Detection of three pear viruses by multiplex RT-PCR assays with co-amplification of an internal control

Bing-gang, MA; Niu, Jianxin; Morley-Bunker, M.; Pan, Lizhong; Zhang, Shaoling; Zhang, Luxia

doi:10.1071/ap07081

Cited by 14 publications

(8 citation statements)

References 19 publications

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“…ASPV has also been isolated from commercial used Pyrus species such as the Japanese pear (P. serotina-pyrifolia) [6,7] and P. sinkiangensis cv. Korla [8]. Recent evidence of ASPV detection in cherry and in sour cherry trees in India [9] and in grapevine (Goszczynski, Genebank EU753978, unpublished data) indicate that the virus host range may be wider than previously considered.…”

mentioning

confidence: 95%

Cydonia japonica, Pyrus calleryana and P. amygdaliformis: three new ornamental or wild hosts of Apple stem pitting virus

Mathioudakis¹,

Candresse²,

Barone³

et al. 2011

Virus Genes

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Japanese quince, ornamental and wild pear symptomless samples were infected with Apple stem pitting virus (ASPV). Identification of ASPV was achieved by different PCR assays that amplified either the RNA polymerase or coat protein gene regions. For further confirmation, 312 bp amplicons within the polymerase gene were sequenced and compared with previously published ASPV sequences and additional sequences of isolates from ancient Italian cultivars. Comparison of the partial sequences isolated from wild/ornamental hosts and from cultivated species revealed significant divergence levels. Among the wild/ornamental isolates, the PCT88 isolate from Pyrus calleryana was the most divergent, having an amino acid deletion and incorporating a unique stretch of amino acids not present in any other isolate. Further to this preliminary partial sequence data, statistical analysis demonstrated that the isolates from wild or ornamental hosts were not more closely related to each other than to isolates from cultivated hosts. These results represent the first report of natural ASPV infection in these novel ornamental and wild Rosaceae hosts.

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mentioning

confidence: 95%

Cydonia japonica, Pyrus calleryana and P. amygdaliformis: three new ornamental or wild hosts of Apple stem pitting virus

Mathioudakis¹,

Candresse²,

Barone³

et al. 2011

Virus Genes

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“…Menzel et al (2003) standardized a protocol to detect apple viruses ACLSV with ASGV and ASPV with ApMV along with internal control. Similarly Ma et al (2008) was able to detect ACLSV with ASGV and ASPV with ASGV in pear. Baswaraj et al (2013) standardize a duplex RT-PCR assay for simultaneous detection of two viruses infecting potato, i.e., Potyvirus (Potato virus Y) and Carlavirus (Potato virus S).…”

Section: Discussionmentioning

confidence: 89%

“…False negative results can be a problem in conventional RT-PCR amplifications because inhibitor in the plant extract can interfere with the amplification process. The use of internal PCR controls targeting the plant genome is one of the methods available to avoid the false negative amplification products in RT-PCR assays and ensure the reliability of diagnosis (Ma et al, 2008). Menzel et al (2002) designed a pair of primers for amplification the nad5 gene, the same was used in the study.…”

Section: Discussionmentioning

confidence: 99%

Simultaneous detection of Apple Chlorotic Leaf Spot Virus and Apple mosaic virus in crab apples and apple rootstocks by duplex RT-PCR

Watpade

Raigond

Pramanick

et al. 2013

Scientia Horticulturae

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“…The precipitated DNA was washed by adding 1 ml of 70% (v/v) ethanol, centrifuged at 14,000×g for 5 min. The pellet was then dissolve in 30 μl of NFW and digested with 1 μl of 10 mg/ml RNAse for 15 min at 37°C (Cuellar et al 2006;Ma et al 2008). Quality of nucleic acid extractions was confirmed under UV light by 0.8% (w/v) TBE agarose gel electrophoresis and staining with 0.5 μg/ml ethidium bromide.…”

Section: Plant Nucleic Acid Isolationmentioning

confidence: 99%

Salicylic-Acid-Induced Self-excision of the Marker Gene nptII from Transgenic Tomato Using the Cre–loxP System

Bing-gang

Duan

et al. 2008

Plant Mol Biol Rep

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The presence of antibiotic-resistant genes in genetically engineered crops together with the target gene has generated a number of environmental and consumer concerns. In order to alleviate public concerns over the safety of food derived from transgenic crops, marker gene elimination is desirable. Marker-free transgenic tomato plants were obtained by using a salicylic-acid-regulated Cre-loxP-mediated site-specific DNA recombination system in which the selectable marker neomycin phosphotransferase nptII and cre genes were flanked by two directly oriented loxP sites. Upon induction by salicylic acid, the cre gene produced a recombinase that eliminated sequences encoding nptII and cre genes, sandwiched by two loxP sites from the tomato genome. Regenerant plants with the Cre-loxP system were obtained by selection on kanamycin media and polymerase chain reaction (PCR) screening. Transgenic plants were screened for excision by PCR using nptII, cre, and PR-1a promoter primers following treatment with salicylic acid. The footprint of the excision was determined by sequencing the T-DNA borders after a perfect recombination event. The excision efficiency was 38.7%. A new plant transformation vector, pBLNSC (Genbank accession number EU327497), was developed, containing six cloning sites and the self-excision system. This provided an effective approach to eliminate the selectable marker gene from transgenic tomato, thus expediting public acceptance of genetically modified tomato.Keywords Cre-loxP recombination system . Marker-free . Self-excision . Solanum lycopersicum . Transgenic tomato Abbreviations 35S cauliflower mosaic virus 35S Cef cefotaxime Plant Mol Biol Rep (

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Detection of three pear viruses by multiplex RT-PCR assays with co-amplification of an internal control

Cited by 14 publications

References 19 publications

Cydonia japonica, Pyrus calleryana and P. amygdaliformis: three new ornamental or wild hosts of Apple stem pitting virus

Cydonia japonica, Pyrus calleryana and P. amygdaliformis: three new ornamental or wild hosts of Apple stem pitting virus

Simultaneous detection of Apple Chlorotic Leaf Spot Virus and Apple mosaic virus in crab apples and apple rootstocks by duplex RT-PCR

Salicylic-Acid-Induced Self-excision of the Marker Gene nptII from Transgenic Tomato Using the Cre–loxP System

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