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Amplified fragment length polymorphism and metabolomic profiles of hairy roots of Psoralea corylifolia L

机译:毛白杨毛状根的扩增片段长度多态性和代谢组学谱

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A reproducible protocol for establishment of hairy root cultures of Psoralea corylifolia L. was developed using Agrobacterium rhizogenes strain ATCC 15834. The hairy root clones exhibited typical sigmoid growth curves. Genomic and metabolomic profiles of hairy root clones along with that of untransformed control were analysed. Hairy root clones, Ps I and Ps II, showed significant differences in their amplified fragment length polymorphism (AFLP) profiles as compared to that of control, besides exhibiting Ri T-DNA-specific bands. These results amply indicate the stable integration of Ri T-DNA into the genomes of these clones. Further, the variations observed between clones in the AFLP profiles suggest the variable lengths and independent nature of Ri T-DNA integrations into their genomes. An isoflavonoid, formononetin, and its glycoside were present only in the hairy root clones while they were absent in the untransformed control. Variations observed in the metabolite profiles of these clones may be attributed to the random T-DNA integrations and associated changes caused by them in the recipient genomes. GC/MS analyses revealed the production of three and six clone-specific compounds in Ps I and Ps II, respectively, suggesting that the clones are dissimilar in their secondary metabolism. HPLC/UV-MS analyses disclosed substantial increases in the total isoflavonoids produced in Ps I (184%) and Ps II (94%) compared to untransformed control.
机译:使用发根农杆菌菌株ATCC 15834,开发了一种可重复的方法,用于建立Psoralea corylifolia L.的毛状根培养物。毛状根克隆表现出典型的乙状结肠生长曲线。分析了毛状根克隆的基因组和代谢组学谱以及未转化的对照。与对照相比,毛状根克隆Ps I和Ps II在扩增片段长度多态性(AFLP)谱中显示出显着差异,除了表现出Ri T-DNA特异性条带。这些结果充分表明Ri T-DNA稳定整合到这些克隆的基因组中。此外,在AFLP图谱中的克隆之间观察到的变异表明Ri T-DNA整合入其基因组的长度可变和独立性。异黄酮,formononetin及其糖苷仅存在于毛状根克隆中,而未转化对照中则不存在。这些克隆的代谢产物谱中观察到的变异可能归因于随机的T-DNA整合以及它们在受体基因组中引起的相关变化。 GC / MS分析显示,分别在Ps I和Ps II中产生了3个和6个克隆特异性化合物,这表明这些克隆的二次代谢不同。 HPLC / UV-MS分析显示,与未转化的对照相比,Ps I(184%)和Ps II(94%)中产生的总异黄酮含量显着增加。

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