[PLANT GENETIC TRANSFORMATION USING CARBON NANOTUBES FOR DNA DELIVERY].

TSitologiia i genetika Pub Date : 2015-11-01
O M Burlaka, Ya V Pirko, A I Yemets, Ya B Blume
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Abstract

The possibility of exploiting carbon nanotubes (CNTs)-based nanocarriers to deliver genes into protoplasts, callus and mesophyll explants of plants was examined. Using single-walled CNTs (SWCNTs) at the concentration of 20 μg/ml and multi-walled CNTs (MWCNTs) at the concentration of 15 μg/ml genetic transformation of Nicotiana tabacum L. mesophyll protoplasts with plasmid pGreen 0029 was carried out and transient expression of reporter yfp gene in the protoplasts was observed. Using SWCNTs at the concentration of 40 μg/ml and MWCNTs at the concentration of 30 μg/ml genetic transformation of N. tabacum callus and leaf explants with nptII gene as a part of plasmid pGreen 0029 was carried out. As a result plant regeneration on selective medium containing 50 mg/lkanamycin was shown. SWCNTs-based nanocarriers de-onstrated their appli-ability to transform protoplasts as well as walled plant cells. Whereas, MWCNTs-based nano-arriers were suitable only for transformation of proto-lasts due to the limiting role of cellulose walls in cell penetration.

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[利用碳纳米管传递 DNA 的植物基因转化]。
研究人员探讨了利用基于碳纳米管(CNTs)的纳米载体向植物原生质体、胼胝体和叶肉外植体传递基因的可能性。使用浓度为 20 μg/ml 的单壁碳纳米管(SWCNTs)和浓度为 15 μg/ml 的多壁碳纳米管(MWCNTs)对烟草叶肉原生质体进行了质粒 pGreen 0029 的遗传转化,并观察到报告基因 yfp 在原生质体中的瞬时表达。使用浓度为 40 μg/ml 的 SWCNTs 和浓度为 30 μg/ml 的 MWCNTs 对 N. tabacum 胼胝体和叶外植体进行了遗传转化,质粒 pGreen 0029 中含有 nptII 基因。结果表明,植物在含有 50 mg/lkanamycin 的选择性培养基上再生。基于 SWCNTs 的纳米载体证明了其转化原生质体和植物贴壁细胞的应用能力。而基于 MWCNTs 的纳米载体只适用于原生质体的转化,这是因为纤维素壁在细胞渗透中起限制作用。
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[PLANT GENETIC TRANSFORMATION USING CARBON NANOTUBES FOR DNA DELIVERY]. OBTAINING OF THE TRANSGENIC HELIANTHUS TUBEROSUS L. PLANTS, CALLUS AND "HAIRY" ROOT CULTURES ABLE TO EXPRESS THE RECOMBINANT HUMAN INTERFERON ALPHA-2b GENE. [Peculiarities of the influence of chemical and physical factors on cytogenetic indices of root meristem in Pisum sativum L]. [Molecular and genetic polymorphism of cultivated soybean detected by AP-PCR, SSRP and SSR]. [Variation of ultrastructural organization of the mitochondria of pea cells in vitro].
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