title: The Development of Simple Methods for the Maintenance and Quantification ofPolymyxa graminis.
abstract: Polymyxa graminis, a root endoparasite of several cereal species, is considered to be non-pathogenic but serves as a vector of various plant viruses belonging to the generaBymovirus,Furovirus, andPecluvirus. Specifically, it reduces barley productivity by transmitting theBarley Yellow Mosaic Virus(BaYMV). To date, due to its obligate biotrophic property, no artificial culturing ofP. graminiswas reported and its quantification was also technically challenging. Here, we developed a novel and simple method to infectP. graminiswithin sterile barley roots in contamination free by preparing nearly pure zoospore inoculum. Such artificial maintenance ofP. graminiswas verified based on the presence of various developmental stages in infected barley roots under microscope. In addition, the population of resting spores in host tissue was determined by establishing standard curve between manually counted number of spores and Ctvalues of 18S rDNA amplification using quantitative real-time PCR. Furthermore, it was validated that standard curve generated was also applicable to estimate the abundance ofP. graminisin soil environments. In conclusion, the present study would help to generate a system to investigate the etiological causes as well as management of plant diseases caused byP. graminisand BaYMV in tissue and soil.
authors: Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, 570-752 Korea.
authors: Swati
authors: S
authors: Tyagi
authors: Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, 570-752 Korea.
authors: Razia
authors: R
authors: Sultana
authors: Department of Agricultural Biology, College of Agriculture and Life Sciences, Chonbuk National University, Jeonju, 561-756 Korea.
authors: Ho-Jong
authors: HJ
authors: Ju
authors: Department of Agricultural Biology, College of Agriculture and Life Sciences, Chonbuk National University, Jeonju, 561-756 Korea.
authors: Wang-Hyu
authors: WH
authors: Lee
authors: Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, 570-752 Korea.
authors: Kangmin
authors: K
authors: Kim
authors: National Institute of Crop Science, RDA, Iseo, 55365 Korea.
authors: Bongchoon
authors: B
authors: Lee
authors: Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, 570-752 Korea.
authors: Kui-Jae
authors: KJ
authors: Lee
keywords: Barley; Polymyxa graminis; Real-time PCR; Resting spore; Zoospore