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Abstract

An efficient system of micropropagation via somatic embryogenesis from root-derived callus was established in Arabica coffee (Coffea arabica L.). Twenty-six callus lines were induced on MS (Murashige and Skoog, 1962) medium supplemented with combinations of NAA (0, 0.1, 0.5, 1 and 2 mg/L) plus BA (0, 1 and 2 mg/L), or 2,4-D (0, 0.1, 0.5, 1 and 2 mg/L) plus TDZ (0, 1 and 2 mg/L). Subsequently, two types of somatic embryos were obtained from callus cultures and named S-type and I-type embryos. The S-type embryos were obtained from an 18-monthold callus line which was induced and maintained at 2 mg/L TDZ and 0.1 mg/L 2,4-D near the end of each period of the subculture. These embryos have a developmental barrier, which did not pass through the torpedo stage and could be overcome by a supplement of 2 or 5 mg/L BA. The I-type embryos were induced from 3-month-old callus when transferred onto induction media, i.e., MS supplemented with TDZ (2 and 5 mg/L) plus 2,4-D (0 and 0.1 mg/L). The significantly highest response, i.e., 13.3 embryos per callus clump was obtained at 2 mg/L TDZ. In this study, the results reveal that TDZ has a crucial effect on embryogenic callus induction, proliferation and subsequent somatic embryogenesis.
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Abstract

Culture gas atmosphere is one of the most important factors affecting embryo development in vitro. The main objective of this study was to compare the effects of CO concentration on the subsequent pre-implantation developmental capacity of pig embryos in vitro, including embryos obtained via parthenogenesis, in vitro fertilization (IVF), and intracytoplasmic sperm injection (ICSI). Pig embryos were developed in four different CO2 concentrations in air: 3%, 5%, 10%, or 15%. The cleavage rate of pig parthenogenetic, IVF, or ICSI embryos developed in CO2 concen- trations under 5% was the highest. There were no significant differences in the oocyte cleavage rate in ICSI embryos in CO2 concentrations under 3% and 5% (p>0.05). However, as CO2 levels increased (up to 15%) the blastocyst output on day 7, from parthenogenetic, IVF, and ICSI em- bryos, decreased to 0%. These findings demonstrate that CO2 positively affects the developmen- tal capacity of pig embryos. However, high or low CO2 levels do not significantly improve the developmental capacity of pig embryos. The best results were obtained for all of the pig embryos at a 5% CO2 concentration.
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