20-hydroxyecdysone, what it can do.
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Biomedical subjects
Publications and source records attributed to F Engelmann.
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A specific protein is Present in serums of all of Leucophaea maderae that mature eggs but not in serums of males, nymphs of either sex, or females that are reproductively inactive. Ablations, microsurgery and remplantation showed that this protein is produced de novo under the influence of the corpus allatum hormone. This protein, essential for egg maturation, is synthesized in isolated abdimens of allatectomized females treated with 0.08 to 0.12 microgram of the t,t,t-isomer of the authentic juvenile hormone. The crops allatum hormone also stimulates toa similar degree the synthesis of other proteins which appear to be essential for egg maturation as well; these are not specific to females.
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For the cockroach species Leucophaea maderae and Periplaneta americana two major juvenile hormone (JH)-binding proteins have been identified: lipophorin (Lp) and vitellogenin (Vg). Each of these macromolecules binds JH with an approximate affinity of K(d) of 10 nM. In Leucophaea the concentration of Lp is augmented by JH during vitellogenesis at the same time when Vg is induced de novo. The circulating levels of each of Lp and Vg at mid-vitellogenesis are in the 10 microM range. Similar values have been determined for Periplaneta. Total JH concentrations (bound and free) can be as high as micromolar in Leucophaea. However, because of the large quantities of the two major JH-binding proteins and their high affinity for JH, we can assume that the amount of free (unbound) JH in circulation is extremely low (the actual values are not know). The JH esterases (JHEs) of the hemolymph in both cockroach species have been isolated by anion exchange chromatography. The JHEs of Leucophaea bound to the anion exchange resin more tightly than the JHE of Periplaneta. The V(max) of the Leucophaea esterases fluctuated by a factor of 2 to 3 during vitellogenesis. The K(m) values for the two distinct esterases of Leucophaea were similar (about 0.15x10(-6) M). On the other hand, k(cat) of the JHEs for Leucophaea at ovulation time was two to three times higher than earlier during vitellogenesis, i.e. 23.30 min(-l) compared to 6.20 min(-1). The JHE of Leucophaea is shown to bind JH III with high affinity: K(d)=3x10(-9) M. However, since there are only very small amounts of JH available for degradation (due to the binding to Lp and Vg), the quantitative removal of JH from circulation, and this includes the release of bound JH, is indeed slow, with a measured half-life of 6-8 h. Classical kinetic assumptions are not met in conditions where the enzyme concentrations exceed by far that of the available substrate. Nonetheless, we attempted to determine the initial velocity of JH hydrolysis under natural conditions, i.e. for undiluted hemolymph, by measuring the initial velocities of JH hydrolysis in serially diluted hemolymph and extrapolating to zero dilution. For in vivo conditions we estimated an initial velocity of JH hydrolysis of <0.1 fmol microl hemolymph(-1) min(-1), i.e. four to five orders of magnitude lower than that measured at substrate saturation in vitro.
In this study, the effect of various factors, including the physiological state of mother-plants and parameters of the encapsulation-dehydration protocol (cryoprotective treatment, freezing protocol) on the regrowth of shoot tips of kiwi in vitro plantlets was studied. The optimal protocol established was the following: shoot tips were sampled on mother-plants after a one month cold-acclimation period at 5 degrees C. For preculture, which was performed at 5 degrees C, encapsulated shoot tips were transferred at 24-hour intervals on solid media with increasing sucrose concentrations, from 0.5 to 1.0M. Beads were then desiccated to 26% moisture content (fresh weight basis), prefrozen from 0 degrees C to -40 degrees C at 0.2 degrees C/min and immersed rapidly in liquid nitrogen. No differences were noted between in vitro plantlets produced from cryopreserved and control shoot tips as regards leaf color, average height, multiplication rate and peroxidase zymogram pattern. Apices of 3 kiwi accessions were frozen using the above protocol with regrowth percentages ranging between 22 and 56%.
Axillary buds sampled from in vitro plants of four grape varieties could withstand cryopreservation using the encapsulation-dehydration technique. Regrowth percentages ranged between 15 and 40%. Buds were encapsulated in alginate beads containing 0.5M sucrose, precultured at 5 degrees C on media containing daily increasing sucrose concentrations from 0.1 to 1.0M, then desiccated to 21% moisture content (fresh weight basis) and frozen slowly (pre-freezing to -40 degrees C at 0.2 degrees C/min followed by immersion in liquid nitrogen). The physiological state of the in vitro mother-plants and the freezing procedure employed dramatically influenced the results. Regrowth after cryopreservation was achieved only when buds were sampled from mother-plants that had been kept without subculture for 3 to 4 months. An additional one-month period of cold-acclimation of mother-plants at 5 degrees C and a two-step freezing procedure improved growth recovery. The multiplication rate of shoots produced from cryopreserved buds was lower than that of controls during the first subcultures after thawing. However, the rooting ability of control and cryopreserved plantlets became similar from the fourth subculture onwards.
Shoot tips excised from apple (Malus domestica Borkh. cv. Golden Delicious) dormant buds immediately before freezing were successfully cryopreserved using three different methods (vitrification, encapsulation-dehydration and two-step freezing). Two-step freezing produced the highest in vitro growth recovery (83%) of cryopreserved shoot tips, followed by vitrification (60%) and encapsulation-dehydration (33%). By contrast, when dormant buds were cryopreserved using the two-step freezing technique, shoot tips excised from buds after rewarming and cultured in vitro displayed only 16% recovery.
The role of pregrowth and preculture treatments in terms of both medium composition and exposure duration on survival of embryonic axes of Citrus madurensis after cryopreservation using the vitrification procedure was investigated. The optimal pregrowth treatment for excised embryonic axes was a 3-day treatment with 0.1M sucrose. Preculture was also essential in increasing survival after cryopreservation. Among the various media and treatment durations evaluated, a 24h-preculture of embryonic axes on medium with 0.3M sucrose and 0.5M glycerol was found to be optimal. Using these pregrowth and preculture conditions followed by treatment at 25 degrees C for 20 min each with a loading solution (0.4M sucrose + 2.0M glycerol) and then the PVS2 vitrification solution, direct immersion in liquid nitrogen, rapid rewarming, unloading in a 1.2M sucrose solution for 20 min and transfer of embryonic axes on recovery medium, 82.5% survival and regrowth without intermediary callus formation were obtained with C. madurensis embryonic axes.
This study compared the field performance of sugarcane plants originating from three different sources: control, non-cryopreserved embryogenic calluses, cryopreserved embryogenic calluses and macropropagated material of the same commercial hybrid. Several agronomic traits were evaluated on 100 plants per treatment over a 27-month period covering the growth of the stool and of the first ratoon. Significant differences between treatments were observed only during the first six months of field growth of sugarcane stools. Stems produced from in vitro cultured material, irrespective of their cryopreservation status, had a smaller diameter and a shorter height than those produced from macropropagated material. These differences disappeared by 12 months of stool field growth.
Osmoconditioning-controlled rehydration of seeds in a solution with low osmotic potential -has been shown to reinvigorate aged seeds. The present work aimed at investigating the effect of osmoconditioning on the germination of cryopreserved seeds of Coffea arabica, whose viability and vigour are drastically affected by cryopreservation. For cryopreservation, seeds were desiccated to 0.21 g H2O/g dw, cooled at 1 degree C/min to -50 degree C, then immersed rapidly in liquid nitrogen. After rapid rewarming, seeds were osmoconditioned for 1 to 6 weeks using solutions with osmotic potentials between -1 and -4 MPa. The time to produce half of the final percentage of normal seedlings, T50, was about three fold lower with osmoconditioned seeds than with non-osmoconditioned seeds (12-14 d vs 36 d). Moreover, after a 6-week osmoconditioning treatment with solutions with osmotic potential of -1 and -1.25 MPa, the percentage of seedlings recovered from cryopreserved seeds was 64-74%, against 13-16% only for cryopreserved seeds which were not osmoconditioned.