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Abstrakt
AFLP Fingerprinting for Identification of Infra-Species Groups of Rhizoctonia solani and Waitea circinata
Bimal S. Amaradasa, Dilip Lakshman and Keenan Amundsen
Patch diseases caused by Thanatephorus cucumeris (Frank) Donk and Waitea circinata Warcup and Talbot varieties (anamorphs: Rhizoctonia species) pose a serious threat to successful maintenance of several important turfgrass species. Reliance on field symptoms to identify Rhizoctonia causal agents can be difficult and misleading. Different Rhizoctonia species and Anastomosis Groups (AGs) vary in sensitivity to commonly applied fungicides and they also have different temperature ranges conducive for causing disease. Thus correct identification of the causal pathogen is important to predict disease progression and make future disease management decisions. Grouping Rhizoctonia species by anastomosis reactions is difficult and time consuming. Identification of Rhizoctonia isolates by sequencing Internal Transcribed Spacer (ITS) region can be cost prohibitive. Some Rhizoctonia isolates are difficult to sequence due to polymorphism of the ITS region. Amplified Fragment Length Polymorphism (AFLP) is a reliable and cost effective fingerprinting method for investigating genetic diversity of many organisms. No detailed analyses have been done to determine the suitability of AFLP for inferring infra-species level of Rhizoctonia isolates. The objective of the present study was to develop AFLP fingerprinting to identify infra-species level of unknown R. solani Kühn and W. circinata isolates. Seventy-nine previously characterized R. solani (n=55) and W. circinata (n=24) isolates were analyzed with AFLP markers generated by four primer pairs. Unweighted Pair Group Method with Arithmetic Mean (UPGMA) correctly grouped R. solani and W.circinata isolates according to their AG, AG subgroup or W.circinata variety. Principle component analysis (PCA) corroborated UPGMA clusters. To our knowledge this is the first time AFLP analysis has been tested as a method to decipher the AG, AG subgroup or W.circinata variety across a wide range of Rhizoctonia isolates.