Introduction: Hyoscyamine, because of its medicinal properties, is an important tropane alkaloid. In order to implement a biotechnological process for its production, hairy roots of Datura species resulting from genetic transformation by Agrobacterium rhizogenes A4 strain have been the subject of this work. In the in vitro alkaloids production programs, optimization of the transformation is a crucial step for obtaining a large number of root lines allowing the selection of efficient lines. Methods: The conditions for hairy roots induction were explored by studying the effect of Datura sp. in vitro seedling etiolation on the genetic transformation. The first step was the establishment of Datura in vitro plantlet cultures followed by the hairy roots induction with A4 strain of A. rhizogenes. The confirmation of the genetic transformation was performed by polymerase chain reaction (PCR) analysis of rolB gene in the roots. After hyoscyamine extraction, it was analyzed (quantitatively and qualitatively) by gas chromatography. Results: 343 root lines were obtained in total, 96 root lines from non-etiolated Datura in vitro seedlings and 247 root lines from etiolated in vitro plantlets. After the selection of six transgenic root lines, tow root lines from each species (D. tramonium, D. tatula and D. innoxia), one from non-etiolated in vitro plantlet and one from etiolated in vitro plantlet, the most hyoscyamine content was 8.43 mg/g D.W. obtained with D. tatula etiolated in vitro seedling. Conclusion: The etiolated in vitro plantlets seem more favorable to hairy roots induction.
Résumé. — La présente étude examine le taux de germination des glands de Chêne vert au laboratoire puis la reprise des plants en pépinière ainsi que le taux de survie en milieu naturel des jeunes individus qui en dérivent. Les glands ont été récoltés sous 5 arbres porteurs dans chacune des 11 provenances réparties dans l’aire algérienne du Chêne vert. Les résultats obtenus montrent que certaines provenances sont caractérisées par des taux de mortalité élevés, pouvant atteindre 60 %. Ces pertes sont plus significatives en phases de levée en pépinière et de reprise en forêt que durant la phase de germination. Ceci suggère que, pour produire des plants vigoureux et viables transplantables en pleine forêt, il faut d’une part sélectionner les meilleures sources de glands et d’autre part maîtriser la phase de développement en pépinière. Ainsi, pour limiter les échecs après transplantation, la sélection des meilleures sources de glands doit se faire tant au niveau provenance qu’au niveau individuel.
Taqerbucht cultivars of date palm are well known by their natural resistance against devastating fungus Bayoud disease. In order to know, if these accessions have the same genetic and morphological profile or each of them constitutes a separate cultivar, we carried out a morphological and molecular characterization and we compared four Taqerbucht (Tq.) date palm cultivars from the southwestern region of Algeria: Tq.hamra cultivar (red fruits), Tq. safra cultivar (yellow fruits), Tq.beïda (white fruits) and Tq.kahla cultivar (black fruits). Seventy one phenotypic characteristics, including 33 quantitative and 38 qualitative traits, have been selected for comparison. Principal component analysis (PCA) and multi-component clustering were used to analyze and compare the data. The results suggest that the four cultivars can be classified into distinct groups. One group contains one cultivar, the Tq.kahla and another group contains the three other cultivars (Tq.safra, Tq.beïda and Tq. hamra). Based on phylogenetic analyses and sequence comparisons, the cultivar Tq. kahla seems to be divergent from the cultivar Tq.hamra, whereas the two cultivars Tq.Safra and Tq.beïda are close to each other. Using 16 Simple Sequence Repeat (SSR) genetic markers to analyze genetic diversity among the cultivars, we found that 13 markers were detectable in 31 allele's loci, and the number of alleles per locus varied from 1–4 with an average of 2.38 alleles per locus. Expected heterozygosity (He) values ranged from 0.375–0.500 and observed heterozygosity (Ho) values from 0.750–1.000.
Morphological diversity and hyoscyamine/scopolamine contents in twelve Algerian samples of Datura stramonium L. of different origin. Morphological diversity was estimated on 15 phenotypic characters of Datura stramonium from twelve samples of different origin ; the Algerian distribution of that species extends from Nedroma (extreme western Algeria : semi-arid bioclimatic stage) to El Kala (at the Algerian-Tunisian border : humid bioclimatic stage). Seeds from these twelve samples were collected and then grown in similar pedoclimatic conditions. Measurements were carried out 60 days after sowing. Results show that these twelve samples were significantly different for the measured traits (leaf area, length of the flowers, number of ramifications, number and biometrics of the capsules and height of the plants). Principal component analysis (PCA) and Factorial discriminant analysis (FDA) were used to discriminate the origins of the samples from their studied characters. The canonical averages of the twelve samples were significantly different, especially in the biometric traits including mature plants, capsules and seeds. The alkaloid content (hyoscyamine and scopolamine) of the twelve samples seems to be influenced by the geographical and bioclimatic distribution. In addition, the most alkaloid productive samples were generally those that invest more in biomass production (leaf area, height and collar diameter of plants).
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