Emre AKSOY, Amir MAQBOOL, Buasimuhan ABUDUREYİMU

Arpa Nikotinamin Sentaz1 (HvNAS1) Genini Yüksek Seviyede İfade Eden Arabidopsis thaliana Bitkileri Demir Eksikliğine Dayanıklılık Gösterir

Demir (Fe) bitki gelişimi için önemli bir eser element olup, Fe eksikliğinde yetişen bitkilerde gelişen kloroza bağlı olarak verim kayıpları yaşanır. Bu kayıpların en aza indirilebilmesi için alınabilecek agronomik önlemlere ilaveten genetik mühendisliği aracılığıyla bitkilerin özellikleri etkin bir şekilde geliştirilebilir. Rizosferde yüksek miktarda bulunan demiri köklerine alabilmek için Arabidopsis thalianagibi dikotlar indirgenme temelli bir stratejiyi kullanırlarken, arpanın (Hordeum vulgare) da yer aldığı graminelerde şelasyon stratejisi evrimleşmiştir. Bu çalışmada, arpada bulunan ve demir ile kompleks oluşturabildiği bilinen nikotinaminin üretiminden sorumlu NİKOTİNAMİN SENTAZ1(HvNAS1) geni klonlanarak, Arabidopsis bitkilerinde konstitütif olarak yüksek seviyede ifade edilmiştir. Elde edilen T3Arabidopsis bitkilerinde HvNAS1ile birlikte demir alımından sorumlu yolakta yer alan Arabidopsis genlerinin de ifade seviyelerinin arttığı belirlenmiştir. Buna bağlı olarak, bitkilerin kök uzunluklarının, kök ve gövde yaş ağırlıklarının, ferrik şelat redüktaz enzim aktivitelerinin de arttığı belirlenmiştir.Ayrıca, transgenik Arabidopsis bitkilerinin kök ve gövdelerinde biriken demir ve çinko seviyelerinde önemli artışlar belirlenmiştir. Sonuç olarak, arpa HvNAS1genini yüksek seviyede ifade eden trangenik Arabidopsis bitkilerinin rizosferden daha fazla demir alabildiği ve bu demiri daha fazla gövdeye taşıyabildiği gösterilmiştir. Bu sayede genetik mühendisliği kullanılarak HvNAS1genini yüksek seviyede ifadeeden ve demir eksikliğine dayanıklı Arabidopsis bitkileri geliştirilmiştir.

Arabidopsis thalianaPlants Overexpressing the Barley Nicotinamine Synthase1(HvNAS1) Gene Show Tolerance to Iron Deficiency

Iron (Fe) is an important trace mineral for plant development, and plants grown in Fe deficiency experience yield losses due to the leaf chlorosis. In addition to agronomic measures that can be taken to minimize these losses, new plant genotypes can be developed effectively through genetic engineering. While dicots such as Arabidopsis thalianause a reduction-based strategy to uptake high amounts of iron from the rhizosphere, the chelation strategy has evolved in Gramineous plants including barley (Hordeum vulgare). In this study, barley NICOTIANAMINE SYNTHASE1(HvNAS1) gene, which is responsible for the production of nicotianamine that can complex with iron, was cloned and expressed at a constitutive high level in Arabidopsis plants. The expression levels of Arabidopsis genes encoding for the proteins involved in iron uptake increased together with HvNAS1in the T3Arabidopsis plants. Moreover, the root lengths, root and stem fresh weights, ferric chelate reductase enzyme activities of the plants also increased in the transgenic Arabidopsis plants under Fe deficiency. In addition, significant increases in iron and zinc levels were determined in the roots and shoots of transgenic Arabidopsis plants. As a result, transgenic Arabidopsis plants overexpressing thebarley HvNAS1gene can take up more iron from the rhizosphere and carry this iron to the shoots. This study demonstrates the power of genetic engineering to develop Arabidopsis plants overexpressing the HvNAS1gene and therefore tolerate iron deficiency.

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