These methods are valuable for identifying latent infections at most stages of infection. Moreover, PCR-based methods are expensive in terms of time and equipment and hence impractical for large-scale use.
These products can be visually detected using agarose gel electrophoresis or through color changes visualized by adding SYBR Green I, ethidium bromide or calcein to the reaction mixture Notomi et al. Both methods have been successfully used in the detection of many DNA- Bhat et al. Within the cultivation areas of Shichuan and Donggang towns, Lanzhou City, China, we collected naturally infected lettuce leaf samples that displayed typical viral symptoms i.
From field experience gained in New Zealand and from employing known tested positives of viruses from New Zealand used here as references Fletcher et al. Other virus reference sources for our experiments, each isolated from lily Lilium oriental cv. PCR was carried out in Sequencing was performed at the Sangon Biotech Shanghai Co.
Shanghai, China. The LAMP reaction was conducted in Betaine has typically been used as an additive for isothermal nucleic acid amplification reactions because it lowers the melting temperature Tm of DNA; however, there is some doubt as to its efficacy for all LAMP reaction systems Ma et al.
Primer pair sequences are provided in Table 1. Water and healthy lettuce leaves were used as the blank and negative controls, respectively. To validate the RT-LAMP assays, 16 leaf samples were collected from lettuces at different growth stages including seedling, rosette or lotus, and heading.
After a min incubation period, reaction products were analyzed by electrophoresis. Typical ladder-like patterns were clearly observed at 60, 80, and min; however, no amplification was observed at 20 and 40 min for either virus Fig. Consequently, the minimum reaction time was determined as 60 min for both viruses. A Temperature. B Time duration.
Lane M: DL marker; lane 1: negative control; lanes 2—6: 20, 40, 60, 80, and min. C MgSO 4. Lane M: DL marker; lane 1: water control; lanes 2—9: 0, 2, 4, 6, 8, 10, 12, and 14 mM. D Betaine. Lane M: DL marker; lane 1: water control; lanes 2—8: 0, 0.
Typical ladder-like patterns were clearly observed by agarose gel electrophoresis without betaine. Ladder-like patterns were observed at 0. The detection limits for LNYV were determined to be 3. The detection limits for CMV were determined to be 2. Both methods yielded similar detection limit results for both viruses Fig. A LNYV. B CMV. Lane M: DL marker; lane 1: negative control; lanes 2— lettuce leaf samples. Although different molecular methods have been developed for LNYV and CMV detection, many require technical expertise and specialized equipment Callaghan and Dietzgen, ; Deyong et al.
Because the field lettuce samples were collected at different growth stages including early seedling, rosette or lotus and heading there may well have been differences in virus concentration. Further survey work needs to be undertaken to verify this conclusion. The negative result in our assays of specimens 3, 6, 14, 16, 17 might be explained either by the presence of other viruses not tested in these experiments e. Plant Sci. Fukuta, S. Development of immunocapture reverse transcription loop-mediated isothermal amplification for the detection of Tomato spotted wilt virus from chrysanthemum.
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Reprints and Permissions. Yang, QQ. A reverse transcription-cross-priming amplification method with lateral flow dipstick assay for the rapid detection of Bean pod mottle virus.
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Advanced search. Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily. Skip to main content Thank you for visiting nature. Download PDF. Subjects Isolation, separation and purification Pathogens. Abstract Bean pod mottle virus BPMV is a destructive virus that causes serious economic losses in many countries every year, highlighting the importance of its effective detection.
Full size table. Figure 1. Full size image. Figure 2. Figure 3. Discussion The isothermal amplification technology CPA has been shown to be effective in the detection of human, animal, and plant pathogens 25 , 29 , References 1. Google Scholar 2. PubMed Google Scholar 3. Google Scholar 4. Google Scholar 5. Google Scholar 9. Google Scholar CAS Google Scholar PubMed Google Scholar PubMed Central Google Scholar View author publications.
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MEGA7: molecular evolutionary genetics analysis version 7. The expression of symptoms in anthurium occurred between days after inoculation DAI , whereas in other experimental hosts symptoms could be observed between 3 and 5 DAI. Among the inoculated genotypes only 'Eidibel' and 'Garoa' showed noticeable local symptoms, i. According to the results obtained in this test Figure 3 , the symptomatic anthuriums collected in the four cities were infected by CMV. Raj et al. The symptoms induced by CMV in anthurium leaves differ from those induced by Tomato spotted wilt virus , which are characterized by brown necrotic leaf spots surrounded by intense yellow Uchida et al.
On the other hand, DsMV induces mosaic on leaves, color alteration and deformation in spathes Lima et al. In spathes presenting vein necrosis, only CMV could be detected data not shown.
The symptoms observed in naturally DsMV-infected plants Rivas et al. Drastic foliar deformation with intense mosaic was frequently observed in different varieties and cultivars. It is possible to avoid or reduce CMV dissemination in anthurium commercial crops through elimination of weeds, mainly Commelina and Tradescantia , those are natural hosts of the virus; removal of symptomatic anthuriums with the destruction of these plants away from the cultivation area; and use of healthy propagative material.
The control of aphid population not only in the production area but also around is important to reduce viral inoculum in medium and long term. Once the CMV is present in anthurium commercial crops, the use of resistant genotypes or that reacted with less drastic symptoms should be considered for replacement of anthuriums already cultivated.
The cultivation of different botanical species in the same area, such as observed in Mogi das Cruzes may have contributed significantly for the sporadic occurrence of CMV-infected anthuriums, since literature data show that the cultivation with a single plant species or species of the same botanical family in large areas may allow an increase in the insect vector population and consequently in viral inoculums; these factors can make impractical the anthurium cultivation in a medium or long term.
The authors wish also to thank Dr. Jesus G. Abrir menu Brasil. Horticultura Brasileira. Abrir menu. Anthurium andraeanum; Araceae; Cucumovirus; diagnosis. Anthurium andraeanum; Araceae; Cucumovirus; diagnose. Keywords: Anthurium andraeanum , Araceae, Cucumovirus , diagnosis. Palavras-chave: Anthurium andraeanum , Araceae, Cucumovirus , diagnose.
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