Potential agronomic options for energy-efficient sugar beet-based bioethanol production in northern Japan

Sugar beet (Beta vulgaris L. subsp. vulgaris) is deemed to be one of the most promising bioethanol feedstock crops in northern Japan. To establish viable sugar beet‐based bioethanol production systems, energy‐efficient protocols in sugar beet cultivation are being intensively sought. On this basis, the effects of alternative agronomic practices for sugar beet production on total energy inputs (from fuels and agricultural materials during cultivation and transportation) and ethanol yields (estimated from sugar yields) were assessed in terms of (i) direct drilling, (ii) reduced tillage (no moldboard plowing), (iii) no‐fungicide application, (iv) using a high‐yielding beet genotype, (v) delayed harvesting and (vi) root+crown harvesting. Compared with the conventional sugar beet production system used in the Tokachi region of Hokkaido, northern Japan, which makes use of transplants, direct drilling and no‐fungicide application contributed to reduced energy inputs from raising seedlings and fungicides, respectively, but sugar (or ethanol) yields were also reduced by these practices, to a greater equivalent extent than the reductions in energy inputs. Consequently, direct drilling (6.84 MJ L^?1) and no‐fungicide application (7.78 MJ L^?1) worsened the energy efficiency (total energy inputs to produce 1 L of ethanol), compared with conventional sugar beet production practices (5.82 MJ L^?1). Sugar yields under conventional plow‐based tillage and reduced tillage practices were similar, but total energy inputs were reduced as a result of reduced fuel consumption from not plowing. Hence, reduced tillage showed improved energy efficiency (5.36 MJ L^?1). The energy efficiency was also improved by using a high‐yielding genotype (5.23 MJ L^?1) and root+crown harvesting (5.21 MJ L^?1). For these practices, no major changes in total energy inputs were noted, but sugar yields were consistently increased. Neither total energy inputs nor ethanol yields were affected by extending the vegetative growing period by delaying harvesting.     

STUDIES ON OXALIC ACID OXIDASE IN GREEN LEAVES

1. The leaves of Chenopodium ambrosioides L. were found to havean intense activity of oxalic acid oxidase. The enzyme was locatedin the chloroplast, being firmly hound to its structure. 2. Properties of this chloroplast oxalic acid oxidase were described.The strict aerobic nature and the stoichiometry of the reactionwere confirmed. 3. Isolation of the enzyme from chloroplasts was performed,rupturing the chloroplasts with a French pressure cell or usingpyridine-water (1:1) as an extracting medium. 4. The enzyme was found to contain flavine and its activitywas enhanced in the presence of flavine added. Accordingly,the enzyme was inferred to be a flavine enzyme. (Received December 23, 1963; )     

PHOTOCHEMICAL INTERCONVERSION BETWEEN PRECURSORS OF PHYCOBILIN CHROMOPROTEIDS IN TOLYPOTHRIX TENUIS

1. The photochemical conversion between the precursors of phycocyaninand phycoerythrin in Tolypothrix tenuis was investigated.
2. Itwas found that the conversion of phycocyanin-precursor intophycoerythrin-precursor was induced by green light, and thereverse reaction by red light. These reactions proceeded exponentially, indicating that the photochemical process was acceleratedautocatalytically by the reaction-product.
3. The rates of thesephotochemical reactions were found to beunaltered by varyingthe incubation temperature (0? to 35?)and the composition ofthe gas atmosphere (presence or absenceof CO₂ and of O₂ orby an inhibitor of photosynthesis, p-chlorophenyldimethylurea.
4. The action spectra of the photochemical interconversions betweenprecursors of phycobilin chromoproteids were found to be distinctlydifferent from the absorption spectra of chlorophyll and carotenoids.The most effective wavelength for inducing the conversion ofphycocyanin- into phycoerythrin-precursor (541 mµ) isnear the absorption maximum of phycoerythrin (565 mµ),and that of the reverse reaction (641 mµ) is near theabsorption maximum of phycocyanin (620 mµ). Additionaldata, indicating that the phycobilin chromoproteids themselvesdo not participate in these processes as light absorber, werealso presented.
5. On the basis of these results, a possiblemechanism of the photochemicalinterconversion between the precursorsof phycobilin chromoproteidsis proposed.

(Received March 13, 1962; )     

Oculocutaneous Albinism in the i6 Mutant of the Medaka Fish is Associated with a Deletion in the Tyrosinase Gene

Three mutant alleles (i¹, i⁴, and i⁵) of the tyrosinase gene in the i locus of the medaka fish Oryzias latipes have hitherto been described, all being associated with transposable element insertion. We have recently identified another allele causing a complete albino phenotype in homozygous carriers and named it i⁶. Sequence comparison between the tyrosinase gene for the i⁶ allele (Tyr-i⁶) and the wild-type gene previously obtained (Tyr-i ⁺) revealed three deletions of 8, 44, and 245 bp. The first two deletions reside in an intron and are differences in the number of tandem tetranucleotide repeats that are polymorphic even among wild-type genes, and, thus, not likely to be responsible for the i⁶ albino phenotype. The largest deletion spans over the last 180 bp of the second intron and the first 65 bp of the third exon. Because of this deletion, the Tyr-i⁶ gene lacks the branch point sequence and the acceptor site for the second intron, both being considered to be necessary for normal RNA splicing. Therefore, the 245-bp deletion is likely to be responsible for the albino phenotype. With a mutant gene of this type, unlike ones bearing transposable element insertions, the possibility of reversion mutations to the wild-type would be negligible. Therefore, fish having the i^e/i⁶ genotype should serve as superior recipients for the tyrosinase gene in rescue experiments.     

Formation of Embryo Thromboblasts in Chick Blastoderm: Morphology, Site of Production and Time of Emergence in the Blood

Thrombocytes in the blood of chick embryos (termed embryo thrombocytes by L ucas and J amroz ) have PAS-positive granules in their cytoplasm. Electron microscopic observations reveal that the embryo thrombocytes contain glycogen granules present singly or in clumps. The presence of these inclusions and other morphological characteristics were used as specific markers to distinguish embryo thrombocytes from primitive erythroid cells. These markers also made it possible to determine the time at which the immature thromboblasts first emerge in blood vessels, and the period of their continued presence in the circulation. In this way we found that thromboblasts were detectable in embryos as early as stage 10⁺ of H amburger and H amilton (after 35 hr incubation) and that the thromboblasts were present in the circulation until day 4 of incubation (stage 23). In ovo and in vitro culture of de-embryonated blastoderm demonstrated that thromboblasts were formed in the area opeca vasculosa. The present observations suggest that embryo thromboblasts are formed at the same time and in the same area as the primitive cells of erythroid line.     

LIGHT-INDUCED REDUCTION OF NITRATE, NITRITE AND HYDROXYLAMINE IN A BLUE-GREEN ALGA, ANABAENA CYLINDRICA

1. Reduction of nitrate, nitrite and hydroxylamine by intact cellsof Anabaena cylindrica was investigated with special referenceto the stimulating effect of light on these processes.
2. Itwas found that in light and under anaerobic condition thesecompounds are reduced to ammonia, with the production of extraoxygen. The stoichiometry of the reactions under these conditionscan be represented as follows: HNO₂+H₂O=NH₂+2O₂ HNO₂+H₂O=NH₂+1O₂ NH₂OH+H₂O=NH₂+O₂+H₂O
3. Reduction of nitrite and hydroxylaminewas markedly suppressedby CMU in the light but not in the dark.KCN inhibited reductionto the same extent both in the lightand in the dark. Reductionin the light was much less sensitiveto the uncoupling agent,DNP, than was that in the dark.
4. Atlow light intensities, CO_2– was suppressed by 20–30per cent by the simultaneous provision of nitrite, but the nitritereduction was not affected at all by CO₂. At high light intensities,reduction of nitrate and nitrite was considerably acceleratedby CO₂
5. On the basis of these findings, a possible mechanismfor thelight stimulation of the reactions in question was brieflydiscussed.

(Received August 22, 1962; )     

FORMATION OF PHYCOERYTHRIN IN PRE-ILLUMINATED CELLS OF TOLYPOTHRIX TENUIS WITH SPECIAL REFERENCE TO NITROGEN METABOLISM

1. The formation, in the dark, of phycoerythrin in the preilluminatedcells of a blue-green alga, Tolypothrix tenuis, was investigatedwith special reference to its nitrogen metabolism.
2. On incubatingthe pre-illuminated algal cells under a darkaerobiccondition,and with nitrate as N-source, the formation of phycoerythrinoccurs after an induction period of about 5 hours. No time-lagis observed in the nitrate-uptake by the organism. Similar resultsare obtained with nitrite, ammonia, urea and arginine as N-sources.
3. The above stated formation of phycoerythrin is suppressedbysubstances such as chloramphenicol and p-fluorophenylalanine,substances known to be potent inhibitors of protein-synthesis.
4. On the basis of these findings, it was inferred that therearetwo consecutive processes involved in the dark-formationofphycoerythrin in the pre-illuminated cells: (i) uptake andconversionof exogenous nitrogen sources into some intermediarynitrogenouscell substances, and (ii) synthesis of the pigment-proteinfromthese substances.

(Received June 27, 1960; )     

EFFECT OF CHROMATIC LIGHTS ON PHYCOBILIN FORMATION IN A BLUE-GREEN ALGA, TOLYPOTHRIX TENUIS

1. The formation of phycobilin pigments in a blue-green alga Tolypothrixtenuis was investigated with special reference to the effectsof preillumination with colored lights.
2. It was discoveredthat the algal cells are capable of formingphycobilin pigmentsin the dark, if they have been previouslyilluminated for severalhours in the presence of CO₂.
3. The color of light applied inthe later period of preillumination(chromatic illumination)was found to affect the ratio of phycoerythrinto phycocyaninformed in the subsequent dark period. A greenlight acceleratesthe dark-formation of phycoerythrin, a redlight that of phycocyanin,and the two lights counteractingwith each other in their effects.
4. These directive effects of the "chromatic illumination" canbe accomplished within a very short period, for instance, in3 minutes if it is preceded by sufficient "preillumination"with an incandescent or day light fluorescent light. The reactionsoccurring during the period of chromatic illumination does notrequire the presence of CO₂ and the aerobic condition.
5. Thealga can be grown heterotrophically when supplied with casaminoacids and glucose. Under such a condition the alga forms phycocyanintogether with chlorophyll and carotenoids, but not phycoerythrin.
6. On the basis of the results obtained, a tentative scheme forthe biosynthesis of phycobilin pigments in the alga was proposed,assuming the light-induced formation of unknown precursors whichare converted into phycocyanin and phycoerythrin in the subsequentdark period.

(Received July 4, 1960; )     

Nitrogen fixation and heterocysts in the blue-green alga Anabaena cylindrica

The site of nitrogen fixation in the blue-green alga Anabaenacylindrica Lemra (Fogg strain) was investigated. Less than 4%of the total nitrogen fixed during a relatively short period(5-15 min) was recovered in heterocysts. When estimated on thecellular nitrogen basis, vegetative cells can fix molecularnitrogen at the same rate as do heterocysts. There was no positivecorrelation between nitrogen fixation and heterocyst formation.Results do not support the hypothesis that the heterocyst isthe main site for nitrogen fixation in blue-green algae. ¹ This work was supported by grant (No. 38814) from the Ministryof Education. (Received July 23, 1971; )     

Web-spinning sawflies of the sylvaticus-group of Pamphilius (Hymenoptera: Pamphiliidae)

Abstract The Pamphilius sylvaticus group is redefined to include seventeen species in the Holarctic: sylvaticus (Linnaeus), fumipennis (Curtis) and nemorum (Gmelin) from Europe; alnicola Ermolenko, daisenus Takeuchi, volatilis (Smith), benesi sp.n., gracilis sp.n., japoni-cus sp.n. and montanus sp.n. from northeastern Asia (mainly Japan); burquei (Provancher), cinctus Harrington, middlekauffi Shinohara & Smith, ocreatus (Say), rileyi (Cresson) and semicinctus (Norton) from eastern N. America; and persicum MacGillivray from eastern N. America and the Caucasus. Each species is described and figured and a key for identification is given.