Molecular characterization of avipoxviruses circulating in Windhoek district,Namibia 2021

Samples from eleven birds (chicken, dove and peacock) with symptoms of fowlpox, caused by the avipoxvirus (APV), were collected in seven different areas of the Windhoek district, Namibia between April and October 2021. A fragment of the 4b core protein and the DNA polymerase gene of APV were amplified by PCR from the DNA of the samples and sequenced. Phylogenetic analysis revealed that the viruses present in the chickens all belonged to clade A1 while the viruses in the doves and peacock were from subclade A3.1. This is the first report of subclade A3.1 avipoxvirus in peacock. In addition, all of the samples obtained from chickens were shown by PCR to be positive for the integration of reticuloendotheliosis virus while those from the doves and peacocks were negative. This study is the first characterization of avipoxvirus in Namibia and provides additional information on the presence of avipoxvirus in southern Africa.     

Identification of Alternaria spp. on wheat by pathogenicity assays and sequencing

The pathogenicity of Alternaria spp. isolated from wheat leaves collected in regions where alternaria leaf blight has been reported was compared with that of IMI reference isolates of A. triticina and A. alternata using two durum and two bread wheat genotypes. To identify isolates putatively corresponding to A. triticina , morphological and DNA sequence analyses based on ribosomal DNA from the internal transcribed spacer (ITS) region (ITS1, 5·8S rRNA gene, ITS2) and toxicity bioassays of culture filtrate were combined. Glasshouse inoculations provided reliable information to assess the pathogenicity of A. triticina isolates on wheat. Alternaria leaf blight symptoms were produced by the A. triticina isolates only on durum wheat cv. Bansi, while A. alternata , A. tenuissima and A. arborescens isolates were found to be nonpathogenic on the wheat cultivars tested. Alternaria triticina isolates were distinguished from other Alternaria species by Simmons and Roberts' sporulation pattern 6 and two to three conidia per sporulation unit associated with primary conidia bearing long (> 7  µ m) apical secondary conidiophores. Phylogenetic analysis also proved effective at discriminating wheat-pathogenic A. triticina from other nonpathogenic Alternaria species. Alternaria triticina isolates yielded longer ITS sequences than A. alternata , A. tenuissima and A. arborescens isolates, leading to clear-cut differences as visualized with agarose gel electrophoresis. Additionally, only culture filtrates of A. triticina isolates caused nonspecific necrotic lesions on leaves of 3-week-old wheat plants.     

Development of specific PCR primers for identification and detection of <Emphasis Type="Italic">Phytophthora capsici</Emphasis> Leon

A PCR-based method was developed for the identification and detection of Phytophthora capsici in pepper plants. Three PCR primers (CAPFW, CAPRV1 and CAPRV2) specific for P. capsiciwere designed based on the sequence of its internal transcribed spacer regions. CAPFW/CAPRV1 amplify a 452 bp product from P. capsici DNA whereas CAPFW/CAPRV2 a 595 bp fragment; neither set amplifies DNA from pepper or several fungi pathogenic to pepper. In conventional (single-round) PCR, the limit of detection was 5 pg DNA for both primer sets, whereas in nested PCR the detection limit for both was of 0.5 fg. However, when the dilution series of target DNA were spiked with plant DNA, amplification declined two-fold in both conventional and nested PCR. The CAPFW/CAPRV2 set in conventional PCR was used to detect P. capsici DNA in inoculated plants. Detection occurred as soon as 8h post-inoculation in stem samples from infected but still symptomless plants. The method was also tested to detect fungal DNA in infected soils.     

Isolation and pathogenicity of Colletotrichum spp. causing anthracnose of strawberry in south west Spain

Anthracnose, caused by Colletotrichum spp., is a major disease of cultivated strawberry, Fragaria × ananassa. This study identifies the Colletotrichum spp. which causes strawberry anthracnose in the southwest of Spain. A survey of the region was carried out, and the strains isolated were identified as C. acutatum by using the polymerase chain reaction (PCR) with genus and species-specific primers, demonstrating that this species is currently the causal agent of strawberry anthracnose in the studied region. The pathogenicity of C. acutatum and C. gloeosporioides strains was evaluated on two principal strawberry cultivars (cvs Camarosa and Ventana) under field conditions, the latter being more pathogenic than the former.     

Effect of distribution and concentration of topically applied neonicotinoid insecticides in buffalograss,Buchloe dactyloides,leaf tissues on the differential mortality of Blissus occiduus under field conditions

BACKGROUND: Neonicotinoid insecticides are generally efficacious against many turfgrass pests, including several important phloem‐feeding insects. However, inconsistencies in control of western chinch bugs, Blissus occiduus, have been documented in field efficacy studies. This research investigated the efficacy of three neonicotinoid insecticides (clothianidin, imidacloprid and thiamethoxam) against B. occiduus in buffalograss under field conditions and detected statistically significant differences in B. occiduus numbers among treatments. A subsequent study documented the relative quantity and degradation rate of these insecticides in buffalograss systemic leaf tissues, using HPLC. RESULTS: Neonicotinoid insecticides initially provided significant reductions in B. occiduus numbers, but mortality diminished over the course of the field studies. Furthermore, while all three neonicotinoids were present in the assayed buffalograss leaf tissues, imidacloprid concentrations were significantly higher than those of clothianidin and thiamethoxam. Over the course of the 28 day study, thiamethoxam concentrations declined 700‐fold, whereas imidacloprid and clothianidin declined only 70‐fold and 60‐fold respectively. CONCLUSIONS: Field studies continued to verify inconsistencies in B. occiduus control with neonicotinoid insecticides. This is the first study to document the relative concentrations of topically applied neonicotinoid insecticides in buffalograss systemic leaf tissues. Copyright © 2012 Society of Chemical Industry     

Spatial and temporal analysis of coffee wilt disease caused by Fusarium xylarioides in Coffea canephora

Coffee wilt disease (CWD) caused by Fusarium xylarioides, considered to be a soil-inhabiting fungus, is endemic in several African countries, affecting commercially important coffee species and causing serious economic losses. Coffee wilt disease development in naturally infected Coffea canephora fields at the Coffee Research Institute in Uganda was assessed from April 2001 to March 2006 to generate information about temporal and spatial spread of the disease. Maps of diseased trees were also generated from the data. Semi-variance analysis was performed on the data to show the spatio-temporal structure of disease. Host influence on the spatio-temporal structure was deduced from the distribution pattern of diseased and healthy trees and analysis of variance. Results show that the temporal disease epidemic progress was slow. The disease was found to spread from initial infections to healthy neighbouring trees, resulting in an aggregated pattern. An infected tree could infect up to three healthy trees away, in any direction. Disease foci formed and expanded with time, coalescing but punctuated in spots planted with resistant hosts. There were varying levels of susceptibility among host genotypes, affecting the rates and levels of epidemic development. The implications of the findings to the control of CWD are discussed.     

Effects of shading and NO3:NH4 ratio on the yield,quality and N metabolism in strawberry

The effect of 50% shading and NO₃:NH₄ ratio (0:100, 75:25, 50:50, and 25:75) in the nutrient solution on growth, yield, quality and N metabolism in hydroponically grown strawberry (Fragaria × ananassa var Camarosa) was evaluated. Both fresh and dry weights of leaves were significantly lower when a high concentration of either NO₃ (100%) or NH₄ (75%) was the sole N source in the nutrient solution. In unshaded plants, increasing of both NH₄ and NO₃ ratio in the nutrient solution reduced photosynthetic (Pn) rate, however in shaded plants the reduction of Pn became more pronounced at a higher ratio of NH₄ in the nutrient solution. The yield in terms of fresh and dry weight of fruit per plant was significantly increased at the 75:25 and 50:50 (NO₃:NH₄) treatments. Fruit size was significantly affected by the treatments, so that the biggest fruits in both shaded and unshaded plants were obtained under the 75:25 and 50:50 (NO₃:NH₄) treatments. Total soluble solid (TSS) in unshaded plants was increased with increasing NH₄ ratio in the nutrient solution, however in shaded plants it was reduced at high NH₄ ratio in the nutrient solution. In both shaded and unshaded plants, higher concentration of NH₄ significantly reduced the post-harvest life of the fruits. The increase of tissue N concentration was nearly proportional to the NH₄ concentration in the nutrient solution. The activity of nitrate reductase (NR) was increased by increasing NH₄ from 0 to 50% and then reduced at a higher ratio of NH₄ in the solution. Shading increased NH₄ concentration so that the shaded plant had nearly twice as high NH₄ concentration in the leaves. The increase of NH₄ concentration induced by shading could be partially the reduction of NH₄ assimilate because of the shortage of carbohydrate.