Reverse transcription loop-mediated isothermal amplification (RT-LAMP) for detection of coconut cadang-cadang viroid (CCCVd) variants in oil palm

Madihah Ahmad Zairun, . and Maizatul Suriza Mohamed, . and Idris Abu Seman, . Reverse transcription loop-mediated isothermal amplification (RT-LAMP) for detection of coconut cadang-cadang viroid (CCCVd) variants in oil palm. pp. 453-463. ISSN 1511-2780

The coconut cadang-cadang viroid (CCCVd) is the causal agent of orange spotting (OS) disease in commercial oil palm (Elaeis guineensis) plantations in Malaysia. At present the methods used to detect OS are time-consuming and can be inaccurate. Here we describe the development of a rapid sensitive and specific method of detecting CCCVd variants in oil palm leaves using a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method. RT-LAMP detected CCCVd variants at a concentration of 2 ng l-1 in less than 1 hr at 63C using total ribonucleic acid (RNA) of leaf extracts; whereas a minimum of 20 ng l-1 was required to detect CCCVd using conventional polymerase chain reaction (PCR). The utility of RT-LAMP as OS detection in plantations was evaluated using symptomatic and asymptomatic oil palm leaf samples from nurseries and field sites. RT-LAMP successfully detected CCCVd in OS-infected samples. Leaves with OS-like symptoms caused by nutrient deficiencies or leaf spot disease other than OS produced a negative result. The RT-LAMP results were confirmed by conventional PCR indicating that RT-LAMP is a valuable tool sensitive and rapid method of diagnosing OS in oil palm plantations that could be performed by plantation personnel for OS disease management in oil palm plantations.

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Abstract

The coconut cadang-cadang viroid (CCCVd) is the causal agent of orange spotting (OS) disease in commercial oil palm (Elaeis guineensis) plantations in Malaysia. At present the methods used to detect OS are time-consuming and can be inaccurate. Here we describe the development of a rapid sensitive and specific method of detecting CCCVd variants in oil palm leaves using a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method. RT-LAMP detected CCCVd variants at a concentration of 2 ng l-1 in less than 1 hr at 63C using total ribonucleic acid (RNA) of leaf extracts; whereas a minimum of 20 ng l-1 was required to detect CCCVd using conventional polymerase chain reaction (PCR). The utility of RT-LAMP as OS detection in plantations was evaluated using symptomatic and asymptomatic oil palm leaf samples from nurseries and field sites. RT-LAMP successfully detected CCCVd in OS-infected samples. Leaves with OS-like symptoms caused by nutrient deficiencies or leaf spot disease other than OS produced a negative result. The RT-LAMP results were confirmed by conventional PCR indicating that RT-LAMP is a valuable tool sensitive and rapid method of diagnosing OS in oil palm plantations that could be performed by plantation personnel for OS disease management in oil palm plantations.

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Item Type:	Article
AGROVOC Term:	Oil palm
AGROVOC Term:	Elaeis guineensis
AGROVOC Term:	Diagnosis
AGROVOC Term:	Molecular biology
AGROVOC Term:	Reverse transcription
AGROVOC Term:	Viroids
AGROVOC Term:	Coconut cadang cadang
AGROVOC Term:	Spots
AGROVOC Term:	Plant diseases
Depositing User:	Mr. AFANDI ABDUL MALEK
Last Modified:	24 Apr 2025 00:54
URI:	http://webagris.upm.edu.my/id/eprint/9196

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