Identification of Heterodera latipons Using PCR with Sequence Characterised Amplified Region (SCAR) Primers

Specific primers and polymerase chain reaction (PCR) assays that identify Fusarium oxysporum f. sp. ciceris and each of the F. oxysporum f. sp. ciceris pathogenic races 0, 1A, 5, and 6 were developed. F. oxysporum f. sp. ciceris- and race-specific random amplified polymorphic DNA (RAPD) markers identified in a previous study were cloned and sequenced, and sequence characterized amplified region (SCAR) primers for specific PCR were developed. Each cloned RAPD marker was characterized by Southern hybridization analysis of Eco RI-digested genomic DNA of a subset of F. oxysporum f. sp. ciceris and nonpathogenic F. oxysporum isolates. All except two cloned RAPD markers consisted of DNA sequences that were found highly repetitive in the genome of all F. oxysporum f. sp. ciceris races. F. oxysporum f. sp. ciceris isolates representing eight reported races from a wide geographic range, nonpathogenic F. oxysporum isolates, isolates of F. oxysporum f. spp. lycopersici, melonis, niveum, phaseoli, and pisi, and isolates of 47 different Fusarium spp. were tested using the SCAR markers developed. The specific primer pairs amplified a single 1,503-bp product from all F. oxysporum f. sp. ciceris isolates; and single 900- and 1,000-bp products were selectively amplified from race 0 and race 6 isolates, respectively. The specificity of these amplifications was confirmed by hybridization analysis of the PCR products. A race 5-specific identification assay was developed using a touchdown-PCR procedure. A joint use of race 0- and race 6-specific SCAR primers in a single-PCR reaction together with a PCR assay using the race 6-specific primer pair correctly identified race 1A isolates for which no RAPD marker had been found previously. All the PCR assays described herein detected up to 0.1 ng of fungal genomic DNA. The specific SCAR primers and PCR assays developed in this study clearly identify and differentiate isolates of F. oxysporum f. sp. ciceris and of each of its pathogenic races 0, 1A, 5, and 6.

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Heterodera Latipons N. Sp., a Cereal Cyst Nematode From the Mediterranean Region

Nematologica ◽

10.1163/187529269x00867 ◽

1969 ◽

Vol 15 (4) ◽

pp. 535-542 ◽

Cited By ~ 21

Author(s):

Mary T. Franklin

Keyword(s):

Mediterranean Region ◽

Cyst Nematode ◽

Cereal Cyst Nematode ◽

The Mediterranean ◽

Heterodera Latipons

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Development of SCAR primers based on a repetitive DNA fingerprint for Escherichia coli detection

The Journal of Microbiology ◽

10.1007/s12275-013-2244-4 ◽

2013 ◽

Vol 51 (1) ◽

pp. 31-35 ◽

Cited By ~ 3

Author(s):

Aphidech Sangdee ◽

Sitakan Natphosuk ◽

Adunwit Srisathan ◽

Kusavadee Sangdee

Keyword(s):

Escherichia Coli ◽

Repetitive Dna ◽

Dna Fingerprint ◽

Scar Primers

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Multiplex PCR assay using SCAR primers to detect Eimeria spp. in chicken

Journal of Parasitic Diseases ◽

10.1007/s12639-012-0142-z ◽

2012 ◽

Author(s):

P. Venkateswara Rao ◽

M. Raman ◽

G. DhinakarRaj ◽

S. Abdul Basith ◽

S. Gomathinayagam

Keyword(s):

Multiplex Pcr ◽

Pcr Assay ◽

Multiplex Pcr Assay ◽

Eimeria Spp ◽

Scar Primers

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BEAUVERIA BASSIANA CHARACTERIZATION AND EFFICACY GRASSHOPPER /ANGARACRIS

Mongolian Journal of Agricultural Sciences ◽

10.5564/mjas.v13i2.525 ◽

2015 ◽

Vol 13 (2) ◽

pp. 96-100

Author(s):

Kh Otgonjargal ◽

S Sugar ◽

E Bazarragchaa ◽

N Enkhbold ◽

B Battur

Keyword(s):

Beauveria Bassiana ◽

Infection Rate ◽

Natural Infection ◽

The Body ◽

Insect Host ◽

Local Strains ◽

As Species ◽

Agricultural Sciences ◽

Scar Primers

The B. bassiana is a fungus of manyarthropods, including more than 200 species of insects and acaridae. When spores of the fungus come into contact with the body of an insect host, they germinate, enter the body, and grow inside, eventually killing the insect. Two local strains, including B. bassiana-G07, which was isolated from grasshopper Oedaleusasiaticus, died on natural infection, and B. bassiana-G10, which was isolated from grasshopper Caliptomus abbreviates, died of soil borne infection, were detected and it was identified as species B. bassiana by PCR. SCAR primers OPB9 F/R677 and OPA15 F/R441 was specific to of B. bassiana. The highest infection rate by B. bassiana-G07and mortality was observed in variants of both concentrations 2.1 x 108 conidia/ml, 2.1 x 109 conidia/ml; where mortality reached 86.3-100%.Mongolian Journal of Agricultural Sciences Vol.13(2) 2014: 96-100

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SCAR MARKER DEVELOPMENT FOR THE CORRECT IDENTIFICATION OF IRIS ENSATA

International Journal of Pharmacy and Pharmaceutical Sciences ◽

10.22159/ijpps.2017v9i12.21677 ◽

2017 ◽

Vol 9 (12) ◽

pp. 201

Author(s):

Mohd Mughees ◽

Shipra Rani Jha ◽

Javed Ahmad ◽

Altaf Ahmad

Keyword(s):

Scar Marker ◽

Rapd Analysis ◽

Geographical Area ◽

Correct Identification ◽

Iris Ensata ◽

Rapd Pattern ◽

Study Results ◽

Ctab Method ◽

High Level ◽

Scar Primers

Objective: The objective of this research was to develop the RAPD based SCAR marker for the correct identification of the Iris ensata Thunb. (I. ensata) plant from its adulterants.Methods: Five samples of I. ensata. from the different geographical area were used in this study. The plant genomic DNA was isolated with the CTAB method with some modification (as dried samples were also used). After that, polymorphism was checked with the help of the 10-mer random primers of OPAA and BG series. Then, the bands of interest were eluted and cloned into pGEMT easy vector for the sequencing. Finally, the sequence is used to develop the SCAR primers (Ir-f andIr-R) specific for I. ensata and the developed primers also validated with respect to the market samples.Results: A putative 580 bp sequence specific for Iris ensata was identified from the randomly amplified polymorphic DNA (RAPD) analysis. To overcome the main limitation of RAPD it has been converted into SCAR markers. So that, this specific band was then eluted, cloned and sequenced. After that, SCAR primers (Ir-F and Ir-R) were synthesized by using this sequence. For the validation of the synthesized SCAR primers, they were tested with respect to the market samples. The amplicon of 260 bp was produced by the SCAR primers in the authentic I. ensata but market samples did not produce any bands with the synthesized SCAR primers.Conclusion: The results of this study show a high level of polymorphism in the RAPD pattern of the different accessions of the plant. Furthermore, this study results in the successful development of the RAPD based SCAR marker for the identification of the I. ensata.

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RAPD Marker Conversion into a SCAR Marker for Rapid Identification of Johnsongrass [Sorghum halepense (L.) Pers.]

Notulae Botanicae Horti Agrobotanici Cluj-Napoca ◽

10.15835/nbha4119001 ◽

2013 ◽

Vol 41 (1) ◽

pp. 306 ◽

Cited By ~ 4

Author(s):

Wenju ZHANG ◽

Shasha WEI ◽

Liping YIN ◽

Zhirui DENG ◽

Jianping YI ◽

...

Keyword(s):

Scar Marker ◽

Rapd Marker ◽

Morphological Characters ◽

Rapid Identification ◽

Sorghum Halepense ◽

Germplasm Identification ◽

Marker Conversion ◽

The World ◽

Sequence Characterized Amplified Regions ◽

Scar Primers

Johnsongrass [Sorghum halepense (L.) Pers.] is a malignant weed in the world, threatening biodiversity at invaded habitats in more than 50 countries. Because of similarity in morphological characters, S. halepense and its relatives, S. almum, S. nitidum, S. propinquum, S. sudanense, and S. bicolor, etc. was difficult to identify. As a supplementary methodolgy for morphology identification, a molecular detection method was established. Sequence Characterized Amplified Regions (SCAR) marker is a recent established, reliabile, and stable molecular marker based on RAPD maker, an effective way for germplasm identification. In this study, one specific band of S.halepense was screened by 163 pairs of RAPD primers. According to the sequences of the band, a pair of special SCAR primers SH1/SH2 was designed and verified by 65 Sorghum DNA samples from all over the world. The results showed SCAR primers SH1/SH2 can be used to distinguish S.halepense and its relatives rapidly with three exceptions of Australia geotypes.

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