Genetic diversity and pathogenic characteristics of Phomopsis asparagi obtained from Asparagus officinalis and A. kiusia
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ORIGINAL ARTICLE
Genetic diversity and pathogenic characteristics of Phomopsis asparagi obtained from Asparagus officinalis and A. kiusianus in Japan Le Dinh Thao1,3 · Masaru Matsumoto1,2 Received: 20 March 2020 / Accepted: 15 July 2020 © Deutsche Phytomedizinische Gesellschaft 2020
Abstract Phomopsis asparagi has been reported as a serious disease, causing stem blight on asparagus cultivars (Asparagus officinalis), and this fungus was also recorded as a common spot disease on wild asparagus (A. kiusianus). In this study, isolates of P. asparagi obtained from A. officinalis and A. kiusianus in different locations of the Kyushu island were successfully assessed the genetic variation through mycelial compatibility grouping (MCG) and random amplified polymorphic DNA (RAPD). Thirty fungal isolates were identified into six MCGs; of these, fifteen isolates were representatively chosen for the RAPD analysis, pathogenicity and extracellular enzyme test. Eight RAPD primers, B62, B66, B68, B71, C83, D22, D32 and E10, generated 100 different patterns from the isolates with 91% of the polymorphic bands. Based on RAPD analysis, P. asparagi isolates were divided into 3 distinct genetic clusters, namely RAPD-I, RAPD-II and RAPD-III at a similarity of 73%. The result indicated that there was a great genetic variation within the isolates from A. kiusianus, presenting in the three clusters. While the isolates from A. officinalis showed high genetic homology and they belonged to the RAPD-II cluster. There is no conflict between genetic clusters distinguished by the RAPD markers and those of MCG. On the other hand, the pathogenicity test revealed that the isolates of RAPD-II and RAPD-III were highly virulent pathogens. In contrast, the isolates of RAPD-I were weak virulent pathogens when inoculated on A. officinalis. Screening of extracellular enzyme activities concluded that all isolates were positive for laccase, lipase and protease activity. Keywords Extracellular enzymes · Mycelial compatibility grouping · RAPD fingerprinting · Wild asparagus
Introduction Asparagus plant (Asparagus officinalis) is an economically important crop in many regions in the world, cultivated at least 62 countries with an estimated total area of 195,819 hectares, and Japan was ranked in the top 10 asparagus producing countries in 2009 (Benson 2012). Kyushu island lying in Southern Japan has a subtropical climate, the warm period from February to October is a favorable condition for growing of asparagus. Therefore, this island is one of * Le Dinh Thao [email protected] 1
Graduate School of Integrated Sciences for Global Society, Kyushu University, Fukuoka 812‑8581, Japan
2
Institute of Tropical Agriculture, Kyushu University, Fukuoka 812‑8581, Japan
3
Division of Pathology and Phyto‑immunology, Plant Protection Research Institute (PPRI), Duc Thang, Bac Tu Liem, Hanoi, Vietnam
the main areas producing asparagus in Japan (Watanabe et al. 2014). However, stem blight caused by Phomopsis asparagi (Sacc.) has recently been the most
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