Assessment of the loose smut fungi (Ustilago nuda and U. tritici) in tissues of barley and wheat by fluorescence microscopy and real-time PCR

Loose smut fungi of barley and wheat (Ustilago nuda and U. tritici, respectively) colonize the plant without causing obvious disease symptoms before heading. The availability of diagnostic methods to detect and follow the growth of these pathogens in the plant would therefore be highly advantageous for both resistance breeding and the development of effective seed treatments. Using seed lots of barley and wheat highly infected with loose smut, we studied the early establishment of the loose smut pathogens in the plant by fluorescence microscopy. In hand-cut sections stained with the fluorochrome Blankophor?, fungal hyphae were observed to invade the shoot apical meristem and leaf primordia during the first days after the onset of germination. At the first node stage the ear and leaf primordia were generally extensively colonized. Hyphae of U. nuda were also regularly observed in high density in the nodes. A protocol was developed for the specific amplification of U. nuda and U. tritici DNA extracted from infected plant tissue. PCR screening of U nuda in seedlings from infected and healthy seed lots was compared to ELISA, microscopy and ultimately head infection of mature plants derived from tillers of the tested seedlings. The results indicated that a prediction of loose smut infection by real-time PCR is possible at the second leaf stage, and that the assay is equally suited for use with spring and winter varieties of barley and wheat.     

The barley mutant emr2 shows enhanced resistance against several fungal leaf pathogens

Homozygous mlo -barley plants are resistant to barley powdery mildew but hypersusceptible to the rice blast fungus Magnaporthe oryzae . A mutational analysis was performed in the barley back-cross line BCIngrid mlo5 which led to the identification of two mutants with enhanced capacity to resist infections by M. oryzae , referred to as enhanced M. oryzae resistance mutants emr1 and emr2 . Here, we report on the characterization of emr2 mutant plants which not only show an almost complete reduction in disease severity after inoculation with M. oryzae but are also resistant to the necrotrophic fungi Drechslera teres and Rhynchosporium secalis . Histological analysis revealed that resistance to M. oryzae was based mainly on the formation of papillae at sites of attempted penetration into epidermal cells. There was no progression of fungal growth into the mesophyll. Additionally, because of the presence of the mlo -allele, emr2 -plants retained resistance to powdery mildew. The emr2 -conditioned broad spectrum resistance was inherited as in a recessive manner. Monitoring of PR -gene expression and enzymatic activity of peroxidases revealed a constitutively activated defence in emr2 .     

Wheat blast caused by Magnaporthe oryzae pathovar Triticum is efficiently controlled by the plant defence inducer isotianil

Journal of Plant Diseases and Protection - Wheat blast caused by Magnaporthe oryzae pv. Triticum is an upcoming threat to wheat cultivation worldwide. The disease crossing over to wheat first...     

Asian grapevine leaf rust caused by Phakopsora euvitis: an important disease in Brazil

Anthropogenic influences such as tourism and climate change affect ecosystems. One consequence is that invasive species can spread to and colonize novel areas. A recent example of this scenario is the appearance, in the early 2000s in Brazil and Australia, of the plant pathogenic fungus Phakopsora euvitis, which causes Asian grapevine leaf rust. It is speculated that the disease was introduced either by long distance transport of spores in the atmosphere or the import of contaminated plant material. While the disease could be eradicated from the Australian continent it is still present in Brazil. The pathogen has spread successfully throughout most Brazilian grapevine‐growing regions, most probably because of the lack of resistant plant cultivars. Today, the disease is endemic in Brazil. This review was written to highlight the increasing impact of the disease in Brazil and summarizes the current knowledge on the disease monocycle under different environmental conditions and the consequences for grapevine management. Gaps in the present state of knowledge are highlighted with the purpose of stimulating further research aimed at identifying efficient strategies to combat the disease.     

Fenhexamid - an efficient and inexpensive fungicide for selection of Magnaporthe oryzae transformants

European Journal of Plant Pathology - Magnaporthe oryzae is one of the most economically important phytopathogenic fungi, and is used as a model organism to study plant-pathogen interactions. To...     

Formation of satellite uredinia as an important trait related to grapevine colonization by Phakopsora meliosmae-myrianthae

Phakopsora meliosmae-myrianthae, the causal agent of Asian grapevine leaf rust, significantly reduces the photosynthetic efficiency of grapevine leaves in green symptomless tissues surrounding lesions. This study took a close look at grapevine leaf colonization kinetics by P. meliosmae-myrianthae and compared it to P. pachyrhizi–soybean and Uromyces appendiculatus–bean colonization. It is already known from the literature that soybean rust, similar to grapevine rust, has a negative effect on leaf photosynthesis greater than would be expected based on visual lesions. However, in contrast to soybean and grapevine rusts, the effect of bean rust on leaf photosynthesis is proportional to the diseased leaf area. Colonization progress was monitored by fungal biomass assessed via histological staining and quantitative polymerase chain reaction (qPCR). Individual lesions of P. meliosmae-myrianthae on grapevine, P. pachyrhizi on soybean and U. appendiculatus on common bean leaves were evaluated every 3–4 days, and the number of uredinia was counted. Staining showed that mycelial colonization did not extend beyond the lesion border. The number of P. pachyrhizi and P. meliosmae-myrianthae uredinia within the lesions increased over time (on average 14-fold), whereas the number of U. appendiculatus uredinia remained the same. These findings were corroborated by qPCR, which revealed a greater increase in fungal biomass for Phakopsora spp. than for U. appendiculatus until 12 days post-inoculation. The high number of satellite uredinia within lesions might be directly related to the impact of this pathogen in photosynthetic efficiency on symptomless areas of diseased grapevine leaves. This study identified accelerated formation of satellite uredinia as an important feature of grapevine colonization by P. meliosmae-myrianthae.     

Tracing seed to seedling transmission of the wheat blast pathogen Magnaporthe oryzae pathotype Triticum

Wheat blast caused by Magnaporthe oryzae pathotype Triticum (MoT), initially restricted to South America, is a global threat for wheat after spreading to Asia in 2016 by the introduction of contaminated seeds, raising the question about transmission of the pathogen from seeds to seedlings, a process so far not well understood. We therefore studied the relationship between seed infection and disease symptoms on seedlings and adult plants. To accomplish this objective, we inoculated spikes of wheat cv. Apogee with a transgenic isolate (MoT-DsRed, with the addition of being resistant to hygromycin). We identified MoT-DsRed in experiments using hygromycin resistance for selection or by observation of DsRed fluorescence. The seeds from infected plants looked either apparently healthy or shrivelled. To evaluate the transmission, two experimental designs were chosen (blotter test and greenhouse) and MoT-DsRed was recovered from both. This revealed that MoT is able to colonize wheat seedlings from infected seeds under the ground. The favourable conditions of temperature and humidity allowed a high recovery rate of MoT from wheat shoots when grown in artificial media. Around 42 days after germination of infected seeds, MoT-DsRed could not be reisolated, indicating that fungal progression, at this time point, did not proceed systemically/endophytically. We hypothesize that spike infection might occur via spore dispersal from infected leaves rather than within the plant. Because MoT-DsRed was not only successfully reisolated from seed coats and germinating seeds with symptoms, but also from apparently healthy seeds, urgent attention is needed to minimize the risks of inadvertent dispersal of inoculum.     

Different Patterns of Host Genes are Induced in Rice by Pseudomonas syringae, a Biological Inducer of Resistance, and the Chemical Inducer Benzothiadiazole (BTH)

Rice seedlings treated with the synthetic compound benzo(1,2,3)thiadiazole-7-carbothioic acid S-methyl ester (BTH) acquired resistance to subsequent attack by the rice blast fungus Magnaporthe grisea (Hebert) Barr. BTH (trade name Bion) has been released to the market as a plant protecting agent for rice. Here, we analysed the pattern of expressed genes in rice plants treated with BTH, and compared this pattern with those induced by the formerly discovered resistance inducer 2,6-dichloroisonicotinic acid (INA) and by Pseudomonas syringae pv. syringae, a non-host pathogen inducing a hypersensitive response. Both INA and BTH induced similar patterns of genes, suggesting that these compounds are functional analogues. In contrast, the patterns induced by the chemical inducers and by P. syringae were clearly dissimilar.