Characterization Of A New Family Of Proteins That Interact With The C-terminal Region Of The Chromatin-remodeling Factor Chd-3

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Characterization of the formation of somatic embryos from mature zygotic embryos of Passiflora ligularis Juss.

Published online: 06 July 2017Passiflora ligularis Juss. is an endemic species from South America with important medicinal and economical properties. The development of improved micropropagation techniques is necessary to provide rapid and efficient clonal propagation of elite genotypes with high resistance and uniform production, as well as a system that can be used for genetic transformation. For this reason, we focused on establishing a protocol for somatic embryogenesis in P. ligularis from mature zygotic embryos. Our results demonstrate that the highest frequencies of somatic embryo formation was observed on a culture medium supplemented with 27.2 µM 2,4-diclorophenoxyacetic acid plus 4.5 µM 6-benzyladenine. Histological analyses of somatic embryogenesis were performed every 7 days after induction over 60 days of exposure to the medium. We present clear evidence for the precise origin of de-differentiation. Initial cell divisions occurred from a group of cells (multicellular origin) on the abaxial surface of the cotyledon in the periphery of the epidermal tissues of mature zygotic embryos after 14 days of incubation. After 21 days, internal segmenting divisions resulted in the embryogenic character of the tissue. Globular embryos contain a protoderm surrounding a mass of vacuolated parenchymatous cells and meristematic regions with an observable procambium zone after 45 days. The complete independence of somatic embryos from the adjacent tissues was histologically confirmed by the absence of vascular continuity between them, after 60 days. We describe for the first time the development of somatic embryos in P. ligularis and demonstrate that somatic embryos develop into plants that can later on be acclimatized

The cryoprotectant PVS2 plays a crucial role in germinating Passiflora ligularis embryos after cryopreservation by influencing the mobilization of lipids and the antioxidant metabolism

Published online: 4 June 2019Cryopreservation is a process whereby biological structures are preserved in liquid nitrogen (−196 °C) without losing their viability. Many cryopreservation techniques use the Plant Vitrification Solution 2 (PVS2) for cryoprotection. This study will therefore evaluate the influence of different exposure times to the cryoprotectant PVS2 and discuss the importance of the mobilization of reserves and the antioxidant metabolism during the germination of cryopreserved Passiflora ligularis embryos. The composition of P. ligularis seeds was analytically determined. We tested the germination capacity and the Germination Speed Index (GSI) of embryos (that is, seeds without external tegument) which were exposed to different PVS2 exposure times (0, 30, 60 and 120 min) at 30 days after thawing. Proline content, hydrogen peroxide, activity of isocitrate lyase (ICL), malate synthase (MSy), lipid peroxidation and antioxidant enzyme activities (SOD, CAT, APX) were measured at 7, 14 and 21 days after cryopreservation. The germination from cryopreserved embryos was maximal (85%) after 60 min PVS2 exposure with a GSI of 0.6. At 60 min, the highest activity of the enzymes involved in the glyoxylate cycle, ICL and MSy were recorded. We hypothesize that a 60 min exposure to PVS2 accelerates the reserve mobilization which correlates positively with germination. Until 60 min, there was a positive correlation between the PVS2 exposure time and the proline content, as well as the activity of antioxidant enzymes (SOD, CAT, APX), and a negative correlation with the lipid peroxidation. This study enables us to optimize the long-term conservation of this species. In conclusion, fundamental research is necessary to optimize the cryopreservation procedure, and this study offers an effective and efficient workflow which can be extrapolated to other (oil-rich) species