Endogenous auxin accumulation/localization during zygotic and somatic embryogenesis of Capsicum chinense Jacq.

Zygotic and somatic embryogenesis in plants is a fascinating event that is finely regulated through the expression of a specific group of genes and dynamic levels of plant hormones whose concerted action determines the fate that specific cells follow towards zygotic or somatic embryo development. This work studied different stages of Capsicum chinense Jacq. zygotic and somatic embryogenesis. HPLC quantification determined that the levels of indole-3-acetic acid (IAA) increase as the zygotic or somatic embryogenesis progresses, being higher at maturity, thus supporting a positive correlation between embryo cell differentiation and IAA increase. A monoclonal anti-IAA-antibody was used to detect IAA levels. Findings revealed a dynamic pattern of auxin distribution along the different embryogenic embryonic stages. In the early stages of zygotic embryos, the IAA gradient was observed in the basal cells of the suspensor and the hypostases, suggesting that they are the initial source of the IAA hormone. As embryogenesis proceeds, the dynamic of the IAA gradient is displaced to the embryo and endosperm cells. In the case of induced somatic embryogenesis, the IAA gradient was detected in the dividing cells of the endodermis, from where pre-embryogenic cells emerge. However, the analysis of somatic embryos revealed that IAA was homogeneously distributed. This study shows evidence supporting a correlation between IAA levels during zygotic or somatic embryogenesis in Capsicum chinense species.

Growth regulators and darkness increase efficiency in in vitro culture of immature embryos from peppers

Common pepper (Capsicum annuum L.) is one of the most important vegetables in the world, and extensive breeding efforts are being made to develop new improved strains of this species. In this regard, in vitro culture of immature embryos may help breeders accelerate breeding cycles and overcome interspecific barriers, among other applications. In this study, we have optimized a protocol for in vitro culture of immature embryos of C. annuum. Levels of indole-3-acetic acid (IAA) and zeatin have been tested to improve the efficiency (germination rates) of this technique in C. annuum embryos at the four main immature stages (i.e. globular, heart, torpedo, and early cotyledonary) from four varietal types of this species (California Wonder, Piquillo, Guindilla, and Bola). The effect of 5-day initial incubation in the dark was also tested on the most efficient hormone formulation. On average, relatively low levels of both IAA and zeatin (0.01 mg L¹ each) (M1) provided the highest germination rates, particularly in the advanced stages (torpedo and cotyledonary). To a lesser extent, the lack of these growth regulators (M0) or high IAA (0.2 mg L¹)/low zeatin (0.01 mg L¹) (M2) combination also had a positive response. On the contrary, high zeatin levels (0.2 mg L¹) produced very low germination rates or callus development (efficiency 0-7 %). Different responses were also found between genotypes. Thus, considering the best media (M0, M1, M2), Bola embryos had the highest rates. M1 plus 5-days of initial dark incubation (M1-D) improved the efficiency rates at all embryo stages, particularly in the earliest (globular) embryos which increased from 3 % to > 20 %.

Growth regulators and darkness increase efficiency in in vitro culture of immature embryos from peppers

Common pepper (Capsicum annuum L.) is one of the most important vegetables in the world, and extensive breeding efforts are being made to develop new improved strains of this species. In this regard, in vitro culture of immature embryos may help breeders accelerate breeding cycles and overcome interspecific barriers, among other applications. In this study, we have optimized a protocol for in vitro culture of immature embryos of C. annuum. Levels of indole-3-acetic acid (IAA) and zeatin have been tested to improve the efficiency (germination rates) of this technique in C. annuum embryos at the four main immature stages (i.e. globular, heart, torpedo, and early cotyledonary) from four varietal types of this species (California Wonder, Piquillo, Guindilla, and Bola). The effect of 5-day initial incubation in the dark was also tested on the most efficient hormone formulation. On average, relatively low levels of both IAA and zeatin (0.01 mg L¹ each) (M1) provided the highest germination rates, particularly in the advanced stages (torpedo and cotyledonary). To a lesser extent, the lack of these growth regulators (M0) or high IAA (0.2 mg L¹)/low zeatin (0.01 mg L¹) (M2) combination also had a positive response. On the contrary, high zeatin levels (0.2 mg L¹) produced very low germination rates or callus development (efficiency 0-7 %). Different responses were also found between genotypes. Thus, considering the best media (M0, M1, M2), Bola embryos had the highest rates. M1 plus 5-days of initial dark incubation (M1-D) improved the efficiency rates at all embryo stages, particularly in the earliest (globular) embryos which increased from 3 % to > 20 %.(AU)