BIOCHEMICAL STUDIES ON PRODUCTION OF SECONDARY METABOLITES FROM DIFFERENT PLANTS BY USING TISSUE CULTURE TECHNIQUES

Abstract

The present study was carried out in Tissue Culture Research Laboratory, Horticulture Research Institute, A.R.C., Giza, Egypt, during 2011 to 2014. Because of the importance of Ginkgo bilobaand industrial needs of ginkgo secondary metabolites significantly directed this study towards elicitation using tissue culture for over production of ginkgo metabolites viausing sodium nitroprusside (SNP)which utilized as the donor of nitric oxide (NO) and using micro algal elicitors (Anabaena Sp. and Spirulina Sp.) to investigate its effect as elicitor onphenols, flavonoids, ginkgo flavone glycosides and ginkgolides accumulation in G. bilobacell suspension cultures and the effect of these elicitors on cells growth. Moreover, studythe effects of different hormone combinations on callus inductionfor production the best callusin Ginkgo biloba cell cultures. The results showed that the optimal medium for inducing leaf-derived callus and the reculture of callus was MS media with 2.0 mg/L naphthalene acetic acid (NAA) + 2.0 mg/L 6-benzylaminopurine (6-BA). It was found that using SNPat concentrations of25 and 50ppm showedthe highestincrease in biomass of cultured Ginkgobiloba on day 13 which was 0.599 and 0.58 g FW, respectively from the initial inoculums of 0.5 g FW. Whereas the highestaccumulation of phenols and flavonoids content with 150 ppm SNP on day 22 showed an increase of 2.8 and 2.2 fold relative to control. Similarly,AnabaenaSp. extract at concentration of150 ppm which showed the highestaccumulation of phenols and flavonoidscontent (2.5 and 2 fold on day 22 of the control,respectively).On the other hand,Anabaenasp. extract at concentration of150 ppm also influenced the highestincrease in biomass G.biloba cultured cells on day 13 day which was 0.69 g FW. Moreover,G.biloba cultures treated with SpirulinaSp. extract at concentrations of150 ppm observed the maximum increase in biomass on day 13 over the control which was 0.705 g FW. Also, the highest accumulation of phenols and flavonoids content increased to 1.7 and 2.4fold, respectively during day 22 in cultures treated with 100 ppmofSpirulinaSp.extract.In addition, the highest contents of ginkgo flavone glycosides and ginkgolidescompounds found in G. biloba cell suspension culture was 1.9 and 1.4 foldof the controlbyusing 150 ppm AnabeanaSp. extractand thehighest contents of the content of these compounds were 1.9 and 2.3 fold with 150ppmSpirulinaSp.extract, respectively.While,the highestcontentof ginkgo flavone glycosides and ginkgolides was increasedto 2.3 and 3.2 foldrelative tocontrol with 150ppm SNP elicitation.