In Vitro Cellular & Developmental Biology - Plant

, Volume 54, Issue 6, pp 658–671 | Cite as

A filter paper-based liquid culture system for citrus shoot organogenesis—a mixture-amount plant growth regulator experiment

  • Randall P. NiedzEmail author
  • Mizuri Marutani-Hert
Plant Tissue Culture


This study determined the effects of a static liquid culture system on shoot regeneration from citrus epicotyl explants. A mixture-amount experiment was used to determine the effects of zeatin riboside (ZR), 6-benzylaminopurine (BA), and indole-3-acetic acid (IAA) on two citrus types—citrange (Citrus sinensis ‘Washington’ L. Osbeck. × Poncirus trifoliata L. Raf var. Carrizo) and sweet orange (Citrus sinensis L. Osbeck var. ‘Ridge Pineapple’). A liquid culture system comprising a Petri dish, cellulose filter paper, and liquid culture medium was used. Shoot regeneration experiments were conducted over 6 wk that included 2 wk in the dark followed by 4 wk in the light. Three responses were measured: (1) number of explants forming buds and/or shoots, (2) number of explants with shoots > 2 mm, and (3) overall explant and shoot quality. The effects of paper disc number, liquid medium volume, and explant size on shoot regeneration were determined. High-quality shoots were produced from explants cultured in 5.25 to 12 mL medium volume and explant sizes ranging from 2 to 15 mm. The effects of the plant growth regulators were similar for the two citrus types and were as follows: (1) use of ZR or BA resulted in high-quality shoot production; (2) ZR and BA were not synergistic; (3) culture in 20 μM ZR resulted in the highest shoot production; (4) BA and IAA were strongly synergistic, with the greatest production with BA when IAA was included in the mixture; and (5) ZR and IAA were antagonistic, particularly with Ridge Pineapple.


Sweet orange Shoot regeneration Mixture-amount Plant growth regulators Experimental design 



We thank Eldridge Wynn for his careful preparation of the media formulations, assistance with the growth of the plant cultures, and collection of the data.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.


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Copyright information

© The Society for In Vitro Biology 2018

Authors and Affiliations

  1. 1.United States Department of Agriculture, Agricultural Research ServiceU.S. Horticultural Research LaboratoryFort PierceUSA

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