Development of specific molecular markers to distinguish and quantify broomrape species in a soil sample

Broomrapes ( Orobanche and Phelipanche spp . ) are destructive obligate plant parasites in Israel and in the Mediterranean basin. Conventional methods for parasitic weeds detection are difficult, since the parasite seeds are extremely small (dust-like seeds) and survive in the soil for several decad...

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Bibliographic Details
Published in:European journal of plant pathology 2019-12, Vol.155 (4), p.1367-1371
Main Authors: Aly, Radi, Bari, Vinay Kumar, Londner, Avishai, Nassar, Jackline Abu, Lati, Ran, Taha-Salaime, Leena, Eizenberg, Hanan
Format: Article
Language:English
Subjects:
Agriculture
Amplification
Biomedical and Life Sciences
Deoxyribonucleic acid
Diagnostic systems
DNA
Ecology
Gene sequencing
Infestation
Life Sciences
Markers
Parasites
Parasitic plants
Plant Pathology
Plant Sciences
Polymerase chain reaction
Ribosomal DNA
Ribulose-bisphosphate carboxylase
Seeds
Soils
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Summary:Broomrapes ( Orobanche and Phelipanche spp . ) are destructive obligate plant parasites in Israel and in the Mediterranean basin. Conventional methods for parasitic weeds detection are difficult, since the parasite seeds are extremely small (dust-like seeds) and survive in the soil for several decades. Here, we report the development of specific molecular markers rbcL 1 based on rbcL (large subunit of the ribulose-bisphosphate carboxylase) gene from Orobanche crenata and ITS 100 based upon unique sequences in the internal transcribed spacer ( ITS ) regions of the nuclear ribosomal DNA of Phelipanche aegyptiaca. Genomic DNA was extracted from soil samples artificially infested with broomrape seeds or tissue of P. aegyptiaca , O. cumana and O. crenata and subjected to PCR analysis. rbcL 1 marker, successfully differentiate between O. crenata and O. cumana, amplified a specific PCR products (1300 bp with O. crenata and 1000 bp with O. cumana ). However, the rbcL 1 marker failed to amplify soil samples with seeds or tissues of P. aegyptiaca or any soil-borne DNA. ITS 100 marker and Real-Time PCR, allowed quantitative diagnostic of the parasite O. cumana in a soil sample; amplified a specific PCR products (100 bp). As expected the universal control primer ( UCP -555) amplified a PCR product (555 bp), when genomic DNA extracted from soil samples with or without broomrape tissues. The development of an efficient, simple and robust molecular marker to detect and distinguish between broomrape species, has a significant insights on the assessment level of infestation and planning eradication program of the parasite in a field crop.
ISSN:0929-1873
1573-8469
DOI:10.1007/s10658-019-01841-9