Plum pox virus detection in dormant plum trees by PCR and ELISA

Adams, Alexandra; Guise,; Crossley,

doi:10.1046/j.1365-3059.1999.00336.x

Cited by 15 publications

(11 citation statements)

References 12 publications

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“…Also, reliable and sensitive detection assays are required for effective plant sanitation. Although ELISA is routinely used to detect plant viruses (Adams et al 1999;Valero et al 2003), it is not sensitive enough to detect plant viruses at very low concentrations in plant tissues. To overcome this problem, RT-PCR has been used for virus detection (Manganaris et al 2003;Sharma et al 2007).…”

Section: Discussionmentioning

confidence: 99%

Virus elimination from in vitro apple by thermotherapy combined with chemotherapy

Dong

Zhang

et al. 2015

Plant Cell Tiss Organ Cult

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We evaluated the effectiveness of thermotherapy at different temperatures, chemotherapy with different concentrations of ribavirin, and combinations of these two methods on virus elimination from apple. We used Malus cv. 'Xinhongjiangjun', an apple cultivar widely grown in China, infected with Apple chlorotic leaf spot virus (ACLSV), Apple stem grooving virus (ASGV), and Apple stem pitting virus (ASPV). The survival and regeneration of plants were evaluated, and the efficiency of virus eradication was determined by reverse-transcription polymerase chain reaction with two primer pairs for each virus. All of the plants treated with 15 and 25 lg/ml ribavirin survived, although their proliferation was slightly inhibited. Ribavirin treatments at 15 and 25 lg/ml resulted in virus elimination rates of 74.4 and 75.0 %, respectively. Higher temperatures significantly affected the growth and proliferation of plants, and almost no axillary shoots regenerated under the highest temperature (38 ± 0.5°C). The average virus elimination rate after 34-36°C treatments for 20 days was no more than 45 %. The virus elimination efficiency was enhanced by combining thermotherapy and chemotherapy treatments. A treatment combining ribavirin (25 lg/ml) and thermotherapy at 36°C (R25 ? T36) resulted in high virus eradication efficiency (95.0 %). Across all of the treatments, the average survival rate of apical shoots (176/273) was 12 % lower than that of axillary shoots (266/349); the average virus elimination efficiency was 65.3 % for apical shoots and 72.7 % for axillary shoots. The results also demonstrated that it was easier to eliminate ASPV than to eliminate ACLSV and ASGV.

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Section: Discussionmentioning

confidence: 99%

Virus elimination from in vitro apple by thermotherapy combined with chemotherapy

Dong

Zhang

et al. 2015

Plant Cell Tiss Organ Cult

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“…DAS-ELISA is routinely used for the detection of fruit-tree viruses such as PPV (Adams 1978;Adams et al 1999) and PNRSV (Scott et al 1989;Pusey and Yadava 1991). Production of virus-free propagation material by meristem-tip culture requires the use of highly specific, sensitive and rapid detection methods to screen the resulting explants.…”

Section: Virus Detectionmentioning

confidence: 99%

“…However, ELISA lacks the sensitivity required for the detection of these viruses, which occur in low concentration in these tissues. In an attempt to overcome this problem RT-PCR using immunocaptured viral particles (IC-RT-PCR) or total RNA as input and nested RT-PCR have been used for detection of PPV (Wetzel et al 1991;Candresse et al 1995;Adams et al 1999) and PNRSV (Spiegel et al 1996;Rosner et al 1998;Helguera et al 2001). Furthermore, methods based on RNA hybridization using radioactive (Varveri et al 1988;Crosslin et al 1992) and non-radioactive (Heuss-La Rosa et al 1995;Heuss et al 1999) cRNA probes have been employed for their detection.…”

Section: Virus Detectionmentioning

confidence: 99%

Elimination of PPV and PNRSV through thermotherapy and meristem-tip culture in nectarine

et al. 2003

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The plum pox virus (PPV) and prunus necrotic ringspot virus (PNRSV) cause serious disease problems in stone-fruit trees. In this work, the possibility of obtaining plant material free from these viruses through thermotherapy and meristem-tip culture from infected nectarine shoots (Prunus persica var. nectarina Max, cv. 'Arm King') was studied. In addition, the detection of these viruses in in vitro cultures and young acclimatized plantlets with double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) and multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) was studied. Meristem-tip explants (0.8-1.3 mm) derived from sprouted buds of winter wood and spring shoots from field grown plants had a 2-5% regeneration response. However, application of thermotherapy to potted nectarine trees (3 weeks at a maximum temperature of 35 degrees C) facilitated excision of longer meristem tips (1.3-2.0 mm) that resulted in a significantly higher regeneration response (38%) in woody plant medium (WPM) without plant growth regulators. Such explants formed multiple shoots with the addition of 8 microM benzylaminopurine and 0.8 microM indoleacetic acid. When they were tested for the presence of PPV and PNRSV, 86% and 81% were found to be virus-free as detected by DAS-ELISA and multiplex RT-PCR, respectively. Individual shoots excised from virus-free cultures readily rooted in vitro (half-strength WPM plus 2 microM indolebutyric acid) and grew to plantlets. The combination of an efficient protocol for virus elimination and the establishment of highly sensitive diagnostics resulted in the production of nectarine plants free from PPV and PNRSV.

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“…The latter were also ELISA-negative whereas the parent plant was ELISA-positive. Enzymelinked immunosorbent assay is routinely used for the detection of plant viruses (Adams et al 1999;Robert et al 1998;Valero et al 2003). It was demonstrated that ELISA lacks the sensitivity required for detection of the viruses, which occur in low concentrations in plant tissues.…”

Section: Resultsmentioning

confidence: 99%

In vitro production of Indian citrus ringspot virus (ICRSV) free kinnow plants employing phytotherapy coupled with shoot tip grafting

Sharma

Rani

Zaidi

et al. 2007

In Vitro Cell.Dev.Biol.-Plant

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This paper reports the elimination of Indian citrus ringspot virus (ICRSV) from "kinnow" (Citrus nobilis Lour × Citrus deliciosa Tenora) employing phytotherapy coupled with shoot tip grafting under in vitro conditions. Nodal segments from infected mother plant (indexed by indirect enzyme-linked immunosorbent assay [ELISA] and reverse transcriptase PCR [RT-PCR]) were cultured on Murashige and Skoog medium containing 2iP (1 mg/l or 4.9 μM) and malt extract (800 mg/l) along with different concentrations of aqueous extracts from leaves of Azadirachta indica (Neem), Sorghum vulgare (Jowar), and roots of Boerhaavia diffusa (Punarnava). Shoot tips were excised from the nodal sprouts and grafted on to rough lemon (Citrus jambhiri) under aseptic conditions. Maximum effect (50% virus elimination) was seen for aqueous leaf extracts of A. indica followed by B. diffusa root extract (42.86%) and S. vulgare leaf extract (31.58%). Plants/plantlets were considered virus-free only when showing negative reactions by both indirect ELISA and RT-PCR.

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Plum pox virus detection in dormant plum trees by PCR and ELISA

Cited by 15 publications

References 12 publications

Virus elimination from in vitro apple by thermotherapy combined with chemotherapy

Virus elimination from in vitro apple by thermotherapy combined with chemotherapy

Elimination of PPV and PNRSV through thermotherapy and meristem-tip culture in nectarine

In vitro production of Indian citrus ringspot virus (ICRSV) free kinnow plants employing phytotherapy coupled with shoot tip grafting

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