Changes in the rotational motion of paramagnetic and fluorescent lipid-soluble probes were used to assess the effects of putrescine, spermidine and spermine on the fluidity of microsomal membranes from primary leaves of bean (Phaseolus vulgaris L.). Surface probes were more strongly immobilized by physiological concentrations of the polyamines than probes that partitioned deep into the bilayer interior. Spermidine and spermine were more effective than putrescine at reducing membrane fluidity, and at equimolar concentrations, the polyamines and calcium had similar effects on the mobility of the membrane probes. Spermine had essentially equivalent effects on the fluidity of native membranes, heat-denatured membranes and liposomes prepared from the total lipid extract of the membranes, indicating that polyamines associate with membrane lipid. These results raise the possibility that some of the physiological effects previously attributed to exogenously added polyamines could reflect membrane rigidification rather than a true physiological response.
The germination of mature somatic embryos of interior spruce was limited by the low frequency of root emergence. In addition, development was abnormal, since elongation and greening of the hypocotyl and cotyledons preceded root emergence by 1–2 weeks. Pretreatment of the embryos on water-saturated Kim-paks increased the frequency of root emergence but did not alter the abnormal pattern of germination. Somatic embryos do not survive desiccation at room humidity, but partial drying at high humidity promoted germination up to 90%. Furthermore, this treatment decreased the time required for root emergence such that elongation of the root and hypocotyl–cotyledon was synchronized over a period of 5–6 days. This germination closely resembled that of excised zygotic embryos. Drying over a range of humidities indicated that humidities of 81% and lower were lethal to the embryos, whereas germination was enhanced following treatment at humidities greater than 95% relative to untreated controls. The best germination and root elongation occurred on one-half strength basal media containing 2–3.4% sucrose. Of the plantlets derived from treated embryos, 50% survived transfer to soil compared with only 5% of the untreated controls. Key words: conifers, desiccation, germination, high relative humidity, partial drying, somatic embryogenesis, spruce.
The ontogenetic course followed by somatic embryos of interior spruce is highly dependent on the media concentration of abscisic acid (ABA). Little or no organized development occurs in the absence of ABA and as the level of ABA is increased, a range of embryo types is produced. “Shooty embryo” structures predominate in many callus lines at low levels of ABA (1‐10 μM), while 10‐20 μM ABA promotes the formation of bipolar embryos that germinate precociously. When ABA is increased to 30‐40 μM, precocious germination is inhibited and opaque cotyledonary embryos characteristic of their zygotic counterparts are formed which enter a period of quiescence. Only “mature” somatic embryos contain significant amounts of storage proteins and the level to which these proteins accumulate is dependent on the concentration of ABA. Indole‐butyric acid (IBA) included with ABA increases the number of mature embryos. Root elongation, which was used as a measure of embryo quality, was never observed from shooty embryo Structures and was 2‐3 fold higher in mature embryos compared to those that germinated precociously.
Lipid and fatty acid analyses were perfornmed on whole leaf extracts and isolated thylakoids from winter rye (Secalk cereak L. cv Punia) grown at 5°C cold-hardened rye (RH) and 20°C nonhardened rye (RNH).Although no significant c ge in total lipid content was observed, growth at low, cold-hardening temperature resulted in a specific 67% (thylakoids) to 74% (whole leaves) decrease in the trans-A3-hexadecenoic acid (trans-16:1) level associated with phosphatidyldiacylglycerol (PG). Electron spin resonance and differential scanning calorimetry (DSC) indicated no significant difference in the fluidity of RH and RNH thylakoids. Separation of chlorophyll-protein complexes by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the ratio of oligomeric light harvesting complex:monomeric light harvesting complex (LHCII,:LHCII3) was 2-fold higher in RNH than RH thylakoids. The ratio of CP1a:CP1 was also 1.5-fold higher in RNH than RH thylakoids. Analyses of winter rye grown at 20, 15, 10, and 5C indicated that both, the tras-16:1 acid levels in PG and the LHCII,:LHCII3 decreased concomitantly with a decrease in growth temperature. Above 40C, differential scanning calorimetry of RNH thylakoids indicated the presence of five major endotherms (47, 60, 67, 73, and 86C
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