Alicyclic acid metabolism in plants 3. Fate of 14C-shikimate and 14C-quinate in mung bean plants

Young mung bean plants (Phaseolus mungo) were exposed to ¹⁴C-shikimateor ¹⁴C-quinate in the light. After 8 or 23.5 hr of incubationat 25°C, radioactivities in free and bound amino acids,organic acids, soluble and insoluble carbohydrates, ether-solublefraction and lignin were determined. Shikimic and quinic acidswere separated by the combined use of paper-chromatography andcolumn chromatography. Specific activity of formed quinate orshikimate was only slightly lower than that of fed shikimateor quinate. Specific activities of phenylalanine, tyrosine andbound tryptophan were high as compared with those of non-aromaticamino acids. Discussion is focused upon the interconversionbetween shikimate and quinate, and their roles in the biosynthesisof aromatic amino acids. (Received November 15, 1968; )     

Partial Synchronization of Carrot Cell Culture by Auxin Deprivation

A strain of carrot cells (Daucus carota cv. Kintoki) grew exponentially in the presence of 2,4-dichlorophenoxyacetic acid (2,4-D, 1 mg/1) with a doubling time of about 2 days. When those cells were transferred to a medium lacking 2,4-D, they continued to grow at almost the same rate for about a week. When the cells were again transferred to the auxin-free medium, the rate of cell division gradually decreased. After the cell division had ceased, cells were returned to the ordinary 2,4-D medium. A burst of cell divisions occurred after about 2 days. Timing of DNA synthesis and of mitosis suggested that the cells had been arrested at G₁ phase. In a medium containing indoleacetic acid instead of 2,4-D, the auxin was rapidly degraded and the culture was similarly synchronized as in the auxin-omitted medium.     

The translocation of photosynthetic products from mesophyll into midrib in tobacco plant III. The transformation of translocated 14C-sugars in the veins

¹⁴C-U-sugars were introduced into tobacco plants through themesophyll, the veins of the first order of branching, and themidrib, and ¹⁴C-compounds in the veins and the midrib whichtranslocated towards the base of the midrib were traced duringthe period of 120 min after the ¹⁴C-sugar introductions. 1) When ¹⁴C-U-sucrose was introduced into the leaf, no matterwhat the means of feeding was, most of the ¹⁴C which translocatedbasipetally in the veins and the midrib was found in the formof sucrose. 2) When ¹⁴C-U-glucose or ¹⁴C-U-fructose was administered tothe leaf dirough the cut vein of the first order of branching,most of the ¹⁴C which translocated basipetally in the veinsand the midrib was found in the form of sucrose. 3) ¹⁴C-U-glucose or ¹⁴C-U-fructose injected into the vascularbundles of the midrib was translocated basipetally, as such,10 and 30 min after injection; and at 30 min, the amount ofthe ¹⁴C-sucrose in the midrib attained 9–22% of the 80%ethanol-soluble ¹⁴C in the midrib. 4) When ¹⁴C-U-glucose or ¹⁴C-U-fructose was supplied to themesophyll, the radioactivities of these hexoses were predominantin the first and second veins soon after application, then decreasingwith a concomitant increase in the radioactivity of the ¹⁴C-sucrose. From these results, it was inferred that in the veins of thefirst and second order of branching, glucose and fructose whichmoved from the mesophyll did not translocate as such, but wereutilized for the synthesis of sucrose available for translocationvia the midrib to the stem. ¹A part of this paper was presented at the Crop Science Societyof Japan, in April, 1969 (Received December 8, 1969; )     

High temperature causes arrest of cell cycle in G2 phase in BmN cells derived from the silkworm, Bombyx mori

Abstract.  The influence of temperature on the insect cell line, BmN, derived from the silkworm, Bombyx mori is investigated. These cells proliferate at an accelerated pace as the temperature increases from 22 to 30 °C, but the growth rate slows at 34 °C, and proliferation stops at 38 °C. At high temperatures, abnormal cellular morphology is observed. Cells treated at 38 °C have cytoplasmic bilateral protrusions and they gradually aggregate and float in the medium. BmN cells without proliferation at 38 °C are viable but have reduced DNA synthesis. At high temperatures, the cell cycle of BmN cells halts at the G₂ phase. After heat treatment of the larvae, an accumulation of larval haemocytes with high DNA content is found, which suggests that the cell cycle arrest at G₂ also occurs in the silkworm at high temperatures.     

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.     

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; )     

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; )