Lily basal rot, caused by Fusarium oxysporum f. sp. lilii, is one of the most serious diseases of lily. Although the lily germplasm which is resistant to F. oxysporum has been used in disease-resistant breeding, few studies on its molecular mechanism of disease resistance have been reported. To comprehensively study the mechanism of resistance to F. oxysporum, transcriptome sequencings of root tissues from Lilium pumilum inoculated with F. oxysporum or sterile water for 6, 12, or 24 h were performed. A total of 50 GB of data were obtained from the transcriptome sequencings of the 6 L. pumilum samples, and 217 098 Unigenes were obtained after the de novo assembly, of which 38.36% Unigenes were annotated. The sequencing results showed that the numbers of differentially expressed genes at 6, 12, and 24 h after inoculation compared with the control were 111, 254, and 2500, respectively. The functional enrichment analysis of the differentially expressed genes showed that several pathways were involved in responses of L. pumilum, mainly including starch and sucrose metabolism, glycolysis/gluconeogenesis, phenylpropanoid biosynthesis, plant hormone signal transduction, flavonoid biosynthesis, vitamin B6 (VB6) biosynthesis, acid biosynthesis, proteasome, and ribosome. Transcription factor analysis revealed that the WRKY and ERF families played important roles in responses of L. pumilum to F. oxysporum. The results of this study elucidate the molecular responses to F. oxysporum in lily and lay a theoretical foundation for improving lily breeding and strategies for lily basal rot resistance.

Ascosphaera apis is the causative agent of honey bee chalkbrood disease, and spores are the only known source of infections. Interference with sporulation is therefore a promising way to manage A. apis. The versicolorin reductase gene (StcU-2) is a ketoreductase protein related to sporulation and melanin biosynthesis. To study the StcU-2 gene in ascospore production of A. apis, CRISPR/Cas9 was used, and eight hygromycin B antibiotic-resistant transformants incorporating enhanced green fluorescent protein (EGFP) were made and analyzed. PCR amplification, gel electrophoresis, and sequence analysis were used for target gene editing analysis and verification. The CRISPR/Cas9 editing successfully knocked out the StcU-2 gene in A. apis. StcU-2 mutants had shown albino and non-functional spore-cyst development and lost effective sporulation. In conclusion, editing of StcU-2 gene has shown direct relation with sporulation and melanin biosynthesis of A. apis; this effective sporulation reduction would reduce the spread and pathogenicity of A. apis to managed honey bee. To the best of our knowledge, this is the first time CRISPR/Cas9-mediated gene editing has been efficiently performed in A. apis, a fungal honey bee brood pathogen, which offers a comprehensive set of procedural references that contributes to A. apis gene function studies and consequent control of chalkbrood disease.

The study was undertaken in Amhara National Regional State, northeastern zones of Ethiopia in 2013/2014. The objectives of the study were to identify the existing local honeybees and their productive characteristics and behavior and generate baseline information for further research and development. Out of two zones, three potential districts have been purposively selected based on their potential. A total of 260 beekeepers personal interviewees were administered using structured and semi-structured questionnaires. The study was designed to address behavioral characteristics and productivities of different local honeybees. About 98.8% of interviewees were male and about 73.9% were at stages of literacy ranging from read and write to diploma level. Nearly 73.4% of beekeepers have more than 5 years of beekeeping experience. The average honeybee colony holding was 4.6 in highlands, 2.8 in mid altitude, and 2.7 in lowlands. About 34% of respondents harvest honey more than two times a year. Based on the honeybees individual color appearance, aggressiveness, and honey productivity, about 36.7% of the respondents have identified three local honeybee types, namely, brown red, black, and mixed. Brown red colored honeybee ecotype has been selected as best due to their better honey production, low aggressiveness, and relatively high reproductive swarming tendency.

Chalkbrood disease is caused by Ascosphaera apis which severely affects honeybee brood. Spore inoculation experiments shown pathogenicity varies among different strains and mutants, however, the molecular mechanism of pathogenicity is unclear. We sequenced, assembled and annotated the transcriptomes of wild type (SPE1) and three mutants (SPE2, SPE3 and SPE4) with reduced pathogenicity that were constructed in our previous study. Illumina sequencing generated a total of 394,910,604 clean reads and de novo Trinity-based assembled into 12,989 unigenes, among these, 9,598 genes were successfully annotated to known proteins in UniProt database. A total of 172, 3,996, and 650 genes were up-regulated and 4,403, 2,845, and 3,016 genes were down-regulated between SPE2-SPE1, SPE3-SPE1, and SPE4-SPE1, respectively. Overall, several genes with a potential role in fungal pathogenicity were detected down-regulated in mutants including 100 hydrolytic enzymes, 117 transcriptional factors, and 47 cell wall related genes. KEGG pathway enrichment analysis reveals 216 genes involved in nine pathways were down-regulated in mutants compared to wild type. The down-regulation of more pathways involved in pathogenicity in SPE2 and SPE4 than SPE3 supports their lower pathogenicity during in-vitro bioassay experiment. Expression of 12 down-regulated genes in mutants was validated by quantitative real time PCR. This study provides valuable information on transcriptome variation caused by mutation for further functional validation of candidate pathogenicity genes in A. apis.

The study was designed to assess the existing beekeeping practices, constraints, and potential of honeybee production in Simada district. The study was carried out in four proportionally selected kebeles of highland, midland, and lowland agro-ecology. Accordingly, a total sample size of 146 beekeepers, depending on their potential, was interviewed using a structured and semi-structured questionnaire. A semi-structured questionnaire, field observation, and focal group discussion were employed to collect primary data. Descriptive statistics such as mean, frequency, and standard deviation were used to analyze the data. The majority of beekeepers in the district are mail-headed, and the majority can read and write. Beekeepers practiced three hive types but mostly used traditional hives. The majority of honeybee colonies are found in midland agro-ecology, but they are not significantly different (P<0.05). About 57.5% of beekeepers obtain their colony through buying, and their colony increases through reproductive swarming. Beekeepers construct both traditional and top-bar hives from the surrounding available material. Frame hives were obtained from GOs on a credit basis. Beekeepers indicated that the majority of honey was harvested in October and November. The second minor harvesting period was from May to June, which depends on the nature of the yearly rainfall conditions. As the respondent's described, they stored honey below one year in a plastic jar, clay jar, and plastic sack when plenty of products were obtained and for medicinal value, unless they used honey during harvesting as a source of income. Predators and pests are major constraints on honey bee production, followed by pesticides and herbicides in the study area. Other identified beekeeping constraints were shown in relative order of importance: drought, death of colony, lack of water, migration, and disease are some of the problems that hinder productivity. Honeybees required feed supplementation during the dry season; about 28% of beekeepers fed their colonies with higher supplements made from February to April. The commonly used supplements were peas and bean flour (Shiro), barley flour (Besso), sugar, honey, and others, including Niger.

Introduction: This study aimed to evaluate effects of ethanol‐extracted honey bee propolis supplementation on the chemical and sensory attributes of raw cow’s milk in the Awi zone, Ethiopia. Methods: Propolis was harvested from three agroecologies in the Awi zone, Banja, Dangla, and Chagni, and extracted using 70% ethanol. Raw cow’s milk samples, collected from Injibara University Dairy Farm, were treated with varying concentrations of propolis (T1 (0), T2 (5), T3 (10), T4 (15), and T5 (20 mL)). The samples were analyzed for physicochemical parameters, including protein, fat, ash, solid non‐fat (SNF), pH, and titratable acidity (TA). Data analysis was conducted using SAS version 9.4 software. Result: Results showed that fat, SNF, ash, pH, TA, and protein contents were significantly ( p < 0.05) influenced by propolis concentration. The effect of propolis treatment on the shelf life and sensory quality of milk was investigated. Propolis‐treated milk demonstrated an extended shelf life of up to 72 h without any signs of deterioration. Sensory evaluation using a 9‐point Hedonic scale indicated that panelists’ preferences were not significantly influenced by the agroecological origin of the propolis harvested ( p > 0.05). However, sensory scores were highly significant with respect to propolis concentration treatment levels ( p < 0.01), suggesting that the concentration of propolis plays a critical role in determining consumer acceptability. These findings highlight the potential of propolis as a natural preservative for milk, enhancing shelf life while maintaining sensory quality. Discussion: The study concludes that the use of 5 mL and 10 mL of propolis concentrations, harvested from Guangua Woreda, extends the shelf life of raw milk without negatively impacting sensory characteristics of raw cow’s milk.