Occurrence, molecular diagnosis and suitable time of detection of citrus greening disease in sweet orange

Gopal, K.; Pradeepthi, E. R.; Gopi, V.; Ahammed, S. Khayum; Sreenivasulu, Y.; Reddy, M. Krishna; Purushotham, K.

doi:10.1556/aphyt.42.2007.1.6

Cited by 8 publications

(8 citation statements)

References 8 publications

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“…Thus, a method was developed by the addition of sodium sulphite to EDTA in extraction of DNA from HLB infected tissue (Gopal et al 2004). The 16S rDNA fragments of HLB were amplified satisfactorily with consistency from the DNA extracted from bark and midrib in a PCR test (Gopal et al 2007). Further, a cool climate period (winter) from September to February was found to be quite suitable for the detection of HLB under Peninsular India conditions (Gopal et al 2007).…”

Section: Introductionmentioning

confidence: 99%

“…The 16S rDNA fragments of HLB were amplified satisfactorily with consistency from the DNA extracted from bark and midrib in a PCR test (Gopal et al 2007). Further, a cool climate period (winter) from September to February was found to be quite suitable for the detection of HLB under Peninsular India conditions (Gopal et al 2007). However, there are no reports on the sampling of particular symptomatic tissues to be taken for HLB detection as the sweet orange and acid lime show a variety of symptoms on infection of HLB which is often confused with micronutrient deficiencies.…”

Section: Introductionmentioning

confidence: 99%

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Symptom-based diagnosis of Huanglongbing (citrus greening) disease by PCR in sweet orange (Citrus sinensisOsbeck) and acid lime (Citrus aurantifoliaSwingle)

Gopal

Gopi

Kalyani

et al. 2010

Archives Of Phytopathology And Plant Protection

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Huanglongbing (HLB), previously known as citrus greening disease, is one of the most destructive diseases of citrus and responsible for the decline of citrus orchards in Andhra Pradesh (AP) and other citrus growing areas in the country. Polymerase chain reaction (PCR) amplification of 1160 bp fragment of 16S rDNA of HLB was observed in mottling symptoms, yellow vein symptoms, symptoms mixed with both citrus yellow mosaic virus and citrus greening and zinc deficiency-like symptoms; however, no amplification was observed in iron deficiency like symptoms. As the disease shows a variety of symptoms in infected trees, PCR detection could be useful for resolving confusion in identification of HLB between actual deficiency and deficiency like symptoms caused by HLB. In acid lime the midrib and veins of the leaves with both mottling and yellow vein symptoms were suitable for isolation of DNA and used as a template in the PCR test.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Symptom-based diagnosis of Huanglongbing (citrus greening) disease by PCR in sweet orange (Citrus sinensisOsbeck) and acid lime (Citrus aurantifoliaSwingle)

Gopal

Gopi

Kalyani

et al. 2010

Archives Of Phytopathology And Plant Protection

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“…Research on new methods such as the isothermal and chimeric primer-initiated amplification of nucleic acids combined with cycling probe technology [7], application of nested PCR [8], comparisons of primers for PCR and nested PCR [9], along with the improvement of DNA isolation for conventional PCR [10] has been reported. However, PCR amplification of the bacteria is very weak during spring and summer seasons [11], increasing the probability of obtaining false negatives during these periods. Therefore, methods that do not rely on the presence of the bacteria in the sample are sought as more reliable throughout the year.…”

Section: Introductionmentioning

confidence: 99%

Untargeted metabolite analysis of healthy and Huanglongbing‐infected orange leaves by CE‐DAD

2009

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Huanglongbing (HLB) is considered the most destructive bacterial citrus disease worldwide. Early detection of HLB is crucial for minimizing its spread. CE was used for the discovery of potential biomarkers for HLB. Optimization of extraction and separation allowed resolving 24 compounds of which 6 were present in significantly higher (p<0.05) concentrations in HLB-infected samples collected monthly for 6 months during the 2007-2008 season. Three of these compounds were identified by mobility and UV spectra as hesperidin, naringenin, and quercetin with mean increase in concentration of 154, 555, and 467%, respectively, above that in healthy leaves. Results support the potential of CE-DAD for untargeted plant metabolomic analysis. CZE, NACE, and MEEKC were compared for metabolic differentiation of healthy and HLB-infected citrus leaves. CZE in a semi-aqueous BGE solution consisting of 8.5 mM of sodium borate (pH 9.3), 15% ACN, and 9% 1-butanol yielded the best peak separation with detection at 190 nm.

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“…The extraction of HLB DNA was carried out by a method wherein sodium sulfi te was added in Tris-EDTA, which yielded improved HLB DNA (Gopal et al, 2007). The presence of high levels of polyphenols and tannins in citrus leaves interferes with obtaining good-quality DNA, and thus affects the reliable detection of pathogens by PCR.…”

Section: Resultsmentioning

confidence: 99%

“…of ethanol and 0.1 vol. of sodium acetate (pH 5.2) at -20 ºC for 2 h. DNA was isolated by the sodium sulfi te EDTA method used for detection of the HLB disease by PCR amplifi cation as reported by Gopal et al (2007).…”

Section: Dna Extractionmentioning

confidence: 99%

Detection of Huanglongbing (Citrus Greening) Disease by Nucleic Acid Spot Hybridization

Gopal

Sudarsan²,

Gopi

et al. 2009

Zeitschrift Für Naturforschung C

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Polymerase chain reaction (PCR) amplifi cation with primers specifi c to the rDNA region successfully amplifi ed the 1160-bp DNA fragment from a Huanglongbing (HLB)-infected sweet orange sample with mottling symptoms leaves, but not from healthy sweet orange plants. The PCR product of 1160-bp was used as probe labeled with biotin for detection of the HLB pathogen in the nucleic acid spot hybridization (NASH) test. It was found that the HLB pathogen could be detected up to 1:100 dilution in HLB-infected tissue. Total DNA extracted from HLB-infected tissue was diluted 2-fold as 900 ng in TE buffer and spotted on a nitrocellulose membrane. Strong signals were observed up to 225 ng of DNA dilution, whereas a moderate signal was recorded at 112 ng. No hybridization signal was observed in the healthy samples, while strong signals were observed in the positive control.

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Occurrence, molecular diagnosis and suitable time of detection of citrus greening disease in sweet orange

Cited by 8 publications

References 8 publications

Symptom-based diagnosis of Huanglongbing (citrus greening) disease by PCR in sweet orange (Citrus sinensisOsbeck) and acid lime (Citrus aurantifoliaSwingle)

Symptom-based diagnosis of Huanglongbing (citrus greening) disease by PCR in sweet orange (Citrus sinensisOsbeck) and acid lime (Citrus aurantifoliaSwingle)

Untargeted metabolite analysis of healthy and Huanglongbing‐infected orange leaves by CE‐DAD

Detection of Huanglongbing (Citrus Greening) Disease by Nucleic Acid Spot Hybridization

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