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Detection of the Chilli Leaf Curl Virus Using an Attenuated Total Reflection-Mediated Localized Surface-Plasmon-Resonance-Based Optical Platform.

ABSTRACT: The development of a nanoparticle-based optical platform has been presented as a biosensor for detecting target-specific plant virus DNA. The binding dynamics of gold nanoparticles has been studied on the amine-functionalized surface by the attenuated total reflection (ATR)-based evanescent wave absorption method monitoring the localized surface plasmon resonance (LSPR). The developed surface was established as a refractive index sensor by monitoring the LSPR absorption peak of gold nanoparticles. This nanoparticle-immobilized surface was explored to establish as a biosensing platform with target-specific immunoglobulin (IgG) antibody-antigen interaction. The IgG concentration-dependent variation of absorbance was correlated with the refractive index change. After successfully establishing this ATR configuration as an LSPR-based biosensor, the single-stranded DNA of the chilli leaf curl virus was detected using its complementary DNA sequence as a receptor. The limit of detection of this sensor was determined to be 1.0 μg/mL for this target viral DNA. This ATR absorption technique has enormous potential as an LSPR based nano-biosensor for the detection of other begomoviruses.

SUBMITTER: Das S

PROVIDER: S-EPMC8280655 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BACKGROUND:In Oman tobacco (Nicotiana tabacum; family Solanaceae) is a minor crop, which is produced only for local consumption. In 2015, tobacco plants exhibiting severe downward leaf curling, leaf thickening, vein swelling, yellowing and stunting were identified in fields of tobacco in Suhar Al-Batina region, Oman. These symptoms are suggestive of begomovirus (genus Begomovirus, family Geminiviridae) infection. METHODS:Circular DNA molecules were amplified from total DNA extracted from tobacco plants by rolling circle amplification (RCA). Viral genomes were cloned from RCA products by restriction digestion and betasatellites were cloned by PCR amplification from RCA product, using universal primers. The sequences of full-length clones were obtained by Sanger sequencing and primer walking. Constructs for the infectivity of virus and betasatellite were produced and introduced into plants by Agrobacterium-mediated inoculation. RESULTS:The full-length sequences of 3 begomovirus and 3 betasatellite clones, isolated from 3 plants, were obtained. Analysis of the full-length sequences determined showed the virus to be a variant of Chilli leaf curl virus (ChiLCV) and the betasatellite to be a variant of Tomato leaf curl betasatellite (ToLCB). Both the virus and the betasatellite isolated from tobacco show the greatest levels of sequence identity to isolates of ChiLCV and ToLCB identified in other hosts in Oman. Additionally clones of ChiLCV and ToLCB were shown, by Agrobacterium-mediated inoculation, to be infectious to 3 Nicotiana species, including N. tabacum. In N. benthamiana the betasatellite was shown to change the upward leaf rolling symptoms to a severe downward leaf curl, as is typical for many monopartite begomoviruses with betasatellites. CONCLUSIONS:The leaf curl disease of tobacco in Oman was shown to be caused by ChiLCV and ToLCB. This is the first identification of ChiLCV with ToLCB infecting tobacco. The study shows that, despite the low diversity of begomoviruses and betasatellites in Oman, the extant viruses/betasatellites are able to fill the niches that present themselves.

Dataset Information

Detection of the Chilli Leaf Curl Virus Using an Attenuated Total Reflection-Mediated Localized Surface-Plasmon-Resonance-Based Optical Platform.

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