INCORPORATION OF AMINO ACIDS INTO SEA URCHIN EGGS STIMULATED INSUFFICIENTLY WITH AN ACTIVATING REAGENT

Investigations were performed on the incorporation of valine-¹⁴C and leucine-³H into sea urchin eggs which were stimulated insufficiently with an activating reagent. Materials used were Pseudocentrotus, Hemicentrotus and Anthocidaris. An insufficient stimulation enhanced the incorporation of amino acids into the eggs, although it provoked no visible cortical changes. The amount of incorporation in this case was 1.6 to 2.7 times as much as that into untreated eggs. In fully activated eggs, the amount of incorporation was more than 4 times that in untreated eggs. The fact that the incorporation of amino acids is increased without accompanying breakdown of the cortical granules indicates that the increase may be linked to an invisible change, probably the fertilizationwave, which is caused by an insufficient stimulation.     

Diversification in flowering time due to tandem FT-like gene duplication, generating novel Mendelian factors in wild and cultivated rice

The complex structure of a single Mendelian factor widespread in the Asian cultivated rice ( Oryza sativa ) and its wild progenitor ( Oryza rufipogon ) that caused diverse phenotypes in the timing of flowering under natural field conditions was investigated in near isogenic lines. These near isogenic lines showed differences in flowering time despite all eight accessions collected from tropical regions possessing a recessive gene allelic to the se-pat gene. Fine mapping in two of these near-isogenic lines revealed that cultivated (Patpaku) and wild (W593) accessions had three and two linked quantitative trait loci (QTL) in the candidate regions, respectively, showing that Patpaku and W593 possessed linked QTLs with different effects in addition to the commonly-observed recessive gene ( se-pat ). Molecular dissection suggested that the tandemly duplicated FT-like genes ( Hd3a and RFT1 ) could be the candidate genes for these QTLs. Interestingly, the linked QTLs differed in their epistases, degree of dominance, and genotype × environment interactions. The nucleotide sequences showed that RFT1 has diverged more rapidly than Hd3a during rice evolution, suggesting phenotypic diversification of the two genes. Phylogenetic analysis implied that the se-pat ⁺ alleles might have emerged in different lineages within O. sativa . The present results strongly suggest that nucleotide divergence and shuffling of the linked QTLs by recombination might have created novel Mendelian factors that probably contribute to responding to local environments.     

Chromosome analysis of five Brazilian species of poison frogs (Anura: Dendrobatidae)

Dendrobatid frogs have undergone an extensive systematic reorganization based on recent molecular findings. The present work describes karyotypes of the Brazilian species Adelphobates castaneoticus, A. quinquevittatus, Ameerega picta, A. galactonotus and Dendrobates tinctorius which were compared to each other and with previously described related species. All karyotypes consisted of 2n = 18 chromosomes, except for A. picta which had 2n = 24. The karyotypes of the Adelphobates and D. tinctorius species were highly similar to each other and to the other 2n = 18 previously studied species, revealing conserved karyotypic characteristics in both genera. In recent phylogenetic studies, all Adelphobates species were grouped in a clade separated from the Dendrobates species. Thus, we hypothesized that their common karyotypic traits may have a distinct origin by chromosome rearrangements and mutations. In A. picta, with 2n = 24, chromosome features of pairs from 1 to 8 are shared with other previously karyotyped species within this genus. Hence, the A. picta data reinforced that the C-banding pattern and the NOR location are species-specific traits in the genus Ameerega. Moreover, the Ameerega monophyletism proposed by previous phylogenetic studies indicates that the karyotypic differences among species in this genus result from a long divergence time.     

Studies on the respiratory system of Aspergillus oryzae I. Development of respiratory activity during germination in the presence and absence of antimycin A

Active growth of Aspergillus oryzae was observed when conidiawere inoculated into a medium containing antimycin A. Immediatelyafter adding antimycin A, to young mycelia germinated in itsabsence, growth stopped, but began again after several hours.This restored growth was antimycin A-insensitive. Percentagegermination was the same in the presence and absence of thisdrug. It seems that drug-resistant germination and growth donot result from selection of resistant cells but result frominduction of antimycin A-insensitive mitochondria in the wholepopulation. Endogenous respiration of cells germinated in theabsence of antimycin A was inhibited by this drug, whereas thatof cells grown in the presence of antimycin A was completelyinsensitive. Antimycin A-sensitivity of cellular respirationseems to determine the effect of this drug on mycelial growth.Mitochondria were isolated from mycelia grown in the presenceand absence of this drug. The difference in antimycin A-sensitivityin endogenous respiration was attributed to a difference inproperties of the mitochondrial respiratory systems. ¹Present address: Department of Chemistry, Institute of MedicalScience, University of Tokyo, Tokyo, Japan (Received December 21, 1969; )     

Change in the indole-3-acetic acid oxidase level in leaves of Begonia evansiana cuttings at the time of aerial tuber formation under short-day conditions

IAA oxidase in Begonia evansiana leaves was partially purifiedby DEAE cellulose chromatography. The enzyme was identifiedas peroxidase from its elution pattern, its Michaelis constantand the effects of various inhibitors and high temperature. In B. evansiana, which requires SD treatment to form aerialtubers, the amount of IAA oxidase in leaves increased to a maximumafter 2 days of SD treatment. When SDs with interrupted nightwere given, a maximum was reached after 6 SDs. Thus, the changein IAA oxidase content had an inverse correlation to that forIAA content observed previously. (Received May 18, 1970; )     

L-Tyrosine carboxy-lyase of barley roots

_L-Tyrosine carboxy-lyase (E.C. 4. 1. 1. 25) was isolated fromroots of germinating barley (Hordeum vulgare). The enzyme requirespyridoxal phosphate for maximum activity. The optimum pH foractivity is about 7.0. The enzyme is inhibited by p-chloromercuribenzoateand hydroxylamine at 10^–3 M. Enzyme activity is foundin extracts from young roots, especially from those in earlystages of development, but not in extracts from shoots of thesame plant. Localization and changes in the amounts of _L-tyrosinecarboxy-lyase and aromatic amines in developing barley seedlingswere measured. Participation of carboxy-lyase in the formationof aromatic amines in barley roots is suggested. (Received July 17, 1970; )     

Ultrastructural Changes of the Two Types of Differentiated Cells during the Migration and Early Culmination Stages of Dictyostelium discoideum

Changes of fine structure during prolonged migration of Dictyostelium discoideum slugs were studied by electronmicroscopy. Prespore specific vacuoles of cells located near the substratum gradually degenerated and the prespore antigen contained in them was lost. During the process, mitochondria in the prespore cells were transformed dramatically: as the mitochondrion elongates, its central part becomes thinner and the cristae become localized at its two ends. Then it bends and its two ends fuse to segregate part of the cytoplasm. The cristae then accumulate in the original ends. Similar mitochondrial transformation was observed in prespore cells of cell masses induced to culminate after a long period of migration.     

Phytochemical studies on tobacco alkaloids XIV. The occurrence and properties of putrescine N-methyltransferase in tobacco roots

Putrescine N-methyltransferase, a new enzyme catalyzing theformation of N-methylputrescine from putrescine and S-adenosyl-L-methioninewas found in roots of tobacco plants. The enzyme was purified30-fold from crude extracts of tobacco roots. NMethylputrescinewas identified as the reaction product by comparison with theauthentic compound. The enzyme had a pH optimum between pH 8and 9, and a molecular weight of about 60,000, as determinedby gel filtration. Km values for putrescine and 5-adenosyl-L-methioninewere 4.0 x 10^–4 M and 1.1 x 10^–4 M, respectively.Enzyme activity was inhibited by N-chloromercuribenzoate andAg⁺. No cofactors were required. Of the various substrates tested,only putrescine served as a methyl acceptor. The enzyme waslocalized exclusively in the roots and its activity was greadyenhanced by decapitation. The presence of putrescine N-methyltransferase in tobacco rootsstrongly suggests that N-methylputrescine participates as anintermediate in nicotine biosynthesis. (Received March 2, 1971; )     

Development of Melanocyte Progenitors in Murine Steel Mutant Neural Crest Explants Cultured With Stem Cell Factor,Endothelin-3, or TPA

Stem cell factor (SCF) has been suggested to be indispensable for the development of neural crest cells into melanocytes because Steel mutant mice (i.e., Sl/Sf¹) have no pig-mented hairs. On the other hand, it has been demonstrated that the addition of endothelin 3 (ET-3) or TPA to neural crest cell cultures can induce melanocyte differentiation without addition of extrinsic SCF. In this study, we excluded the influence of intrinsic SCF by using SI/SI mouse embryos to study more precisely the effects of natural cytokines, such as extrinsic soluble SCF or ET-3, or chemical reagents, such as TPA or cholera toxin. We found that SCF is supplied within the wild-type neural crest explants and that ET-3 cannot induce melanocyte differentiation or proliferation without SCF. These results indicate that SCF plays a critical role in survival or G1/S entry of melanocyte progenitors and that SCF initially stimulates their proliferation and then ET-3 accelerates their proliferation and differentiation. TPA has the ability to elicit neural crest cell differentiation into melanocytes without exogenously added SCF but it is not as effective as SCF because many more melanocytes developed in the wild-type neural crest explants cultured with TPA.