In vitro production of autotetraploid Ponkan mandarin (Citrus reticulata Blanco) using cell suspension cultures

• Published in: Euphytica Vol. 173; no. 2; pp. 235 - 242
• Main Authors: Dutt, M., Vasconcellos, M., Song, K. J., Gmitter, F. G., Grosser, J. W.
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.05.2010; Springer; Springer Nature B.V
• Subjects: Agronomy. Soil science and plant productions; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; Cell culture; Cell differentiation; Citrus; Citrus fruits; Colchicine; Enzymes; Ethylenediaminetetraacetic acid; Fundamental and applied biological sciences. Psychology; Genetics and breeding of economic plants; Life Sciences; Plant Genetics and Genomics; Plant Pathology; Plant Physiology; Plant reproduction; Plant Sciences; Sugars; Tetrahedra; Autotetraploid; Flow cytometry; Colchicine; Enzymatic maceration; Cell suspension; Cell culture; Enzyme; Citrus fruit; Citrus reticulata; Rutaceae; Rootstock; In vitro; Tetraploidy; Suspension culture; Autotetraploidy; Dicotyledones; Angiospermae; Genetic improvement; Autoploidy; Spermatophyta; Mandarin; Maceration
• ISSN: 0014-2336; 1573-5060
• DOI: 10.1007/s10681-009-0098-y

An efficient protocol for colchicine mediated production of in vitro autotetetraploids from Ponkan mandarin using cell suspension cultures is described. Cells were treated with 1 g l −1 colchicine for 4 or 8 days before transfer into solid EME medium supplemented with 5% maltose. Colchicine treated cells were placed in medium with or without an overlay of 1:2 medium–mixture of liquid 0.6 M BH3 medium and 0.15 M EME + maltose liquid media. It was observed that modifying the immediate cell environment by addition of the liquid overlay played a positive role in cell differentiation and subsequent plant regeneration. Ploidy levels were determined with a flow cytometer and confirmed by chromosome staining using the enzymatic maceration method. A large number of non-chimeric autotetraploids were generated using this method. Such plants have great value in a breeding program for the development of seedless triploid citrus, as very few available tetraploid breeding parents are easy to peel.