Abstract
Background/objectives: Orobanche is one of the important parasites of solanaceae crops and causes heavy yield losses. This study aimed to use DNA barcoding technique for identification of Orobanche species. Methods: Surveillance study was conducted in the year 2018 on tomato crops grown in four different districts in Balochistan. From these tomato fields, 15 Orobanche plant samples were collected and put in polyethylene bags containing silica gel for drying. The dried samples were processed for DNA extraction using CTAB method. The extracted DNA was confirmed through gel electrophoresis which were further amplified on PCR using rbcL, matK primers. Then, the PCR products were processed for DNA sequencing using Sanger method. Further, the DNA sequences were edited and aligned with BioEdit and MegaX software and the aligned sequences were matched on NCBI for BLAST. The FASTA results of both partial and/or complete genome were run on MegaX for phylogenetic analysis. Findings: The success rates for PCR amplification was 100% for rbcl primer, while it was 68% for matK primer. However, the resolution power of matK was higher than that obtained from rbcL as manifested in the rate of DNA acquisition sequence. The DNA barcode results of 15 samples revealed 99.16% rate of DNA acquisition sequence for identification of P. ramosa and 96.41 for P. purpurea. While, Rbcl rate of DNA sequencing show no results. Application/Improvement: from this study it was inferred that matK gene performed better than rbcL in Orobanche species identification and proved as a potential gene for other pseudogene plants species identification. Keywords: Orobanche; species; DNA barcode; rbcL; matK; tomato
Highlights
The family Orobanchaceae contains 15 obligate parasitic genera with 250 species and is considered closely related to the Scrophulariaceae [1,2]
The plant parasite Orobanche was considered as medicinal plant in Europe and Russia during 16th century which is used for the treatment of diseases like kidney and bladder’s stone and further this plant was utilized for detoxification of wine
This study was conducted to identify the Orobanche species occurring in tomato crop in four district of Balochistan such as Mastung, Pishin Killa Saifulla and Quetta
Summary
The family Orobanchaceae contains 15 obligate parasitic genera with 250 species and is considered closely related to the Scrophulariaceae [1,2]. Orobanche species parasitize and cause invasive damage to a wide range of economically important plants, e.g. legumes, sunflower, tomato and tobacco [5], enhancing production of some very useful constituents like phenylpropanoids and derivatives; such as coumarins, lignin, suberin, cutin, and tannin. These constituents are involved in the resistance of the host, as a defense mechanism against broomrape [6]. The plant parasite Orobanche was considered as medicinal plant in Europe and Russia during 16th century which is used for the treatment of diseases like kidney and bladder’s stone and further this plant was utilized for detoxification of wine
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