The intersectional hybrid between Weigela hortensis and W. maximowiczii (Caprifoliaceae)

Morphologically intermediate plants between Weigela hortensis (Siebold & Zucc.) K.Koch and W. maximowiczii (S.Moore) Rehder have been found in Miyagi and Yamagata Pref., northern Japan. Quantitative character analyses of flowers, pollen stainability and molecular analyses indicated that the intermediate plants were hybrids of those two species. This is the first record of an intersectional hybrid with W. maximowiczii (sect. Weigelastrum ) as one of the parent species. The morphological differences among hybrid individuals imply the possibility of backcrosses or formation of second or later generations of hybrids, although those may be quite rare because of a low frequency of viable pollen grains. Causes of hybridization between two distantly-related species in Weigela are discussed. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 138 , 369–380.     

Respiration in Eggs of the Echiuroid, Urechis unicinctus, Before and After Fertilization

In eggs of the echiuroid Urechis unicinctus the respiration rate, which is not altered by fertilization, is inhibited by rotenone, antimycin A and cyanide. The respiration in echiuroid eggs is probably mediated by the mitochondrial respiratory chain. In fertilized eggs, the respiration was inhibited by oligomycin and stimulated by the uncouplers of oxidative phosphorylation 2,4-dinitrophenol and carbonylcyanide p-trifluoromethoxyphenylhydrazone, whereas respiration in unfertilized eggs was insensitive to these compounds. Insemination increased the respiratory rate in eggs in the presence of uncouplers and reduced it in the presence of oligomycin. These findings suggest that the capacity of electron transport in mitochondira is elevated by fertilization but becomes latent on fertilization-induced coupling of respiration with oxidative phosphorylation. Strong stimulation of the respiration in unfertilized eggs was induced by dichlorophenol indophenol, phenazine methosulfate and tetramethyl p-phenylenediamine, suggesting possible sites at which electron transport is regulated in unfertilized eggs. The resulting stimulation of respiration in unfertilized eggs was insensitive to uncouplers and oligomycin, but became sensitive to them after fertilization simultaneously with considerable decrease in its rate. Fertilization-induced coupling of the respiration seemed to reduce the respiratory rate enhanced artificially by these redox compounds.     

Inhibitory Effect of Calmodulin Antagonists and Calcium Antagonists on Fertilization-Induced Cyanide-Insensitive Respiration in Sea Urchin Eggs

During initial several minutes after fertilization, sea urchin eggs exhibited high rate of respiration which was only slightly inhibited by cyanide. This cyanide-insensitive respiration was inhibited by calcium antagonists, diltiazem and verapamil, and calmodulin antagonists, N-(6-aminohexyl)-5-chloro-1-naphthalensulfonamide hydrochloride (W-7), N-(6-aminohexyl)-1-naphthalenesulfonamide hydrochloride (W-5) and chlorpromazine, which were added within 1 min after insemination. The inhibitory effect of W-7 on cyanide-insensitive respiration was higher than that of W-5. Cyanide-sensitive respiration of fertilized eggs observed after this initial period was not inhibited by these compounds. Ca²⁺ influx in eggs just after fertilization was inhibited by calcium antagonists but was rather enhanced by calmodulin antagonists. Fertilization-induced stimulation of cyanide-insensitive respiration probably results from calmodulin-dependent reactions which are activated by Ca²⁺ influx.     

Utilization of Seed Reserves and Currently Produced Photosynthates by Embryonic Tissues of Pine Seedlings

The importance of seed reserves for growth of Pinus resinosaAit. during and shortly after seed germination was studied undercontrolled conditions. Tissues in the resting embryo were notcompletely differentiated. Many small, presumably reserve particleswere present in the embryo in addition to reserves in the megagametophyte.During seed germination, procambia in the embryo first differentiatedprotophloem 2 days after seeds were sown. The radicle beganto emerge from the seed coat at 5 days, at which time initialxylem formation was observed. Also, at approximately the sametime, primordia of primary needles were forming in the peripheralzone of the apex. Elements of the photosynthetic apparatus,including stomata and mesophyll with chloroplasts, were differentiatedfirst in the hypocotyl and then in cotyledons between 5 and8 days after seeds were sown. Photosynthetic rates of youngseedlings were correlated with rates of cotyledon expansion.During early developmental stages, reserve particles in megagametophytecells and embryo cells gradually disappeared. Surgical removalof megagametophytes at various stages of seed germination resultedin subsequent growth inhibition of the hypocotyl-radicle axis,with early removal of cotyledons suppressing most growth. Growthof primary needles appeared to be influenced indirectly by megagametophytereserves, probably by changes in amount of photosynthetic tissue.The embryo alone possessed capacity to differentiate such tissuesas primary needle primordia, stomata, and primary and secondaryvascular systems. Megagametophyte reserves appeared to contributeto growth of embryonic tissues only after the embryo itselfinitiated growth. Both current photosynthesis of seedlings andseed reserves contributed importantly to seedling development.     

A new method in growth-electrophysiology: Pressurized intra-organ perfusion

Abstract A new experimental system was devised for the simultaneous measurement of elongation rate and the activity of the spatially separate electrogenic ion pumps of a hypocotyl segment excised from a seedling of Vigna unguiculata L. Walp. under enforced intra-organ perfusion by artificial solutions. The pathway of the perfusion medium was apoplastic space, including xylem vessels as main routes. The elongation rate of the segment was highly dependent on the perfusion pressure applied. It was possible to increase the growth rate under pressurized perfusion by 10-30 times as much as that without perfusion. Elongation rate was also dependent on respiration under perfusion, being retarded reversibly by anoxia a few minutes after the activities of the electrogenic ion pumps were stopped. Perfusion pressure had a little influence on the membrane potential (V_px) below a breakdown level (c. 130 kPa). Perfusion of mannitol or sorbitol solution of appropriate concentration reduced the elongation rate reversibly.     

Effect of Aminopterin and Deoxyribonucleosides on the Cleavage and Embryogenesis of the Sea Urchin, Hemicentrotus pulcherrimus

In the embryos of the sea urchin, Hemicentrotus pulcherrimus , reared with 150 μM aminopterin from the time of fertilization, cessation of the development occurred at the blastula stage, at which the dTTP level became quite low. Another addition of thymidine to the embryo culture containing aminopterin resulted in an elevation of dTTP concentration in the embryos and allowed them to develop normally. Decrease in the dTTP level, resulting from the inhibition of thymidylate synthesis by aminopterin, probably causes a failure of egg cleavage and development. 5-Bromo-2'-deoxyuridine (BUdR) also released the aminopterin-inhibition of egg cleavage and allowed the treated embryos to develop to early gastrulae. Thereafter, the degeneration of archenteron occurred and these embryos became large permanent blastulae. Other deoxyribonucleosides failed to cancel the inhibition by aminopterin of egg cleavage. In the embryos kept with both BUdR and aminopterin, BUdR incorporation into DNA occurred at a similar rate as in thymidine incorporation in the embryos kept with thymidine and aminopterin, and was inhibited by another addition of thymidine. Without aminopterin treatment, BUdR incorporation hardly occurred and the embryos developed normally. BUdR incorporation into DNA in place of thymidine probably occurs in aminopterin-treated embryos, resulting in abnormal development.     

Melittin, a Component of Bee Venom, Activates Unfertilized Sea Urchin Eggs

Melittin, which is known to stimulate phospholipase A , in many cells, caused as much elevation of fertilization membranes and increase in respiration of unfertilized eggs of the sea urchins Anthocidaris crassispina and Hemicentrotus pulcherrimus as normal fertilization.
In melittin-activated eggs, amino acid transport was decreased to less than that of unfertilized eggs, nucleoside transport was only slightly, activated, protein synthesis was rather inhibited and neither DNA synthesis nor cleavage was observed. It is concluded that although melittin induces the cortical reaction and activation of respiration in unfertilized eggs, its cytotoxicity prevents any "late changes".     

STAGING OF NEWT BLASTULA EMBRYOS AND FIRST APPEARANCE OF PRIMARY MESODERMAL COMPETENCE

Using the swimbladder of the crusian carp ( Carrasius auratus ) as an inductor, the first appearance of mesodermal competence in the presumptive ectoderm of the newt ( Triturus pyrrhogaster ) blastula was investigated. The time course of embryonic development before the gastrula stage was determined by counting the number of surface cells on a 0.25 mm line at the animal pole. Pregastrula embryos with 2–3, 4–5, 6–7 and 7–8 cells roughly correspond to those at 14, 14–12, 8–6 and 4–0 hr before the beginning of gastrulation. Using presumptive ectoderm of the early gastrula stage, 15 min was found to be the minimum time of contact necessary for the realization of induction. The reactivity of the presumptive ectoderm from pregastrula embryos was tested by 30 min contact. Presumptive ectoderm up to the 4–5 cell stage did not react to the inductor. It may become competent within the next 4–8 hr, since the ectoderm from embryos in the 6–7 cell stage was reactive.     

Studies on the metabolism of a sulfur-oxidizing bacterium IV.1 Growth and oxidation of sulfur compounds in Thiobacillus thiooxidans

By immersing a few small cellophane bags containing BaCO³ powderin STARKEY's medium, the duration of lag phase in the growthof Thiobacillus thiooxidans is minimized and the yield of cellsis increased ten times that of the previous method. The activitiesof oxidation for sulfur and sulfite change with growth. Sulfiteis oxidized at a comparable rate to that of sulfur oxidationat pH values between 6.0 and 6.5. In the presence of cysteineor glutathione, thiosulfate can be oxidized at a pH above 5.0.At pH values below 4.5, apparent oxidation of thiosulfate andtetrathionate to sulfate is observed. This result is accountedfor by the facts that thiosulfate is decomposed to sulfur andsulfite under the acidic condition at pH values below 4.5, andthat tetrathionate is reduced to thiosulfate enzymatically.In the oxidation of tetrathionate, oxygen uptake begins aftera lag phase, the duration of which depends on the concentrationsof cells and of tetrathionate. Cysteine is oxidized to cystine.The oxidation is strongly inhibited by metal-chelating agents.The cysteine oxidizing activity is, however, quite stable andis not lost by treating cells with organic solvents, sonic oscillation,by heating or lyophilization. ¹III=References (11). ²Partly supported by a grant from the Ministry of Education.