Scald diagnosis, PCR is considered the most sensitive, however this technique has limitations for use in large-scale analysis, particularly because of the difficulty/complexity associated with DNA extraction. Detection success largely depends upon the host species and the protocol used for DNA extraction. Here we describe a comparison of five DNA extraction methods and determine which one is more efficient for large scale detection of Leaf Scald through PCR. DNA extraction was performed from: a. Bacterial culture; b. Bacterial culture added to the vascular fluid from sugarcane; and c. From infected plant stalks. Of the five methods tested, three were effective for large-scale detection by PCR.
“…Few studies have been conducted regarding PCR feasibility directly from vascular fluid over the years Comstock and Irey [4] Lopes et al [15] Wang et al [7]. Results from this work shows that fast methods of DNA extraction, which took approximately 2 hours for a total of 100 samples, may be as effective as more time-demanding and elaborated protocols that require around 24 hours to complete [16][17][18][19][20]. Therefore, large scale diagnosis of leaf scald in sugarcane may be performed through the DNA extraction techniques M2, M4 or M5, since these methods are fast, simple and do not use phenol as one of its reagents [21][22][23].…”
Among the techniques used for Leaf Scald diagnosis, PCR is considered the most sensitive, however this technique has limitations for use in large-scale analysis, particularly because of the difficulty/complexity associated with DNA extraction. Detection success largely depends upon the host species and the protocol used for DNA extraction. Here we describe a comparison of five DNA extraction methods and determine which one is more efficient for large scale detection of Leaf Scald through PCR. DNA extraction was performed from: a. Bacterial culture; b. Bacterial culture added to the vascular fluid from sugarcane; and c. From infected plant stalks. Of the five methods tested, three were effective for large-scale detection by PCR.
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