Abstract
Lupins anthracnose is a destructive seed and airborne disease caused by Colletotrichum lupini, affecting stems and pods. Primary seed infections as low as 0.01–0.1% can cause very severe yield losses. One of the most effective management strategies is the development of a robust and sensitive seed detection assay to screen seed lots before planting. PCR-based detection systems exhibit higher levels of sensitivity than conventional techniques, but when applied to seed tests they require the extraction of PCR-quality DNA from target organisms in backgrounds of saprophytic organisms and inhibitory seed-derived compounds. To overcome these limitations, a new detection protocol for C. lupini based on a biological enrichment step followed by a PCR assay was developed. Several enrichment protocols were compared with Yeast Malt Broth amended with ampicillin, streptomycin, and lactic acid were the most efficient. A species-specific C. lupini primer pair was developed based on rDNA IGS sequences. The specificity was evaluated against 17 strains of C. lupini, 23 different Colletotrichum species, and 21 different organisms isolated from seeds of Lupinus albus cv. Multitalia, L. luteus cv. Mister, and L. angustifolius cv. Tango. The protocol described here enabled the detection of C. lupini in samples artificially infected with less than 1/10,000 infected seed.
Highlights
Lupinus spp. are important agronomic crops worldwide, with several species that are grown as ornamentals or as pioneer plants in soil maintenance programmes [1,2]
The most common fungi isolated from the three lupin species were Alternaria alternata, Aspergillus spp., Cladosporium oxysporum, Mucor sp., and Penicillium spp
Fungi of 16 genera and 10 species were recovered from lupin seeds, in addition to one yeast and one sterile mycelium, as shown in
Summary
Lupinus spp. are important agronomic crops worldwide, with several species that are grown as ornamentals or as pioneer plants in soil maintenance programmes [1,2]. Lupin plants are key components of a wide range of soils and climates, mainly around the Mediterranean basin, East Africa, and the entire American continent. Lupin is an important crop mainly due to its elevated seed protein content, its adaptability to low fertility soils, and its valuable influence on crop rotation [1,4,5,6]. The seed composition of specific cultivars of white lupin (Lupinus albus) is very similar to soybean (35% proteins and 0.5% starch), with some species that are able to reach high levels of up to 18% of oil content. The vividly colored flowers of lupins contribute to their importance as ornamental plants (e.g., L. polyphyllus, known as large-leaved lupine), and are amongst the most popular ornamental plants in temperate climate
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