In this study an antifungal protein from Urginea indica bulbs was purified to homogeneity by acid precipitation, Diol 300 Gelfiltration, and C 18 reverse phase HPLC. Its molecular mass was estimated to be 29 kDa and periodic acid-Schiff (PAS) staining showed that identified antifungal molecule is a glycoprotein. The neutralization of antifungal activity after periodate oxidation of 29 kDa glycoprotein suggests that the glycan part of the molecule appears to be involved in antifungal activity. N-terminal amino acid sequence of the purified protein was determined as SQLKAXIXDF. This sequence had no sequence similarity with any antifungal proteins. A polyclonal antiserum was raised against purified protein and used in immunolocalization analysis. Results suggest that it is localized to the cell wall of the bulb. Antifungal tests have demonstrated that U. indica protein exerts a fungistatic effect. It completely inhibits the germination of spores and hyphal growth of Fusarium oxysporum.
Summary Urginea indica Kunth. Hyacinthaceae, commonly called Indian Squill, represents a species complex. Intraspecies variation is common. Karyological studies were made in six accessions of Urginea collected from six different places in India, of which four accessions of Urginea indica were collected from Kanakapura, Trichendur, Kashmir valley, and Kerala. Two accessions of Urginea wightii were from Gubbi and Bellary. Variations in ploidy were noted. Among the Urginea indica accessions, diploids (2n=20) were found in Kanakapura, Trichendur, and Kashmir Valley; a pentaploid (2n=50) was noticed in the Kerala accession, whereas two different ploidies were noticed in the Urginea wightii accessions of Gubbi and Bellary (2n=20, 2n=40) within the same plant. There is close homology between Kanakapura and Tiruchendur with subtelocentric being a major component.
The wild plants of Cymbopogon flexuosus (Nees ex Steud) Wats were subjected to sUV-B treatment to analyze essential oil variation and DNA polymorphism. Significant difference in essential oil yield and citral content was observed. Plants treated with sUV-B for different intervals of time (15min, 30min, 1h, 1.5h, 2.0h, 2.5h, 3.0h, 3.5h and 4.0h) along with control were subjected to RAPD analysis with 10 random decamer primers. Five primers (OPA-01, OPA-09, OPY-18, OPY-09 and OPG-10) produced polymorphism generating 57 amplicons, of which 27 were polymorphic and 30 were monomorphic in nature. Primer OPA-01 showed highest polymorphism (60%) and OPA-09 showed lowest polymorphism (12.5%). High genetic similarity was observed in treatment with 1.5h and 2.5h and least genetic similarity in treatment with 3.5h and control plants. Dendrogram constructed on genetic similarity coefficient showed two clusters viz., cluster I and II. The study indicates the significant variation at intraspecies level with respect to essential oil yield, citral content and DNA polymorphism in sUV-B treated plants which can be further exploited for developing high yielding chemotypes in C. flexuosus.
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