ABSTRACTS New Biotechnology · Volume 25S · September 2009 (0 or 600 mg/seedling). Inoculation with a mixture of mycelium from S. bovinus, L. laccata and L. determinus and with a mixture of spores of P. tinctorius and S. citrinum enhanced plant development, measured by shoot height, and plant biomass, without the need of fertiliser. Results indicate that selected ECM fungi can be beneficial biotechnological tools in nursery production of P. pinaster. doi:10.1016/j.nbt.2009.06.880 3.4.05 Biofertilizers and their significance to environmental and sustainable agriculture S. Mohamed Said Ali ∗ , H. Soliman, A. Abdallah, T. Moharram, S. Ahmed Minia University, Cairo, Egypt The use of biofertilizers has become a great hope for Egyptian agri- culture, particularly in the field of production of medicinal plants, especially when the economic and environmental points of view are considered, since they reduce the environmental pollution and production costs, in addition to improving the quality. Therefore, the influence of using locally isolated nitrogen-fixing bacteria as biofertilizers were studied. The isolates used were three represen- tative strains of either Azotobacter and Azospirillum (two isolates of Azotobacter out of 60 isolates, these isolates were Az.NP7-Az.PR1, and one isolate of Azospirillum out of 60 isolates, this isolate was As.Pp1) which were screened for their activity in nitrogen fixing and stimulation effect by used root exudates of black cumin plants and used for further inoculation studies as biofertilizers. These iso- lates were isolated at random from rhizosphere and rhizoplane of black cumin, parsley and fenugreek cultivars at the different stages of growth. The effect of inoculation with N2-fixing bacteria. (Biofertilizer) on growth plant characteristics of tested medicinal plants (Nigella sativa, Trigonella foenum-graecum and Pet- roselinum sativum) plants were studied. Inoculation was conjugated with the application of four doses of mineral N, i.e. 0, 50%, 75% and one full dose (100%) from the recommended doses. Results obtained showed that the application of biofertilizers (N2-fixers) produced better growth and reduced the N requirement in many medicinal plants such as Nigella sativa, Trigonella foenum and Pet- roselinum sativum compared with untreated plants. doi:10.1016/j.nbt.2009.06.881 3.4.06 Genetic transformation and elicitation as yield enhance- ment strategy for glycyrrhizin production by cell cultures of Abrus precatorius L. V.S. Karwasara ∗ , V.K. Dixit Department of Pharmaceutical Sciences, Sagar, India Abrus precatorius L. (family Fabaceae) known as Indian liquorice which is common bedding deciduous vine. It is a very good source of sweet principle compound glycyrrhizin (a triterpenoid saponin). It can be used as a very good substitute for Liquorice . Genetic transformation and elicitation has proved to be an effec- tive way to enhance secondary metabolites in plant cell cultures. In this study, we describe the amalgamation of Ti and Ri plasmid- mediated transformation of Abrus precatorius and elicitation for the improved production of glycyrrhizin in the established crown gall and hairy root culture of A. precatorius. Agrobacterium tumefaciens MTCC 431 and MTCC 2250 and Agrobacterium rhizogenes MTCC 532 and NCIM5140 were used for the Abrus genetic transforma- tion. Opine assay and polymerase chain reaction confirmed the Abrus genetic transformation. High yielding transformed cell line of A. precatorius was developed from leaf explant taken from asepti- cally germinated Abrus plantlets, cultured on solid Murashige and Skoog medium (0.8%agar) without growth regulators. Suspension cultures were developed on optimized media composition and culture conditions (photoperiod of 12/12 h light/dark cycle, 5.8 pH, 27 ± 1 ◦ C temperature and 120 rpm). The Agrobacterium trans- formed A. precatorius cell culture which further elicited with biotic (dried cell powder and culture filtrate of Aspergillus niger and Rhi- zopus stolonifer) and abiotic elicitors (yeast extract, salicylic acid, ascorbic acid) at optimum concentration. This resulted in syner- gistic promotion of glycyrrhizin accumulation compared with the untransformed cell culture. Glycyrrhizin productivity of 32.5 mg/L was obtained in 20 days of cultivation. The Abrus transformed cell culture showed 2.5-fold increase in glycyrrhizin content when compared with untransformed callus (0.0126%DW). Maximum enhancements of 3.02-fold and 2.83-fold in glycyrrhizin content were obtained with Aspergillus niger (10%v/v) and R. stolonifer (7.5%v/v), respectively, 3rd and 6th day after elicitor treatment. The application of yeast extract (10 mg/L), ascorbic acid (10 M) and salicylic acid (10 M) resulted in 1.88, 1.92 and 1.65-fold increase in glycyrrhizin content respectively when compared with the control culture. Present study reports very first successful genetic transformation of A. precatorius and study on cell cultures of A. precatorius capable to produce glycyrrhizin. These results also indicate potential to develop a suitable methodology to produce glycyrrhizin at large scale. doi:10.1016/j.nbt.2009.06.882 3.4.07 Indole-3-acetic acid production by plant associated bac- teria: potential to alter endogenous IAA content and growth of Triticum aestivum L. B. Ali ∗ , A. Nasim Sabri, S. Hasnain University of the Punjab, Lahore, Pakistan Bacterial strains of Bacillus, Pseudomonas, Escherichia, Micro- coccus and Staphylococcus genera isolated from rhizosphere, histoplane and phyllosphere of different plant species were iden- tified by 16S rDNA gene sequencing. Strains were evaluated to enhance endogenous indole-3-acetic acid (IAA) content and growth of Triticum aestivum var. Inqalab-91. Gas chromatography and mass spectrometric (GC—MS) analysis revealed that bacte- S308 www.elsevier.com/locate/nbt