In: Sydowia 72, (2020): 0021-0033, ISSN 0082-0598, DOI 10.12905/0380.sydowia72-2020-0021, Published online on January 28, 2020

Microsatellite-primed PCR assessed the genetic diversity of Leucostoma cinctum (Diaporthales) in Iran

Somayeh Bakhshi, Khalil-Berdi Fotouhifar & Mohammad Javan-Nikkhah

Bakhshi S., Fotouhifar Kh.-B. & Javan-Nikkhah M. (2020) Microsatellite-primed PCR assessed the genetic diversity of Leucostoma cinctum (Diaporthales) in Iran. – Sydowia 72: 21–33.

For investigation of genetic diversity of Leucostoma cinctum (Valsaceae) isolates, using MP-PCR assay, 63 isolates of the fungus were recovered from infected different tree samples collected from 11 provinces in Iran during 2004–2011. Four of eight tested primers, including (CGA)5, (GACA)4, (ATC)7 and (AC)8, were selected based on the generated polymorphism, detectable banding pattern in agarose gel and the reproducibility of results. Overall, 134 DNA bands were obtained using these four primers, with five monomorphic loci (3.7 %) and 129 polymorphic loci (96.3 %). The high level of polymorphism was generated by (GACA)4 and (ATC)7 primers, which showed 100 % polymorphism. The polymorphisms revealed by (AC)8 and (CGA)5 primers were 94.3 % and 91.7 %, respectively. Cluster analysis of data based on Jaccard’s similarity coefficient was done by UPGMA method using PAST software version 2.17c. The results showed that the similarity level of all tested isolates was at least 30 %, and at this level of similarity, five main clusters and 15 groups were identified, showing a high genetic diversity among populations of L. cinctum. Matrix correlation coefficient for all dendrograms varied from r = 0.59 – 0.7. In the present study, significant polymorphisms were identified in 14 ascospore progenies of two separate asci that were recovered from two separate perithecia on plum and cherry
trees. In general, there was no relationship between groups of dendrogram and geographic origins of the isolates. However, most of the isolates were separated according to their host plant.

Keywords: Canker disease, Cytospora cincta, Sordariomycetes, plant pathogen, polymorphism, biodiversity.