RESUMEN
The marshmallow plant (Althaea officinalis L.) has been used for centuries in medicine and other applications. Valuable secondary metabolites have previously been identified in Agrobacterium rhizogenes-generated transgenic 'hairy' roots in this species. In the present study, transgenic roots were produced in A. officinalis using A. rhizogenes. In addition to wild-type lines, roots expressing the anti-human immunodeficiency virus microbicide candidate, cyanovirin-N (CV-N), were generated. Wild-type and CV-N root lines were transferred to liquid culture and increased in mass by 49 and 19 % respectively over a 7 day culture period. In the latter, the concentration of CV-N present in the root tissue was 2.4 µg/g fresh weight, with an average secretion rate into the growth medium of 0.02 µg/ml/24 h. A. officinalis transgenic roots may therefore in the future be used not only as a source of therapeutic secondary metabolites, but also as an expression system for the production of recombinant pharmaceuticals.
Asunto(s)
Agrobacterium/genética , Proteínas Bacterianas/biosíntesis , Proteínas Portadoras/biosíntesis , Regulación de la Expresión Génica de las Plantas , Raíces de Plantas/genética , Plantas Modificadas Genéticamente/genética , Agrobacterium/metabolismo , Althaea/genética , Fármacos Anti-VIH/química , Fármacos Anti-VIH/metabolismo , Antiinfecciosos/química , Antiinfecciosos/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Reactores Biológicos , Proteínas Portadoras/química , Proteínas Portadoras/genética , Técnicas de Cultivo de Célula , Medios de Cultivo , Humanos , Plantas Modificadas Genéticamente/metabolismoRESUMEN
OBJECTIVE: To explore a new method for identification Astragali Radix from its adulterants by using ITS sequence. METHOD: Thirteen samples of the different Astragali Radix materials and 6 samples of the adulterants of the roots of Hedysarum polybotrys, Medicago sativa and Althaea rosea were collected. ITS sequence was amplified by PCR and sequenced unidirectionally. The interspecific K-2-P distances of Astragali Radix and its adulterants were calculated, and NJ tree and UPGMA tree were constructed by MEGA 4. RESULT: ITS sequences were obtained from 19 samples respectively, there were Astragali Radix 646-650 bp, H. polybotrys 664 bp, Medicago sativa 659 bp, Althaea rosea 728 bp, which were registered in the GenBank. Phylogeny trees reconstruction using NJ and UPGMA analysis based on ITS nucleotide sequences can effectively distinguish Astragali Radix from adulterants. CONCLUSION: ITS sequence can be used to identify Astragali Radix from its adulterants successfully and is an efficient molecular marker for authentication of Astragali Radix and its adulterants.