ABSTRACT
Plinia cauliflora is an important Brazilian species that produces highly appreciated fruits, with a great potential of commercialization. However, the high cost of seedlings is a bottleneck for the expansion of commercial orchards. The present study aimed to investigate somatic embryogenesis as a propagation method for P. cauliflora using seeds as explants. To induce embryogenic mass (EM) and somatic embryo (SE) development we evaluated the supplementation of culture medium with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), combined or not with activated charcoal (AC). For the embryo maturation, we investigated the effects of AC, polyethylene glycol (PEG), Gelzan®, 6-benzylaminopurine and gibberellin supplementation. For the EM induction, the best results were obtained in MS culture medium supplemented with 300 µM 2,4-D and 1 g L-1 AC. During the first maturation phase, the supplementation of 30 g L-1 PEG improved the somatic embryo formation at the torpedo and cotyledonary stages, whereas the maturation treatments did not result in the conversion of the embryos into plantlets. The anatomical analysis showed that the 2,4-D presence for 60 days may have been deleterious for embryonic development. These results represent the first report of P. cauliflora somatic embryogenesis and its feasibility for mass propagation.
Subject(s)
Embryonic Development , BrazilABSTRACT
This chapter deals with the features of somatic embryogenesis (SE) in Araucaria angustifolia, an endangered and native conifer from south Brazil. In this species SE includes the induction and proliferation of embryogenic cultures composed of pro-embryogenic masses (PEMs), which precede somatic embryos development. A. angustifolia SE model encompasses induction, proliferation, pre-maturation, and maturation steps. Double-staining with acetocarmine and Evan's blue is useful to evaluate the embryonic somatic structures. In this chapter we describe A. angustifolia SE protocols and analyzes morphological features in the different SE developmental stages.
Subject(s)
Plant Development/genetics , Plant Somatic Embryogenesis Techniques/methods , Tissue Culture Techniques/methods , Tracheophyta/growth & development , Brazil , Flowers/genetics , Flowers/growth & development , Germination/genetics , Seeds/genetics , Seeds/growth & development , Tracheophyta/geneticsABSTRACT
UNLABELLED: DNA methylation is an epigenetic regulatory mechanism of gene expression which can be associated with developmental phases and in vitro morphogenetic competence in plants. The present work evaluated the effects of 5-azacytidine (AzaC) and 2,4-dichlorophenoxyacetic acid (2,4-D) on Acca sellowiana somatic embryogenesis (SE) and global DNA methylation levels by high-performance liquid chromatography mass spectrometry (HPLC/MS/MS). 2,4-D-free treatments revealed no somatic embryo formation in both accessions tested. Treatments supplemented with 2,4-D pulse plus AzaC in the culture medium resulted in increased embryo formation. In AzaC-free treatment, HPLC/MS/MS analysis showed a gradual increase in methylation levels in cultures of both accessions tested during SE induction. Treatment with AzaC and 2,4-D-free resulted in a marked decrease in methylation for both accessions, ranging from 37.6 to 20.8 %. In treatment with 2,4-D and AzaC combined, the 85 accession showed increasing global methylation levels. Otherwise, the 101X458 accession, in the same treatment, showed a decrease between 10 and 20 days, followed by an increase after 30 days (39.5, 36.2 and 41.6 %). These results indicate that 2,4-D pulse combined with AzaC improves SE induction. However, the conversion phase showed that although positively influencing SE induction, AzaC had a dysregulatory effect on the stage of autotrophic plant formation, resulting in significantly lower conversion rates. The results suggest that DNA methylation dramatically influences SE in Acca sellowiana, and global DNA methylation dynamics are related to morphogenetic response. KEY MESSAGE: 5-Azacytidine combined with 2,4-D increases the number of Acca sellowiana somatic embryos. Global DNA methylation is directly affected by these compounds.