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    Doubled Haploid Production – Mechanism and Utilization in Plant Breeding
    (Springer International Publishing, 2023) Yel, Ilknur; Dönmez, Betül Ayça; Yeşil, Binnur; Tekinsoy, Merve; Saeed, Faisal; Bakhsh, Allah
    The discovery of haploids in plants led plant breeders to help to produce double haploids. The chromosome numbers of double haploids are different from haploids. This technique shortens the time to produce homozygous plants as compared to the conventional breeding method. After doubling the chromosome, it produces two identical homologous chromosomes. This helps to achieve homozygosity of crop up to one generation early as compared to conventional breeding. Once an interesting gene combination is achieved through conventional breeding; further mixing of genes is considered a real challenge. The production of double haploids is dependent on haploid inducer lines. The current breakthroughs of molecular actors triggering induction of haploid in plants have an important role of processes related to the development of gamete, interactions, and stability of the genome. These findings allowed translation of induction of haploid, and genome editing technologies can be helpful to produce haploid inducer lines. These recent discoveries can be helpful for improved breeding strategies. Besides that, it also provides deeper information regarding sexual reproduction in plants. In this chapter, we discussed how we can produce haploid inducer lines and with the help of new biotechnological tools to produce double haploid plants. © The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Switzerland AG 2023.
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    Expression of Jaburetox 2-Ec in Potato Encodes Resistance Against Tuta absoluta Meyrick (Lepidoptera: Gelechiidae)
    (Springer, 2024) Tekinsoy, Merve; Bakhsh, Allah; Caliskan, Mehmet Emin
    The tomato leaf miner Tuta absoluta Meyrick is the most serious pest of potato worldwide. In the present study, we explored an alternate approach to induce resistance in potatoes against T. absoluta by engineering Jaburetox-2Ec, an insecticidal peptide derived from an isoform of urease from Jack bean (Canavalia ensiformis L.). The construct was developed by cloning a PCR-amplified fragment of Jaburetox-2Ec (270 bp) in a pBIN61 vector under the control of a 35S promoter and was named pBIN-JBTX. Furthermore, the recombinant plasmid expressing Jaburetox-2Ec was transformed into potato cv. Lady Olympia using Agrobacterium tumefaciens strain EHA105. The results showed the proper integration of the T-DNA region in primary transformants. Quantitative PCR assays showed the independent transgenic lines had enhanced transcript levels of Jaburetox. The leaf biotoxicity assays were performed by releasing pre-starved 2nd, 3rd, and 4th instar larvae of T. absoluta on transgenic potato plants. As a result, depending on the larval stage, the mortality rate of larvae feeding on transgenic plants ranged from 11 to 33%. Additionally, it was found that larvae feeding on control plants gained more weight than those feeding on transgenic plants. Besides that, deformities in larvae feeding on transgenic plants were also recorded. Overall, the results exhibited the insecticidal activity of Jaburetox 2-Ec against T. absoluta. The transgenic lines can be used as a source of germplasm for an efficient potato breeding programme.
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    Overexpression of jaburetox 2-Ec in potato (Solanum tuberosum L.)
    (Niğde Ömer Halisdemir Üniversitesi, 2022) Tekinsoy, Merve; Çalışkan, Mehmet Emin
    Bu çalışmada patates bitkisinde Jaburetox 2-Ec peptidinin ifadesinin artırılarak Tuta absoluta zararlısına karşı dirençli transgenik patates genotiplerinin geliştirilmesi amaçlanmıştır. Bu amaçla, 35S promotoru kontrolündeki hedef peptit dizisini içeren gen kaseti, Agrobakterium-aracılı gen transferi yöntemiyle Lady Olympia çeşidine aktarılmıştır. Deney yürütülürken bitkinin yaprak ve interned kısımları kullanılarak Agrobakterium (EHA 105) ile enfeksiyon sağlanmıştır.Transgenik bitki ve bakteri seçici markörü olarak 50 mg L-1 konsantrasyonlu Neomisin fosfotransferaz (nptII) geni kullanılmıştır.Yapılan moleküler analizler sonucunda hedef diziyi içeren T-DNA bölgesinin bitki genomuna entegre olduğu gösterilmiştir. Ardından yaprak besleme yöntemi ile transgenik bitkilerin ikinci, üçüncü ve dördüncü evre Tuta absoluta larvalarına karşı direnci test edilmiştir. Sonuçta transgenik bitkilerle beslenen larvalarda, larval döneme bağlı olarak 11-33 % arasında ölüm oranı tespit edilmiştir. Ayrıca control bitkilerle beslenen larvalarda ağırlık artışının transgenik bitkilerle beslenen larvalara kıyasla daha yüksek olduğu belirlenmiştir. Bu tez çalışması sonucunda, Jaburetox 2-Ec' nin Tuta absoluta zararlısına karşı insektisidal aktiviteye sahip olduğu, ancak bu etkinin doğrulanması açısından daha fazla araştırmaya ihtiyaç olduğu sonucuna varılmıştır.