Mass propagation of Juniperus procera Hoechst. Ex Endl. From seedling and screening of bioactive compounds in shoot and callus extract

Abstract

Background Juniperus procera Hoechst. ex Endl. is a medicinal tree in Saudi Arabia, primarily in the Enemas region, but it is locally threatened due to die-back disease and difficulties regarding seed reproduction (seed dormancy and underdeveloped embryonic anatomy, and germination rate < 40%). Hence, the alternative methods for reproduction of Juniperus procera are really needed for conservation and getting mass propagation for pharmaceutical uses. Results In this manuscript, we articulated the successful in vitro shoot multiplication and callus induction of J. procera by using young seedling as explants and detected an important antibacterial and antitumor product. Explants were grown on different types of media with the supplement of different combinations of Plant Growth Regulators (PGRs) at different concentrations. The best media for shoot multiplication was Woody Plant Media (WPM) supplemented with PGRs (0.5 μM of IAA and 0.5 μM BAP or 0.5 μM IBA and 0.5 μM BAP). Whereas for callus induction and formation Woody Plant Media (WPM) with the addition of PGRs (0.5 μM 2,4-D and 0.5 μM BAP) was better than the Chu Basal Salt Mixture (N6), Gamborg’s B-5 Basal Medium (B5), and Murashige and Skoog media. The possibility of multiplication of J. procera in vitro creates significant advantages to overcome the difficulties of seeds dormancy for the reproduction of plants, conservation of trees, and getting mass propagation material for pharmaceutical studies. The shoot and callus extract of J. procera was detected using gas chromatography-mass spectrometry analysis and revealed more than 20 compounds related to secondary metabolites, which contained antibacterial and antitumor agents, such as ferruginol, Retinol, and Quinolone as well as confirmed by Direct Analysis in Real Time, Time of Flight Mass Spectrometry (DART-ToF-MS). Podophyllotoxin (PTOX) was detected in callus material by HPLC with sigma standard and confirmed by DART-ToF-MS and UV spectra. Conclusion We successfully conducted in vitro shoot multiplication and callus induction from J. procera seedlings using WPM and a different combination of PGRs and, detected an important antibacterial and antitumor product such as ferruginol and podophyllotoxin. According to our findings, J. procera has become a new natural source of novel bioactive compounds.

Biogenic silver nanoparticles improve bioactive compounds in medicinal plant Juniperus procera in vitro

“…Moreover, it has been reported that ferruginol inhibits the growth of cancer cells ( González et al, 2014 ). Recently, we detected ferruginol in the different parts of J. procera using GC-MS, DART-MS, and HPLC ( Salih et al, 2021a , b ; Figure 8A ), and it is a dominant compound in different parts of this plant. This study separated ferruginol and identified it using HPLC, with ferruginol standard ( Figures 8B,C ).…”

“…7 g/L of agar was added, and the pH was maintained at 5.7. Following the protocol, we recently reported ( Salih et al, 2021a ). Next, different concentrations of biogenic AgNPs (0.0, 5, 10, 20, and 50 mg/L) were added to the WPM before sterilizing at 121°C for 20 min.…”

High-frequency adventitious shoot organogenesis from in vitro stem explants of Scutellaria araxensis Grossh

“…Ex Endl. (Salih et al, 2021), Ficus religiosa (Hesami et al, 2018), and Securidaca longipedunculata (Lijalem and Feyissa, 2020). Bisht et al (2012) investigated the in vitro propagation of Polygonatum verticillatum and suggested that the favorable impact of cytokinin and auxin combination is due to increased RNA synthesis.…”

Optimization Method for Phenolic Compounds Extraction from Medicinal Plant (Juniperus procera) and Phytochemicals Screening

“…Since the ethanol extract from seed contained a higher value of phenolic compounds, it was submitted to GC-MS analysis to identify the phytochemical compounds and resulted in very important bioactive compounds such as ferruginol, phenanthrene, and n-hexadecanoic acid related to phenolic compounds (Table 5 and Figure 5). The bioactive compounds in the leaf extract of J. procera were screened previously [43]; however, some variation has been recorded between the seed and leaf extract (Table 5) in the screened bioactive compounds. Moreover, different parts of the plant occupy a pool of bioactive compounds containing potential chemical groups [30].…”

“…The injection temperature was 280 • C and the column temperature was adjusted to 300 • C. Helium gas was used as the carrier with a flow rate of 1 mL/min. The electron ionization energy was 70 eV while the GC-MS analysis leaf extract of J. procera was undertaken and published recently [43].…”