A Comparative Study of Apigenin Content and Antioxidant Potential of CosmosBipinnatus Transgenic Root Culture

Background: Flavonoid-derived components have been studied for their therapeutic properties. Objectives: Apigenin has shown remarkable antioxidant and anti-inflammatory features, so we should have a reliable source of apigenin. Methods: In this study, we used high-performance liquid chromatography method to compare the amount of apigenin in flower, root, leaf, and stem of three varieties of osmos bipinnatus, i.e., ‘Dazzler,’ ‘Xanthos,’ ‘Sensation Pinkie’, and in transgenic root culture of C. bipinnatus ‘Dazzler’. Besides, the antioxidant activity of C. bipinnatus ‘Dazzler’ transgenic root culture was evaluated using Ferric Reducing Antioxidant Power (FRAP) assay. Results: Dazzler variety flowers showed the highest recovery of apigenin with 0.799 mg/100 mg Dry Weight (DW). However, the Sensation pinkie variety leafs had the lowest recovery with 0.089 mg/100mg. Apigenin content in transformed roots (0.797 mg/100 mg DW) of C. bipinnatus ‘Dazzler’ was significantly higher than non-transformed roots (0.42 mg/100 mg DW). The ethanolic extract of hairy root showed the FRAP value of 668.1 µM Fe2+/mg that was comparatively more than the wild root FRAP value (426.2 µM Fe2+/mg). Conclusion: In conclusion, the presence of apigenin in high amounts in hairy root cultures of C. bipinnatus ‘Dazzler’ indicates its great potential for the future pharmaceutical industry.

Determination of some conditions for hairy root induction in soybean by Agrobacterium rhizogenes

This study was conducted to determine some conditions for transformation via Agrobacterium rhizogenes on soybean cultivars HLDN29 and DT84. Cotyledon explants were more efficient in hairy root induction compared with hypocotyl explants in both cultivars of soybean. The highest root induction rate and average root number were observed in HLDN29 explants infected with ATCC11325 and ATCC15834 strains (approx. 96% - 100% and 8 roots per explant) and in DT84 explants infected with ATCC15834. Six to eight day - old cotyledonary leaves after sowing were optimal and appropriate for hairy root induction. Direct inoculation and immersion methods showed no significant difference in root induction rate and average root number in both HLDN29 and DT84 cultivars. Transgenic root lines were identified based on PCR analysis with virD and rolC sequences - specific primers.

Transgenic root cultures of Gentiana punctata L.

Shoot cultures of <em>Gentiana punctata</em> L. were inoculated with suspension of <em>Agrobacterium rhizogenes</em> strain A4 M70GUS. Hairy roots which appeared 2-3 weeks later were cultured on hormone-free, liquid, WPM (Lloyd and McCown 1980) basal medium for more than 5 years (60 subcultures). Growth rate of transformed roots was higher than the growth rate of nontransformed roots. Spontaneous shoot regeneration occured only in three culture vessels in subcultures No. 40 and 42. Plants had phenotype characteristics typical for <em>A. rhizogenes</em> transformed plants including: wrincled leaves, short internodes, plagiotropic roots and in general their growth rate was reduced. These plants also manifested precocious formation of flower buds without vernalization and flowering under in vitro conditions. Flowers were pale yellow, the same as in the standard phenotype.

Nodule-Expressed Cyp15a Cysteine Protease Genes Map to Syntenic Genome Regions in Pisum and Medicago spp.

PsCyp15a is a gene that encodes a vacuolar cysteine protease expressed in wilt-induced shoots of Pisum sativum (pea) and in root nodules. To further the understanding of nodular PsCyp15a expression, a region 5′ to the coding sequence of the gene was cloned. Varying lengths of 5′ untranslated sequence were fused with the uidA coding region and introduced from Agrobacterium rhizogenes into “hairy roots” of Vicia hirsuta. In this transgenic root nodulation assay, a promoter sequence of 900 bp was sufficient to give an expression pattern indistinguishable from that obtained in pea nodules by in situ hybridization. An orthologue of PsCyp15a was cloned from nodule mRNA of Medicago sativa and a corresponding gene identified in M. truncatula was also shown to express strongly in nodules. With molecular mapping techniques, it was demonstrated that these genes map to a syntenic genome location in pea and Medicago spp., but the map positions of the Cyp15a genes cannot be correlated with existing nodulation mutants.