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BSTRACTS

0 or 600 mg/seedling). Inoculation with a mixture of myceliumrom S. bovinus, L. laccata and L. determinus and with a mixture ofpores of P. tinctorius and S. citrinum enhanced plant development,easured by shoot height, and plant biomass, without the need of

ertiliser. Results indicate that selected ECM fungi can be beneficialiotechnological tools in nursery production of P. pinaster.

oi:10.1016/j.nbt.2009.06.880

.4.05

iofertilizers and their significance to environmentalnd sustainable agriculture

. Mohamed Said Ali ∗ , H. Soliman, A. Abdallah, T. Moharram, S.hmed

Minia University, Cairo, Egypt

he use of biofertilizers has become a great hope for Egyptian agri-ulture, particularly in the field of production of medicinal plants,specially when the economic and environmental points of viewre considered, since they reduce the environmental pollution androduction costs, in addition to improving the quality. Therefore,he influence of using locally isolated nitrogen-fixing bacteria asiofertilizers were studied. The isolates used were three represen-ative strains of either Azotobacter and Azospirillum (two isolates ofzotobacter out of 60 isolates, these isolates were Az.NP7-Az.PR1,nd one isolate of Azospirillum out of 60 isolates, this isolate wass.Pp1) which were screened for their activity in nitrogen fixingnd stimulation effect by used root exudates of black cumin plantsnd used for further inoculation studies as biofertilizers. These iso-ates were isolated at random from rhizosphere and rhizoplane oflack cumin, parsley and fenugreek cultivars at the different stagesf growth. The effect of inoculation with N2-fixing bacteria.

(Biofertilizer) on growth plant characteristics of testededicinal plants (Nigella sativa, Trigonella foenum-graecum and Pet-

oselinum sativum) plants were studied. Inoculation was conjugatedith the application of four doses of mineral N, i.e. 0, 50%, 75%nd one full dose (100%) from the recommended doses. Resultsbtained showed that the application of biofertilizers (N2-fixers)roduced better growth and reduced the N requirement in manyedicinal plants such as Nigella sativa, Trigonella foenum and Pet-

oselinum sativum compared with untreated plants.

oi:10.1016/j.nbt.2009.06.881

.4.06

enetic transformation and elicitation as yield enhance-ent strategy for glycyrrhizin production by cell

ultures of Abrus precatorius L.

.S. Karwasara ∗ , V.K. Dixit

Department of Pharmaceutical Sciences, Sagar, India

brus precatorius L. (family Fabaceae) known as Indian liquoricehich is common bedding deciduous vine. It is a very good

ttga

308 www.elsevier.com/locate/nbt

New Biotechnology · Volume 25S · September 2009

ource of sweet principle compound glycyrrhizin (a triterpenoidaponin). It can be used as a very good substitute for Liquorice.

enetic transformation and elicitation has proved to be an effec-ive way to enhance secondary metabolites in plant cell cultures.n this study, we describe the amalgamation of Ti and Ri plasmid-

ediated transformation of Abrus precatorius and elicitation for themproved production of glycyrrhizin in the established crown gallnd hairy root culture of A. precatorius. Agrobacterium tumefaciensTCC 431 and MTCC 2250 and Agrobacterium rhizogenes MTCC

32 and NCIM5140 were used for the Abrus genetic transforma-ion. Opine assay and polymerase chain reaction confirmed thebrus genetic transformation. High yielding transformed cell linef A. precatorius was developed from leaf explant taken from asepti-ally germinated Abrus plantlets, cultured on solid Murashige andkoog medium (0.8%agar) without growth regulators. Suspensionultures were developed on optimized media composition andulture conditions (photoperiod of 12/12 h light/dark cycle, 5.8H, 27 ± 1 ◦C temperature and 120 rpm). The Agrobacterium trans-ormed A. precatorius cell culture which further elicited with bioticdried cell powder and culture filtrate of Aspergillus niger and Rhi-opus stolonifer) and abiotic elicitors (yeast extract, salicylic acid,scorbic acid) at optimum concentration. This resulted in syner-istic promotion of glycyrrhizin accumulation compared with thentransformed cell culture. Glycyrrhizin productivity of 32.5 mg/Las obtained in 20 days of cultivation. The Abrus transformed

ell culture showed 2.5-fold increase in glycyrrhizin content whenompared with untransformed callus (0.0126%DW). Maximumnhancements of 3.02-fold and 2.83-fold in glycyrrhizin contentere obtained with Aspergillus niger (10%v/v) and R. stolonifer

7.5%v/v), respectively, 3rd and 6th day after elicitor treatment.he application of yeast extract (10 mg/L), ascorbic acid (10 �M)nd salicylic acid (10 �M) resulted in 1.88, 1.92 and 1.65-foldncrease in glycyrrhizin content respectively when compared withhe control culture. Present study reports very first successfulenetic transformation of A. precatorius and study on cell culturesf A. precatorius capable to produce glycyrrhizin. These results alsondicate potential to develop a suitable methodology to producelycyrrhizin at large scale.

oi:10.1016/j.nbt.2009.06.882

.4.07

ndole-3-acetic acid production by plant associated bac-eria: potential to alter endogenous IAA content androwth of Triticum aestivum L.

. Ali ∗ , A. Nasim Sabri, S. Hasnain

University of the Punjab, Lahore, Pakistan

acterial strains of Bacillus, Pseudomonas, Escherichia, Micro-occus and Staphylococcus genera isolated from rhizosphere,istoplane and phyllosphere of different plant species were iden-

ified by 16S rDNA gene sequencing. Strains were evaluatedo enhance endogenous indole-3-acetic acid (IAA) content androwth of Triticum aestivum var. Inqalab-91. Gas chromatographynd mass spectrometric (GC—MS) analysis revealed that bacte-