The current study was carried out during the two consecutive winter seasons of 2019/2020 and 2020/2021 at the Experimental Farm of Ash-Shoubak University College, Jordan, as a pot experiment. This experiment was planned to evaluate the impact of various soil salinity levels (1.17, 3.34, 6.51, and 9.68 ds/m) and amino acid types (control, L-tryptophan acid at 100 ppm, glutamine acid at 200 ppm, and L-tryptophan acid + glutamine acid) as well as their combinations on growth, salt resistance index, and some of the chemical constituents of rosemary plants. The obtained results indicated that plant height and the total herb dry weight of rosemary, the salt resistance index (SRI), and the total chlorophyll in leaves were discernably reduced with increasing soil salinity levels compared with the control. However, salinity enhanced leaf proline content. Each amino acid or its mixture improved plant growth, chlorophyll content, and SRI parameters. The SRI percentage of R. officinalis enhanced to more than 100% under a soil salinity level of 1.17 ds/m, combined with amino acids at any type, when compared with the other combination treatments. Furthermore, GC/MS showed that the identified compounds ranged from 98.39% to 99.18% and the unidentified compounds from 0.82% to 1.61% from the volatile oil of rosemary plants. The major constituents of volatile oil samples were camphor (34.95% to 40.21%), D-verbenone (13.74% to 15.23%), and α-pinene (13.21% to 16.73%).

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Solanum villosum is a wild distinguished medicinal herb that has few studies describing its medicinal properties with no sustainable production and conservation studies. In this study, in vitro cultures of microshoots and callus of Solanum villosum were multiplied in vitro on Murashige and Skoog media supplemented with different growth regulators. The MS media that have 2,4 Dichlorophenoxy acetic acid (2,4-D) and Kinetin at the levels of (2.0, 1.2 mg/L; respectively) were used for the induction and multiplication of callus that yielded the highest fresh weight of (6.03 ± 0.15) g. Cryopreservation was applied successfully to seeds using the dehydration technique and for both microshoots and callus using v-cryoplate technique. The highest regrowth rate was obtained in callus (95 ± 0.13) after pre-culturing callus on 0.3 M sucrose for 5 days. The most inhibited bacteria strains by microshoots extracts were, Klebsiella pneumoniae and Staphylococcus epidermidis (30 ± 0.25 and 25 ± 0.17 mm; respectively) using the disk diffusion assay. In MIC method; The extract obtained from microshoots achieved best results of MIC values (9.77 and 39.06 (µg/ml) against (Micrococcus luteus and Klebsiella pneumonia; respectively). A sustainable protocol of callus culture and cryopreservation protocol for in vitro Solanum villosum has been set for the first time which makes this research novel. Furthermore, promising results of antimicrobial activity for both in vitro and wild Solanum villosum were obtained.

Solanum villosum is a wild distinguished medicinal herb that has few studies describing its medicinal properties with no sustainable production and conservation studies. In this study, in vitro cultures of microshoots and callus of Solanum villosum were multiplied in vitro on Murashige and Skoog media supplemented with different growth regulators. The MS media that have 2,4 Dichlorophenoxy acetic acid (2,4-D) and Kinetin at the levels of (2.0, 1.2 mg/L; respectively) were used for the induction and multiplication of callus that yielded the highest fresh weight of (6.03 ± 0.15) g. Cryopreservation was applied successfully to seeds, while microshoots and callus were cryopreserved via v-cryoplate technique. The highest regrowth rate was obtained in callus (95± 0.13) after pre-culturing callus on 0.3 M sucrose for 5 days. The most inhibited bacteria strains by microshoots extracts were, Klebsiella pneumoniae and Staphylococcus epidermidis (30±0.25 and 25±0.17 mm; respectively) using the disk diffusion assay. In MIC method; The extract obtained from microshoots achieved best results of MIC values (9.77 and 39.06 (µg/ml) against (Micrococcus luteus and Klebsiella pneumonia; respectively). A sustainable protocol of callus culture and cryopreservation protocol for in vitro Solanum villosum has been set for the first time which makes this research novel. Furthermore, promising results of antimicrobial activity for both in vitro and wild Solanum villosum were obtained.