PCR-based detection of<i>Phomopsis azadirachtae</i>in die-back affected neem seeds

Girish, K.; Bhat, Sanjay P.; Raveesha, K. A.

doi:10.1080/03235400701287628

Cited by 4 publications

(5 citation statements)

References 21 publications

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“…) and PF1/PF2 (Girish et al. ) were validated in the present study and found to amplify DNA from other species of Phomopsis, indicating they are not specific to P. azadirachtae . Many studies have reported the occurrence of wide host range for a number of Phomopsis species (Ash et al.…”

Section: Discussionsupporting

confidence: 60%

“…) and PF1/PF2 (Girish et al. ) were validated across DNA of different species of Phomopsis . These primer sets Phf/Phr and PF/PR amplified DNA from different species of Phomopsis used in the study (Table ) and resulted in an amplicon of 141 and 154 bp (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…) and PF1/PF2 (Girish et al. ) were validated using DNA of different Phomopsis species (Table ) including P. azadirachate . The PCR conditions were same as described by Prasad et al.…”

Section: Methodsmentioning

confidence: 99%

“…) and PF1/PF2 (Girish et al. ) has been reported. In the genus Phomopsis, more than 1000 species have been described (Uecker ).…”

Section: Introductionmentioning

confidence: 95%

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Species‐Specific PCR‐Based Assay for Identification and Detection of Phomopsis (Diaporthe) azadirachtae Causing Die‐Back Disease in Azadirachta indica

Vedashree

Sateesh

Chowdappa

et al. 2015

Journal of Phytopathology

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Die-back disease caused by Phomopsis (Diaporthe) azadirachtae is the devastating disease of Azadirachta indica. Accurate identification of P. azadirachtae is always problematic due to morphological plasticity and delayed appearance of conidia. A species-specific PCR-based assay was developed for rapid and reliable identification of P. azadirachtae by designing a species-specific primer-targeting ITS region of P. azadirachtae isolates. The assay was validated with DNA isolated from different Phomopsis species and other fungal isolates. The PCR assay amplified 313-bp product from all the isolates of P. azadirachtae and not from any other Phomopsis species or any genera indicating its specificity. The assay successfully detected the pathogen DNA in naturally and artificially infected neem seeds and twigs indicating its applicability in seed quarantine and seed health testing. The sensitivity of the assay was 100 fg when genomic DNA of all isolates was analysed. The PCR-based assay was 92% effective in comparison with seed plating technique in detecting the pathogen. This is the first report on the development of species-specific PCR assay for identification and detection of P. azadirachtae. Thus, PCR-based assay developed is very specific, rapid, confirmatory and sensitive tool for detection of pathogen P. azadirachtae at early stages.

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Section: Discussionsupporting

confidence: 60%

Section: Resultsmentioning

confidence: 99%

“…) and PF1/PF2 (Girish et al. ) were validated using DNA of different Phomopsis species (Table ) including P. azadirachate . The PCR conditions were same as described by Prasad et al.…”

Section: Methodsmentioning

confidence: 99%

“…) and PF1/PF2 (Girish et al. ) has been reported. In the genus Phomopsis, more than 1000 species have been described (Uecker ).…”

Section: Introductionmentioning

confidence: 95%

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Species‐Specific PCR‐Based Assay for Identification and Detection of Phomopsis (Diaporthe) azadirachtae Causing Die‐Back Disease in Azadirachta indica

Vedashree

Sateesh

Chowdappa

et al. 2015

Journal of Phytopathology

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“…This species has been drawn considerable attention for studying epidemiology and management of the disease, phytotoxicity, crude toxin extraction and biocontrol by microbial antagonism (Girish and Bhat 2008;Girish et al 2009;Nagendra Prasad et al 2010).…”

Section: Names Of Common Phytopathogens In Current Usementioning

confidence: 99%

The genus Phomopsis: biology, applications, species concepts and names of common phytopathogens

et al. 2011

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The genus Phomopsis (teleomorph Diaporthe) comprises phytopathologically important microfungi with diverse host associations and a worldwide distribution. Species concepts in Phomopsis have been based historically on morphology, cultural characteristics and host affiliation. This paper serves to provide an overview of the current status of the taxonomy in Phomopsis with special reference to biology, applications of various species, species concepts, future research perspectives and names of common pathogens, the latter being given taxonomic reappraisal. Accurate species identification is critical to understanding disease epidemiology and in developing effective control measures for plant diseases. Difficulties in accurate species identification using morphology have led to the application of alternative approaches to differentiate species, including virulence and pathogenicity, biochemistry, metabolites, physiology, antagonism, molecular phylogenetics and mating experiments. Redefinition of Phomopsis/Diaporthe species has been ongoing, and some species have been redefined based on a combination of molecular, morphological, cultural, phytopathological and mating type data. Rapid progress in molecular identification has in particular revolutionized taxonomic studies, providing persuasive genetic evidence to define the species boundaries. A backbone ITS based phylogenetic tree is here in generated using the sequences derived from 46 type, epitype cultures, and vouchers and is presented as a rough and quick identification guide for species of Phomopsis. The need for epitypification of taxonomic entities and the need to use multiple loci in phylogenies that better reflect species limits are suggested. The account of names of phytopathogens currently in use are listed alphabetically and annotated with a taxonomic entry, teleomorph, associated hosts and disease symptoms, including brief summaries of taxonomic and phylogenetic research. Available type culture information and details of gene sequences derived from type cultures are also summarized and tabulated.

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A survey of die-back disease of neem in Tamil Nadu, India and PCR-based confirmation of the isolates

Prasad

Bhat

Girish

2011

Archives Of Phytopathology And Plant Protection

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A survey of die-back disease of neem was done in different agro climatic regions of Tamil Nadu, India using Global Positioning System (GARMIN 12). Twigs of Azadirachta indica (Neem) infected with die-back were collected from different regions of Tamil Nadu, India and they were further analyzed to determine the pathogen. Phomopsis azadirachtae the causal organism was isolated on malt extract agar from die-back infected neem twigs. They were identified by conventional and molecular methods. Phomopsis genus specific primers (5.8S r-DNA) were then used for the confirmation of P. azadirachtae -the causative agent of die-back of neem by Polymerase chain reaction (PCR). Studies revealed the amplification of expected 141bp DNA in P. azadirachtae isolated from the diseased trees of different regions of Tamil Nadu confirming the causal organism of die-back of neem. Studies revealed a very high incidence of die-back in most of the places of Tamil Nadu. Hand held GPS was used in the study which would help in continuous monitoring of the diseased trees.

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PCR-based detection ofPhomopsis azadirachtaein die-back affected neem seeds

Cited by 4 publications

References 21 publications

Species‐Specific PCR‐Based Assay for Identification and Detection of Phomopsis (Diaporthe) azadirachtae Causing Die‐Back Disease in Azadirachta indica

Species‐Specific PCR‐Based Assay for Identification and Detection of Phomopsis (Diaporthe) azadirachtae Causing Die‐Back Disease in Azadirachta indica

The genus Phomopsis: biology, applications, species concepts and names of common phytopathogens

A survey of die-back disease of neem in Tamil Nadu, India and PCR-based confirmation of the isolates

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