Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 103, Issue 1, pp 93–101 | Cite as

Influence of plant growth regulators and water stress on ramet induction, rosette engrossment, and fructan accumulation in Agave tequilana Weber var. Azul

  • Rita Barreto
  • Jorge Nieto-Sotelo
  • Gladys I. Cassab
Original Paper


Agave tequilana Weber var. Azul plants reproduce asexually by producing ramets. Continuous production of ramets throughout the vegetative cycle of the parent delays the time of harvesting of heads for tequila production. Little is known about the factors influencing their emergence. Heads are engrossed rosettes where fructans are stored. We show here that, in plantlets grown in vitro, growth regulators such as 2,4-dichlorophenoxyacetic acid (2,4-D), a combination of 1-naphthaleneacetic acid (NAA)/6-benzyladenine (BA), or abscisic acid (ABA) increased the production of ramets, whereas BA, NAA, gibberellic acid (GA3), glycerol, or a combination of glycerol/ABA decreased ramet production. Plantlets that developed ramets did not form heads. Head formation was improved on solid media in the presence of BA, NAA, the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC), or the water stress inducer polyethylene glycol (PEG). Basal Murashige–Skoog (MS) liquid media also enhanced rosette engrossment, which was further increased by addition of ACC or PEG. In contrast, CoCl2, an ethylene biosynthesis inhibitor, reduced rosette engrossment. Furthermore, heads from A. tequilana plantlets grown in tissue culture in MS media, or in MS media supplemented with NAA, ACC or PEG, showed fructan concentrations 10–30 times higher than in leaves from greenhouse-grown plants. Our results indicated that BA, NAA, water stress, and ethylene are critical regulators of rosette engrossment, whereas asexual reproduction in A. tequilana seems to be controlled by a complex hormonal network.


ABA Asexual reproduction Auxin Cytokinin Ethylene Gibberellin Reserve carbohydrates 



2,4-Dichlorophenoxyacetic acid


Abscisic acid


1-Aminocyclopropane-1-carboxylic acid






Fresh weight


Gibberellic acid


Murashige–Skoog media


1-Naphthaleneacetic acid


Polyethylene glycol



We thank Dr. I. del Real and M.C.R. Ayala (Sauza-Pedro Domecq) for providing A. tequilana Azul var. Weber plants and their committed support to the project. We appreciate the generous help of F. González and Dr. A. López-Munguía in the determination of fructose levels in A. tequilana samples and Dr. M.A. Rosales for statistical analysis of the data. We also warmly thank all the members of the laboratory for their continuous support and discussion of the data. The technical expertise of M. Saucedo Ramírez, computer support of R. Bahena, A. Martínez Valle, and J.M. Hurtado, and library support of Shirley Ainsworth are fully appreciated. This work was supported by a grant from Pedro Domecq (P-150) to J.N.-S. and G.I.C.

Supplementary material

11240_2010_9758_MOESM1_ESM.pdf (36 kb)
Supplemental Fig. 1.Effect of ACC and PEG on rosette development in plantlets of A. tequilana grown for 90 days in solid media. Rosette engrossment is described as the percentage of plants that showed this response. Concentrations of chemicals were: ACC-1 (1 μM), ACC-10 (10 μM), PEG-5 (5% PEG [v/v]), PEG-8 (8% PEG), and PEG-10 (10% PEG). MS, ACC-10, and PEG-5 data were obtained from six, three, and two separate experiments, respectively. Data for ACC-1, PEG-8, and PEG-10 were obtained from one single experiment. Data represent average ± SD values. Bars with different letters differ significantly. (PDF 35 kb)


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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.Departamento de Biología Molecular de Plantas, Instituto de BiotecnologíaUniversidad Nacional Autónoma de MéxicoCuernavacaMexico
  2. 2.Facultad de CienciasUniversidad Autónoma del Estado de MorelosCuernavacaMexico

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