Abstract
Aim of study: A review on the propagation methods of four Quercus species, namely Q. suber, Q. robur, Q. ilex and Q. canariensis, through somatic embryogenesis and anther embryogenesis are presented.Area of study: The study comprises both Mediterranean and Atlantic oak species located in Spain.Material and Methods: Somatic embryogenesis was induced on immature zygotic embryos of diverse oak species, permitting the multiplication of half-sib families. Induction of haploid embryos and doubled haploids was assayed in both Q. suber and Q. ilex by temperature stress treatments of anthers containing late vacuolated microspores. The haploid origin of the anther embryos has been evaluated by quantitative nuclear DNA analysis through flow cytometry and by DNA microsatellite markers. Genetic transformation of cork oak has also been performed by means of Agrobacterium tumefaciens vectors. Proteomic analysis has been conducted to screen the diverse protein profiles followed by in vitro derived embryos during their development.Research highlights: Successful plant regeneration from both somatic and haploid embryos has been achieved. In the particular case of cork oak, doubled-haploid plants were obtained. Plantlets regenerated from selected parent trees through somatic embryogenesis were acclimated in the greenhouse and in the nursery, and were planted in an experimental plot in the field. Preliminary evaluation of the cork quality of the plants showed a good heritability correlation with the parent trees. This article revises the work of and is dedicated to Dr. M.A. Bueno, who devoted much of her professional life to the research on Biotechnology and Genetics of forest species, leading the Laboratory of Forest Biotechnology at the Spanish Institute of Agronomic Research (INIA).Key words: anther embryogenesis; microspore; pollen; Quercus canariensis; Quercus ilex; Quercus robur; Quercus suber; somatic embryogenesis.
Highlights
Area of study: The study comprises both Mediterranean and Atlantic oak species located in Spain
Induction of haploid embryos and doubled haploids was assayed in both Q. suber and Q. ilex by temperature stress treatments of anthers containing late vacuolated microspores
Somatic embryogenesis has been assayed in this oak. First results in this species were obtained by Chalupa (1990), who reported successful plantlet regeneration from immature embryos cultured on Murashige and Skoog (MS) or Woody Plant Medium (WPM; Lloyd and McCown, 1981), supplemented with benzyl adenine (BA) 4.4 μM, alone or combined with 2.9 μM gibberellic acid (GA3) or indole-butyric acid (IBA) 0.5 to 5 μM
Summary
Cork oak (Quercus suber L.) is a forest species playing a major role in many Mediterranean ecosystems. The transformation of 5.8% embryos selected on hygromycin was confirmed by expression of β-glucuronidase four months after co-cultivation (Fig. 2) and by the presence of nopaline synthase terminator, amplified by polymerase chain reaction This method is one of the few reports on the genetic transformation of cork oak somatic embryos (Sanchez et al, 2005). Somatic embryos germinated into plantlets, producing more than 900 individuals which were transferred to 100 ml nursery pots filled with peat: perlite: vermiculite (1:1:1). The percentage of virgin cork thickness of the main shoot of the somatic embryo-derived plantlets showed a moderately strong association with the cork thickness in twigs of their parent trees This progeny test permitted the estimation of a high
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