Immature Axes Research Articles

The fern Stauropteris oldhamia Binney: New data on branch development and adaptive significance of the hypodermal aerenchyma

Well-preserved specimens of Stauropteris oldhamia are described. The material was collected in the early 1920s from the Lower Westphalian (Early Pennsylvanian) Saurue seam from Belgium. The fossil plants occur as permineralized axes fragments within a coal ball. This study confirms most of the interpretations made by previous researchers. The observation of immature axis however suggests a less regular organization than previously interpreted beyond the three first branching orders. We also highlight the presence of profusely and dichotomously branched aphlebiae, the lack of laminate organs as well as the presence of hypodermal aerenchyma in all plant parts. We interpret these features as part of a very specialized assimilatory apparatus indicating an adaptation to a humid swamp environment.

Discrete levels of desiccation sensitivity in various seeds as determined by the equilibrium dehydration method

This study examined the hypothesis that desiccation sensitivities of recalcitrant and intermediate seeds can be categorized into discrete levels of critical water potential. The equilibrium dehydration method was used to determine the critical water potential (CWP) below which desiccation damage started to occur. The CWP values of Bruguiera cylindrica, Lansium domesticum, Litchi chinensis and Lumitzera racemosa are approximately ‐4 MPa. The CWP values of Andira inermis, Avicennia alba, Castanea sinensis (from New Zealand), Citrus aurantifolia, Ginkgo biloba, Nephelium lappaceum and Theobroma cacao (immature axis) are approximately ‐8 MPa. The CWP values of Acer pseudoplatanus, Castanea sinensis (from China), Quercus rubra and Theobroma cacao (mature axis) are approximately ‐12 MPa. The CWP values of Artocarpus heterophyllus and Hevea brasiliensis are approximately � 23 MPa, while the CWP values of Acer platanoides, Azadirachta indica, Carica papaya and Coffea arabica are approximately ‐73 MPa. Together with data available in earlier literature, these CWP values suggest that there are five discrete levels of critical water potential among desiccation-sensitive seed tissues. These data support the hypothesis that discrete levels of desiccation sensitivity occur among recalcitrant and intermediate seeds, and suggest that specific damaging and protective mechanisms exist at certain hydration levels.

Desiccation sensitivity and activities of free radical-scavenging enzymes in recalcitrant Theobroma cacao seeds

AbstractMature and immature axes of Theobroma cacao (cocoa) seeds tolerated desiccation under a rapid-drying regime to critical water contents of 1.0 and 1.7 g g-1 dw, respectively. These critical water contents corresponded to water contents below which activities of free radical-scavenging enzymes (ascorbate peroxidase, peroxidase and superoxide dismutase) decreased rapidly during desiccation. The decline in axis viability below the critical water content was correlated with sharp increases in lipid peroxidation and cellular leakage. Cotyledon tissues were more desiccation-tolerant than axes, with a low critical water content of 0.24 g g–1dw. Desiccation sensitivity in cotyledon tissues was also correlated with the decrease in superoxide dismutase activity and increased lipid peroxidation products. However, in the cotyledons, no ascorbate peroxidase activity was detected at any water content, and peroxidase activity was gradually reduced as desiccation proceeded. Cocoa embryonic axes contained large amounts of sucrose, raffinose and stachyose but only traces of reducing monosaccharides. Desiccation sensitivity of recalcitrant cocoa axes did not appear to be due to the lack of sugar-related protective mechanisms during desiccation, and it was more likely related to the decrease of enzymic protection against desiccation-induced oxidative stresses.

Fourier transform infrared microspectroscopy detects changes in protein secondary structure associated with desiccation tolerance in developing maize embryos

Isolated immature maize (Zea mays L.) embryos have been shown to acquire tolerance to rapid drying between 22 and 25 d after pollination (DAP) and to slow drying from 18 DAP onward. To investigate adaptations in protein profile in association with the acquisition of desiccation tolerance in isolated, immature maize embryos, we applied in situ Fourier transform infrared microspectroscopy. In fresh, viable, 20- and 25-DAP embryo axes, the shapes of the different amide-I bands were identical, and this was maintained after flash drying. On rapid drying, the 20-DAP axes had a reduced relative proportion of alpha-helical protein structure and lost viability. Rapidly dried 25-DAP embryos germinated (74%) and had a protein profile similar to the fresh control axes. On slow drying, the alpha-helical contribution in both the 20- and 25-DAP embryo axes increased compared with that in the fresh control axes, and survival of desiccation was high. The protein profile in dry, mature axes resembled that after slow drying of the immature axes. Rapid drying resulted in an almost complete loss of membrane integrity in the 20-DAP embryo axes and much less so in the 25-DAP axes. After slow drying, low plasma membrane permeability ensued in both the 20- and 25-DAP axes. We conclude that slow drying of excised, immature embryos leads to an increased proportion of alpha-helical protein structures in their axes, which coincides with additional tolerance of desiccation stress.

Effects of developmental status and dehydration rate on characteristics of water and desiccation-sensitivity in recalcitrant seeds of Camellia sinensis

AbstractThe effect of rate of dehydration was assessed for embryonic axes from mature seeds of Camellia sinensis and the desiccation sensitivity of axes of different developmental stages was estimated using electrolyte leakage. Rapidly (flash) dried excised axes suffered desiccation damage at lower water contents (0.4 g H2O (g DW)−1) than axes dried more slowly in the whole seed (0.9 g H2O (g DW)−1). It is possible that flash drying of isolated axes imposes a stasis on deteriorative reactions that does not occur during slower dehydration. Differential scanning calorimetry (DSC) of the axes indicated that the enthalpy of the melting and the amount of non-freezable water were similar, irrespective of the drying rate.Very immature axes that had completed morphogenesis and histodifferentiation only were more sensitive to desiccation (damage at 0.7 g H2O (g DW)−1) than mature axes or axes that were in the growth and reserve accumulation phase (damage at 0.4 g H2O (g DW)−1). As axes developed from maturity to germination, their threshold desiccation sensitivity increased to a higher level (1.3−1.4 g H2O (g DW)−1). For the very immature axes, enthalpy of the melting of tissue water was much lower, and the level of non-freezable water considerably higher, than for any other developmental stage studied.There were no marked correlations between desiccation sensitivity and thermal properties of water. Desiccation sensitivity appears to be related more to the degree of metabolic activity evidenced by ultrastructural characteristics than to the physical properties of water.

Homoiohydrous (recalcitrant) seeds: Developmental status, desiccation sensitivity and the state of water in axes of Landolphia kirkii Dyer

The desiccation sensitivity in relation to the stage of development was investigated in embryonic axes from the homoiohydrous (recalcitrant) seeds of Landolphia kirkii. Electrolyte leakage, used to assess membrane damage after flash (very rapid) drying, indicated that axes from immature (non-germinable) seeds were the most desiccation-tolerant, followed by those from mature seeds, while axes from seeds germinated for increasing times were progressively more desiccation-sensitive. Differential scanning calorimetry was used to study the relationship between desiccation sensitivity and the properties of water in the tissues. Axes from immature seeds had a lower content of non-freezable water than that of any other developmental stage and a higher enthalpy of melting of freezable water. For mature and immature axes electrolyte leakage increased at the point of loss of freezable water. At other developmental stages the water content at which electrolyte leakage increased markedly correlated with the other properties of the water, such as the change in the shape of the melting endotherm and the onset temperature. Ultrastructural studies of axes at the various developmental stages showed changes in the degree and pattern of vacuolation, the presence and quantities of lipid and starch, and the degree of endomembrane development. The results are discussed in relation to current hypotheses on the basis of desiccation tolerance.

Maturation and germination of Phaseolus vulgaris embryonic axes in culture

In this study of embryo development in Phaseolus vulgaris L., we found that immature embryonic axes placed in culture show a growth lag before germinating. The length of this lag phase varies according to axis age at excision, but is not affected by transfer to fresh medium, alteration of sucrose concentration between 0.5 and 2%, or whether the culture medium is liquid or agar-solidified. The lag phase was shortened by both actinomycin D and cordycepin treatment, and by treatment with 10(-5) to 10(-6) M benzyladenine. The effect of abscisic acid (ABA) varied with concentration: below a certain level, it had no effect on the lag phase, but above that level it inhibited, germination. This threshold concentration was 10(-7) M for 20-d-old axes but increased to 10(-5) M by the time the axes were 32 to 34 d old. To determine whether the axes were continuing their embryonic development during the lag phase, we tested them for desiccation-tolerance and for synthesis of phaseolin, a seed storage protein which is specific for embryos of P. vulgaris. The ability to germinate after rapid desiccation was acquired by axes at 26 d past anthesis; when axes younger than this were placed in culture, they developed desiccation-tolerance during the lag phase of growth, indicating that they were continuing embryonic maturation. Phaseolin was present in isolated axes, although at lower levels than in cotyledons. It accumulated during axis development in parallel with total protein, staying at about 1% of total protein content. When isolated immature axes were pulsed with (3)H-or (14)C-amino acids, they incorporated label into phaseolin, shown by precipitation with anti-phaseolin antibody. Isolated axes from mature seeds, however, did not synthesize detectable amounts of phaseolin. Immature axes cultured in vitro for a period of one to several days continued synthesizing phaseolin until the day prior to visible germination. Treatment of cultured axes with ABA increased the amount of precursor amino acids incorporated into protein, but had a small or no effect on the relative proportion of phaseolin synthesized. We conclude that P. vulgaris axes in culture continue to develop embryonically for a period of time which seems to be under intrinisc control by the axis. This contrasts with "precocious germanation", a pattern of embryo behavior seen in many other species. When such embryos are excised from seeds while immature and placed in culture, they switch promptly from embryo development into germination. If ABA or water stress is responsible for preventing precocious germination, it may be that a high level of ABA is maintained or synthesized internally by embryonic axes of Phaseolus, while in other embryos the maternal environment supplies ABA and/or causes water stress.