Rapid detection of Piper yellow mottle virus and Cucumber mosaic virus infecting black pepper (Piper nigrum) by loop-mediated isothermal amplification (LAMP)

Bhat, Alangar Ishwara; Siljo, A.; Deeshma, K.P.

doi:10.1016/j.jviromet.2013.06.012

Cited by 60 publications

(33 citation statements)

References 18 publications

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“…PCR has been used for the rapid, sensitive, and reliable detection of badnaviruses infecting different crops and vectors [ 58 , 63 , 64 , 65 , 66 , 67 ]. Real-time PCR and loop-mediated isothermal based assays have been reported for the detection of a few badnaviruses [ 68 , 69 , 70 ].…”

Section: Diagnosis and Cure Of Badnavirusesmentioning

confidence: 99%

“…PYMoV-free cuttings are important for planting of black pepper, a perennial, vegetatively propagated crop. Due to the symptomless nature of certain infected plants, sensitive assays such as PCR, real-time PCR, and loop-mediated isothermal amplification (LAMP) were developed for identification of virus-free mother plants for propagation [ 69 , 70 , 157 ].…”

Section: Virus Characterizationmentioning

confidence: 99%

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Badnaviruses: The Current Global Scenario

Bhat

Höhn²,

Selvarajan

2016

Viruses

149

105

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Badnaviruses (Family: Caulimoviridae; Genus: Badnavirus) are non-enveloped bacilliform DNA viruses with a monopartite genome containing about 7.2 to 9.2 kb of dsDNA with three to seven open reading frames. They are transmitted by mealybugs and a few species by aphids in a semi-persistent manner. They are one of the most important plant virus groups and have emerged as serious pathogens affecting the cultivation of several horticultural crops in the tropics, especially banana, black pepper, cocoa, citrus, sugarcane, taro, and yam. Some badnaviruses are also known as endogenous viruses integrated into their host genomes and a few such endogenous viruses can be awakened, e.g., through abiotic stress, giving rise to infective episomal forms. The presence of endogenous badnaviruses poses a new challenge for the fool-proof diagnosis, taxonomy, and management of the diseases. The present review aims to highlight emerging disease problems, virus characteristics, transmission, and diagnosis of badnaviruses.

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Section: Diagnosis and Cure Of Badnavirusesmentioning

confidence: 99%

Section: Virus Characterizationmentioning

confidence: 99%

Badnaviruses: The Current Global Scenario

Bhat

Höhn²,

Selvarajan

2016

Viruses

149

105

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“…To diagnose several important DNA and RNA plant viruses, LAMP and reverse transcription LAMP (RT-LAMP) have been used. These viruses include Squash leaf curl virus (SLCV) [35], Tomato chlorosis virus (ToCV) [36], Cucurbit chlorotic yellows virus (CCYV) [37,38], Tomato yellow leaf curl virus (TYLCV) [39], Tomato spotted wilt virus (TSWV) [40], Cucumber mosaic virus (CMV) [41], Potato virus Y (PYV) [42], Potato leafroll virus (PRLV) [43], Cucumber green mottle mosaic virus (CGMMV) [44], Chinese wheat mosaic virus (WCHMV0) [45], Barley yellow dwarf viruses (BYDV) [46], Japanese yam mosaic virus (JYMV) [47], Plum pox virus (PPV) [48], Tobacco mosaic virus (TMV) [49], nine rice-infecting viruses [50], and three begomoviruses infecting tomato in Panama, i.e., Potato yellow mosaic Panama virus (PYMPV), Tomato leaf curl Sinaloa virus (STLCV) and Tomato yellow mottle virus (TYMV) [51]. However, no LAMP technique for the identification of CuLCrV has been previously reported.…”

Section: Introductionmentioning

confidence: 99%

Development of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cucurbit Leaf Crumple Virus

Waliullah

Ling

Cieniewicz

et al. 2020

IJMS

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A loop-mediated isothermal amplification (LAMP) assay was developed for simple, rapid and efficient detection of Cucurbit leaf crumple virus (CuLCrV), one of the most important begomoviruses that infects cucurbits worldwide. A set of six specific primers targeting a total 240 nt sequence regions in the DNA A of CuLCrV were designed and synthesized for detection of CuLCrV from infected leaf tissues using real-time LAMP amplification with the Genie® III system, which was further confirmed by gel electrophoresis and SYBR™ Green I DNA staining for visual observation. The optimum reaction temperature and time were determined, and no cross-reactivity was seen with other begomoviruses. The LAMP assay could amplify CuLCrV from a mixed virus assay. The sensitivity assay demonstrated that the LAMP reaction was more sensitive than conventional PCR, but less sensitive than qPCR. However, it was simpler and faster than the other assays evaluated. The LAMP assay also amplified CuLCrV-infected symptomatic and asymptomatic samples more efficiently than PCR. Successful LAMP amplification was observed in mixed virus-infected field samples. This simple, rapid, and sensitive method has the capacity to detect CuLCrV in samples collected in the field and is therefore suitable for early detection of the disease to reduce the risk of epidemics.

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“…Oleh karena itu, penggunaan benih sehat dalam budidaya tanaman harus dilakukan, terutama benih bebas dari CMV dan PYMoV (Deeshma dan Bhat 2014). Salah satu cara untuk memastikan apakah benih lada yang akan ditanam bebas dari CMV atau PYMoV, maka dapat digunakan cara deteksi secara serologi (Bhat dan Siljo 2014;Bhat et al 2013). Teknik EnzymeLinked Immunosorbent Assay (ELISA) merupakan salah satu teknik deteksi yang aplikatif dan umum digunakan untuk virus tanaman.…”

Section: Pendahuluanunclassified

DETEKSI CMV DAN PYMoV PADA BENIH LADA (Piper nigrum) DENGAN TEKNIK ELISA

Mariana

Miftakhurohmah

2017

bul. littro

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ABSTRAKSalah satu kendala penting pada budidaya tanaman lada yaitu penyakit kerdil yang disebabkan oleh Piper yellow mottle virus (PYMoV) dan Cucumber mosaic virus (CMV). Tindakan pencegahan penyebaran penyakit dapat dilakukan salah satunya dengan penggunaan benih sehat bebas dari infeksi PYMoV dan CMV. Tujuan penelitian adalah mendeteksi keberadaan PYMoV dan CMV pada benih lada siap tanam secara serologi menggunakan teknik ELISA. Benih lada yang dideteksi berumur 5 bulan, berasal dari penangkar benih di Sukabumi (Jawa Barat) dan Purbalingga (Jawa Tengah). Varietas lada yang diambil di Sukabumi adalah Natar 1, Petaling, dan Lampung Daun Kecil., masingmasing sebanyak 10 sampel, sedangkan di Purbalingga hanya ada Natar 1, diambil 30 sampel. Di setiap lokasi pembibitan juga dilakukan pengamatan terhadap gejala infeksi virus yang ditemukan. Deteksi virus dilakukan secara Double Antibody Sandwich (DAS)-ELISA menggunakan antiserum Banana streak virus (BSV) untuk PYMoV dan antiserum CMV untuk deteksi CMV. Hasil ELISA dibaca nilai absorbannya dengan elisa reader pada panjang gelombang 405 nm. Sampel dinilai positif, jika nilai absorbansinya 1,5 kali lebih besar daripada kontrol negatif. Gejala infeksi virus pada benih bervariasi, yaitu klorotik, belang, dan belang disertai perubahan bentuk daun. Hasil pengujian ELISA menunjukkan bahwa 66% benih lada dari Sukabumi dan 46% dari Purbalingga terinfeksi virus CMV. Namun, tidak ada satu pun yang menunjukkan reaksi positif terhadap antiserum PYMoV. Hal ini diduga karena konsentrasi PYMoV terlalu rendah sehingga tidak terdeteksi secara ELISA. Untuk mencegah penyebaran virus, deteksi virus pada benih lada penting dilakukan sebelum digunakan sebagai bahan tanaman. Kata kunci: Piper nigrum, ELISA, virus ABSTRACT One important problem on the cultivation of black pepper plants is dwarf disease caused by Piper yellow mottle virus (PYMoV) and Cucumber mosaic virus (CMV). The purpose of this research was to detect the presence of CMV and

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Rapid detection of Piper yellow mottle virus and Cucumber mosaic virus infecting black pepper (Piper nigrum) by loop-mediated isothermal amplification (LAMP)

Cited by 60 publications

References 18 publications

Badnaviruses: The Current Global Scenario

Badnaviruses: The Current Global Scenario

Development of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cucurbit Leaf Crumple Virus

DETEKSI CMV DAN PYMoV PADA BENIH LADA (Piper nigrum) DENGAN TEKNIK ELISA

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