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Production of chlorogenic acid and isoorientin hypoglycemic compounds in Cecropia obtusifolia calli and in cell suspension cultures with nitrate deficiency

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Abstract

The antidiabetic properties of Cecropia obtusifolia are attributed to chlorogenic acid (CGA) and isoorientin (ISO) phenolic compounds; both compounds possess hypoglycemic, hypolipidemic, and antioxidant properties. As a potential strategy for an adequate supply of authentic plant raw material, the aim of this study was to establish in vitro conditions for the development of cell suspension cultures that produce these bioactive compounds. Callus cultures of leaf explants from acclimatized tree and in vitro plantlets were set up using different auxin levels; treatments with 2,4-dichlorophenoxyacetic acid (2,4-D) and α-naphthalene acetic acid (NAA) to 8.92 µM with 6-benzylaminopurine (BAP) at 2.22 μM stimulate highest callus production. Seedling cotyledon, hypocotyl, leaf, and stem explants developed calli bearing roots with 2,4-D. With NAA, hypocotyl, cotyledon, and leaf explants developed morphogenic calli; 75% of stem explants formed calli, and the remaining calli developed shoots. Determined CGA concentrations in calli were similar to those detected in the leaves of wild trees, and ISO was not produced. Cell suspension cultures were established from leaf explants friable calli with 8.92 μM 2,4-D in combination with 2.22 μM BAP, employing 4 and 5% inocula in fresh weight; CGA levels were maintained and ISO was produced only at the end of logarithmic growth. On diminishing nitrate content in Murashige and Skoog (MS) medium to 8.0 mM, maximum cell biomasses diminished, CGA production is increased and twice with 16.0 and, instead of CGA production is tripled and quadrupled with 16.0 and 8.0 mM nitrates, respectively, and ISO synthesis was induced earlier and for a longer time period, increasing its levels at the end of culture. Two compounds with ultraviolet spectra similar to those of caffeic and ferulic acids were formed. Our results offer a protocol of cell suspension cultures for C. obtusifolia bioactive production and hypoglycemic property conservation.

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Abbreviations

BAP:

6-Benzylaminopurine

CGA:

Chlorogenic acid

2,4-D:

2,4-Dichlorophenoxyacetic acid

DM:

Diabetes mellitus

DW:

Dry weight

IBA:

Indolebutyric acid

MSD:

Minimal significant difference

IAA:

Indole-3-acetic acid

ISO:

Isoorientin

MS:

Murashige and Skoog

NAA:

α-Naphthalene acetic acid

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Acknowledgments

MP Nicasio-Torres is grateful for the financial support (2004/077) provided for this research by FOFOI-IMSS, and to Prof. Abigail Aguilar of the Medicinal Plant Herbarium of the Mexican Social Security Institute (IMSSM) for plant material certification. Further appreciation is expressed, especially to my friend, Dr. Eduardo Aranda-Escobar† for his invaluable help.

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Authors and Affiliations

  1. Laboratorios de Biotecnología y Farmacología, Centro de Investigación Biomédica del Sur, Instituto Mexicano del Seguro Social, Argentina No. 1 Centro, 62790, Xochitepec, Morelos, México

    María del Pilar Nicasio-Torres & Lucía Aguilar-Santamaría

  2. Centro de Diagnóstico en Metabolismo Energético y Medicina Mitocondrial, S.C. 03100, México, Distrito Federal, México

    Mariana Meckes-Fischer

  3. Centro de Investigaciones Químicas, Universidad Autónoma del Estado de Morelos, 62210, Cuernavaca, Morelos, México

    María Luisa Garduño-Ramírez

  4. Jardín Botánico, Instituto de Biología, Universidad Nacional Autónoma de México, 04510, México, Distrito Federal, México

    Víctor Manuel Chávez-Ávila

  5. Departamento de Biotecnología, Universidad Autónoma Metropolitana-Iztapalapa, 09340, México, Distrito Federal, México

    Francisco Cruz-Sosa

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  1. María del Pilar Nicasio-Torres
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  2. Mariana Meckes-Fischer
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  3. Lucía Aguilar-Santamaría
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Correspondence to María del Pilar Nicasio-Torres.

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Communicated by E. Lojkowska.

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Nicasio-Torres, M.P., Meckes-Fischer, M., Aguilar-Santamaría, L. et al. Production of chlorogenic acid and isoorientin hypoglycemic compounds in Cecropia obtusifolia calli and in cell suspension cultures with nitrate deficiency. Acta Physiol Plant 34, 307–316 (2012). https://doi.org/10.1007/s11738-011-0830-9

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  • DOI: https://doi.org/10.1007/s11738-011-0830-9

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