Molecular detection of nine rice viruses by a reverse-transcription loop-mediated isothermal amplification assay

Le, Dung Tien; Netsu, Osamu; Uehara-Ichiki, Tamaki; Shimizu, Takumi; Choi, Il-Ryong; Omura, Toshihiro; Sasaya, Takahide

doi:10.1016/j.jviromet.2010.09.004

Cited by 64 publications

(33 citation statements)

References 14 publications

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“…This means that LAMP reaction with the loop primers provides greater sensitivity than LAMP reaction without the loop primers and is in agreement with a previous study carried out by Nagamine et al (2002). By adding a fluorescent agent during the RT-LAMP reaction, the RT-LAMP product could be detected visually by observing fluorescence emitted by the reaction solution under UV irradiation (Le et al, 2010;Tomita et al, 2008). In order to visualize the product of RT-LAMP in this study, 1 µl of fluorescent detection reagent (Eiken Chemicals Co., Tokyo, Japan) was added into the RT-LAMP reaction tubes.…”

supporting

confidence: 90%

“…LAMP does not need the extra step of visualization by gel electrophoresis (Goto et al, 2009). Some plant viruses have been detected by LAMP, such as Tomato spotted wilt virus (Fukuta et al, 2004), Plum pox virus (Varga and James, 2006) , Peach latent mosaic viroid (Boubourakas et al, 2009), Japanese yam mosaic virus (Fukuta et al, 2003), Rice black-streaked dwarf virus, Rice dwarf virus, Rice gall dwarf virus, Rice ragged stunt virus, Rice transitory yellowing virus, Rice stripe virus, Rice grassy stunt virus, Rice tungro spherical virus, Rice tungro bacilliform virus (Le et al, 2010), and Potato virus Y (Nie, 2005). The objective of this study was to evaluate feasibility of RT-LAMP method for detection of PLRV.…”

mentioning

confidence: 99%

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Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Ju¹

2011

The Plant Pathology Journal

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A new reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for the Potato leafroll virus (PLRV) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to address its advantages over RT-PCR. RT-LAMP primers were designed from the open reading frame 3 (ORF3) sequence of PLRV. The RT-LAMP reactions were conducted without or with a set of loop primers. By real-time monitoring using Turbimeter, the RT-LAMP (with loop primers) detects PLRV in less than 30 min, compared to 120 min of RT-PCR. By adding fluorescent reagent during the reaction, final products of the RT-LAMP were fluorescently visualized under UV light or could be differentiated by naked-eye inspection under normal light. The RT-LAMP was extremely sensitive, about 2000-fold more sensitive than RT-PCR. This study presents great potential of the RT-LAMP for diagnosis and PLRV epidemiology because RT-LAMP method is speedy, sensitive, inexpensive, and convenient.

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supporting

confidence: 90%

mentioning

confidence: 99%

Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Ju¹

2011

The Plant Pathology Journal

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“…Serangan virus penyebab kerdil rumput akan menyebabkan gejala yang sangat khas pada tanaman padi yang terinfeksi sehingga untuk deteksinya akan mudah dilakukan berdasarkan pengamatan visual (Le et al 2010). Menurut Heong dan Hardy (2009), tanaman padi terinfeksi RGSV menunjukkan gejala stunting dan proliferasi pendek, tegak, dan daun sempit yang berwarna hijau pucat atau kuning pucat dalam warna dan daun yang terinfeksi dapat menunjukkan gejala mottling pada daun muda dan muncul bintik-bintik berkarat pada daun tua.…”

Section: Pembahasanunclassified

Respons lima varietas padi terhadap infeksi virus penyebab penyakit kerdil rumput (Rice Grassy Stunt Virus)

Rahmawati

Sulandari

Hartono

2015

Prosiding Seminar Nasional Masyarakat Biodiversitas Indonesia

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Abstrak. Rahmawati Y, Sulandari S, Hartono S. 2015. Respons lima varietas padi terhadap infeksi virus penyebab penyakit kerdil rumput (Rice Grassy Stunt Virus). Pros Sem Nas Masy Biodiv Indon 1: 1123-1126. Rice Grassy Stunt Virus (RGSV) merupakan virus penyebab penyakit kerdil rumput padi yang merupakan anggota dari genus Tenuivirus. RGSV ditularkan secara persisten oleh wereng coklat. Serangan wereng coklat selain menyebabkan padi kering seperti bekas terbakar (hopperburn) juga berperan sebagai vektor penyakit kerdil yang disebabkan oleh virus. Saat ini di DI Yogyakarta dan Jawa Tengah dilaporkan terjadi outbreak wereng coklat dan penyakit kerdil rumput. Penyakit kerdil sangat sulit dikendalikan sehingga sangat merugikan karena dapat menyebabkan puso. Pengendalian menggunakan varietas masih terkendala karena selama ini belum ada varietas padi yang toleran terhadap RGSV. Pada umumnya semua varietas padi yang ditanam oleh petani hanya tahan terhadap vektor (wereng coklat) saja. Penelitian ini bertujuan untuk mengetahui respons beberapa varietas padi yang banyak ditanam petani terhadap infeksi virus penyebab penyakit kerdil. Parameter yang diamati meliputi masa inkubasi, variasi gejala, kejadian dan intensitas penyakit yang ditimbulkan. Penelitian ini dilaksanakan di Laboratorium Virologi Tumbuhan dan rumah kaca Jurusan Hama dan Penyakit Tumbuhan, Fakultas Pertanian, Universitas Gadjah Mada (UGM) Yogyakarta. Sumber inokulum diambil dari Desa Sumber Rahayu, Moyudan, Sleman, Yogyakarta. Wereng coklat yang digunakan sebagai vektor merupakan hasil rearing di laboratorium. Inokulasi virus dilakukan terhadap lima varietas padi meliputi: Membramo, Ciherang, Inpari 13, IR64, dan Situ Bagendit. Berdasarkan hasil penelitian diketahui setiap varietas memiliki respons yang berbeda terhadap penyakit kerdil padi. Insidensi penyakit yang ditimbulkan varietas Membramo (85,19%), Situ Bagendit dan Inpari 13 (92,59%), Ciherang (96,3%), dan IR64 (100%). Setiap varietas memiliki perbedaan masa inkubasi. Variasi gejala juga berbeda antar varieta,s antara lain berupa anakan tegak, daun kaku pucat atau menguning, serta terdapat bercak pada daun. Pada varietas Situ Bagendit tanaman tidak terlalu kerdil, tetapi pada Ciherang gejala yang ditimbulkan sangat parah, yaitu: tanaman menjadi sangat kerdil. Kata kunci: kerdil rumput, padi, RGSV, varietas, wereng coklat Abstract. Rahmawati Y, Sulandari S, Hartono S. 2015. Response of five rice varieties to viral infections by Rice Grassy Stunt Virus that causes rice grassy disease. Pros Sem Nas Masy Biodiv Indon 1: 1123-1126. Rice grassy stunt virus (RGSV), a member of the genusTenuivirus, is the virus that causes rice grassy disease in rice. RGSV is persistently transmitted by the brown planthopper (BPH). Besides attack, BPH also induces hopper burn disease by drying leaves and acts as vectors of stunt disease caused by a virus. The occurrence of an outbreak of BPH and grassy stunt disease has been reported in Yogyakarta and Central Java. Stunt disease is very difficult to control. Moreover, it is...

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“…Pns6 is required for cell-to-cell movement of RDV (Li et al, 2004). Some assays have been developed for the detection of RDV in rice or its insect vector: RT-PCR (Le et al, 2010), reversetranscription loop-mediated isothermal amplification assay (RT-LAMP) (Le et al, 2010), transmission electron microscopy (TEM) (Lu et al, 1998;Mizuno and Kano, 1991;Wei et al, 2007), nucleic acid hybridization (Yu et al, 1999), Western blot (Suzuki et al, 1994), polyclonal antibody (pAb)-based double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) (Takahashi et al, 1991). Among these assays, enzyme-linked immunosorbent assay (ELISA) was most suitable for highthroughput detection of RDV in rice plants and leafhopper vectors.…”

Section: Introductionmentioning

confidence: 99%

Monoclonal antibody-based serological assays and immunocapture-RT-PCR for detecting Rice dwarf virus in field rice plants and leafhopper vectors

Wu¹,

Ni²,

Liu³

et al. 2014

Journal of Virological Methods

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Molecular detection of nine rice viruses by a reverse-transcription loop-mediated isothermal amplification assay

Cited by 64 publications

References 14 publications

Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Respons lima varietas padi terhadap infeksi virus penyebab penyakit kerdil rumput (Rice Grassy Stunt Virus)

Monoclonal antibody-based serological assays and immunocapture-RT-PCR for detecting Rice dwarf virus in field rice plants and leafhopper vectors

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