An efficient hairy root induction system for an important endangered medicinal plant, Dracocephalum kotschyi, was developed through Agrobacterium rhizogenesmediated transformation by modifying the co-cultivation medium using five bacterial strains, A4, ATCC15834, LBA9402, MSU440, and A13 (MAFF-02-10266). A drastic increase in transformation frequency was observed when a Murashige and Skoog medium lacking NH 4 NO 3 KH 2 PO 4, KNO 3 and CaCl 2 was used, resulting in hairy root induction frequencies of 52.3 %, 69.6 %, 48.6 %, 89.0 %, and 80.0 % by A4, A13, LBA9402, MSU440, and ATCC15834 strains, respectively. For shoot induction, hairy roots and unorganized tumors induced by strain ATCC15834 were placed on an MS media supplemented with 0.1, 0.25, 0.5, and 1 mg/l BA plus 0.1 mg/l NAA. The high frequency of shoot regeneration and number of shoot were obtained in the medium containing 0.25 mg/l BA and 0.1 mg/l NAA. Root induction occurred from the base of regenerated shoots on the MS medium supplemented with 0.5 mg/l IBA after 10 days.
Iran has a long history of acid lime cultivation and propagation. In this study, genetic variation in 28 acid lime accessions from five regions of south of Iran, and their relatedness with other 19 citrus cultivars were analyzed using Simple Sequence Repeat (SSR) and Inter-Simple Sequence Repeat (ISSR) molecular markers. Nine primers for SSR and nine ISSR primers were used for allele scoring. In total, 49 SSR and 131 ISSR polymorphic alleles were detected. Cluster analysis of SSR and ISSR data showed that most of the acid lime accessions (19 genotypes) have hybrid origin and genetically distance with nucellar of Mexican lime (9 genotypes). As nucellar of Mexican lime are susceptible to phytoplasma, these acid lime genotypes can be used to evaluate their tolerance against biotic constricts like lime Bwitches' broom disease^.
En este estudio, se analizó la diversidad genética en 19 cultivares de cítricos mediante la repetición de secuencia simple (SSR), la repetición de secuencia inter-simple (ISSR) y los marcadores de secuencia polimórfica amplificada segmentada (CAPS). Se utilizaron nueve cebadores para SSR, nueve cebadores ISSR y dos cebadores para CAPS para la puntuación del alelo. Se analizaron una región de ADN de cloroplasto (rbcL-ORF106) y una región de ADN mitocondrial (18S-5S) usando marcador de secuencia polimórfica amplificada escindida (CAPS) en 19 accesiones de cítricos cultivadas en Irán. En total, se detectaron 45 genes SSR y 131 ISSR polimórficos alelos y organelos del genoma del árbol. El análisis de conglomerados de los datos SSR y ISSR se realizó utilizando el método UPGMA y se basó en el coeficiente de Jaccard. El resultado de esta investigación mostró que los cebadores SSR e ISSR eran altamente informativos y eficientes para detectar la variabilidad genética y las relaciones de las accesiones de los cítricos. Y el análisis de marcadores de CAPS Los resultados mostraron que Bakraee y uno de tipo off cal mexicana tenían un patrón de bandas similar al Clementine Mandarin, mientras que Pummelo se consideraba como padre materno de otros genotipos estudiados Citron considerado como padre padre mostró un patrón de bandas definido entre 19 genotipos estudiados que confirmó Citoplasma herencia de orgánulos celulares de la madre.
Background: Cosmos bipinnatus is an important medicinal plant with antioxidative, antigenotoxic, anti-inflammatory, and antiproliferative effects on several cancer cell lines. It has great potential for development as a promising cancer chemo-preventive agent. Hairy root-based culture technology is a new sustainable production platform for producing specific pharmaceutical secondary metabolites. Objectives: The current study developed and introduced a reliable transformation system for C. bipinnatus by optimization of aspects important in transformation frequency using Agrobacterium rhizogenes. Methods: Five bacterial strains, including ATCC 15834, ATCC 31798, A7, MAFF-02-10266, and MSU440, 2 explant types (leaf and stem), and 2 co-cultivation media (full MS and ½ MS) were examined. Genomic DNA was extracted using a modified CTAB protocol from putative transgenic root lines and the control root. Transgenic hairy root lines were approved by means of Polymerase Chain Reaction (PCR) using specific rolB gene primers. Results:The highest ratio of genetically transformed root induction was found from leaf explants using A. rhisogenes strains ATCC15834 and MSU440 (72% to 73%). When ½ MS medium was used as a co-cultivation medium, a significant increase in transformation frequency (84%) was observed. Conclusions: The MSU440 Agrobacterium strain and ½ MS co-cultivation medium could significantly improve genetic transformation efficiency for establishment of hairy root-based cultures for C. bipinnatus.
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