Study on genetic variation of Hyrcanian Calonectria pseudonaviculata (boxwood blight agent) isolates using RAPD and ISSR markers

Subject Areas : Plant Microbiology

Parisa Khazaeli ¹ , Saeed Rezaee ² , Mansoureh Mirabolfathy ³ , Hamidreza Zamanizadeh ⁴ , Hadi Kiadaliri ⁵

1 - Ph.D. student, Department of Plant Pathology, College of Agriculture and Natural Resources, Science and Research branch, Islamic Azad University, Tehran, Iran.
2 - Assistant Professor, Department of Plant Pathology, College of Agriculture and Natural Resources, Science and Research branch, Islamic Azad University, Tehran, Iran.
3 - Professor, Iranian Research Institute of Plant Protection, Agricultural Research Education and Extension Organization (AREEO), Tehran, Iran.
4 - Professor, Department of Plant Pathology, College of Agriculture and Natural Resources, Science and Research branch, Islamic Azad University, Tehran, Iran.
5 - Assistant Professor, Department of Forestry, College of Agriculture and Natural Resources, Science and Research branch, Islamic Azad University, Tehran, Iran.

Received: 2017-06-08 Accepted : 2017-08-17 Published : 2017-08-23

Keywords: Calonectria pseudonaviculata, ISSR marker, RAPD marker, &beta, -tubulin gene,

Abstract :

Background & Objectives: Boxwood tree is one of the Iranian endemic trees expanded throughout Hyrcanian forests. They are of particular importance among the world's forest reserves. Recently, Calonectria pseudonaviculata has been considered as one of the most important causes of blight and leaf defoliation of the boxwood in Iran. For the first time, the present study was aimed to evaluate the genetic variation of Hyrcanian C. pseudonaviculata isolates using RAPD and ISSR markers. Material & Methods: In this study, 75 fungal isolates were collected from the infected boxwoods throughout Hyrcanian forests in the North of Iran. The isolates were assessed based on morphology and colony color. Genetic diversity of the isolates was studied using RAPD and ISSR markers. A part of the beta-tubulin gene was sequenced and deposited at NCBI. Phylogenetic analysis was carried out using PAUP* v. 4.0b10. Results: The colony color of isolates on potato dextrose agar (PDA) medium was brown with pale hale, conidia were cylindrical, rounded at both ends, 1-septate, 48- 68 µm. Stipe was long (90-14 µm) and hyaline, with the extension terminating in a broadly ellipsoid papillate vesicle, the widest part above the middle. Phylogenetic tree based on the β-tubulin gene showed that all isolates are placed into the same group. Our results indicated that the population of this fungus has a low genetic diversity in Iran. Conclusion: It seems that the variation of this pathogen is started to change genetically in some areas due to selection pressure phenomenon.