Detection of Two Peronospora spp., Responsible for Downy Mildew, in Opium Poppy Seed

Author:

Thangavel Tamilarasan1ORCID,Jones Suzanne2,Scott Jason B.2ORCID,Livermore Mark3,Wilson Calum R.4ORCID

Affiliation:

1. Tasmanian Institute of Agriculture (TIA), University of Tasmania, New Town, Tasmania 7008, Australia

2. TIA, University of Tasmania, Burnie TAS 7320, Australia

3. Tasmanian Alkaloids Pty Ltd, P.O. Box 130, Westbury, TAS 7303, Australia

4. TIA, University of Tasmania, New Town, Tasmania 7008, Australia

Abstract

Downy mildew is a serious threat to opium poppy production globally. In recent years, two pathogen species, Peronospora somniferi and Peronospora meconopsidis, which induce distinct symptoms, have been confirmed in Australia. In order to manage the spread of these pathogens, identifying the sources of inoculum is essential. In this study, we assessed pathogen presence associated with poppy seed. We developed PCR and qPCR assays targeting the coxI and coxII gene regions, for the detection, differentiation, and quantification of P. somniferi and P. meconopsidis in poppy seed. These results were complemented and compared with direct seed histological examination and a seed washing combined with viability staining for oospore detection. The majority of seed lots from all harvest years contained detectable P. meconopsidis, the earliest (1987) predating the first official record of the disease in Tasmania (1996). In contrast, only seed lots harvested in 2012 or later contained P. somniferi, evidence of its more recent introduction. P. meconopsidis contamination was estimated to be as high as 33.04 pg DNA/g of seed and P. somniferi as high as 35.17 pg DNA/g of seed. Incidence of pathogen contamination of seeds, estimated via a group testing protocol, ranged from 0 to 9% (P. meconopsidis) or 0 to 11% (P. somniferi). Mycelia were predominately found external to the seed coat. Seed washing and viability staining demonstrated that putatively viable oospores were present in the majority of seed lots. Transmission testing confirmed both pathogens can be successfully transmitted from infested seed to infected seedling. PCR and qPCR pathogen assays were found to be reliable and offer a routine test for determining pathogen inoculum in poppy seeds.

Funder

Australian Research Council

Publisher

Scientific Societies

Subject

Plant Science,Agronomy and Crop Science

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