In Vitro Culture of <i>Rosmarinus officinalis</i> L. in a Temporary Immersion System: Influence of Two Phytohormones on Plant Growth and Carnosol Production
Emerging infectious diseases have become a major global problem with public health and economic consequences. It is an urgent need to develop new anti-infective therapies. The natural diterpene carnosol exhibit a wide variety of interesting antibacterial and antiviral properties, and it is considere...
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MDPI AG
2021-07-01
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| Series: | Pharmaceuticals |
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| Online Access: | https://www.mdpi.com/1424-8247/14/8/747 |
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| author | Eder Villegas-Sánchez Mariana Macías-Alonso Soraya Osegueda-Robles Lisset Herrera-Isidrón Hector Nuñez-Palenius Joaquín González-Marrero |
| author_facet | Eder Villegas-Sánchez Mariana Macías-Alonso Soraya Osegueda-Robles Lisset Herrera-Isidrón Hector Nuñez-Palenius Joaquín González-Marrero |
| author_sort | Eder Villegas-Sánchez |
| collection | DOAJ |
| description | Emerging infectious diseases have become a major global problem with public health and economic consequences. It is an urgent need to develop new anti-infective therapies. The natural diterpene carnosol exhibit a wide variety of interesting antibacterial and antiviral properties, and it is considered a theoretical inhibitor of COVID-19 M<sup>pro</sup>. However, this compound is present in the family Lamiaceae in low quantities. To obtain carnosol in concentrations high enough to develop pharmacological studies, we evaluated the efficiency of a micropropagation protocol of <i>Rosmarinus officinalis</i> using a solid medium and a temporary immersion system (TIS), as well as the effect of 6-benzylaminopurine (6-BAP) and α-naphthaleneacetic acid (NAA) on the growth of shoots. Moreover, we developed and validated an analytical method to quantify carnosol using the H-point standard additions method in the high-performance liquid chromatography diode array detector (HPLC-DAD). After 30 days of culture, TIS produced the maximum number of shoots per explant (24.33 ± 1.15) on a liquid medium supplemented with 6-BAP at 5.0 mg L<sup>−1</sup>. Next, we also evaluated the effect of immersion time and frequency for TIS. After 72 days of culture, the best results were obtained with an immersion cycle of 1 min every 12 h, yielding 170.33 ± 29.40 shoots. The quantification of carnosol on the samples was performed at a flow rate of 1.2 mL min<sup>−1</sup> using binary isocratic mobile phase system 60:40 (<i>v</i>/<i>v</i>) 10 mM formic acid (pH 3.0) (A) and acetonitrile (B) on a reverse-phase column. The content of carnosol in the in vitro cultures was around 8-fold higher than in the wild plant. The present study represents an efficient alternative method to obtain carnosol for its pre-clinical and clinical development. |
| first_indexed | 2024-03-10T08:29:11Z |
| format | Article |
| id | doaj.art-4e89627eacaf4e03b0098b7b388a35f7 |
| institution | Directory Open Access Journal |
| issn | 1424-8247 |
| language | English |
| last_indexed | 2024-03-10T08:29:11Z |
| publishDate | 2021-07-01 |
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| record_format | Article |
| series | Pharmaceuticals |
| spelling | doaj.art-4e89627eacaf4e03b0098b7b388a35f72023-11-22T09:11:10ZengMDPI AGPharmaceuticals1424-82472021-07-0114874710.3390/ph14080747In Vitro Culture of <i>Rosmarinus officinalis</i> L. in a Temporary Immersion System: Influence of Two Phytohormones on Plant Growth and Carnosol ProductionEder Villegas-Sánchez0Mariana Macías-Alonso1Soraya Osegueda-Robles2Lisset Herrera-Isidrón3Hector Nuñez-Palenius4Joaquín González-Marrero5Instituto Politécnico Nacional, UPIIG, Av. Mineral de Valenciana, No. 200, Col. Fracc, Industrial Puerto Interior, Silao 36275, MexicoInstituto Politécnico Nacional, UPIIG, Av. Mineral de Valenciana, No. 200, Col. Fracc, Industrial Puerto Interior, Silao 36275, MexicoInstituto Politécnico Nacional, UPIIG, Av. Mineral de Valenciana, No. 200, Col. Fracc, Industrial Puerto Interior, Silao 36275, MexicoInstituto Politécnico Nacional, UPIIG, Av. Mineral de Valenciana, No. 200, Col. Fracc, Industrial Puerto Interior, Silao 36275, MexicoDivisión de Ciencias de la Vida, Campus Irapuato-Salamanca, Universidad de Guanajuato, Ex-Hacienda El Copal. Km. 9 Carr. Irapuato-Silao, Irapuato 36824, MexicoInstituto Politécnico Nacional, UPIIG, Av. Mineral de Valenciana, No. 200, Col. Fracc, Industrial Puerto Interior, Silao 36275, MexicoEmerging infectious diseases have become a major global problem with public health and economic consequences. It is an urgent need to develop new anti-infective therapies. The natural diterpene carnosol exhibit a wide variety of interesting antibacterial and antiviral properties, and it is considered a theoretical inhibitor of COVID-19 M<sup>pro</sup>. However, this compound is present in the family Lamiaceae in low quantities. To obtain carnosol in concentrations high enough to develop pharmacological studies, we evaluated the efficiency of a micropropagation protocol of <i>Rosmarinus officinalis</i> using a solid medium and a temporary immersion system (TIS), as well as the effect of 6-benzylaminopurine (6-BAP) and α-naphthaleneacetic acid (NAA) on the growth of shoots. Moreover, we developed and validated an analytical method to quantify carnosol using the H-point standard additions method in the high-performance liquid chromatography diode array detector (HPLC-DAD). After 30 days of culture, TIS produced the maximum number of shoots per explant (24.33 ± 1.15) on a liquid medium supplemented with 6-BAP at 5.0 mg L<sup>−1</sup>. Next, we also evaluated the effect of immersion time and frequency for TIS. After 72 days of culture, the best results were obtained with an immersion cycle of 1 min every 12 h, yielding 170.33 ± 29.40 shoots. The quantification of carnosol on the samples was performed at a flow rate of 1.2 mL min<sup>−1</sup> using binary isocratic mobile phase system 60:40 (<i>v</i>/<i>v</i>) 10 mM formic acid (pH 3.0) (A) and acetonitrile (B) on a reverse-phase column. The content of carnosol in the in vitro cultures was around 8-fold higher than in the wild plant. The present study represents an efficient alternative method to obtain carnosol for its pre-clinical and clinical development.https://www.mdpi.com/1424-8247/14/8/747infectious diseasescarnosolin vitrocultures<i>Rosmarinus officinalis</i>rosemary |
| spellingShingle | Eder Villegas-Sánchez Mariana Macías-Alonso Soraya Osegueda-Robles Lisset Herrera-Isidrón Hector Nuñez-Palenius Joaquín González-Marrero In Vitro Culture of <i>Rosmarinus officinalis</i> L. in a Temporary Immersion System: Influence of Two Phytohormones on Plant Growth and Carnosol Production Pharmaceuticals infectious diseases carnosol in vitro cultures <i>Rosmarinus officinalis</i> rosemary |
| title | In Vitro Culture of <i>Rosmarinus officinalis</i> L. in a Temporary Immersion System: Influence of Two Phytohormones on Plant Growth and Carnosol Production |
| title_full | In Vitro Culture of <i>Rosmarinus officinalis</i> L. in a Temporary Immersion System: Influence of Two Phytohormones on Plant Growth and Carnosol Production |
| title_fullStr | In Vitro Culture of <i>Rosmarinus officinalis</i> L. in a Temporary Immersion System: Influence of Two Phytohormones on Plant Growth and Carnosol Production |
| title_full_unstemmed | In Vitro Culture of <i>Rosmarinus officinalis</i> L. in a Temporary Immersion System: Influence of Two Phytohormones on Plant Growth and Carnosol Production |
| title_short | In Vitro Culture of <i>Rosmarinus officinalis</i> L. in a Temporary Immersion System: Influence of Two Phytohormones on Plant Growth and Carnosol Production |
| title_sort | in vitro culture of i rosmarinus officinalis i l in a temporary immersion system influence of two phytohormones on plant growth and carnosol production |
| topic | infectious diseases carnosol in vitro cultures <i>Rosmarinus officinalis</i> rosemary |
| url | https://www.mdpi.com/1424-8247/14/8/747 |
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