Development of high-resolution melting PCR (HRM-PCR) assay to identify native fungal species associated with the wheat e
- PDF / 904,377 Bytes
- 7 Pages / 595.276 x 790.866 pts Page_size
- 45 Downloads / 192 Views
MICROBIAL GENETICS • ORIGINAL PAPER
Development of high-resolution melting PCR (HRM-PCR) assay to identify native fungal species associated with the wheat endosphere Tomasz Cłapa 1
&
Katarzyna Mikołajczak 2 & Lidia Błaszczyk 2 & Dorota Narożna 1
Received: 20 May 2020 / Revised: 9 August 2020 / Accepted: 18 August 2020 # The Author(s) 2020
Abstract Understanding the complexity and biodiversity of fungal communities associated with the wheat endosphere can facilitate the identification of novel strains that might be beneficial to the host plant. However, the differentiation and taxonomic classification of the endosphere-associated fungi with respect to various cultivars and plant organs are challenging, time-consuming, and expensive, even with the use of molecular techniques. In the present work, we describe a fast, simple, and low-cost method based on high-resolution melting PCR (HRM-PCR) for the identification and differentiation of wheat endogenous fungal isolates. Using this approach, we differentiated 28 fungal isolates, which belonged to five different genera, namely Alternaria, Penicillium, Epicoccum, Fusarium, and Trichoderma. Furthermore, the results of the study revealed that this method can allow large-scale screening of cultured samples. Keywords High-resolution melting PCR . Identification assay . fungal species . Wheat endosphere
Introduction Wheat is the second most widely grown cereal worldwide after rice. According to statistics, the global wheat production in 2017–2018 was 771.7 million tons (FAOSTAT, http:// www.fao.org/statistics/en/, 2020). Regarding quantity and the area of cultivation, wheat is the most popular among the cereals grown in the EU, accounting for nearly half of the total production of crops (EU, Eurostat, https://ec.europa.eu/ eurostat/, 2020). However, in Europe, wheat production is affected by a number of abiotic and biotic stress factors (Trnka et al. 2014). Various strategies, based on chemical, genetic, and agricultural principles, are recommended for improving the production of wheat under combined unfavorable Communicated by: Agnieszka Szalewska-Palasz * Tomasz Cłapa [email protected] 1
Department of Biochemistry and Biotechnology, Poznań University of Life Sciences, Dojazd 11 60-632 Poznań Poland
2
Department of Pathogen Genetics and Plant Resistance, Institute of Plant Genetics, Polish Academy of Sciences, Strzeszyńska 34 60-479 Poznań Poland
environmental conditions. Among them, the use of biological control agents (BCAs) is the most promising and environmental-friendly approach. Fungi associated with the plant endosphere that live inside the plant tissues throughout or at least for a part of their life cycle without causing any disease symptoms their host are considered to be good candidates as BCAs (Rodriguez et al. 2009; Comby et al. 2016; Gdanetz and Trail 2017). The presence of these fungi, which are classified as endophytes, in wheat has already been demonstrated by several authors (Crous et al. 1995; Larran et al. 2002; Istifadah and McGee 2006;
Data Loading...
