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    Crispr/cas9-mediated immunity in plants against pathogens
    (Caister Academic Press, 2018) Sameeullah, Muhammad; Khan, Faheem Ahmed; Özer, Göksel; Aslam, Noreen; Gurel, Ekrem; Waheed, Mohammad Tahir; Karadeniz, Turan
    Global crop production is highly threatened due to pathogen invasion. The huge quantity of pesticides application, although harmful to the environment and human health, is carried out to prevent the crop losses worldwide, every year. Therefore, understanding the molecular mechanisms of path-ogenicity and plant resistance against pathogens is important. The resistance against pathogens is regulated by three important phytohormones, viz. salicylic acid (SA), jasmonic acid (JA) and ethylene (ET). Here we review the possible role of CRISPR technology to understand the plant pathogenicity by mutating genes responsible for pathogen invasion or up-regulating the phytohormones genes or resistant genes. Thus hormone biosynthesis genes, receptor and feeding genes of pathogens could be important targets for modifications using CRISPR/Cas9 following multiplexing tool box strategy in order to edit multiple genes simultane-ously to produce super plants. Here we put forward our idea that the genes would be either mutated in case of plant receptor protein targets of pathogens or up-regulation of resistant genes or hormone bio-synthesis genes will be better choice for resistance against pathogens. © 2018, Caister Academic Press. All rights reserved.
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    CRISPR/Cas9-mediated Immunity in Plants Against Pathogens
    (Mdpi, 2018) Sameeullah, Muhammad; Khan, Faheem Ahmed; Ozer, Goksel; Aslam, Noreen; Gurel, Ekrem; Waheed, Mohammad Tahir; Karadeniz, Turan
    Global crop production is highly threatened due to pathogen invasion. The huge quantity of pesticides application, although harmful to the environment and human health, is carried out to prevent the crop losses worldwide, every year. Therefore, understanding the molecular mechanisms of pathogenicity and plant resistance against pathogens is important. The resistance against pathogens is regulated by three important phytohormones, viz. salicylic acid (SA), jasmonic acid (JA) and ethylene (ET). Here we review the possible role of CRISPR technology to understand the plant pathogenicity by mutating genes responsible for pathogen invasion or up-regulating the phytohormones genes or resistant genes. Thus hormone biosynthesis genes, receptor and feeding genes of pathogens could be important targets for modifications using CRISPR/Cas9 following multiplexing tool box strategy in order to edit multiple genes simultaneously to produce super plants. Here we put forward our idea that the genes would be either mutated in case of plant receptor protein targets of pathogens or up-regulation of resistant genes or hormone biosynthesis genes will be better choice for resistance against pathogens.
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    Enhanced somatic embryogenesis, plant regeneration and total phenolic content estimation in Lycium barbarum L.: a highly nutritive and medicinal plant
    (Springer, 2022) Verma, Sandeep Kumar; Gantait, Saikat; Mukherjee, Epsita; Gurel, Ekrem
    Lycium barbarum (Solanaceae), commonly known as Goji (or wolfberry), is popular for its nutritive and medicinal properties and is called a “super fruit” or “super food”. Considering the importance of the economic utility, the potential of different explants (hypocotyl, leaf and root) of L. barbarum for plant regeneration and somatic embryogenesis has been evaluated in the present study. Two sets of experiments were carried out; the first compared Murashige and Skoog (MS) medium supplemented with different concentrations of 2,4-dichlorophenoxy acetic acid (2,4-D), N6-benzylaminopurine, thidiazuron (TDZ), kinetin, and zeatin alone, while the second set tested the combinations of TDZ with 2,4-D to induce callus and subsequent shoot or embryo formation, respectively. For callus and subsequent shoot induction (for the first set of the experiment), 2,4-D was the most effective for callus induction (100%), while TDZ at 1 mg/L produced a mean of 5 shoots per callus. For the second set of experiment spontaneous induction of somatic embryos as well as subsequent maximum shoot regeneration was recorded in 0.25 mg/L 2,4-D+ 1 mg/L TDZ supplemented MS medium. Hypocotyl explant proved to be the most responsive organ that induced as many as 6 somatic embryos and subsequent 9 regenerated shoots per callus. Later, the plantlets were successfully acclimatized (100%) and finally transferred to the greenhouse. Total phenolic content was measured from shoot, brown callus and white callus of L. barbarum L. grown in MS medium supplemented with auxin alone (for callus) and auxin-cytokinin (for shoots). The highest amount of total phenolic content (640 mg GAE/g DW) was obtained in the shoot. The described protocol provides a simple way to regenerate plants through direct and indirect organogenesis as well as somatic embryogenesis, which would be useful for mass propagation, large-scale production of secondary metabolites, germplasm conservation, and genetic transformation studies in this medicinally important species. © 2022, The Author(s), under exclusive licence to Korean Society of Crop Science (KSCS).
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    In silico designing and characterization of outer membrane protein K (OmpK) from Vibrio anguillarum and its expression in Nicotiana tabacum for the development of a plant-based vaccine against fish vibriosis
    (Elsevier, 2024) Malik, Muhammad Suleman; Elahi, Iqra; Sameeullah, Muhammad; Ijaz, Fatima; Batool, Neelam; Khalid, Fatima; Gurel, Ekrem
    Vibriosis is caused by Vibrio anguillarum in various species of aquaculture. A novel, secure, and stable vaccine is needed to eradicate vibriosis. Here, for reverse vaccinology and plant -based expression, the outer membrane protein K (OmpK) of V. anguillarum was chosen due to its conserved nature in all Vibrio species. OmpK, an ideal vaccine candidate against vibriosis, demonstrated immunogenic, non -allergic, and non-toxic behavior by using various bioinformatics tools. Docking showed the interaction of the OmpK model with TLR-5. In comparison to costly platforms, plants can be used as alternative and economic bio-factories to produce vaccine antigens. We expressed OmpK antigen in Nicotiana tabacum using Agrobacterium-mediated transformation. The expression vector was constructed using Gateway (R) cloning. Transgene integration was verified by polymerase chain reaction (PCR), and the copy number via qRT-PCR, which showed two copies of transgenes. Western blotting detected monomeric form of OmpK protein. The total soluble protein (TSP) fraction of OmpK was equivalent to 0.38% as detected by ELISA. Mice and fish were immunized with plant -derived OmpK antigen, which showed a significantly high level of anti-OmpK antibodies. The present study is the first report of OmpK antigen expression in higher plants for the potential use as vaccine in aquaculture against vibriosis, which could provide protection against multiple Vibrio species due to the conserved nature OmpK antigen.
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    Nanoelicitation: A Promising and Emerging Technology for Triggering the Sustainable In Vitro Production of Secondary Metabolites in Medicinal Plants
    (Springer Nature, 2022) Javed, Rabia; Yucesan, Buhara; Zia, Muhammad; Gurel, Ekrem
    To obtain larger amounts of secondary metabolites is essentially needed by the global industrial market that is not feasible by traditional methods because these are time-consuming and result in eradication of plant stock by overexploitation. Therefore, in vitro culturing techniques are adopted to obtain the maximum quantity of secondary metabolites in a minimum time. Elicitors are key players for getting desired yields of secondary metabolites in plants. Eliciting the in vitro cell and tissue cultures is an efficient approach for the production of medicinally important plant secondary metabolites. Different parameters of optimized micropropagation protocols are exploited to be used as elicitors for further enhancement of secondary metabolites from medicinal plant species. The secondary metabolites produced by the plant cells include phenolics, flavonoids, alkaloids, terpenoids, and tannins. These metabolites are boosted under stressful conditions, whether biotic or abiotic in origin. The potential role of nanoparticles in the enhancement of secondary metabolic products in medicinal plants is a recent hot topic in the field of medicinal plant biotechnology. Nanoparticles have evolved as potent novel elicitors that significantly stimulate medicinal plant secondary metabolism. Various kinds of nanoparticles including metallic and metallic oxide nanoparticles and carbon-based nanomaterials are believed to induce abiotic stress to medicinal plants under in vitro conditions by which plant defense system is elicited, triggering biochemical as well as physiological responses, consequently producing enhanced and sustainable quantities of secondary metabolites. These industrially important bioactive metabolites are beneficial for the prevention of multiple diseases in the health-care system. Therefore, nanoelicitation should be applied as an effective tool for ameliorated stimulation and accumulation of secondary metabolites. However, in some cases, after the efficient uptake and translocation, nanoparticles produce deleterious effects causing phytotoxicity. © The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2022.