Loop-mediated isothermal amplification assay: A specific and sensitive tool for the detection of Bipolaris oryzae causing brown spot disease in rice
Rice brown spot caused by Bipolaris oryzae (Breda de Haan) is the predominant disease in rice, infecting millions of hectares of cultivated rice, and greatly reduces yields. The various available diagnostic methods for B. oryzae disease detection are time consuming and require sophisticated instrume...
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| Veröffentlicht in: | Phytoparasitica 2022-07, Vol.50 (3), p.543-553 |
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| creator | Lakshmi, K. R. Swarna Kamalakannan, A. Gopalakrishnan, C. Rajesh, S. Panneerselvam, S. Ganapati, Patil Santosh |
| description | Rice brown spot caused by
Bipolaris oryzae
(Breda de Haan) is the predominant disease in rice, infecting millions of hectares of cultivated rice, and greatly reduces yields. The various available diagnostic methods for
B. oryzae
disease detection are time consuming and require sophisticated instruments. Thus, an affordable, rapid and highly specific assay is needed. Here, we report a loop-mediated isothermal amplification (LAMP) assay protocol for specifically detecting
B. oryzae
by targeting a gene encoding a
glycoside hydrolase
(GH) family 13 protein. A set of six specific, sensitive LAMP primers were designed, allowing the effective LAMP detection of pathogens after incubation at 62°C for 60 min. Visual detection was performed by the addition of hydroxynapthol blue dye. In a positive reaction, the color changed to intense sky blue, while in a negative reaction, the color remained violet. The developed LAMP assay was highly sensitive, with a detection limit of 100 femtograms. The high specificity of the assay for the detection of
B. oryzae
was validated. Thus, the LAMP assay protocol provides a rapid, reliable tool for the early detection of
B. oryzae
under laboratory conditions. |
| doi_str_mv | 10.1007/s12600-022-00979-3 |
| format | Article |
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Bipolaris oryzae
(Breda de Haan) is the predominant disease in rice, infecting millions of hectares of cultivated rice, and greatly reduces yields. The various available diagnostic methods for
B. oryzae
disease detection are time consuming and require sophisticated instruments. Thus, an affordable, rapid and highly specific assay is needed. Here, we report a loop-mediated isothermal amplification (LAMP) assay protocol for specifically detecting
B. oryzae
by targeting a gene encoding a
glycoside hydrolase
(GH) family 13 protein. A set of six specific, sensitive LAMP primers were designed, allowing the effective LAMP detection of pathogens after incubation at 62°C for 60 min. Visual detection was performed by the addition of hydroxynapthol blue dye. In a positive reaction, the color changed to intense sky blue, while in a negative reaction, the color remained violet. The developed LAMP assay was highly sensitive, with a detection limit of 100 femtograms. The high specificity of the assay for the detection of
B. oryzae
was validated. Thus, the LAMP assay protocol provides a rapid, reliable tool for the early detection of
B. oryzae
under laboratory conditions.</description><identifier>ISSN: 0334-2123</identifier><identifier>EISSN: 1876-7184</identifier><identifier>DOI: 10.1007/s12600-022-00979-3</identifier><language>eng</language><publisher>Dordrecht: Springer Netherlands</publisher><subject>Agricultural production ; Agriculture ; Assaying ; Biomedical and Life Sciences ; Bipolaris oryzae ; Brown spot ; Color ; Cultivation ; Disease detection ; Ecology ; Gene amplification ; Glycosidases ; Glycoside hydrolase ; Grain cultivation ; Hydrolase ; Life Sciences ; Original Article ; Pathogens ; Plant diseases ; Plant Pathology ; Plant Sciences ; Proteins ; Rice</subject><ispartof>Phytoparasitica, 2022-07, Vol.50 (3), p.543-553</ispartof><rights>The Author(s), under exclusive licence to Springer Nature B.V. 2022</rights><rights>The Author(s), under exclusive licence to Springer Nature B.V. 2022.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c319t-4d306cf1d064a102125ae96c9d6b27b2106d90cb21ebbc62cf8f8b891d5a7add3</citedby><cites>FETCH-LOGICAL-c319t-4d306cf1d064a102125ae96c9d6b27b2106d90cb21ebbc62cf8f8b891d5a7add3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s12600-022-00979-3$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s12600-022-00979-3$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids></links><search><creatorcontrib>Lakshmi, K. R. Swarna</creatorcontrib><creatorcontrib>Kamalakannan, A.</creatorcontrib><creatorcontrib>Gopalakrishnan, C.</creatorcontrib><creatorcontrib>Rajesh, S.</creatorcontrib><creatorcontrib>Panneerselvam, S.</creatorcontrib><creatorcontrib>Ganapati, Patil Santosh</creatorcontrib><title>Loop-mediated isothermal amplification assay: A specific and sensitive tool for the detection of Bipolaris oryzae causing brown spot disease in rice</title><title>Phytoparasitica</title><addtitle>Phytoparasitica</addtitle><description>Rice brown spot caused by
Bipolaris oryzae
(Breda de Haan) is the predominant disease in rice, infecting millions of hectares of cultivated rice, and greatly reduces yields. The various available diagnostic methods for
B. oryzae
disease detection are time consuming and require sophisticated instruments. Thus, an affordable, rapid and highly specific assay is needed. Here, we report a loop-mediated isothermal amplification (LAMP) assay protocol for specifically detecting
B. oryzae
by targeting a gene encoding a
glycoside hydrolase
(GH) family 13 protein. A set of six specific, sensitive LAMP primers were designed, allowing the effective LAMP detection of pathogens after incubation at 62°C for 60 min. Visual detection was performed by the addition of hydroxynapthol blue dye. In a positive reaction, the color changed to intense sky blue, while in a negative reaction, the color remained violet. The developed LAMP assay was highly sensitive, with a detection limit of 100 femtograms. The high specificity of the assay for the detection of
B. oryzae
was validated. Thus, the LAMP assay protocol provides a rapid, reliable tool for the early detection of
B. oryzae
under laboratory conditions.</description><subject>Agricultural production</subject><subject>Agriculture</subject><subject>Assaying</subject><subject>Biomedical and Life Sciences</subject><subject>Bipolaris oryzae</subject><subject>Brown spot</subject><subject>Color</subject><subject>Cultivation</subject><subject>Disease detection</subject><subject>Ecology</subject><subject>Gene amplification</subject><subject>Glycosidases</subject><subject>Glycoside hydrolase</subject><subject>Grain cultivation</subject><subject>Hydrolase</subject><subject>Life Sciences</subject><subject>Original Article</subject><subject>Pathogens</subject><subject>Plant diseases</subject><subject>Plant Pathology</subject><subject>Plant Sciences</subject><subject>Proteins</subject><subject>Rice</subject><issn>0334-2123</issn><issn>1876-7184</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>BENPR</sourceid><recordid>eNp9kMGKFDEQhoMoOK6-gKcCz62VZCbd8bYuri4MeNFzSCfVa5aeTpvKKONz-MBmdwRve0oRvu8v6hfitcS3ErF_x1IZxA6V6hBtbzv9RGzk0Juul8P2qdig1ttOSaWfixfMd4gNl7gRf_Y5r92BYvKVIiTO9TuVg5_BH9Y5TSn4mvICntmf3sMl8Erh_hv8EoFp4VTTT4Ka8wxTLtB0iFQpPGh5gg9pzbMviSGX029PEPyR03ILY8m_lpaXK8TE5JkgLVBSoJfi2eRnplf_3gvx7frj16vP3f7Lp5ury30XtLS120aNJkwyotl6ie26nSdrgo1mVP2oJJpoMbSBxjEYFaZhGsbByrjzvY9RX4g359y15B9H4uru8rEsbaVTpjfaDoPdNUqdqVAyc6HJrSUdfDk5ie6-fXdu37X23UP7TjdJnyVu8HJL5X_0I9Zf7z6K-A</recordid><startdate>20220701</startdate><enddate>20220701</enddate><creator>Lakshmi, K. 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R. Swarna</au><au>Kamalakannan, A.</au><au>Gopalakrishnan, C.</au><au>Rajesh, S.</au><au>Panneerselvam, S.</au><au>Ganapati, Patil Santosh</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Loop-mediated isothermal amplification assay: A specific and sensitive tool for the detection of Bipolaris oryzae causing brown spot disease in rice</atitle><jtitle>Phytoparasitica</jtitle><stitle>Phytoparasitica</stitle><date>2022-07-01</date><risdate>2022</risdate><volume>50</volume><issue>3</issue><spage>543</spage><epage>553</epage><pages>543-553</pages><issn>0334-2123</issn><eissn>1876-7184</eissn><abstract>Rice brown spot caused by
Bipolaris oryzae
(Breda de Haan) is the predominant disease in rice, infecting millions of hectares of cultivated rice, and greatly reduces yields. The various available diagnostic methods for
B. oryzae
disease detection are time consuming and require sophisticated instruments. Thus, an affordable, rapid and highly specific assay is needed. Here, we report a loop-mediated isothermal amplification (LAMP) assay protocol for specifically detecting
B. oryzae
by targeting a gene encoding a
glycoside hydrolase
(GH) family 13 protein. A set of six specific, sensitive LAMP primers were designed, allowing the effective LAMP detection of pathogens after incubation at 62°C for 60 min. Visual detection was performed by the addition of hydroxynapthol blue dye. In a positive reaction, the color changed to intense sky blue, while in a negative reaction, the color remained violet. The developed LAMP assay was highly sensitive, with a detection limit of 100 femtograms. The high specificity of the assay for the detection of
B. oryzae
was validated. Thus, the LAMP assay protocol provides a rapid, reliable tool for the early detection of
B. oryzae
under laboratory conditions.</abstract><cop>Dordrecht</cop><pub>Springer Netherlands</pub><doi>10.1007/s12600-022-00979-3</doi><tpages>11</tpages></addata></record> |
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| ispartof | Phytoparasitica, 2022-07, Vol.50 (3), p.543-553 |
| issn | 0334-2123 1876-7184 |
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| subjects | Agricultural production Agriculture Assaying Biomedical and Life Sciences Bipolaris oryzae Brown spot Color Cultivation Disease detection Ecology Gene amplification Glycosidases Glycoside hydrolase Grain cultivation Hydrolase Life Sciences Original Article Pathogens Plant diseases Plant Pathology Plant Sciences Proteins Rice |
| title | Loop-mediated isothermal amplification assay: A specific and sensitive tool for the detection of Bipolaris oryzae causing brown spot disease in rice |
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