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Mixture screening and mixture-amount designs to determine plant growth regulator effects on shoot regeneration from grapefruit (Citrus paradisi macf.) epicotyls

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Abstract

The objective of this study was to improve shoot regeneration from grapefruit. Because many commercially grown citrus types are apomictic, important in vitro applications such as Agrobacterium-mediated transformation commonly use epicotyl explants from in vitro seedlings; thus, adequate adventitious shoot production is an important prerequisite for efficient use of these applications. Eight plant growth regulators were studied—six cytokinins (6-benzylaminopurine, kinetin, zeatin trans-isomer, 6-[γ,γ-dimethylallylamino] purine, zeatin riboside trans-isomer and meta-topolin) and two auxins (α-naphthalene acetic acid and indole-3-acetic acid). An iterative design strategy was followed that included mixture and mixture-amount experimental designs suitable for resolving proportional and concentration effects; in vitro effects of cytokinins and auxins are affected by both proportion and concentration. One-centimeter-long explants were excised from the epicotyl of etiolated, in vitro-grown seedlings. Explants were placed onto experimental formulations and cultured in growth cabinets at 27°C over 6 wk, which included 2 wk in the dark followed by 4 wk in the light. The results indicated that (1) 6-benzylaminopurine or zeatin riboside were the most effective cytokinins for inducing shoot regeneration in citrus; (2) zeatin riboside singly or in combination with indole-3-acetic acid resulted in the highest quality, the greatest number of explants with buds/shoots, and the greatest shoot number; and (3) 6-benzylaminopurine and indole-3-acetic acid improved shoot regeneration vs. 6-benzylaminopurine at a considerably lesser cost than zeatin riboside and indole-3-acetic acid.

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Acknowledgment

We thank Mr. Eldridge Wynn for his careful preparation of the media formulations, growth of the plant cultures, and setup and collection of the data for this study.

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Authors and Affiliations

  1. Agricultural Research Service, U.S. Horticultural Research Laboratory, 2001 South Rock Road, Ft. Pierce, FL, 34945-3030, USA

    Randall P. Niedz & Terence J. Evens

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  1. Randall P. Niedz
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  2. Terence J. Evens
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Correspondence to Randall P. Niedz.

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Editor: J. Finer

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Niedz, R.P., Evens, T.J. Mixture screening and mixture-amount designs to determine plant growth regulator effects on shoot regeneration from grapefruit (Citrus paradisi macf.) epicotyls. In Vitro Cell.Dev.Biol.-Plant 47, 682–694 (2011). https://doi.org/10.1007/s11627-011-9381-4

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  • DOI: https://doi.org/10.1007/s11627-011-9381-4

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