Erysiphe corylacearum as a new pathogen of hazelnut in the Czech Republic.

In July 2021, leaves and shoot tops of the common hazel (Corylus avellana L.), with a whitish coating, were found in the Czech Republic (southern Moravia region). The infected hazel bushes were found along a road in a deciduous forest and in an urban garden. In most European countries, Phyllactinia guttata is found on the abaxial surface of the leaves in the form of a continuous whitish to light grey mycelium, possibly with large black chasmothecia. In our case, the mycelium was present on both sides of the leaves, but the symptoms and the incidence were much stronger on the adaxial side. The first symptoms usually appeared on the adaxial side of the leaves as small white radially expanding patches of mycelium. In the final stage, the spots merged and covered a substantial part of the leaf blade (50-85 % on the adaxial side, 5-25 % on the abaxial side). When the abaxial side of the leaves was infected, chlorotic spots were evident on the adaxial side. The spots of powdery mildew were small (3-15 mm), whitish, rounded to irregular, effuse eventually becoming confluent, and occurred primarily on the adaxial side of the leaves. Conidiophores (30-53×4-6 µm) grew on the amphigenic mycelium, were erect, consisted of 1-3 cells, i.e. cylindrical foot cell and followed 1-2 cells, from which hyaline ellipsoid to doliform-limoniform conidia (17-34 ×15-21) (n = 50) were individually detached. Single or in groups dark brown chasmothecia (77-116 µm in diameter) had up to hyaline 8-15 aseptate straight appendages (50-102 µm) with multiple (3-5×) dichotomously branched apexes and recurved tips. Chasmothecia contained 3-6 asci (42-62 × 34-55 µm) with 4-8 obovoid to broadly ellipsoidal hyaline ascospores (14-22 × 75 µm). Based on morphological characters, the powdery mildew was identified as Erysiphe corylacearum (2). Morphological identification was confirmed by molecular analysis of samples. DNA was extracted from symptomatic leaves tissue using the DNeasy Plant Mini Kit, (Quiagen, Hilden, Germany) and the rDNA internal transcribed spacer region of 2 isolates was amplified using primers PMITS1 and PMITS2 (Cunnington et al. 2003) and sequenced. BLAST analysis of our 720bp fragments (both identical and represented by GenBank accession no. OR432526) showed 100% sequence identity to ITS rDNA sequences of E. corylacearum other countries of Central Europe for example from Austria (MW031866), Italy (MW045428), Hungary (OQ411007), Germany (OP554268) or Slovakia (MT176105). Pathogenicity was verified on two-year-old plants of Corylus avellana. Healthy leaves were artificially infected by dusting conidia from infected leaves. Inoculated plants were incubated under controlled conditions (21-23 °C, 70-80 % relative humidity). Characteristic symptoms of powdery mildew appeared on the adaxial side of the leaves 9-12 days after inoculation. Control plants treated with distilled water remained symptomless. Powdery mildew isolated from inoculated leaves was morphologically identical to isolates from naturally infected leaves. The first record of E. corylacearum in Europe on cultivated hazelnut species was reported by Sezer et al. (2017) in Turkey in 2013. Within a few years, the E. corylacearum spread and was recorded on various species of Corylus in other European countries (for example Mezzalama et al., 2020; Rosati et al., 2021; Beenken et al., 2022; Boneva et al., 2023), East Asia (Arzanlou et al., 2018) and the USA (Meparishvili 2019). To our knowledge, this is the first report of Erysiphe corylacearum in the Czech Republic.

Relationship Between Dehydrin Accumulation and Winter Survival in Winter Wheat and Barley Grown in the Field.

Low temperatures represent a crucial environmental factor determining winter survival (WS) of barley and wheat winter-type varieties. In laboratory experiments, low temperatures induce an active plant acclimation response, which is associated with an enhanced accumulation of several stress-inducible proteins including dehydrins. Here, dehydrin accumulations in sampled wheat (WCS120 protein family, or WCS120 and WDHN13 transcripts) and barley (DHN5 protein) varieties grown in two locations for two winters were compared with the variety WS evaluated by a provocation wooden-box test. A high correlation between dehydrin transcripts or protein relative accumulation and variety WS score was found only in samples taken prior vernalization fulfillment, when high tolerant varieties accumulated dehydrins earlier and to higher level than less tolerant varieties, and the plants have not yet been vernalized. After vernalization fulfillment, the correlation was weak, and the apical development indicated that plants reached double ridge (DR) in barley or stayed before DR in wheat. Dehydrin proteins and transcripts can be thus used as reliable markers of wheat or barley variety winter hardiness in the field conditions; however, only at the beginning of winter, when the plants have not yet finished vernalization. In wheat, a higher correlation was obtained for the total amount of dehydrins than for the individual dehydrin proteins. HIGHLIGHTS: -More tolerant winter-type wheat and barley plants reveal higher threshold induction temperatures for dehydrin accumulation in comparison to less tolerant varieties. Thus, more tolerant winter cereals have higher dehydrin levels than the less tolerant ones upon the same ambient temperature in November samplings.-A significant correlation between dehydrin transcript/protein accumulation and winter survival was found in both winter wheat and winter barley plants in the field conditions, but only prior to vernalization fulfillment.

Comparative analysis of ELISA, one step RT-PCR and IC-RT-PCR for the detection of bean yellow Mosaic virus in gladiolus.

To overcome the problematic detection of Bean yellow mosaic virus (BYMV) concentration in gladiolus corms or cormlets, we tested suitability of different diagnostic methods such as ELISA, one step RT-PCR and IC-RT-PCR. Among these methods, DAS-ELISA and one step RT-PCR method was able to detect virus in leaves and it failed to detect in corms or cormlets whereas IC-RT-PCR method was very sensitive and able to detect virus in both leaves and corms or cormlets of gladiolus plants this is due to during an RNA isolation step and viral particles are enriched by antibody or plastic affinity capture whilst PCR inhibitors are eliminated from the sample during the immunocpature stage of the test. Therefore, IC-RT-PCR was the most reliable method of diagnosing BYMV in gladiolus corms or cormlets.

Expression of dehydrin 5 during the development of frost tolerance in barley (Hordeum vulgare).

The Dhn5 gene is the major cold-inducible dehydrin gene in barley. This study deals with the relationship between Dhn5 gene expression and its protein product accumulation, and the development of frost tolerance (FT) upon cold acclimation (CA) in 10 barley cultivars of different growth habits and geographical origins. The activation of Dhn5 gene expression was determined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), the accumulation of DHN5 protein was evaluated by protein gel blot analysis using a specific anti-dehydrin antibody, and the acquired level of FT was determined by a direct frost test. During the first 2 weeks of CA, there was a rapid increase in Dhn5 gene expression, DHN5 protein accumulation and FT in all cultivars examined. After 2 weeks of CA, differences in DHN5 accumulation and in FT measured as lethal temperature (LT(50)) were observed between the cultivars belonging to different growth habits. Specifically, intermediate (I) and winter (W) cultivars showed a higher level of DHN5 accumulation and FT than the spring (S) cultivars, which exhibited a lower level of accumulated DHN5 and FT. (Intermediate cultivars do not have vernalization requirement, but they are able to induce a relatively high level of FT upon CA.) In contrast, no differences between the cultivars belonging to different growth habits in Dhn5 mRNA accumulation were found. After 3 weeks of CA, the differences in accumulated DHN5 and FT between the individual growth habits became evident due to different developmental regulation of FT. The amount of accumulated DHN5 corresponded well with the level of FT of individual cultivars. We conclude that the amount of accumulated DHN5 after a certain period of CA differed according to the growth habits of cultivars and can be used as a marker for determination of FT in barley.