Understanding the molecular basis of Zymoseptoria tritici-wheat interactions

Zymoseptoria tritici is causal to one of the most devastating diseases of wheat, Septoria tritici blotch, STB, in Europe. An estimated 70% of fungicides deployed in Europe is for managing STB. Fundamental research has been undertaken with a focus on understanding the mechanistic basis of STB disease with partial success. However, lack of molecular tools has withheld functional genomics leading to extensive gaps in knowledge of infection biology of Z.tritici. Current research was conducted to develop molecular tools to facilitate forward and reverse genetic screens in Z.tritici for characterizing putative effector genes involved in STB disease. As a part of this study, a genome editing tool, CRISPR/Cas9, was employed to facilitate high frequency targeted genome manipulations in Z.tritici. Protoplasts were transformed with Cas9 protein and gRNA in form of preassembled ribonucleoprotein (RNP) complex targeting nitrate reductase (NiaD) and Orotidine 5/-phosphate-decarboxylase (PyrG). Subsequent screening revealed no mutations in either gene. In attempt to enhance efficacy, RNP complex was co-transformed with homologous directed repair (HDR) cassette with 1kb homologous DNA flanking targeted cut site in NiaD. Again though, no CRISPR-induced mutations were identified questioning suitability of CRISPR/Cas9 in Z.tritici. In contrast, transformation of RNP into Parastagonospora nodorum targeting Tox3 effector gene resulted in 100% editing of mutants. Efficacy of RNP was further tested when co-transformed with a Tox3-HDR cassette harbouring 1kb of homologous flanks which demonstrated homologous recombination (HR) efficiencies exceeding 70%. Further transformation of Tox3-HDR cassette with 50bp microhomology flanks was also successful with 25% HR efficacy. These data highlight a significant potential of CRISPR/Cas9 in expediting transgene-free gene knockouts in P.nodorum. However, its efficacy is dependent on targeted pathogen. Contrary to expectations set by successful application of CRISPR/Cas9 for targeted gene deletions in Z.tritici, four effector candidates were identified based on previous RNA sequencing data in Solomon lab. Agrobacterium-mediated transformation was carried out for successful mutant generation. Two candidate genes, Zt10 and Zt61 were found to play a potential role in pathogenicity inplanta. It was found that Zt10 gene might be important for necrotic lesion development as deletion of this gene resulted in significantly reduced necrotic lesions over time across different wheat cultivars. Once mutated, Zt61 candidate gene showed reduction in disease symptoms inplanta. Zt61 has a predicted cellulose binding domain and shares high structure similarity to bacterial expansin protein, BsEXLX1. Hence, Zt61 gene was cloned and expressed in E.coli, to purify Zt61 protein, further confirmed via mass spectrometry. Upon wheat infiltration, it caused a general necrotic and chlorotic activity, NCA. Mutation in its three amino acids led to decrease NCA suggesting their partial role. Discovery of candidates like Zt10 and Zt61 provide an insight into disease mechanism of Z.tritici. Complementary to these reverse genetics approaches, a forward genetics approach was also employed to screen for Z.tritici potential avirulence genes that are recognized by Stb19 gene in Lorikeet. For this, UV-mutagenesis was performed to generate a library of Z.tritici mutants and screened for pycnidia development. Four mutants were able to develop pycnidia on Lorikeet indicating a potential mutation in the encoding AvrStb19 gene. Subsequent analysis of these four mutants using Illumina sequencing failed to identify a potential AvrStb19. Besides due to a number of unforeseen challenges, a list of nine putative candidates was obtained to be characterized functionally in future studies. Hence, this project did shed light on the technical limitations of this approach and provided important information on how to improve such experiment in Z. tritici in future

Greening of supply chain to drive performance through logical integration of supply chain resources

Sustainability has always been a concern of humankind in one form or the other. Still, it has come into sharper focus after the promulgation of the sustainable development goals in 2015 and the disruptive forces unleashed by the COVID‐19 pandemic. Supply chain management is an aspect of business operations wherein the need for sustainability has been felt more keenly. The academic and business understanding of various nuances of incorporating sustainability, particularly environmental concerns in the supply chain, is still evolving. Our study seeks to enrich the growing literature in the area by proposing to uncover a novel, logical sequence of intangible supply chain resources that can amplify the impact of green supply chain management practices (GSCMPs) on business performance. We use the dual‐theoretical lens of a resource‐based view and stakeholder theories to conceptualize the sequential mediational role of supply chain visibility, resilience, and robustness between GSCMP and performance. Analyzing data collected from 318 individuals working in the manufacturing sector in the United Kingdom, we found a positive direct association of GSCMP with performance and the serial mediational role of visibility and robustness between the two. The findings of our study are pertinent for theorists as well as managers

Empirical examination of societal, financial and technology-related challenges amid COVID-19 in service supply chains: evidence from emerging market

Purpose: This paper reports an empirical examination of the societal, financial and technology-related challenges amid the ongoing pandemic (COVID-19) in the service supply chain network. Design/methodology/approach: A combination of qualitative approach (for items generation pertinent to the constructs involved) and quantitative approach (self-administered questionnaires from the top and middle management of the sampled companies) was used for data collection. In total, 272 complete responses were received and analyzed through structural equation modeling. Findings: The results provided empirical evidence that social and physical distancing, travel restrictions, work from home and lockdown practices have two conflicting effects: On one hand, these practices have contributed to the reduction of economic activities, including the low economic outlook, low productivity, high unemployment, poverty, fall in customer demands, dissatisfaction and mental health, that ultimately impacts rise financial and societal issues. On the other hand, the results revealed an insignificant influence of COVID-19 on creating technology-related challenges in the service sector. It shows that the organizations are doing well in combating the technology-related challenges amidst the current pandemic. Research limitations/implications: Findings of the inquiry recommend implications for the services industry to harmonize a comprehensive strategy and revisit the global norms in sustainable supply chain management activities that have been the backdrop in their operations for a long time. Practical implications: Findings of the inquiry recommend implications for the services industry to harmonize a comprehensive strategy and revisit the global norms in supply chain management activities that have been the backdrop in their operations for a long time. Originality/value: Prior studies in the context of the COVID-19 outbreak and its implications have given more attention to the exploratory and theoretical discussion than to empirical evidence. This paper contributes to filling this knowledge gap by empirically exploring the societal, financial and technology-related challenges created by COVID-19. The analysis in this paper covers three dimensions of the PEST model, namely economic, societal and technological factors. This study also helps in laying out a platform for investigating the PEST (political, economic, social and technological) model for guiding the services industry in strategic decision-making in a new era due to COVID-19

Socio-economic and technological new normal in supply chain management : lessons from COVID-19 pandemic

Purpose This paper explores the new normal activities and strategic responses of the service industry towards the challenges created by the coronavirus disease 2019 (COVID-19) outbreak and other constructs and validates the measurement scale for socio-economic and technological new normal activities following lockdown and social distancing practices. Design/methodology/approach First, structured interviews with 28 participants helped us generate items and develop survey instruments for cross-sectional data collection in the second phase. So, the authors received 256 complete responses from the top and middle management of the services industry. Exploratory factor analysis helped us explore the factors and reliability of the items. Confirmatory factor analysis aided us in generating and confirming the factorial structure of the constructs. Findings Results indicated that amid COVID-19's pandemic, new normal activities are emerging in which organizations are deploying crisis strategies to safeguard their business and stakeholders. Organizations are re-opening swiftly, focusing on digital transformation, developing digital platforms for ease in working and improved consumer services, to name a few operational changes. Practical implications Discussion on empirical analysis revolves around the guidelines to service industry's managers and top management to improve shortcomings in combating the challenges they face in their operations. Originality/value Prior studies have provided substantial insights on the COVID-19 pandemic, but relatively little research exists on new normal activities in the supply chain network of the service industry. Among other reasons for such less empirical evidence on new normal activities is the unavailability of a comprehensive tool for measuring the socio-economic and technological new normal activities. This paper is a contribution to bridging this knowledge gap.©2022 Emerald Publishing Limited. This manuscript version is made available under the Creative Commons Attribution–NonCommercial 4.0 International (CC BY–NC 4.0) license, https://creativecommons.org/licenses/by-nc/4.0/fi=vertaisarvioitu|en=peerReviewed

Understanding Influencers of College Major Decision: The UAE Case

This study aims to understand and analyze what influences female students to choose a college major in the United Arab Emirates (UAE). To accomplish our target, we conducted a survey with mostly female first-year undergraduate students (N = 496) at Zayed University to understand the personal, social, and financial factors influencing students’ major choices. Further, this study also asked students to specify their actions before deciding on their major and assessed the information that could be helpful for future students to decide on their majors. Last, the study investigated how Science, Technology, Engineering, and Mathematics (STEM) students differ from other students in their major decision. The results show that financial factors such as income and business opportunities related to the major are crucial. Further, gender suitability for the job and passion are influential. Students conduct internet searches, use social media, and read brochures in the process of major decisions. Moreover, students think job alignment with the UAE vision and information related to job availability, income, and skills are critical for future students to decide on their major. Finally, STEM students are more influenced by business opportunities, prestige, and career advancement than others

Socio-economic and technological new normal in supply chain management: lessons from COVID-19 pandemic

Purpose: This paper explores the new normal activities and strategic responses of the service industry towards the challenges created by the coronavirus disease 2019 (COVID-19) outbreak and other constructs and validates the measurement scale for socio-economic and technological new normal activities following lockdown and social distancing practices. Design/methodology/approach: First, structured interviews with 28 participants helped us generate items and develop survey instruments for cross-sectional data collection in the second phase. So, the authors received 256 complete responses from the top and middle management of the services industry. Exploratory factor analysis helped us explore the factors and reliability of the items. Confirmatory factor analysis aided us in generating and confirming the factorial structure of the constructs. Findings: Results indicated that amid COVID-19\u27s pandemic, new normal activities are emerging in which organizations are deploying crisis strategies to safeguard their business and stakeholders. Organizations are re-opening swiftly, focusing on digital transformation, developing digital platforms for ease in working and improved consumer services, to name a few operational changes. Practical implications: Discussion on empirical analysis revolves around the guidelines to service industry\u27s managers and top management to improve shortcomings in combating the challenges they face in their operations. Originality/value: Prior studies have provided substantial insights on the COVID-19 pandemic, but relatively little research exists on new normal activities in the supply chain network of the service industry. Among other reasons for such less empirical evidence on new normal activities is the unavailability of a comprehensive tool for measuring the socio-economic and technological new normal activities. This paper is a contribution to bridging this knowledge gap

Combination of two rare mutations causes β-thalassaemia in a Bangladeshi patient

Screening of mutations that cause β-thalassaemia in the Bangladeshi population led to the identification of a patient with a combination of two rare mutations, Hb Monroe and HBB: −92 C > G. The β-thalassaemia major male individual was transfusion-dependent and had an atypical β-globin gene cluster haplotype. Of the two mutations, Hb Monroe has been characterized in detail. Clinical effects of the other mutation, HBB: −92 C > G, are unknown so far. Bioinformatics analyses were carried out to predict the possible effect of this mutation. These analyses revealed the presence of a putative binding site for Egr1, a transcription factor, within the HBB: −92 region. Our literature survey suggests a close relationship between different phenotypic manifestations of β-thalassaemia and Egr1 expression

Assessing the efficacy of CRISPR/Cas9 genome editing in the wheat pathogen Parastagonspora nodorum

Background The genome-editing tool CRISPR/Cas9 has revolutionized gene manipulation by providing an efficient method to generate targeted mutations. This technique deploys the Cas9 endonuclease and a guide RNA (sgRNA) which interact to form a Cas9-sgRNA complex that initiates gene editing through the introduction of double stranded DNA breaks. We tested the efficacy of the CRISPR/Cas9 approach as a means of facilitating a variety of reverse genetic approaches in the wheat pathogenic fungus Parastagonospora nodorum. Results Parastagonospora nodorum protoplasts were transformed with the Cas9 protein and sgRNA in the form of a preassembled ribonuclear protein (RNP) complex targeting the Tox3 effector gene. Subsequent screening of the P. nodorum transformants revealed 100% editing of those mutants screened. We further tested the efficacy of RNP complex when co-transformed with a Tox3-Homology Directed Repair cassette harbouring 1 kb of homologous flanking DNA. Subsequent screening of resulting transformants demonstrated homologous recombination efficiencies exceeding 70%. A further transformation with a Tox3-Homology Directed Repair cassette harbouring a selectable marker with 50 bp micro-homology flanks was also achieved with 25% homologous recombination efficiency. The success of these homology directed repair approaches demonstrate that CRISPR/Cas9 is amenable to other in vivo DNA manipulation approaches such as the insertion of DNA and generating point mutations. Conclusion These data highlight the significant potential that CRISPR/Cas9 has in expediting transgene-free gene knockouts in Parastagonospora nodorum and also in facilitating other gene manipulation approaches. Access to these tools will significantly decrease the time required to assess the requirement of gene for disease and to undertake functional studies to determine its role.HK was supported by an Australian Government Research Training Program (RTP) Scholarship. SW was supported by Australian Research Council Grant DE16010089

Characterization and Antibacterial Evaluation of Biodegradable Mannose‐Conjugated Fe‐MIL‐88NH2 Composites Containing Vancomycin against Methicillin‐Resistant Staphylococcus aureus Strains

The emergence of bacterial resistance has increased the economic burden of infectious diseases dramatically during the previous few decades. Multidrug resistance (MDR) is difficult to cure in both Gram‐negative and positive bacteria and is often incurable with traditional and broad-range antibiotics. Therefore, developing techniques to increase the antibacterial activity of therapeutic drugs is essential. Metal‐organic frameworks (MOFs) are extremely versatile hybrid materials made of metal ions coupled via organic bridging ligands. They have been widely used as an excellent vehicle for drug delivery due to their low toxicity, biodegradability, and structural stability upon loading and functionalization. The present study focused on the synthesis of mannose (MNS)‐coated MOFs with enhanced surface contact with S. aureus cells. The MNS coating on the surface of MOFs enhances their adherence to bacteria by binding to lectins present on the bacterial cell, resulting in improved VCM cellular penetration and activity against resistant bacteria. Various techniques, including atomic force microscopy, DLS, TGA, FT‐IR, and DSC, were employed to analyze MNS‐coated MOFs. They were also evaluated for their efficacy against resistant S. aureus. The results indicated that when VCM was loaded into MNS‐coated MOFs, their bactericidal activity rose dramatically, resulting in the greater suppression of resistant S. aureus. AFM investigation of S. aureus strains demonstrated total morphological distortion after treatment with MNS‐coated drug-loaded MOFs. The results of this work suggest that MNS‐coated MOFs may be effective in reversing bacterial resistance to VCM and open new pathways for improving antibiotic therapy for diseases associated with MDR