Pathotype identification of Leptosphaeria maculans with PCR and oligonucleotide primers from ribosomal internal transcribed spacer sequences

Xue, Bingye; Goodwin, Paul H.; Annis, Seanna L.

doi:10.1016/0885-5765(92)90009-k

Cited by 50 publications

(15 citation statements)

References 15 publications

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“…Considerably greater sequence variation is found in the internal transcribed spacer (ITS) regions between the rRNA genes within a rRNA repeat unit (rDNA; 70,102,106,164,166). Even more sequence differences are in the nontranscribed spacer (NTS) regions between the rDNA repeat units and still more in the intergenic spacer (IGS or IGR) regions, or noncoding sequences that occur within the rDNA repeat unit of some fungi (75).…”

Section: < Zmentioning

confidence: 99%

The Polymerase Chain Reaction and Plant Disease Diagnosis

Henson¹

1993

Annual Review of Phytopathology

122

127

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Section: < Zmentioning

confidence: 99%

The Polymerase Chain Reaction and Plant Disease Diagnosis

Henson¹

1993

Annual Review of Phytopathology

122

127

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“…Nazar et al (102), for example, found adequate sequence differences in the ITS regions of the wilt fungi, Verticillium dahliae and V. albo-atrum, to design primers that specifically amplify the DNA of each species (Figure 2). Primers based on differences in ITS 1 sequences Leptosphaeria maculans allow specific amplification of either weakly or highly virulent isolates of this fungal pathogen, the latter being detectable in infected canola leaves even before symptom development (164).…”

Section: < Zmentioning

confidence: 99%

The Polymerase Chain Reaction and Plant Disease Diagnosis

Henson¹,

French²

1993

Annu. Rev. Phytopathol.

374

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“…These include those present in many of the house keeping genes viz., ITS regions (Iacomi-Vasilescu et al 2002;Pavon et al 2011Pavon et al , 2012, β-tubulin (Mostert et al 2006;Aroca et al 2008), translation elongation factor 1 alpha, TEF1 (Yadav et al 2011), calmodulin (Mule et al 2006), avirulence genes (Lievens et al 2009), mitochondrial genes such as the multicopy cox Iand cox II (Martin and Tooley 2003;Tooley et al 2006;Seifert et al 2007;Nguyen and Seifert 2008), etc. The ITS regions of ribosomal RNA often contains pathogen specific conserved sequences that enable designing of genera and species consensus primers (White et al 1990;Xue et al 1992). Also the high copy numbers of rRNA genes in a fungal genome serve as an (Jasalavich et al 1995;Kusaba and Tsuge 1995;Pavlina et al 2002;Udayashankar et al 2012).…”

Section: Discussionmentioning

confidence: 98%

Rapid, specific and sensitive molecular detection assay for Alternaria helianthi that causes leaf blight disease in sunflower

Chavhan¹,

Hinge²,

Chinchole³

et al. 2015

Eur J Plant Pathol

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The leaf blight disease of sunflower (Helianthus annuus) caused by Alternaria helianthi is a serious threat to its cultivation worldwide. Early and accurate detection of the pathogen is critical to efficient disease management and in avoiding further losses due to epidemics in sunflower. Conventional methods of detection and identification are time consuming, labour intensive, and lack specificity and sensitivity. A realtime PCR based TaqMan® probe assay was developed to target 156 bp Internal Transcribed Spacer (ITS) region of A. helianthi for its detection using fungal genomic DNA and infected plant tissues and seeds of sunflower. The specificity of the probe and primers was confirmed by testing their cross reactivity using genomic DNA of closely related Alternaria species isolated from 17 crop plants and 15 fungal species of other genera. No cross-reactivity could be detected with any of the other non-target fungal strains used in this study. The assay successfully detected as low as 1.0 pg fungal genomic DNA and up to 1 % infection in sunflower seed lots. To the best of our knowledge, this is the only probe based real-time PCR assay that enable high specificity and sensitivity for rapid detection of A. helianthi in infected seeds and plant tissues. The assay may also hold promise for application in effective bio-threat and risk mitigation program by early and accurate detection of the pathogen for effective management.

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Pathotype identification of Leptosphaeria maculans with PCR and oligonucleotide primers from ribosomal internal transcribed spacer sequences

Cited by 50 publications

References 15 publications

The Polymerase Chain Reaction and Plant Disease Diagnosis

The Polymerase Chain Reaction and Plant Disease Diagnosis

The Polymerase Chain Reaction and Plant Disease Diagnosis

Rapid, specific and sensitive molecular detection assay for Alternaria helianthi that causes leaf blight disease in sunflower

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