农杆菌介导的海藻糖生物合成酵母基因转化春亚麻荠

A. Kvasko, A. S. Lazarets, S. Isayenkov, A. Yemets
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摘要

的目标。本研究的目的是获得亚麻荠(Camelina sativa, L.)添加酵母海藻糖合成基因TPS1和TPS2的克兰茨系提高抗旱性。方法。以苜蓿FEORZhYaF-1基因型种子进行离体培养。在3种不同的营养培养基上添加不同的激素组合,培养5日龄的亚麻荠幼苗的下胚轴节段和芽分生组织进行再生。利用TPS1和TPS2基因构建载体pGWB2-TPS1和pGWB2-TPS2进行遗传转化。结果。下胚轴外植体在添加1mg /l BAP和0.1 mg/l NAA的培养基上再生效率最高,分生组织外植体在添加1.5 mg/l BAP和0.5 mg/l NAA的培养基上再生效率最高。通过农杆菌介导的转化,在相应浓度的湿霉素培养基上获得茶树品系。通过pcr分析证实了所获得植株的转基因性质。结论。研究了苜蓿FEORZhYaF-1基因型的离体植株再生效率。采用两种外植体和两种载体构建pGWB2-TPS1和pGWB2-TPS2,分别植入TPS1和TPS2酵母海藻糖合成基因,获得了转基因亚麻荠系。
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Agrobacterium-mediated transformation of spring camelina with yeast genes of trehalose biosynthesis
Aim. The aim of the study was the obtaining of Camelina sativa (L.) Crantz lines with yeast genes of trehalose synthesis TPS1 and TPS2 to increase their resistance to drought. Methods. Seeds of C. sativa genotype FEORZhYaF-1 were used for in vitro culture establishment. For this hypocotyl segments and shoot meristems of 5-days-old camelina seedlings were cultivated on three different nutrient media for regeneration supplemented with various hormone combinations. Vector constructions pGWB2-TPS1 and pGWB2-TPS2 with TPS1 and TPS2 genes have been used for genetic transformation. Results. The highest efficiency of plant regeneration from hypocotyl explants was found on medium supplemented with 1 mg/l BAP and 0.1 mg/l NAA, and from meristem explants – on medium with 1.5 mg/l BAP and 0.5 mg/l NAA. Agrobacterium-mediated transformation was conducted out, and camelina lines were picked up on corresponding medium with selective concentration of hygromycin. Transgenic nature of obtained plants was confirmed by PCR-analysis. Conclusions. The efficiency of in vitro plant regeneration of C. sativa genotype FEORZhYaF-1 has been investigated. Two types of explants and two vector constructions pGWB2-TPS1 and pGWB2-TPS2 with TPS1 and TPS2 yeast trehalose synthesis genes have been used for obtaining of transgenic camelina lines.
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