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

Abstract

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.

Morphological changes and self-incompatibility breakdown associated with autopolyploidization in Hylocereus species (Cactaceae)

“…There is a lot of evidence of the successful induction of autopolyploids, which exhibit improved traits compared with those of their donor lines, using anti-mitotic reagents (Kadota and Niimi 2002;Eeckhaut et al 2004;Liu et al 2007;Zhang et al 2008;Campos et al 2009;Dhooghe et al 2009;Viehmannova et al 2009;Dutt et al 2010;Glowaka et al 2010;Kulkarni and Borse 2010;Xu et al 2010;Ye et al 2010;Zhang et al 2010). Indeed, successful artificial autopolyploidization in Hylocereus species was recently reported (Tel-Zur et al 2011b).…”

Enhancement of artemisinin content in tetraploidArtemisia annuaplants by modulating the expression of genes in artemisinin biosynthetic pathway

“…Polyploids often show novel phenotypes, such as broader leaves, good quality, high yield, and increased resistance to environment stress and diseases 9,10. To induce autopolyploids, colchicine treatments are widely used 11–15. With the basic genetic material remain the same, chromosome doubling in autopolyploids is often accompanied by conspicuous changes in secondary metabolism.…”

Breeding triploid plants: a review

“…But colchicine-induced tetraploids ex vitro were confronted with low mutation rate, high chimeric rate and reverse mutation, were difficult to select and vulnerable to environmental disturbance. Fusion of protoplasts together with colchicine-induction created homogenous tetraploid of Citrus reticulata Blanco (Dutt et al 2010). But fusion of protoplasts is technically demanding (Eeckhaut et al 2013).…”