Determination of different growth media and Agrobacterium rhizogenes strains on the hairy root growth of Valerian (Valeriana officinalis L.) | ||
| تحقیقات گیاهان دارویی و معطر ایران | ||
| Article 8, Volume 41, Issue 1 - Serial Number 123, April 2025, Pages 96-111 PDF (862.88 K) | ||
| Document Type: Research Paper | ||
| DOI: 10.22092/ijmapr.2025.352320.2886 | ||
| Authors | ||
| Mehdi Heydari-Rahni1; Yousef Filizadeh* 2; Mohammad Nasri3 | ||
| 11Department of Agronomy, Varamin-Pishva Branch, Islamic Azad University, Varamin, Iran. | ||
| 2Department of Agronomy and Plant Breeding, Agricultural College, Shahed University, Tehran, Iran | ||
| 3Department of Agronomy, Varamin-Pishva Branch, Islamic Azad University, Varamin, Iran. | ||
| Abstract | ||
| Background and Objectives: Valeriana, a genus in the Caprifoliaceae family, includes several species, notably Valeriana officinalis L. (valerian). Valerian root extract contains various bioactive compounds, including valerenic acid, valeranone, valepotriates, and gamma-aminobutyric acid, which have been widely used as sedatives to treat sleep disorders such as insomnia. This study aimed to evaluate the induction of valerian hairy roots using three different strains of Agrobacterium rhizogenes (A13, R1601, and LBA9402) under four different growth media compositions: (1) MS full strength, (2) MS supplemented with KH2PO4, NH4NO3, and KNO3, (3) MS supplemented with KH2PO4, NH4NO3, KNO3, and CaCl2, and (4) a medium containing MgSO4, microelements, Na2EDTA, FeSO4, and vitamins. Methodology: This study was a factorial experiment based on a completely randomized design with eight replications under laboratory conditions. Seeds of V. officinalis were obtained from a commercial supplier (Pakan Seed Company, Isfahan, Iran) and germinated under greenhouse conditions. Before sowing, seeds were washed with tap water and distilled water. Seeds that settled at the bottom of the washing vessel were selected as they were expected to be the most viable. They were soaked overnight in water before being washed with a 10% detergent solution for 10 minutes, followed by surface disinfection with 70% ethanol for 30 seconds and 6% NaOCl for 5 minutes. After rinsing four times with sterile water, the seeds were germinated on solid Schulz medium, without growth regulators, at 25°C under a 16-hour light/8-hour dark photoperiod (45 µE m⁻² S⁻¹). Hairy root induction was performed using spray and leaf disc methods, and transformation was confirmed by polymerase chain reaction (PCR) analysis for the presence of rolB and virD genes. Hairy root induction rate, lateral branches per centimeter, number of hairy roots per explant, and dry weight of hairy roots were measured after 60 days. Essential oil was extracted by hydro-distillation and analyzed using gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS). Results: The study confirmed that the tested A. rhizogenes strains could induce hairy roots in valerian. The fastest hairy root emergence was observed in explants inoculated with strain A13, occurring within 10 days. The highest induction rate (92%), hairy root dry weight (272 mg), and number of hairy roots per explant (19.2) were obtained in leaf explants inoculated with the A13 strain and cultured in MS medium supplemented with KH2PO4, NH4NO3, KNO3, and CaCl2. No significant differences were observed in the morphological traits of hairy roots induced by different A. rhizogenes strains. Conclusion: The findings indicate that growth media composition, appropriate A. rhizogenes strain, and optimized growth conditions enhanced hairy root biomass production. The A13 strain was the most effective in promoting mass hairy root production, particularly in MS medium supplemented with KH2PO4, NH4NO3, KNO3, and CaCl2. Given the complex physiological processes involved, valerian hairy root cultures can preserve valeric acid found in vegetative tissues. Future research should focus on identifying the optimal growth media conditions that maximize vegetative growth and secondary metabolite production in valerian. | ||
| Keywords | ||
| Valeriana officinalis; Agrobacterium rhizogense; culture media; hairy root dry weight | ||
| References | ||
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