Isolation and identification of Sclerotinia sclerotiorum the casual agents of white mold in eggplants by polymerase chain reaction (PCR) technique

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Research Paper 01/04/2018
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Isolation and identification of Sclerotinia sclerotiorum the casual agents of white mold in eggplants by polymerase chain reaction (PCR) technique

Haneen Abd-alhaleem Ali , Neran Salem Aljarah
J. Bio. Env. Sci.12( 4), 30-38, April 2018.
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This study was carried out at Plant Protection Dept. / College of Agriculture/ University of Baghdad, in 2016-2017. Due to phenotypic differences between four Sclerotinia sclerotiorum isolates, the polymerase chain reaction (PCR) technique using the universal fungal primer pairs of the Internal transcribed spacer 4 and 5 (ITS4⁄ITS5) was depended to identify. The four S. sclerotiorum isolates (SS1, SS2, SS3 and SS4) were isolated from infected eggplants cultivated in plastic houses in Karbala (one isolate) and Baghdad (three isolates). DNA of each isolate was extracted from the sclerotia. Results of sequences analysis revealed that the four isolates shared 99-100% identities with the equivalent sequences of the fungal isolates conserved international Gen Bank, using MEGA6 program variations in morphological characteristics (mycelium growth rate, sclerotia numbers and their distribution in the plate), were found between the isolates on PDA at 25±2°c. Isolates pathogenicity were evaluated on egg plants, 30 days old, in pots under plastic house conditions by inoculating the stem under meristem, without injury, with 5 mm disk of fungal mycelium growth from edge of isolate cultural, 5 days old, and measuring the spot length after 7 days of inoculation. Significant difference in the pathogenicity between the isolates was recorded. SS1 from Karbala was found the more active with lesion length attained to 4.25 cm. Variations in oxalic acid production were manifested between isolates in potato sucrose broth medium (PSB) as determined by high performance liquid chromatography (HPLC) which found to be 10.95, 8.70, 4.90, 8.60 mg/L. of PSB medium  for SS1, SS2, SS3, SS4 respectively.


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