磷有效性对大豆根冠中碳分配的影响

在水培条件下,研究不同浓度磷影响大豆根冠中碳分配的结果表明：磷有效性对大豆根冠中碳分配的影响依赖于磷浓度与胁迫时间。磷浓度高于0．125mmol．L^-1或低磷胁迫7d以内,大豆根冠中碳分配受到的影响不显著。低磷胁迫14d的大豆的净光合速率和根呼吸速率均显著下降,根冠比显著提高。这显示长期低磷胁迫下大豆碳同化总量和根呼吸消耗的碳量虽然减少,但根系生长的碳消耗则增加,光合碳同化形成的碳水化合物向根部的分配是受到促进的。     

Subunit Structure of Soybean 11S Globulin

The acidic and the basic subunits were shown to be present in equimolar amounts in the 11S globulin molecule by the densitometric scanning of the SDS gel and the molecular weight consideration. The four acidic subunits (A₁, A₂, A₃ and A₄) were found to be present in the approximate molar ratio of 1:1:2:2. Four basic subunits separated and designated as B₁, B₂, B₃ and B₄ based on the relative mobilities in the acidic gel in 7 m urea were found to be present in the approximate molar ratio of 1:1:2:2. The four basic subunits were fractionated in approximately same amounts into three different peaks, peak I (B₁ and B₂), peak II (B₃) and peak III (B₄) by CM-Sephadex C–50 column chromatography in the presence of 6 m urea. Three kinds of intermediary subunits of 11S globulin were fractionated with DEAE-Sephadex A–50 in the absence of reducing agents in 6 m urea, and disulfide bonds appeared to participate in the binding between the acidic and the basic subunits in the molar ratio of 1: 1 with the following combinations; A₁ and A₂ combined with B₃, A₃ with B₁ and B₂, and A₄ with B₄. In view of the above results and molecular weight consideration, a new model of subunit structure was proposed for 11S globulin.     

Food Processing Characteristics of Soybean 11S and 7S Proteins

This paper deals with the contribution of protein components in soybean seeds to the physical properties of tofu-gel. Results obtained in tofu-making, using crude 11S and 7S components from defatted soybean meal, indicated that there presented significant difference between tofu-gels from crude 11S and 7S, namely, the tofu-gel from crude 11S was remarkably harder than that from crude 7S. And it has been recognized that the proportion of 11S to 7S in total protein of soybean seeds considerably differed among varieties and that the difference of the proportion might be related to the physical properties of tofu-gel prepared.     

Crosslinking of Soybean 7S and 11S Proteins by Transglutaminase

Transglutaminase catalyzes the formation of intermolecular and intramolecular ε-(γ-glutamyl)lysyl crosslinks in proteins. The study here examined the substrate effectiveness of soybean 7S and 11S proteins in the intermolecular-crosslinking reaction catalyzed by guinea pig liver transglutaminase.

Both 7S and 11S proteins could act as the substrate for the transglutaminase reaction. The reaction with 11S protein was faster than that of 7S protein. Analyses of the reaction products by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that three main subunit groups of 7S protein and two acidic subunit groups of 11S protein were polymerized through the formation of intermolecular crosslinks by transglutaminase. Interestingly enough, no intermolecular crosslink was formed between the basic subunits of 11S protein. The possible significance of the intermolecular crosslinking catalyzed by transglutaminase is discussed, including the use of this enzyme reaction to improve the properties of food protein.     

Carbohydrate Component in 7S Protein of Soybean Casein Fraction

Carbohydrate components in the 7S protein from soybean casein fraction were found to be mannose and hexosamine. The former was identified by paper and starch-column chromatographies and its content was approximately 4% per protein. The latter, hexosamine, was contained about 1.2% per protein.

Mannose was considered as an integral constituent of the 7S protein from the data of heat and acid denaturation, paper electrophoresis and column chromatography with Sephadex G-200.     

Distribution and Redistribution of Sulfur Supplied as [35S]Sulfate to Roots during Vegetative Growth of Soybean

Soybean (Glycine max L.) plants were grown in nutrient solution containing 10 [mu]M sulfate and were treated at various times with [35S]sulfate for 48 h. Growth was then continued in unlabeled solution. The sulfur content of each leaf increased rapidly until it was about 40% expanded; small, additional increases occurred until the leaf was about 70% expanded after which the sulfur content decreased by about 50%. Leaves that were about 60 to 70% expanded during the pulse were strongly labeled but then underwent a significant loss of 35S label. Leaves that were in the early stages of expansion imported little 35S label during the pulse but acquired 35S label during the chase period as they expanded (i.e. redistribution). Most of the redistributed 35S label was derived from other leaves. The rates of both sulfur import and sulfur export by a leaf were greatest at about 70% expansion. Leaves that acquired 35S label during early development retained a much higher proportion of their label than leaves that were more developed, suggesting that the sulfur acquired by leaves during early development is preferentially incorporated into a pool that is less mobile than the sulfur acquired in the later stages of leaf growth.     

Association of Phosphatidylcholine with Soybean 11S Globulin

To address the multiplicity of aromatic ring hydroxylation dioxygenases, we used PCR amplification and denaturing gradient gel electrophoresis (DGGE). The amplified DNA fragments separated into five bands, A to E. Southern hybridization analysis of RHA1 total DNA using the probes for each band showed that band C originated from a couple of homologous genes. The nucleotide sequences of the bands showed that bands A, C, and E would be parts of new dioxygenase genes in RHA1. That of band B agreed with the bphA1 gene, which was characterized previously. That of band D did not correspond to any known gene sequences. The regions including the entire open reading frames (ORFs) were cloned and sequenced. The nucleotide sequences of ORFs suggested that the genes of bands A,C, and E may respectively encode benzoate, biphenyl, and polyhydrocarbon dioxygenases. Northern hybridization indicated the induction of the gene of band A by benzoate and biphenyl, and that of the gene of band C by biphenyl and ethylbenzene, supporting the above notions. The gene of band E was not induced by any of these substrates. Thus the combination of DGGE and Southern hybridization enable us to address the multiplicity of the ring hydroxylation dioxygenase genes and to isolate some of them.     

Growth and Metabolism of Soybean as Affected by Paclobutrazol

Paclobutrazol, an experimental growth retardant, was soil-appliedat the rate of 125 or 250 µg active ingredient per 10cm pot to 19 day-old soybean plants. This compound considerablyreduced plant height, leaf area, and stem dry weight. In addition,paclobutrazol-treated plants had numerous thickened lateralroots at the soil surface and had increased chlorophyll andsoluble protein contents compared to controls. During the first14 days after treatment, paclobutrazol increased the activitiesof NAD- and NADP-glyceraldehyde-3-phosphate dehydrogenase andaminotransferases but decreased the activity of nitrate reductase.Net photosynthesis (P_n) of the first and second trifoliatesof treated plants remained fairly constant throughout the studywhile control P_n declined during the latter portion of the experimentalperiod presumably due to leaf senescence. This decline of P_nin controls was accompanied by a decrease in the activitiesof NAD- and NADP-glyceraldehyde-3-phosphate dehydrogenase, nitratereductase, aminotransferases, and NAD malate dehydrogenase.Activities of these enzymes also tended to decline in paclobutrazol-treatedplants, but were still considerably higher than in controlsat the end of the experiment. The activities of RNase, protease,and glutamic dehydrogenase were higher in controls than in treatedplants. Our results suggest that paclobutrazol not only modifiesthe activity of a number of soybean enzymes but also delaysthe onset of senescence, thereby prolonging the period of normalmetabolic activity in a given leaf. ¹ Permanent address: Department of Botany, University of Jodhpur,India.     

The Effect of Source-Sink Alterations on Soybean Seed Growth

Soybeans (Glycine max L. Merrill) were grown in the greenhouseand in the field to investigate the effect of variations inthe assimilate supply during the linear phase of seed developmenton the rate and duration of growth of individual seeds. Increasedassimilate supplies, created by partial fruit removal, increasedrates of dry matter accumulation, duration of seed growth, andfinal seed size (weight per seed). Reductions in the supplyof assimilate to the developing seed, created by shading (60per cent) the plants during the linear phase of seed development,lowered seed growth rate but did not affect final seed sizebecause of a longer duration of seed growth. Nitrogen stressduring seed development, created by removing N from the nutrientmedium, did not affect seed growth rate but shortened the durationof seed growth and reduced final seed size. The data indicatethat the growth characteristics of soybean seed are influencedby the supply of assimilate to the seed during the linear phaseof seed development. Glycine max L., soybean, seed growth rate, duration of seed growth, effective filling period     

Interaction of Kinetin and Various Inhibitors in the Growth of Soybean Tissue

The growth of kinetin-requiring soybean callus was inhibited by the purine analogs 2,6-diaminopnrine, 8-azaguanine and 8-azaadenine. The purine analog 6-mercaptopnrine and the antibiotic puromycin were not effective as inhibitors at the concentrations employed. Reversal of the inhibition was attempted by raising the kinetin concentration. Kinetin did not reverse the inhibition due to 2,6-dianiinopurine but it did lessen to some extent the inhibition due to 8-azagnanine. It is suggested the kinetin affects RNA metabolism.     

Gelation Phenomena Induced by Alkali-alcohol Treatment of 7S and 11S Components in Soybean Globulins

Transparent gels containing about 2% protein were obtained by mixing alkaline dope solution of 7S or 11S soybean proteins with alcohol. The 7S component showed the ability to form a stronger gel than the 11S. This phenomenon depended on pH and alcohol concentration. In 66 % ethanol, the viscosity of the 7S and 11S reached maxima at pH 11.4 and 11.2, respectively. Above these pH levels where further unfolding and dissociation into subunits of the protein molecules occur, the viscosity decreased rather. The effectiveness of alcohol to increase viscosity increased in the order; n-butanol < tert-butanol < n-propanol < iso-propanol < ethanol < methanol. Alcohols having minor hydrophobicity were more effective for increasing viscosity, but ethylene glycol was ineffective. The addition of NaCl or 2-mercaptoethanol to ethanol-mixed alkaline dope solutions resulted in the remarkable increment of the viscosity, especially for the 7S.     

Growth and N Allocation in Rice Plants under CO2 Enrichment

The effects of CO2 enrichment on growth and N allocation of rice (Oryza sativa L.) were examined. The plants were grown hydroponically in growth chambers with a 14-h photoperiod (1000 [mu]mol quanta m-2 s-1) and a day/night temperature of 25/20[deg]C. From the 28th to 70th d after germination, the plants were exposed to two CO2 partial pressures, namely 36 and 100 Pa. The CO2 enrichment increased the final biomass, but this was caused by a stimulation of the growth rate during the first week of the exposure to elevated CO2 partial pressures. The disappearance of the initial stimulation of the growth rate was associated with a decreased leaf area ratio. Furthermore, CO2 enrichment decreased the investment of N in the leaf blades, whereas the N allocation into the leaf sheaths and roots increased. Thus, the decrease in leaf N content by CO2 enrichment was not due to dilution of N caused by a relative increase in the plant biomass but was due to the change in N allocation at the whole-plant level. We conclude that the growth responses of rice to CO2 enrichment are mainly controlled by leaf area expansion and N allocation into leaf blades at the whole-plant level.     

抗生素对大豆愈伤组织的诱导和生长的影响

用红霉素、头孢唑唑钠、头孢拉定、头孢霉素（国产和进口）等5种抗生素对农杆菌LBA4404进行抑菌试验,以头孢霉素的抑菌效果最好。头孢霉素作为抑菌剂用大于豆遗传转化试验时,在下胚轴浓度以300mg/L,在子叶节以500mg/L。大豆品种对卡那霉素的反应在出愈率上表现相似,在褐化率上表现有些不同。大豆不同外植体对卡那霉素的反应存在较大差异,以真叶反应最敏感,下胚轴反应最迟钝。在以卡那霉素作为抗性选择标记时,选择压力真叶和子叶节以50－100mg/L为好,下胚轴以100－200mg/L为宜。     

Relationship between Gelation and Aggregation of Alkali-treated Soybean 7S and 11S Globulins

Gel was obtained when alkaline dope solutions of the 7S and 11S globulins (8% protein concentration) prepared at pH above 11 were dialyzed against phosphate buffer, pH 7.6, µ= 0.3. To make clear the mechanism of gelation, the relationship between changes in viscosity and aggregation phenomena of the neutralized dope solutions was investigated by means of viscosity measurement, disc electrophoresis and gel filtration, comparing the 7S and 11S fractions. In conclusion, it is revealed that the gel is constituted with macromolecule aggregates, and to form the aggregates which are suitable for gelation, all of the following conditions must be satisfied at least : 1); Unfolding and dissociation into subunits once (above pH 11), 2); High ionic strength in the media (µ=0.3), 3); Formation of hydrogen, hydrophobic and disulfide bonds, 4); High protein concentration (above 8%).     

Anaerobic Growth and Denitrification among Different Serogroups of Soybean Rhizobia

We screened soybean rhizobia originating from three germplasm collections for the ability to grow anaerobically in the presence of NO₃⁻ and for differences in final product formation from anaerobic NO₃⁻ metabolism. Denitrification abilities of selected strains as free-living bacteria and as bacteroids were compared. Anaerobic growth in the presence of NO₃⁻ was observed in 270 of 321 strains of soybean rhizobia. All strains belonging to the 135 serogroup did not grow anaerobically in the presence of NO₃⁻. An investigation with several strains indicated that bacteria not growing anaerobically in the presence of NO₃⁻ also did not utilize NO₃⁻ as the sole N source aerobically. An exception was strain USDA 33, which grew on NO₃⁻ but failed to denitrify. Dissimilation of NO₃⁻ by the free-living cultures proceeded without the significant release of intermediate products. Nitrous oxide reductase was inhibited by C₂H₂, but preceding steps of denitrification were not affected. Final products of denitrification were NO₂⁻, N₂O, or N₂; serogroups 31, 46, 76, and 94 predominantly liberated NO₂⁻, whereas evolution of N₂ was prevalent in serogroups 110 and 122, and all three were formed as final products by strains belonging to serogroups 6 and 123. Anaerobic metabolism of NO₃⁻ by bacteroid preparations of Bradyrhizobium japonicum proceeded without delay and was evident by NO₂⁻ accumulation irrespective of which final product was formed by the strain as free-living bacteria. Anaerobic C₂H₂ reduction in the presence of NO₃⁻ was observed in bacteroid preparations capable of NO₃⁻ respiration but was absent in bacteria that were determined to be deficient in dissimilatory nitrate reductase.     

Homology between the Acidic Subunits of Soybean 11S Globulin

The formaldehyde dehydrogenase (EC 1.2.1.1) from the yeast Pichia pastoris IFP 206 was purified to homogeneity. The enzyme had a molecular weight of 84,000 daltons and was composed of two identical subunits of a molecular weight of 39,000 daltons. The N-terminal end of the subunits is blocked. The protein showed 6,3 free -SH groups per mole and 12,5 in the presence of NAD⁺. Enzyme stability was increased by addition of glycerol during the purification.

The enzyme activity is NAD⁺ and glutathione dependent. The reaction product is S formylglutathione. The presence of an S-formylglutathione hydrolase (EC 3.1.2.12) in the cell free extract was detected. The formaldehyde dehydrogenase showed an optimum pH of 7.9 and an optimum temperature of 47°C. The activation energy was 3.2 kcal/mol. The Michaelis constants for NAD⁺ and S-hydroxymethyl glutathione were respectively 0.24 mM and 0.26 mM.