Abstract
We selected two cultivars of Brassica campestris ssp. pekinensis (Chinese cabbage), ‘Futian 50’ and ‘Changkuai,’ to investigate the factors that influence microspore embryogenesis and plantlet regeneration. We have also discussed some protocols for the induction culture of microspore-derived embryos and for rooting and transplantation of microspore-derived plantlets. We obtained the following findings in our study. Although NAA promotes the development of embryo and reduces the percentage of abnormal embryos, it inhibits the formation of microspore-derived embryos. 2,4-D also inhibits the formation of microspore-derived embryos. Low concentrations of cytokinins facilitate embryogenesis, while high concentrations inhibit embryogenesis; BA has a stronger influence than zeatin (Z) on embryo induction. The combined effects of auxin and cytokinin are synergistic. Organic compounds increase the rate of formation of microspore-derived embryos. However, activated charcoal (AC) inhibits embryo development. The better the development of the embryos, the higher is the plantlet regeneration rate. The plant regeneration rate increased significantly on the MS culture medium supplemented with 200 mg·L-1 AC. The MS medium s suitable for the subculture of the regenerated plantlets. MS medium containing 0.1 mg·L-1 NAA is the optimal medium for rooting of microspore-derived plantlets. Key words: Chinese cabbage, microspore culture, embryogenesis, plant regeneration.
Highlights
Doubled haploids (DHs) are being commonly used in plant breeding as an important means of accelerating the development of new cultivars (Yao et al, 2008) or as one of the favored mapping populations for constructing linkage maps to identify quantitative trait loci (Pilet et al., 2001; Zhao et al, 2005; Yang et al, 2008)
In recent years, isolated microspore culture has attracted a lot of interest because of its great potential for DH production
(1982) first reported the embryogenesis in microspore cultures of Brassica napus, embryogenesis has been induced in microspore cultures of different Brassica species (Babbar et al, 2004)
Summary
Doubled haploids (DHs) are being commonly used in plant breeding as an important means of accelerating the development of new cultivars (Yao et al, 2008) or as one of the favored mapping populations for constructing linkage maps to identify quantitative trait loci (Pilet et al., 2001; Zhao et al, 2005; Yang et al, 2008). A microspore-derived embryo is a suitable system for gene mapping (Graner, 1996; Zhang et al 2005; Geng et al, 2007; Yu et al, 2008), genetic transformation (Stǒger et al, 1995) and selection of dominant and recessive traits (Polsoni et al, 1988; Swanson, 1989; Liu et al., 2003; Wang et al, 2008; Zou et al, 2009). Since Lichter (1982) first reported the embryogenesis in microspore cultures of Brassica napus, embryogenesis has been induced in microspore cultures of different Brassica species (Babbar et al, 2004).
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