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Adventitious shoot formation in decapitated dicotyledonous seedlings starts with regeneration of abnormal leaves from cells not located in a shoot apical meristem

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Abstract

Regeneration of new shoots in plant tissue culture is often associated with appearance of abnormally shaped leaves. We used the adventitious shoot regeneration response induced by decapitation (removal of all preformed shoot apical meristems, leaving a single cotyledon) of greenhouse-grown cotyledon-stage seedlings to test the hypothesis that such abnormal leaf formation is a normal regeneration progression following wounding and is not conditioned by tissue culture. To understand why shoot regeneration starts with defective organogenesis, the regeneration response was characterized by morphology and scanning electron and light microscopy in decapitated cotyledon-stage Cucurbita pepo seedlings. Several leaf primordia were observed to regenerate prior to differentiation of a de novo shoot apical meristem from dividing cells on the wound surface. Early regenerating primordia have a greatly distorted structure with dramatically altered dorsoventrality. Aberrant leaf morphogenesis in C. pepo gradually disappears as leaves eventually originate from a de novo adventitious shoot apical meristem, recovering normal phyllotaxis. Similarly, following comparable decapitation of seedlings from a number of families (Chenopodiaceae, Compositae, Convolvulaceae, Cucurbitaceae, Cruciferae, Fabaceae, Malvaceae, Papaveraceae, and Solanaceae) of several dicotyledonous clades (Ranunculales, Caryophyllales, Asterids, and Rosids), stems are regenerated bearing abnormal leaves; the normal leaf shape is gradually recovered. Some of the transient leaf developmental defects observed are similar to responses to mutations in leaf shape or shoot apical meristem function. Many species temporarily express this leaf development pathway, which is manifest in exceptional circumstances such as during recovery from excision of all preformed shoot meristems of a seedling.

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Acknowledgments

Contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel, No. 506/06. This work was partially supported by a Research Grant from the Ministry of Agriculture and Rural Development, Israel, to V.G. K. Kathiravan received a BOYSCAST Fellowship from the Ministry of Science and Technology, Government of India. S.A. was supported by the MASHAV program of the Ministry of Foreign Affairs, Israel. The authors thank Dr. Moshe Lapidot, ARO Volcani Center, for the use of greenhouse facilities and Naomi Bahat, EM Unit, Faculty of Agriculture, the Hebrew University of Jerusalem, Rehovot, for skilled technical assistance. The authors thank Prof. Simcha Lev-Yadun for critical comments on the ms.

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  1. Krishnan Kathiravan

    Present address: Dept of Biotechnology, University of Madras, Guindy Campus, Chennai, 600 025, India

Authors and Affiliations

  1. Department of Plant Pathology and Weed Science, Agricultural Research Organization, The Volcani Center, P. O. Box 6, Bet Dagan, 50250, Israel

    Sampath Amutha, Krishnan Kathiravan, Sima Singer & Victor Gaba

  2. Department of Food Science, Agricultural Research Organization, The Volcani Center, P. O. Box 6, Bet Dagan, 50250, Israel

    Liana Jashi & Ilan Shomer

  3. Department of Vegetable Crops and Plant Genetics, Agricultural Research Organization The Volcani Center, P. O. Box 6, Bet Dagan, 50250, Israel

    Benjamin Steinitz

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  1. Sampath Amutha
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Correspondence to Victor Gaba.

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Amutha, S., Kathiravan, K., Singer, S. et al. Adventitious shoot formation in decapitated dicotyledonous seedlings starts with regeneration of abnormal leaves from cells not located in a shoot apical meristem. In Vitro Cell.Dev.Biol.-Plant 45, 758–768 (2009). https://doi.org/10.1007/s11627-009-9232-8

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