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In-vitro Callus Induction and Rosmarinic Acid Quantification in Callus Culture of Satureja khuzistanica Jamzad (Lamiaceae).

Sahraroo A, Babalar M, Mirjalili MH, Fattahi Moghaddam MR, Nejad Ebrahimi S - Iran J Pharm Res (2014)

Bottom Line: Determination and quantification of RA in cultured calli were performed by HPLC UV/MS analysis.Calli induced from the plant and maintained on supplements of IBA and BAP in the absence of light produced RA 7.5% based on dry weight (DW).No differentiation was observed in any callus during the course of this study.

View Article: PubMed Central - PubMed

Affiliation: Department of Horticulture, Faculty of Agriculture, University of Tehran, Karaj, Iran.

ABSTRACT
In the present study, an efficient protocol has been developed for callus induction and production of RA in callus culture of Satureja khuzistanica for the first time. In-vitro callus induction was achieved from young shoot tip explants cultured on MS and B5 media supplemented with different concentrations of IBA (0.1, 1.0, 2.0 and 5.0 mgL(-1)) solely or in combination with cytokinins BAP and KIN (1.0, 2.0 and 5.0 mgL(-1)). B5 medium supplemented with 1.0 mgL(-1) IBA plus 5.0 mgL(-1) BAP and MS medium fortified with 2.0 mgL(-1) IBA and 2.0 mgL(-1) BAP were the most favorable media for callus formation with the highest induction rate (96%). Maximum growth index (2.89 and 2.63) and maximum callus biomass (2.34 and 2.33 g fresh weight) were obtained from the callus cultured on B5 medium supplemented with 1.0 mgL(-1) IBA plus 5.0 mgL(-1) BAP and MS medium fortified with 1.0 mgL(-1) IBA plus 1.0 mgL(-1) KIN, respectively. Determination and quantification of RA in cultured calli were performed by HPLC UV/MS analysis. Calli induced from the plant and maintained on supplements of IBA and BAP in the absence of light produced RA 7.5% based on dry weight (DW). No differentiation was observed in any callus during the course of this study.

No MeSH data available.


Related in: MedlinePlus

Callus induction at the basal end of Satureja khuzistanica shoot tip explants after two weeks of inoculation
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Figure 2: Callus induction at the basal end of Satureja khuzistanica shoot tip explants after two weeks of inoculation

Mentions: A preliminary experiment was carried out using in-vitro young leaves, internodes and shoot tip as explants in the callus induction experiment. However, young leaves and internodes cultured with either PGRs or without them began to turn brown after one week of culture. Only shoot tip explants survived and used for further study (data not shown). During the present set of experiment, different PGRs (IBA, BAP and KIN) on MS and B5 media were also tried for callus induction from shoot tip explants. Visible calli were produced at the basal ends of shoot tip explants after two weeks of inoculation (Figure 2). Depending on the PGRs treatments and type of medium culture used, a wide range of variation in frequency of callus formation and nature of callus was observed. No calli were induced in the media without the plant growth regulators (Table 1), indicating that PGRs are required for callus induction. Karam et al. (2003) also recorded the same results and emphasized the importance of exogenous PGRs for dividing cells and thus callus formation in Salvia fruticosa (23). Our results revealed that the highest percentage of callus induction (96%) was achieved from the B5 medium supplemented by 1 mgL-1 IBA + 5 mgL-1 BAP as well as MS medium containing 2 mgL-1 IBA + 2 mgL-1 BAP (Table 1). Similar observations with IBA and BAP, at different concentrations to support callus induction and proliferation were reported earlier in S. hortensis (17) and in other Lamiaceae member, Agastache rugosa (24). However, optimal concentration of these compounds may depend on many factors, such as a genotype of original plant, explants origin and etc. In relation to the callus nature, the calli obtained from B5 medium was friable and white green in color. Friable and light green colored calli were also induced in nearly all media containing 1 mgL-1 IBA. In our study, callus biomass progressively increased with an increase in the BAP concentration (Table 2). In this case, the maximum growth index (2.89 and 2.63) and maximum callus biomass (2.34 and 2.33 g fresh weight) were obtained from the callus cultured on B5 medium supplemented with 1.0 mgL-1 IBA plus 5.0 mgL-1 BAP and MS medium fortified with 1.0 mgL-1 IBA plus 1.0 mgL-1 KIN, respectively (Table 2).


In-vitro Callus Induction and Rosmarinic Acid Quantification in Callus Culture of Satureja khuzistanica Jamzad (Lamiaceae).

Sahraroo A, Babalar M, Mirjalili MH, Fattahi Moghaddam MR, Nejad Ebrahimi S - Iran J Pharm Res (2014)

Callus induction at the basal end of Satureja khuzistanica shoot tip explants after two weeks of inoculation
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4232813&req=5

Figure 2: Callus induction at the basal end of Satureja khuzistanica shoot tip explants after two weeks of inoculation
Mentions: A preliminary experiment was carried out using in-vitro young leaves, internodes and shoot tip as explants in the callus induction experiment. However, young leaves and internodes cultured with either PGRs or without them began to turn brown after one week of culture. Only shoot tip explants survived and used for further study (data not shown). During the present set of experiment, different PGRs (IBA, BAP and KIN) on MS and B5 media were also tried for callus induction from shoot tip explants. Visible calli were produced at the basal ends of shoot tip explants after two weeks of inoculation (Figure 2). Depending on the PGRs treatments and type of medium culture used, a wide range of variation in frequency of callus formation and nature of callus was observed. No calli were induced in the media without the plant growth regulators (Table 1), indicating that PGRs are required for callus induction. Karam et al. (2003) also recorded the same results and emphasized the importance of exogenous PGRs for dividing cells and thus callus formation in Salvia fruticosa (23). Our results revealed that the highest percentage of callus induction (96%) was achieved from the B5 medium supplemented by 1 mgL-1 IBA + 5 mgL-1 BAP as well as MS medium containing 2 mgL-1 IBA + 2 mgL-1 BAP (Table 1). Similar observations with IBA and BAP, at different concentrations to support callus induction and proliferation were reported earlier in S. hortensis (17) and in other Lamiaceae member, Agastache rugosa (24). However, optimal concentration of these compounds may depend on many factors, such as a genotype of original plant, explants origin and etc. In relation to the callus nature, the calli obtained from B5 medium was friable and white green in color. Friable and light green colored calli were also induced in nearly all media containing 1 mgL-1 IBA. In our study, callus biomass progressively increased with an increase in the BAP concentration (Table 2). In this case, the maximum growth index (2.89 and 2.63) and maximum callus biomass (2.34 and 2.33 g fresh weight) were obtained from the callus cultured on B5 medium supplemented with 1.0 mgL-1 IBA plus 5.0 mgL-1 BAP and MS medium fortified with 1.0 mgL-1 IBA plus 1.0 mgL-1 KIN, respectively (Table 2).

Bottom Line: Determination and quantification of RA in cultured calli were performed by HPLC UV/MS analysis.Calli induced from the plant and maintained on supplements of IBA and BAP in the absence of light produced RA 7.5% based on dry weight (DW).No differentiation was observed in any callus during the course of this study.

View Article: PubMed Central - PubMed

Affiliation: Department of Horticulture, Faculty of Agriculture, University of Tehran, Karaj, Iran.

ABSTRACT
In the present study, an efficient protocol has been developed for callus induction and production of RA in callus culture of Satureja khuzistanica for the first time. In-vitro callus induction was achieved from young shoot tip explants cultured on MS and B5 media supplemented with different concentrations of IBA (0.1, 1.0, 2.0 and 5.0 mgL(-1)) solely or in combination with cytokinins BAP and KIN (1.0, 2.0 and 5.0 mgL(-1)). B5 medium supplemented with 1.0 mgL(-1) IBA plus 5.0 mgL(-1) BAP and MS medium fortified with 2.0 mgL(-1) IBA and 2.0 mgL(-1) BAP were the most favorable media for callus formation with the highest induction rate (96%). Maximum growth index (2.89 and 2.63) and maximum callus biomass (2.34 and 2.33 g fresh weight) were obtained from the callus cultured on B5 medium supplemented with 1.0 mgL(-1) IBA plus 5.0 mgL(-1) BAP and MS medium fortified with 1.0 mgL(-1) IBA plus 1.0 mgL(-1) KIN, respectively. Determination and quantification of RA in cultured calli were performed by HPLC UV/MS analysis. Calli induced from the plant and maintained on supplements of IBA and BAP in the absence of light produced RA 7.5% based on dry weight (DW). No differentiation was observed in any callus during the course of this study.

No MeSH data available.


Related in: MedlinePlus