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    Artificial intelligence-based approaches to evaluate and optimize phytoremediation potential of in vitro regenerated aquatic macrophyte Ceratophyllum demersum L.
    (Springer Heidelberg, 2023) Aasim, Muhammad; Ali, Seyid Amjad; Aydin, Senar; Bakhsh, Allah; Sogukpinar, Canan; Karatas, Mehmet; Khawar, Khalid Mahmood
    Water bodies or aquatic ecosystem are susceptible to heavy metal accumulation and can adversely affect the environment and human health especially in underdeveloped nations. Phytoremediation techniques of water bodies using aquatic plants or macrophytes are well established and are recognized as eco-friendly world over. Phytoremediation of heavy metals and other pollutants in aquatic environments can be achieved by using Ceratophyllum demersum L. - a well-known floating macrophyte. In vitro regenerated plants of C. demersum (7.5 g/L) were exposed to 24, 72, and 120 h to 0, 0.5, 1.0, 2.0, and 4.0 mg/L of cadmium (CdSO4 & BULL;8H(2)O) in water. Results revealed significantly different relationship in terms of Cd in water, Cd uptake by plants, bioconcentration factor (BCF), and Cd removal (%) from water. The study showed that Cd uptake by plants and BCF values increased significantly with exposure time. The highest BCF value (3776.50) was recorded for plant samples exposed to 2 mg/L Cd for 72 h. Application of all Cd concentrations and various exposure duration yielded Cd removal (%) between the ranges of 93.8 and 98.7%. These results were predicted through artificial intelligence-based models, namely, random forest (RF), extreme gradient boosting (XGBoost), and multilayer perceptron (MLP). The tested models predicted the results accurately, and the attained results were further validated via three different performance metrics. The optimal regression coefficient (R-2) for the models was recorded as 0.7970 (Cd water, mg/L), 0.9661 (Cd plants, mg/kg), 0.9797 bioconcentration factor (BCF), and 0.9996 (Cd removal, %), respectively. These achieved results suggest that in vitro regenerated C. demersum can be efficaciously used for phytoremediation of Cd-contaminated aquatic environments. Likewise, the proposed modeling of phytoremediation studies can further be employed more comprehensively in future studies aimed at data prediction and optimization.
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    Development of Transgenic Potato Lines Expressing Ecdysone Receptor Gene of Colorado Potato Beetle
    (Springer, 2018) Bakhsh, Allah; Hussain, Tahira; Aasim, Muhammad; Pirlak, Ugur; Aksoy, Emre; Caliskan, Mehmet Emin
    [Abstract Not Availabe]
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    Development of transgenic tobacco lines with pyramided insect resistant genes
    (Tubitak Scientific & Technological Research Council Turkey, 2018) Bakhsh, Allah; Dinc, Tolga; Hussain, Tahira; Demirel, Ufuk; Aasim, Muhammad; Caliskan, Mehmet Emin
    Insect pests are among the major constraints rendering drastic decreases in crop yield. 'I he expression of stacked insecticidal genes in crops can lead to resistance durability and can delay the development of resistance in target insect pests. The present study was designed to introduce an insect resistance trait in locally cultivated Turkish tobacco cultivars (Basma and Nail) with pyramided insecticidal genes. Agrobacterium strain LBA4404 harboring plasmid pKGH4 with cry1Ac and cry2A genes under the control of 35S promoter was used to infect leaf discs of both cultivars; plasmid also contained uidA within the T-DNA region for earlier screening of putative transformants. The overall transformation efficiency was calculated as 30.7% and 18.8% in Basma and Nail, respectively. PCR results confirmed the integration of cry1Ac, cry2A, uidA, and nptII genes in 40 plants of Basma and 16 plants of Nail. ELISA results showed variation in expression of cry1Ac protein among transgenic plants varying from 0.017 to 0.607 mu g/g of fresh tissue. Bioassay results with potato tuber moth (Phthorimea operculella Zeller) showed significant mortality of the targeted pest on primary transfonnants. Furthermore, T1 transgenic progeny exhibited the inheritance of T-DNA in Mendelian as well as non-Mendelian fashion. The results revealed that lines can serve as a source of germplasm in tobacco breeding programs.
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    Targeted expression of insecticidal hybrid SN19 gene in potato leads to enhanced resistance against Colorado potato beetle (Leptinotarsa decemlineata Say) and tomato leafminer (Tuta absoluta Meyrick)
    (Springer, 2017) Ahmed, Hussein Abdullah Ahmed; Onarici, Selma; Bakhsh, Allah; Akdogan, Guray; Karakoc, Omer Cem; Ozcan, Sancar Fatih; Aydin, Gulsum
    The expression of insecticidal genes must be induced at appropriate time and in sufficient amount to confer protection against targeted pests. However, the increased scientific reports of resistance development in insect pest against insecticidal delta-endotoxins, produced by Bacillus thuringiensis, provide impetus for the development of alternative insect management strategies. The present study was conducted to investigate the importance of targeted expression of a hybrid insecticidal gene (SN19) in potatoes. For this purpose, two plant expression vectors were constructed by cloning hybrid SN19 gene (cry1Ba-domain I-III and cry1Ia-domain II) under the control of a wound-inducible promoter isolated from Asparagus officinalis (AoPR1) and CaMV 35S promoter, and were transferred to Agrobacterium tumefaciens strain EHA 105. Four potato genotypes (Marabel, Innovator, Tokat 10/1 and Tokat 6/24) were transformed with EHA 105 strain harboring pTF101.1 35S-SN19 and pTF101.1 AoPR1-SN19 constructs. Phosphinothricin (PPT) was used at concentration of 1 mg/l for selection of primary transformants. PCR results showed the presence of both introduced SN19 and bar genes in 43 plants out of total 154 putative transgenics. Expression of SN19 protein in primary transformants was confirmed by Western blot assays. The mechanical wounding of transgenic plants exhibited more accumulated levels of SN19 proteins during post wounding period. Leaf biotoxicity assays with Colorado potato beetle (Coleoptera) and tomato leafminer (Lepidoptera) exhibited 100% mortality of the pests in primary transformants. Based on our mortality results with both constructs, we concluded that the potato transgenic lines exhibited targeted expression of insecticidal gene under the control of AoPR1 promoter upon insect wounding with eliminated toxicity of Cry protein and hence can be further used effectively in potato breeding programme.