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Plant Tissue Culture (plant + tissue_culture)
Selected AbstractsDevelopment of Auxotrophic Agrobacterium tumefaciens for Gene Transfer in Plant Tissue CultureBIOTECHNOLOGY PROGRESS, Issue 3 2004Jason I. Collens Auxotrophic strains of Agrobacterium tumefaciens were generated for use in liquid co-culture with plant tissue for transient gene expression. Twenty-one auxotrophs were recovered from 1,900 tetracycline-resistant insertional mutants generated with a suicide vector transposon mutagenesis system. Twelve of these auxotrophs were characterized on a nutrient matrix. Isolates were screened for growth in plant cell and root culture, and three auxotrophs were identified that had limited growth: adenine (ade-24), leucine (leu-27), and cysteine (cys-32). Ade-24 displayed poor T-DNA delivery in a transient expression test delivering GUS from a binary vector, while cys-32 displayed the best ability to deliver DNA of these three auxotrophs. The growth yield of cys-32 on cysteine was assessed to provide a quantitative basis for co-culture nutrient supplementation. The utility of cys-32 for delivering T-DNA to plant tissues is demonstrated, where an 85-fold enhancement in GUS expression over wild-type A. tumefacienswas achieved. [source] Potential of bacterial indoleacetic acid to induce adventitious shoots in plant tissue cultureLETTERS IN APPLIED MICROBIOLOGY, Issue 2 2007B. Ali Abstract Aims:, The main aim of this study was to investigate the possible role of indoleacetic acid (IAA) from bacteria to induce in vitro adventitious shoots in internodal explants of Brassica oleracea L. Methods and Results:, Culture supernatant of Halomonas sp. RE1 and Halomonas sp. HT1 that contain 21 and 40 ,g ml,1 IAA, respectively, was used to supplement Murashige and Skoog (MS) medium. Two combinations that were supplemented with bacterial supernatant (BS) are MS + BS and MS + BS + 10%CW (coconut water) while basal MS medium was used as control. The amounts of BS used in this experiment were 50, 100, 150 and 200 ,l in 5 ml MS medium in each combination. In vitro -grown internodal explants of B. oleracea were inoculated on these media combinations and incubated in a growth chamber at 25 ± 1°C and exposed to 16-h cool fluorescent light. After 5,6 weeks of incubation adventitious shoot induction was observed in all treatments that were supplemented with BS as compared with the controls where very low response was observed. The frequency of shoot induction was high in media that were supplemented with 10%CW in the presence of bacterial auxin. Conclusions:, It was concluded that IAA of microbial origin has the potential to induce adventitious shoots in internodal explants. Significance and Impact of the Study:, IAA from bacteria can be effectively used in plant tissue culture; especially a combination of MS + BS + 10%CW is very cost-effective as compared with synthetic phytohormones for in vitro studies. [source] HPLC-DAD in identification and quantification of selected coumarins in crude extracts from plant cultures of Ammi majus and Ruta graveolensJOURNAL OF SEPARATION SCIENCE, JSS, Issue 14 2003Marian Kami Abstract This paper describes a method for the separation and determination of selected coumarins and furanocoumarins in the crude extracts from plant tissue cultures of Ammi majus hairy roots and Ruta graveolens cell suspensions, cultured in vitro, separately or together as co-cultures. The usefulness of the three main components of the eluent used in reversed-phase high performance liquid chromatographic analysis, namely: methanol (MeOH), acetonitrile (ACN), and tetrahydrofuran (THF), and different elution programs, was assessed. In the optimal analytical method a Lichrospher® RP-18e 5-,m column, a THF-MeOH elution gradient, and a UV/VIS DAD detector were used. Due to the presence of many different compounds in the investigated plant extracts, the use of a UV/VIS DAD detector was essential. Coumarins were identified by comparison of their UV spectra with those of the analytical standards, and characterization of peak purity. [source] Preparative isolation and purification of alkannin/shikonin derivatives from natural products by high-speed counter-current chromatography,BIOMEDICAL CHROMATOGRAPHY, Issue 2 2009Andreana N. Assimopoulou Abstract Alkannin and shikonin (A/S) and their derivatives have been found in the roots of several Boraginaceous species and are also produced through plant tissue cultures. The chiral compounds A/S are potent pharmaceutical substances with a wide spectrum of biological and pharmacological activities like wound healing, antimicrobial, anti-inflammatory, anticancer and antioxidant activity. High-speed counter-current chromatography (HSCCC) was applied for the first time to the separation, preparative isolation and purification of A/S and their esters from extracts of Alkanna tinctoria roots, as well as commercial samples. The constituents of HSCCC fractions and their purity were determined by high-performance liquid chromatography,diode array detection,mass spectrometry (HPLC-DAD-MS), since DAD cannot detect oligomeric A/S derivatives that are present in most of the samples containing the respective monomeric derivatives. The purity of HSCCC fractions was compared with the one of fractions isolated by column chromatography (CC) using as stationary phases silica gel and Sephadex LH-20. As shown, the purity of monomeric alkannin/shikonin was greater by HSCCC than CC separation of commercial A/S samples. Copyright © 2008 John Wiley & Sons, Ltd. [source] | |||||||||||||