PECULIARITIES OF THE GROWTH OF Artemisia tilesii Ledeb. “HAIRY” ROOTS WITH DIFFERENT FOREIGN GENES

Т. А. Bohdanovych
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Abstract

Aim. To compare Artemisia tilesii “hairy” root lines with different transferred genes in terms of the relationship between the total content of flavonoids, the levels of antioxidant activity (AOA) and reducing power (RP), as well as the activity of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), rolB and rolC genes. Methods. We compared the root lines Nos. 10 and 16, obtained by transformation with the wild Agrobacterium rhizogenes strain A4, lines Nos. 2 and 4, obtained using A. rhizogenes carrying pSV124 vector with ifn-α2b and nptII genes, as well as the roots of non-transformed plants that were cultivated in vitro. The presence and activity of rolB, rolC, PAL, and CHS genes were determined by PCR and real-time PCR, respectively. The content of flavonoids, AOA and RP were determined by standard tests with AlCl3, DPPH (2,2-diphenyl-1-picrylhydrazyl) and K3[Fe(CN)6] accordingly. Results. The content of flavonoids in most of the lines was higher than in the control, and correlated with AOA and RP. Roots No. 10 were characterized by the fastest growth, which coincided with higher activity of rolB and rolC genes. The activities of PAL and CHS in “hairy” roots were lower than those in non-transformed ones. Conclusions. Root lines carrying only rolB and rolC and lines with additional ifn-α2b and nptII genes had similar ranges of flavonoids concentration, AOA and RP levels that exceeded those in the control. The dependence of the root growth rate, and lack of the dependence of the flavonoids content with the activity of the rol genes were demonstrated. PAL activity inversely correlated with flavonoids content in all experimental lines, which may be the result of overproduction of compounds in transgenic roots.
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黄花蒿生长的特点。带有不同外源基因的“毛茸茸”根
的目标。比较不同转移基因的蒿“毛状”根系总黄酮含量、抗氧化活性(AOA)和还原力(RP)水平以及苯丙氨酸解氨酶(PAL)、查尔酮合成酶(CHS)、rolB和rolC基因活性之间的关系。方法。我们将转化获得的10号和16号根系与野生根农杆菌A4、2号和4号根系(根农杆菌携带带有ifn-α2b和nptII基因的pSV124载体)以及未转化的离体培养植株的根系进行了比较。采用PCR和real-time PCR分别检测rolB、rolC、PAL和CHS基因的存在和活性。采用AlCl3、DPPH(2,2-二苯基-1-苦基肼基)和K3[Fe(CN)6]为标准试剂测定黄酮类化合物、AOA和RP的含量。结果。大部分品系黄酮类化合物含量均高于对照,且与AOA和RP呈正相关。10号根生长最快,其rolB和rolC基因活性较高。毛状根PAL和CHS活性低于未转化根。结论。仅携带rolB和rolC基因的根系和添加ifn-α2b和nptII基因的根系的黄酮类化合物浓度、AOA和RP水平均超过对照。结果表明,黄酮类化合物含量与这些基因的活性无关,而与根系生长速率有关。在所有试验品系中,PAL活性与黄酮类化合物含量呈负相关,这可能是转基因根中化合物过量产生的结果。
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审稿时长
20 weeks
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