Investigation of epidemiology of soil-borne viruses in triticale with the objective of development virus resistant cultivars with high biomass yields for extraction of biogas and ethanol, sub project 1: Investigation of of the epidemiology of soil-borne viruses in triticale

Project

Project code: JKI-EP-08-1128
Contract period: 01.03.2012 - 31.03.2016
Budget: 521,108 Euro
Purpose of research: Applied research

Triticale exhibits a high yield potential and is a potential cereal for the efficiently production of biomass for the extraction of biogas and ethanol in line with using conceptions of peripheral bio energy. The production of triticale is heavy at risk by soil-borne virus in many growing areas. The plant health is a main precondition for the stable use of triticale as renewable resource. The JKI undertake the task to clarify epidemiological aspects of pathogens and to develop pest control proposals. The project promotion could make a contribution to establishment basics for breeding of high-output energy plants for the cultivation in infested by soil-borne viruses fields. This project supports the BMELV efforts in the respect to the development of measures to ensure the further utilization of triticale growing areas, heavily infested with soil-borne viruses, to stabilize the cultivation of triticale in such locations with the aim to produce profitable renewable raw. In the second year of the project a continuous collection of data regarding the pathogen populations in different infection sites took place. The nucleotide sequences of SBWMV-N from isolates from wheat and barley were compared for the coat protein/read through protein region. There were no differences between the SBWMV-N sequences from wheat, barley and the SBWMV De (strain Nebraska) originally published by König et al. For WSSMV up to now only the nucleotide sequence for the 3’ half of the RNA1 was deposited in Genbank. We have extended the available sequence information by a region in the 5’ half and submitted the sequence information to Genbank (NCBI Genbank accession KJ609243). Based on sequence homology with WYMV we can reason that the sequence information analysed by us codes for the 7K protein and part of the CI protein of WSSMV. On the basis of our investigations into the diversity of the plasmodiophorid vector Polymyxa graminis we did not find preferences of certain formae speciales for particular host plants. Also, there seems to be no specialization as to which virus is transmitted by which forma specialis. Polymyxa sequences unambiguously assigned to f. sp. temperata or tepida were cloned and used as standards for Polymyxa real time PCR. Alignments of Polymyxa sequences available from the database were carried out in order to design specific primer pairs for real time PCR. In 2013, we have tested a large number of triticale field samples by real time PCR for the presence of SBCMV and WSSMV. For the total RNA preparation we used leaf extracts that had been generated with a sap press for ELISA, thereby limiting the amount of additional work and at the same ensuring comparability of samples. In order to prevent non-specific amplification products, an assay using a FAM-hydrolysis probe was developed for the detection of WSSMV. This allowed the clear detection of WSSMV-infected samples and the quantification of the amount of virus RNA in the samples. A manuscript regarding quantification of SBCMV and WSSMV in field samples of triticale by Real Time PCR will be submitted for publication. Furthermore, the resistance evaluation of promising varieties and breeding material was carried out in 4 different locations.

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