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[PLANT GENETIC TRANSFORMATION USING CARBON NANOTUBES FOR DNA DELIVERY]. [利用碳纳米管传递 DNA 的植物基因转化]。
Pub Date : 2015-11-01
O M Burlaka, Ya V Pirko, A I Yemets, Ya B Blume

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.

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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引用次数: 0
OBTAINING OF THE TRANSGENIC HELIANTHUS TUBEROSUS L. PLANTS, CALLUS AND "HAIRY" ROOT CULTURES ABLE TO EXPRESS THE RECOMBINANT HUMAN INTERFERON ALPHA-2b GENE. 获得能够表达重组人类干扰素 ALPHA-2b 基因的转基因 HELIANTHUS TUBEROSUS L. 植物、胼胝体和 "毛状 "根培养物。
Pub Date : 2015-09-01
O M Maistrenko, Yu S Luchakivska, N M Zholobak, M Ya Spivak, M V Kuchuk

This work is the first to our knowledge to describe the successful attempt of Agrobacterium rhizogenes-mediated transformation of topinambour in order to obtain the transgenic H. tuberosus plants, callus and "hairy" root cultures. The plasmid vectors contained the sequence of interferon gene fused with Nicotiana plumbagenifolia L. calreticulin apoplast targeting signal driven by 35S CaMV promoter or root-specific Mll promoter. Nearly 75% isolated Ri-root lines and callus cultures were proved (by PCR analysis) to contain HuINFa-2b transgene. We also managed to obtain H. tuberosus transgenic plants through somatic embryogenesis on the transgenic "hairy" root culture. The obtained transgenic H. tuberosus cultures exhibited high-level antiviral activity that ranged from 2000 to 54500 IU/g FW that makes this crop considered a promising source of recombinant interferon alpha 2b protein.

据我们所知,这项研究首次成功地描述了根瘤农杆菌介导的topinambour转化,从而获得转基因H. tuberosus植株、胼胝体和 "多毛 "根培养物。质粒载体包含由 35S CaMV 启动子或根部特异性 Mll 启动子驱动的干扰素基因序列与 Nicotiana plumbagenifolia L. calreticulin apoplast targeting signal 融合。近 75% 分离出的 Ri-root 株系和胼胝体培养物被证明(通过 PCR 分析)含有 HuINFa-2b 转基因。我们还设法通过转基因 "多毛 "根培养物上的体细胞胚胎发生获得了转基因 H. tuberosus 植物。所获得的转基因 H. tuberosus 培养物表现出高水平的抗病毒活性,从 2000 IU/g FW 到 54500 IU/g FW 不等,这使得该作物被认为是重组干扰素 alpha 2b 蛋白的一个有前途的来源。
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引用次数: 0
[Peculiarities of the influence of chemical and physical factors on cytogenetic indices of root meristem in Pisum sativum L]. [化学和物理因素对豌豆根分生组织细胞遗传指数影响的特殊性]。
Pub Date : 2002-05-01
S S Rudenko, T V Morozova, S S Kostyshyn, Ie D Strel'chenko

The results of comparative analysis of cytogenetic effects of various chemical and physical factors on meristemic cells of field pea (Pisum sativum L.) are presented. Cytogenetic effects of aluminum, cadmium and selenium compounds as well as X-ray irradiation were studied. Cytogenetic indices were investigated for different doses of the above factors and peculiarities of their influence on the ratio of mitotic phases were analyzed. The increasing in the proportion of cells at the specific mitotic stage was revealed. There were anaphase stage for aluminum chloride, prophase stage for cadmium chloride, meta- and anaphase stages for sodium selenate, and telophase stage for X-rays. Investigated chemical elements may be ranged according to their ability to induce the aberrant anaphase frequency in the row: Se (from 3.75 x 10(-6) M) > A1 (from 3.86 x 10(-5) M) > Cd (from 8.44 x 10(-5) M). Maximal experimental dose of X-ray irradiation (9.03 x 10(-3) C/kg) induces the frequency of chromosome aberration similar to those induced by maximal experimental doses of aluminum and selenium (3.86 x 10(-4) M and 8.34 x 10(-6) M).

本文介绍了各种化学和物理因素对大田豌豆(Pisum sativum L.)分生组织细胞的细胞遗传效应的比较分析结果。研究了铝、镉和硒化合物以及 X 射线照射对细胞遗传的影响。研究了不同剂量上述因素的细胞遗传指数,并分析了它们对有丝分裂期比例影响的特殊性。结果表明,处于特定有丝分裂阶段的细胞比例在增加。氯化铝的有丝分裂期为无丝分裂期,氯化镉的有丝分裂期为前期,硒酸钠的有丝分裂期为中期和无丝分裂期,X 射线的有丝分裂期为端期。所研究的化学元素可根据其诱导行间无丝分裂异常频率的能力进行划分:硒(从 3.75 x 10(-6) M)> A1(从 3.86 x 10(-5) M 起)> 镉(8.44 x 10(-5) M)。X 射线辐照的最大实验剂量(9.03 x 10(-3) C/kg)诱发染色体畸变的频率与铝和硒的最大实验剂量(3.86 x 10(-4) M 和 8.34 x 10(-6) M)诱发的频率相似。
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引用次数: 0
[Molecular and genetic polymorphism of cultivated soybean detected by AP-PCR, SSRP and SSR]. [利用 AP-PCR、SSRP 和 SSR 检测栽培大豆的分子和遗传多态性]。
Pub Date : 2001-09-01
A F Brik, Iu M Sivolap

Molecular-genetic polymorphism of cultivated soybean from different geographic regions was analysed using three PCR methods (AP-PCR, SSRP and ISSR). The dendrogram of phenogenetic relationship was constructed on the base of obtained data. The allocation of varieties are in a good agreement with pedigree information. Main parameters such as polymorphism level, expected heterozygosity, and marker index were calculated. It was shown that ISSR is the method of choice for the differentiation of soybean varieties.

利用三种 PCR 方法(AP-PCR、SSRP 和 ISSR)分析了不同地理区域栽培大豆的分子遗传多态性。根据获得的数据构建了表型遗传关系树枝图。品种分配与血统信息十分吻合。计算了多态性水平、预期杂合度和标记指数等主要参数。结果表明,ISSR 是区分大豆品种的首选方法。
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引用次数: 0
[Variation of ultrastructural organization of the mitochondria of pea cells in vitro]. [体外豌豆细胞线粒体超微结构组织的变化]。
Pub Date : 2001-03-01
D O Klimchuk

Unusual huge organelles together with mitochondria of typical organization were found in a part of population of callus cells of pea. They like mitochondria are surrounded by two biologic membranes, the inner one has invaginations. Inclusions in the matrix are a specific character of structural organization of these unusual mitochondria. These inclusions look as electronically dense bars that extend parallel to longitudinal axis of organelle and consist of fibril-like elements. Organization of inclusions and cristae as well as a shape of organelles are changing during cell development. Possible nature of the unusual mitochondria is discussed.

在豌豆的部分胼胝体细胞中发现了异常巨大的细胞器,以及具有典型组织结构的线粒体。它们和线粒体一样被两层生物膜包围,内层膜有内陷。基质中的内含物是这些不同寻常的线粒体结构组织的一个特殊特征。这些内含物看起来像电子致密条,平行于细胞器的纵轴延伸,由纤维状元素组成。内含物和嵴的组织以及细胞器的形状在细胞发育过程中不断变化。讨论了不寻常线粒体的可能性质。
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引用次数: 0
[T-DNA induced anomalies of flowers and male sterility in transgenic tobacco plants: morphometric and cytological analysis]. [T-DNA诱导的转基因烟草植物花朵异常和雄性不育:形态计量学和细胞学分析]。
Pub Date : 2000-11-01
Iu V Sidorchuk, A A Zagorskaia, E V Deĭneko, N V Shamina, V K Shumnyĭ

Morphometric and cytological analysis have been carried out in transgenic tobacco (Nicotiana tabacum L.) plants possessing male sterility and abnormalities of flowers. Mutation affected flower structure and conducted to long pistil and male sterility. It was established that development of microspores was blockaded on different stages of gametogenesis but the first injuries were detected for tetrad stage.

对具有雄性不育和花异常的转基因烟草(Nicotiana tabacum L.)植株进行了形态计量和细胞学分析。突变影响了花的结构,导致长雌蕊和雄性不育。结果表明,小孢子的发育在配子发生的不同阶段都受到了阻碍,但在四分体阶段首次检测到了损伤。
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引用次数: 0
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