GenBank Accession Numbers featured prominently in the work of Weir et al. (2012) and Silva et al (2012). (Z)-4-Hydroxytamoxifen Kindly ensure that you return both OQ509805-808 and OQ507698-724. Phylogenetic analyses of multiple loci, utilizing both newly obtained and GenBank sequences, established that three isolates (UBOCC-A-116036, -116038, and -116039) grouped with the species *C. gloeosporioides* sensu stricto, whereas the remaining isolate (UBOCC-A-116037) clustered within *C. karsti*. Incubation at 20°C for a duration of ten days led to the development of symptoms, indistinguishable from the initial presentation, localized around the inoculation point; meanwhile, control samples inoculated with water remained unaffected. The re-isolated fungal colonies, derived from the lesions, exhibited a morphology identical to the original isolates' form. Infections due to Colletotrichum species have significantly impaired citrus production in several Mediterranean countries, including Italy (Aiello et al., 2015), Portugal (Ramos et al., 2016), Tunisia (Ben Hadj Daoud et al., 2019), and Turkey (Uysal et al., 2022). C. gloeosporioides s.s. and C. karsti were identified through these investigations as the agents of causation. These two Colletotrichum species exhibited the highest frequency. Citrus and its related European genera exhibit an association, as reported in the study by Guarnaccia et al. (2017). Our research, as far as we are aware, reveals the inaugural account of C. gloeosporioides and C. karsti leading to anthracnose in grapefruit crops in France, thereby confirming the presence of these pathogens in the Mediterranean fringe. Due to the crucial economic position of citrus production in the Mediterranean region, the presence of Colletotrichum species is noteworthy. For 'should', continuous monitoring is essential, and a well-devised control strategy must be put in place.
The beverage known as tea, a plant species of Camellia sinensis, has been enjoyed globally for its purported health-enhancing properties since its origins in southwestern China 60 to 70 million years ago, with a high concentration of polyphenols, as detailed by Pan et al. (2022). The Puer tea (10273 'E, 2507' N) in Yunnan, China, experienced a decline in yield and quality during the period from October to December 2021, due to a disease presenting symptoms similar to leaf spot. The survey, performed in a 5700 m^2 field, revealed leaf spot symptoms on an approximate 60% prevalence of tea plants. The onset of symptoms included shrinking and yellowing, later progressing to the formation of circular or irregular brown spots. Ten symptomatic leaves, selected from ten distinct trees, were obtained for pathogen isolation. Portions of diseased tissue, 0.5 cm in length, were excised at the point where infected and healthy tissue joined. pathology of thalamus nuclei After sterilizing their surfaces (5 minutes in 75% ethanol, 2 minutes in 3% NaOCl, and three rinses with sterile distilled water), the pieces were dried and transferred to potato dextrose agar (PDA) plates, which were kept in darkness at 25 degrees Celsius for five days. Single-spore isolates FH-1, FH-5, FH-6, and FH-7 were isolated, and their morphological structures and internal transcribed spacer (ITS) and translation elongation factor 1-alpha (TEF) gene sequences proved identical. Consequently, the FH-5 representative isolate was selected for subsequent investigation. After 7 days of incubation at 28°C, fungal colonies exhibited a white or light yellow pigmentation on PDA. Hyaline, aseptate conidia, round or oval in shape, were found singly or in clusters on hyphae or conidia stalks, and their dimensions were 294, 179, 182, and 02 µm (n = 50). Primary conidiophores, appearing early and having a verticillium-like structure (Figure 1.K, L), typically exhibit a 1-3 level verticillate arrangement, predominantly branching divergently, with accompanying phialides, and measuring 1667 ± 439 µm in length (n=50). After one week, secondary conidiophores (Figure 1I, J), typically exhibiting a penicillate structure, often further branched, emerge, reaching an average length of 1602 ± 383 μm (n = 50). The descriptions of Clonostachys rosea Schroers H.J. in Schroers et al. (1999) precisely matched the observed morphological characteristics. The amplification and sequencing of the internal transcribed spacer (ITS) region and the translation elongation factor 1-alpha (TEF) gene, employing primers ITS1/ITS4 and EF1-728F/EF1-986R, respectively, resulted in the identification of C. rosea as the pathogen, in line with the findings of Fu Rongtao's 2019 research. The PCR product sequences, corresponding to accession numbers ON332533 (ITS) and OP080234 (TEF), were archived in GenBank. BLAST searches of the derived sequences revealed a 99.22% similarity (510 nucleotides out of 514) and a 98.37% similarity (241 nucleotides out of 245) with C. rosea HQ-9-1 sequences in GenBank, accession numbers MZ433177 and MZ451399, respectively. MEGA 70's maximum likelihood phylogenetic analysis successfully placed isolate FH-5 in a well-supported cluster that included C. rosea. The pathogenicity of the FH-5 strain was tested employing a pot assay. A sterilized needle was used to mark the leaves of ten healthy tea plants. By spraying a spore suspension of FH-5 (105 spores per milliliter) onto the leaves until the excess drained away, plants were inoculated; control leaves were sprayed with sterile water. In a climate-controlled box set at 25 degrees Celsius and 70% relative humidity, inoculated plants were placed. A total of three pathogenicity tests were executed. Symptoms emerged on all inoculated leaves; conversely, the control leaves displayed no symptoms. At 72 hours post-inoculation, pale yellow lesions appeared at the wound's edge, accompanied by the initial appearance of brown spots. Subsequently, typical lesions analogous to those on field plants emerged two weeks later. Following re-isolation, the identical fungus was characterized morphologically and molecularly (using ITS and TEF markers) from the infected leaves, but not from the non-inoculated leaves. Subsequently, *C. rosea* has also been observed to be involved in the pathogenesis of diseases in broad bean (Vicia faba) crops. Exploring studies on Afshari et al. (2017) work and Diaz et al. (2022)'s research on garlic, alongside Haque M.E et al. (2020) findings on beets, and other plant species. Based on our research, this is the pioneering account of C. rosea-related leaf spot affecting tea in China. The leaf spot on tea is effectively addressed through the valuable information presented in this study.
Gray mold in strawberries is attributable to a multitude of Botrytis species, encompassing Botrytis cinerea, B. pseudocinerea, B. fragariae, and B. mali. Widespread in the production regions of the eastern United States and Germany are the species B. cinerea and B. fragariae; their distinction is pivotal for formulating efficacious disease management strategies. Polymerase chain reaction (PCR) currently constitutes the sole means of differentiating these species in field specimens, a method that is time-consuming, laborious, and costly. This study's development of a loop-mediated isothermal amplification (LAMP) method relied on the nucleotide sequences of the species-specific NEP2 gene. Only B. fragariae DNA was selectively amplified by the specially designed primer set; no other Botrytis species were amplified. Intein mediated purification The identified plant pathogens included B. cinerea, B. mali, and B. pseudocinerea, along with others. A rapid DNA extraction technique proved successful in enabling the LAMP assay to amplify fragments from DNA extracted from the infected fruit, validating its capability to detect small amounts of B. fragaria DNA in field-infected specimens. Additionally, a masked assay was undertaken to identify B. fragariae within 51 samples extracted from strawberry cultivation areas in the eastern United States, using the LAMP method. In the testing of B. fragariae samples, a reliability of 935% (29 out of 32) was achieved. Conversely, no amplification occurred for B. cinerea, B. pseudocinerea, or B. mali samples within the 10-minute reaction time. The results of our study indicate a specific and reliable application of LAMP for isolating B. fragariae from afflicted fruit, promising effective control strategies in the field.
Chilli peppers (Capsicum annuum), a globally significant vegetable and spice, are widely cultivated, especially in China. Chili pepper plants in Guilin, Guangxi, China, at the geographical location of 24 degrees 18 minutes North and 109 degrees 45 minutes East, showed signs of fruit rot in October 2019. Spots of irregular shape, dark green in color, initially appearing near the middle or base of the fruit, gradually enlarged and changed to larger grayish-brown lesions, resulting in the fruit's decay. The fruit's eventual demise came when the water within it evaporated away, causing a complete drying-out. Three towns in various counties of Guilin yielded three disease samples, characterized by a chilli fruit disease incidence percentage fluctuating between 15% and 30%. To disinfect, 33 mm pieces of diseased fruit margins were initially treated with 75% ethanol for 10 seconds, followed by 2% NaOCl for one minute, and lastly rinsed in sterile distilled water three times. Tissue fragments were placed on separate plates of potato dextrose agar (PDA) and kept at 25 degrees Celsius for seven days of incubation. From three fruits with diseased tissues, a uniform isolation rate of 100% was observed for fifty-four fungal isolates that shared similar morphology. Among the selections, GC1-1, GC2-1, and PLX1-1 were selected for detailed analysis proceeding. Following 7 days of incubation at 25°C in complete darkness, the colonies cultivated on PDA displayed a considerable amount of whitish to yellowish aerial mycelium. Cultured on carnation leaf agar (CLA) for 7 days, macroconidia displayed a long, hyaline, and falcate structure. Dorsal and ventral lines gradually widened toward the apex, with a curved apical cell and a foot-shaped basal cell. Generally containing two to five septa, the strains exhibited varying dimensions. GC1-1 macroconidia showed a length range from 2416 to 3888 µm and a width range from 336 to 655 µm (average 3139448 µm). GC2-1 macroconidia demonstrated lengths from 1944 to 2868 µm and widths from 302 to 499 µm (average 2302389 µm). PLX1-1 macroconidia exhibited a length range from 2096 to 3505 µm, and widths from 330 to 606 µm (average 2624451 µm).