Two 5 mm × 5 mm segments of infected plant tissue were surface-sterilized by treating them with 95% ethanol for one minute, subsequently with 70% ethanol for one minute, and lastly with 1% sodium hypochlorite for one minute, to isolate the causal pathogen. Following this, the samples underwent a triple rinsing with distilled water, were subsequently air-dried using sterile filter paper, and were then placed into a 15% water agar medium containing 100 ppm streptomycin, followed by incubation in the dark at 25 degrees Celsius. After single-hypha-tip purification, three independent isolates (HNO-1, HNO-2, HNO-3) from Haenam, and three others (KJO1-1, KJO1-2, KJO1-3) from Ganjin were produced. The process involved subculturing hyphae originating from randomly selected independent tissue samples from each location on potato dextrose agar (PDA, Sparks, MD 21152, USA). The PDA colonies were initially pigmented white, later exhibiting a transition to a light brown color after two weeks. Within two weeks on PDA, all collected isolates displayed the formation of dark brown to black, irregular and globose sclerotia. Isolates characterized by binuclear hyphae, displaying a color gradient from white to dark brown, and branching orthogonally, with a septum positioned near the branch point, as well as the presence of multinucleate cells, are consistent with the species Ceratobasidium cereale, according to previous studies by Boerema et al. (1977), Burpee (1980), and Sharon et al. (2008). The ITS sequence (with GenBank accession numbers provided) serves as a key element in molecular identification. In order to amplify the MW691851-53 (HNO-1 to HNO-3) and MW691857-59 (KJO1-1 to KJO1-3) genes, along with LSU (OQ397530-35), rpb2 (OQ409878-83), tef1 (OQ409884-89), and atp6 (OQ409890-95) from six isolates, the corresponding primer sets, ITS4/5 (White et al., 1990), LROR/LR5 (Vilgalys and Hester, 1990), bRPB2-6F/bRPB2-71R (Matheny, 2005; Reeb et al., 2004), TEF1-F/TEF1-R (Litvintseva et al., 2006), and ATP61/ATP62 (Kretzer and Bruns, 1999), respectively, were utilized. The ITS region's genetic sequence displayed 99.7% identity to the C. cereale strain WK137-56 (KY379365) and 99.8% to the Ceratobasidium sp. sequence. learn more AG-D (KP171639). Employing the maximum likelihood method within the MEGA X software package (Kumar et al., 2018), the concatenated ITS-LSU, rpb2, tef1, and atp6 sequences demonstrated that the six isolates were grouped inside a clade encompassing C. cereale (Gonzalez et al., 2016; Ji et al., 2017; Tomioka et al., 2021; Li et al., 2014). The Korean Agricultural Culture Collection received isolates HNO-1 (accession number KACC 49887) and KJO1-1 (accession number KACC 410268) as two representative samples. Six isolates were cultivated on sterilized ray grains at 25°C in complete darkness for three weeks, producing the inoculum necessary for pathogenicity studies. Five oat varieties ( Within pots containing a mixture of 80 grams of infected ray grains, 150 grams of composite soil, and 150 milliliters of water (Baroker Garden Soil, Seoul Bio Co., LTD), Choyang seeds were planted. A combination of 150 grams of composite soil, 150 milliliters of water, and 80 grams of sterilized ray grains was applied to the control. To ensure uniform growth conditions, all inoculated and control pots were placed within a 20°C growth chamber, illuminated by a 12-hour photoperiod and maintaining 65% humidity. Seedlings' oat sheaths, three weeks after inoculation, displayed the characteristic symptoms of sharp eyespots. The control seedlings exhibited no symptoms whatsoever. The infection assays were carried out in triplicate, demonstrating similar results. Following successful re-isolation, the pathogen's identity was confirmed using both morphological and molecular analysis techniques. Etiological studies on oats are relatively scarce in Korea, due to their lesser economic appeal when compared to barley and wheat. Sharp eyespot disease, a consequence of C. cereale infection, has been previously recorded in barley and wheat (Kim et al., 1991); however, this current report details the first identification of this disease in oats in Korea.
The oomycete Phytopythium vexans (de Bary et al.) is a significant pathogen impacting the root and crown systems of a diverse range of plants, such as woody ornamentals, fruit trees, and forest trees, as it inhabits both water and soil environments. For successful nursery production, early and accurate identification of Phytophthora is critical, as this pathogen is quickly transported to neighboring plants via the irrigation system. Unfortunately, conventional strategies for the detection of this pathogen are frequently characterized by time-consuming procedures, ambiguous outcomes, and substantial financial burdens. Therefore, a focused, sensitive, and timely molecular diagnostic methodology is requisite for overcoming the deficiencies of conventional identification strategies. For the purpose of identifying *P. vexans*, this current investigation established a loop-mediated isothermal amplification (LAMP) assay. In the process of designing and evaluating LAMP primer sets, PVLSU2 was identified as specific for P. vexans, exhibiting no amplification of other closely related oomycetes, fungi, and bacteria. The developed assays, moreover, were sufficiently sensitive to amplify DNA quantities up to 102 femtograms per reaction. Real-time LAMP assays proved more sensitive in identifying infected plant samples than traditional PCR and culture-based methods. Simultaneously, both LAMP-based assessments pinpointed a minimum of 100 zoospores suspended in 100 milliliters of water. P. vexans detection in disease diagnostic laboratories and research institutions is anticipated to be expedited by LAMP assays, enabling timely preparedness responses to disease outbreaks.
Infestations of powdery mildew are directly linked to the fungal species Blumeria graminis f. sp. The wheat crops in China are vulnerable to the destructive tritici (Bgt) strain. Developing mildew-resistant cultivars requires as an initial step the mapping of quantitative trait loci (QTL) associated with powdery mildew resistance and the creation of markers easily employed by breeders. Researchers identified an all-stage resistance gene, along with several quantitative trait loci (QTLs), within a population of 254 recombinant inbred lines (RILs), generated by crossing Jingdong 8 and Aikang 58. Across three consecutive growing seasons and in six distinct field environments, the population's resistance to powdery mildew was assessed using two unique Bgt isolate mixtures, designated #Bgt-HB and #Bgt-BJ. The study of genotypic data from the Wheat TraitBreed 50K SNP array revealed the presence of seven stable quantitative trait loci (QTLs) on chromosome arms 1DL, 2AL, 2DS, 4DL, 5AL, 6BL.1, and 6BL.2. Resistance conferred by the QTL on 2AL extended to all stages of Bgt race E20, as demonstrated in greenhouse experiments, and its contribution to explaining up to 52% of the phenotypic variance in field trials was observed, but this effect was specific to the #Bgt-HB strain. The gene Pm4a was predicted as the contributor to this QTL, determined by its placement within the genome and its genetic sequence. QPmja.caas-1DL, a complex entity, requires careful consideration. QPmja.caas-4DL and QPmja.caas-6BL.1 were tentatively identified as novel QTL potentially conferring resistance to powdery mildew. In their action against both Bgt mixtures, QPmja.caas-2DS and QPmja.caas-6BL.1 showcased a potential for broad-spectrum resistance. Using a comprehensive panel of 286 wheat cultivars, a KASP marker tightly linked to QPmja.caas-2DS was developed and validated. The QTL and marker findings are highly valuable for wheat researchers and breeders, considering the prominent roles Jingdong 8 and Aikang 58 play as cultivars and breeding parents.
China's Yangtze River basin is home to the perennial herbaceous plant, Bletilla striata, a species belonging to the Orchidaceae family, with widespread distribution. role in oncology care The medicinal properties of B. striata, a plant found in China, are commonly harnessed to reduce wound bleeding and inflammation. A noticeable prevalence (over 50%) of leaf spot symptoms was observed on B. striata plants in a traditional Chinese medicine plantation (approximately 10 hectares) located in Xianju City, Zhejiang Province, China, during September 2021. Initial observations revealed small, round, pale brown, necrotic lesions on the foliage. Later, the lesions' centers turned grayish-brown, while their margins darkened with subtle bumps, ultimately growing to 5-8 mm across on the leaves. Subsequently, the minuscule patches extended and consolidated, developing into necrotic lines measuring approximately 1 to 2 centimeters. Leaves displaying disease symptoms were surgically removed, surface-sterilized, and planted on potato dextrose agar (PDA). After 3 days of incubation at 26°C, fungal colonies (2828 mm) manifested grayish-black mycelia spreading throughout the tissues. The color of basal conidia ranged from pale to dark brown, contrasting with the pale brown color of apical conidia. Central cells of apical conidia possessed both increased size and a darker pigmentation than those in basal conidia. Smooth conidia, displaying either a fusiform, cylindrical, or slightly curved shape, terminated with rounded tips. Extending from 2234 meters to 3682 meters, the items' lengths averaged 2863 meters, alongside 2 to 4 septations. These septations showed subtle constrictions. In order to obtain a pure culture, the isolation of monospores was carried out. Following its creation, strain BJ2Y5 was deposited in the Strain Preservation Center at Wuhan University, China, where it received the designation CCTCC M 2023123. From PDA plates incubated at 26 degrees Celsius for a period of seven days, the newly grown mycelia and conidia were gathered. Employing the Ezup Column Fungi Genomic DNA Purification Kit (Sangon Biotech Co., Shanghai, China), DNA was extracted. Medical Resources Based on an examination of DNA sequences from three genes – glyceraldehyde 3-phosphate dehydrogenase (GAPDH), the internal transcribed spacer (ITS), and the partial second largest subunit of RNA polymerase II (RPB2) – the phylogenetic position of isolate BJ2-Y5 was determined. Upon performing a BLAST search using GenBank accession numbers, the results. Isolates OP913168, OP743380, and OP913171 exhibited a genetic similarity of 99% when compared to the reference isolate CBS 22052.