Recombinase Polymerase Amplification Assay for Simultaneous Detection of Maize Chlorotic Mottle Virus and Sugarcane Mosaic Virus in Maize

Abstract

Maize chlorotic mottle virus (MCMV) can cause maize lethal necrosis (MLN) when coinfected with potyvirids, such as sugarcane mosaic virus (SCMV), maize dwarf mosaic virus, or wheat streak mosaic virus. MLN is often caused by coinfection of MCMV and SCMV, which has been reported in China and several countries of Africa. In this study, a recombinase polymerase amplification (RPA) assay was established for simultaneous detection of MCMV and SCMV in maize. The RPA assay can be completed within 30 min at 38 °C. The primers for the RPA assay were specific since no crossreaction was detected with other selected viruses that infected maize in China. The detection limit of the RPA method was 10 2 copies μL −1 , which was about 10-fold more sensitive than that of the conventional PCR method. Moreover, the RPA assay can be successfully applied to detect maize samples collected in the field. These results demonstrated that the established RPA assay is a rapid and efficient method to conduct simultaneous detection of MCMV and SCMV, which provides an alternative technology for MLN diagnosis.

ZmmiR398b negatively regulates maize resistance to sugarcane mosaic virus infection by targeting ZmCSD2/4/9

“…In the present study, we characterized the ZmmiR398b- seedlings as described previously (Gao et al, 2021). The first systemically infected leaves were harvested to determine the accumulations of ZmmiR398b, ZmCSD2, ZmCSD4, ZmCSD9, and SCMV RNA and CP.…”

“…Maize ( Zea mays ) inbred lines Chang7‐2 and Mo17 and transgenic B104 lines were grown in growth chambers (28°C/22°C, 16 h/8 h, day/night). SCMV crude extracts were prepared and inoculated on the first true leaves of 1‐week‐old maize seedlings as described previously (Gao et al., 2021 ). The first systemically infected leaves were harvested to determine the accumulations of ZmmiR398b, ZmCSD2 , ZmCSD4 , ZmCSD9 , and SCMV RNA and CP.…”

Development of recombinase polymerase amplification-based colorimetric detection assay for rapid identification of invasive cassava mealybug, Phenacoccus manihoti Matile-Ferrero

“…A thermal cycler is a must in PCR for the amplification of DNA, whereas RPA protocol can be completed by holding the reaction mixture in hands, which eliminates the requirement of a costly thermal cycler; this feature of the technique enhances its utilization in the identification of invasive pests in point of entry as well as field. The recombinase polymerase amplification can be achieved in a simple water bath which is much cheaper than a thermal cycler for higher efficacy of results ( Gao et al, 2021 ). The addition of SYBR Green I dye to the RPA amplified products makes the assay as colorimetric since SYBR Green I is an asymmetric cyanine dye known for its remarkable sensitivity.…”

Entailing the Next-Generation Sequencing and Metabolome for Sustainable Agriculture by Improving Plant Tolerance

“…Then, the advent of genomic science assisted scientists to reveal the genetic composition of crops and the disease-causing pathogens and provided the key clues to monitor plant and pathogen interaction for crop improvements by employing the NGS and metabolomics to determine the host genetic factors that the disease-causing agent engineers during attack or infection and the ability to gain defense through reprogramming the genetic makeup [ 185 ]. For example, the sugarcane mosaic virus (SCMV) is a major threat to the maize-cultivating farming community in China’s maize-cultivating farming community; it was revealed by analyzing transcriptomic data that SCMV actively participates in downregulating the photosynthesis-responsible genes, leading to the phenotype of chlorotic lesions [ 186 ].…”