LACTATE AND GLUCOSE OXIDATION SYSTEMS IN ACETOBACTER SUBOXYDANS

1. From a strain of Acetobacter suboxydans, a glucose and a lacticenzyme were obtained in cell-free states. The lactic enzymeshows as strong activity as the glucose enzyme but is more stablethan the latter toward various purification procedures: bothare sensitive to high temperature treatment. Activities of thetwo enzymes and the MICHAELIS constants of the glucose enzymewere determined under both aerobic and anaerobic conditions.
2. Carbon monoxide inhibits the oxygen-uptake in both glucoseandlactate oxidation. WARBURG's distribution constant for lactateoxidation is 6.7. These results suggest the participation ofan heme enzyme in the oxidation system.
3. Effects of copperreagents, narcotics and PCMB were also examined.
4. The dehydrogenaseactivities (reduction of dye) of the enzymesare more sensitiveto high temperature than the correspondingactivities in oxygen-uptake.
5. By combining a dehydrogenase preparation which has lost itsoxygen-absorbing activity through acetone treatment, with aheated extract, a partial recovery of oxygen-uptake can be realizedin lactate oxidation.
6. L-Cysteine is utilized as hydrogen donorby the bacterium. Thisoxidative reaction, unlike the oxidationof lactate, is notinhibited by surface active reagents.

(Received May 16, 1960; )     

A gibberellin-regulated protein phosphorylated by a putative Ca2+-dependent protein kinase is G-protein mediated in rice root

The GA-signal transduction pathways downstream to the Gα protein in rice seedling root were investigated using in-gel kinase assay and in vitro protein phosphorylation techniques with a Gα protein defective mutant, d1. A 50-kDa protein kinase was detected downstream to Gα protein in the membrane fraction of rice seedling roots using an in-gel kinase assay with histone III-S as a substrate. The activity of a 50-kDa protein kinase increased in the wild-type rice by gibberellin (GA₃) treatment, but did not change in the d1 mutant. This protein kinase activity was inhibited by the Ca²⁺ chelator ethyleneglycol-bis-(beta-aminoethylether)-N,N,N ¹,N ¹-tetraacetic acid (EGTA), protein kinase inhibitors, staurosporine and H7, and calmodulin antagonist, trifluoperazine, suggesting that the 50-kDa protein kinase is a putative plant Ca²⁺-dependent protein kinase (CDPK). The activity of the 50-kDa putative CDPK reached its highest level at 3 h after GA₃ treatment and then gradually declined with time. In order to identify the endogenous substrate for 50-kDa putative CDPK, two-dimensional polyacrylamide gel electrophoresis followed by in vitro protein phosphorylation was carried out. The phosphorylation activity of an endogenous protein PP30, identified as an unknown protein having molecular weight 30 kDa and isoelectric point 5.8 was increased in the wild-type rice by GA₃ treatment, compared with the d1 mutant. The addition of GA₃ treated membrane fraction, which predominantly represent a 50-kDa putative CDPK further increased the phosphorylation of PP30. Almost similar to GA₃ treatment, phosphorylation activity of PP30 was also increased by the treatment with cholera toxin in the wild-type rice but not in d1 mutant. These results suggest that the 50-kDa putative CDPK and an unknown protein, PP30 promoted by GA₃ treatment are G-protein mediated in rice seedling roots.     

COMPONENTS OF THE ELECTRON-TRANSFERRING SYSTEM IN ACETOBACTER SUBOXYDANS AND RECONSTRUCTION OF THE LACTATE OXIDATION SYSTEM

1. Cytochromes a₁⁵⁹⁰, b⁵⁶⁰, c₁⁵⁵⁴ and c₁⁵⁵² were isolated andpurifiedfrom a strain of Acetobacter suboxydans. The proceduresusedwere described in detail.
2. The main cytochrome band at550-560 mµ in intact cellssplitted at liquid air temperatureinto two bands, 551 mµ(strong) and 559 mµ (weak).
3. Optical and physiological properties of the four cytochromeswere investigated. Lactic dehydrogenase activity was found tobe associated with cytochrome c₁⁵⁵⁴. The two c₁-type cytochromes,especially cytochrome c₁⁵⁵⁴, persisted in their reduced formafter the purification through many steps.
4. By some combinationsof isolated components reconstruction ofthe oxygen uptake systemcould be realized.
5. The oxygen-consuming activity of purifiedoxidase preparationswas accelerated by a-tocopherol but notby Emasoll 4130 andTween 80.
6. Some discussions were made onthe nature of terminal oxidase,the role of cytochrome c₁⁵⁵²in the electron-transport system,and persistence of reducedstate of c₁-type cytochromes.
7. A possible scheme of the electron-transferringsystem of Acetobactersuboxydans was presented.

(Received May 16, 1960; )     

Crystallization and some properties of cryptocytochrome c from aerobically-grown Pseudomonas denitrificans

1. 1. Analyses of cytochrome types in intact cells of aerobically-and anaerobically-grownPs. denitrificans indicated a higherratio of cytochrome c to cytochrome b in the former than inthe latter.
2. 2. Anaerobically-grown cells contained about twotimes as muchcryptocytochrome c as did aerobically-grown cells.
3. 3. Crystalline cryptocytochrome c obtained from the solublefraction of cell-free extracts of aerobically-grown cells manifestedthe same properties as cryptocytochrome c from anaerobically-growncells, i. e., absorption maxima, autooxidizability, redox potential,molecular weight, haem content, etc.
4. 4. Cryptocytochrome cwas reversibly converted to a true haemochrometype spectrumby alcohols, detergents, carboxylic acid salts,guanidine saltor high pH values.

(Received December 16, 1968; )     

Seed germination time course and seedling development of Clintonia udensis Trautv. et Mey. (Uvulariaceae): a typical cool temperate woodland perennial in Japan

Seed germination time course and seedling development mechanisms of Clintonia udensis Trautv. et Mey. (Uvulariaceae) were investigated under experimental condition. Seed germination tests were carried out under four thermal regimes, i. e. 10, 15, 20, and 25°C, after seeds were harvested, and stored at 5°C in wet conditions for 6 months under light‐exposed or shaded conditions. Approximately 63% of all seeds produced had the potential to germinate beyond 4 years and 6 months. The developmental process after germination continued for over 2 years. Phase I: the radicle first breaks through the seed coat 2 years after fructification. Phase II: the radicle becomes much larger with a hypocotyle. Phase III: part of the cotyledon elongates over 20 mm. Phase IV: the plumule further develops in two steps, i. e. the plumule is first formed, while cotyledon is disappearing, and then the plumule appears with second and third radicles, growing with cotyledon.     

太平洋日本沿岸斑点原海豚的年龄鉴定与生长

本文利用改良的石蜡包埋切片染色技术,对太平洋日本沿岸收集到的286头斑点原海豚(Stenella attenuata)的牙齿标本进行了年龄鉴定,定义了斑点原海豚的牙齿质和牙骨质生长层组(GLGs)。从年龄和生长的角度对斑点原海豚的一些种群生物学特征进行了分析,并推算了部分种群参数。斑点原海豚在约18龄时,牙齿质停止生长,牙髓腔封闭。牙齿质中计数到的GLGs最多为18层,牙骨质中计数到的最多为42层。斑点原海豚的年龄与生长在一定范围内遵循一定的幂函数关系。5—6龄以前,斑点原海豚生长迅速,雌雄生长无明显差异。8—14龄之间,雄性的生长快于雌性。雌性约17龄时达到饱和体长191cm,雄性约22龄达到饱和体长201cm[动物学报51(3)：476-485,2005]。     

Alterations by a defect in a rice G protein α subunit in probenazole and pathogen-induced responses

The rice dwarf1 (d1) mutant, which is deficient in an α subunit (Gα) of heterotrimeric G protein, was used to obtain specific evidence on the functions of Gα protein in defence signalling in rice. Using proteome analysis, a probenazole‐inducible protein (PBZ1) was detected in the cytosolic fraction of leaf blade of the wild type, but not the d1 mutant. After treatment with probenazol, PBZ1 reached maximal levels at 72 h in the wild type but 96 h in the d1 mutant. The induction of PBZ1 by probenazole treatment was inhibited by protein kinase inhibitors. A 48‐kDa putative mitogen‐activated protein kinase (MAPK) and a 55‐kDa putative Ca²⁺‐dependent protein kinase (CDPK) showed lower activities in the cytosolic fraction of the d1 mutant than that of the wild type. The activities of these protein kinases were enhanced at 24 h in the wild type and 48 h in the d1 mutant after probenazole treatment. Although the d1 mutant responded to the rice blast fungus similarly to the wild type, the d1 mutant developed rice blight symptoms earlier than the wild type when infected with Xoo. In addition, the blight symptoms were more severe on the mutant than on the wild type, and wilting was frequently observed in the d1 mutant. Furthermore, induction by the bacterial infection of the 48‐kDa putative MAPK and PBZ1 was delayed by 2 and 4 d, respectively, in the d1 mutant compared with the wild type. These results indicate that the Gα protein plays a role in the induction of PBZ1 and protein kinases by probenazole and Xoo, and suggest that the 48‐kDa putative MAPK may be involved in a signalling pathway for resistance to bacterial infection.     

NUTRITIONAL STUDIES OF THE EDIBLE SEAWEED PORPHYRA TENERA I. THE INFLUENCE OF DIFFERENT B12 ANALOGUES, PLANT HORMONES, PURINES AND PYRIMIDINES ON THE GROWTH OF CONCHOCELIS

The red alga Porphyra tenera has been obtained in axenic cultureby the "dip and drag" technique in an agarized medium containingantibiotics. In the axenic culture, the Conchocelis of P. teneraneeds vitamin B₁₂ for growth. The addition of other vitaminsdoes not increase growth further. The pattern of specificityis similar to that of Escherichia coli 113-3. Factor B, pseudovitaminB₁₂ and Factor Z₂ support as much growth as vitamin B₁₂, while2-methylmercaptoadenine and 5-methyl benzimidazole cobalamineincrease the growth more than B₁₂. All the analogues containingbenzimidazole can replace B₁₂. Kinetin, adenine, indoleaceticacid and especially gibberellic acid increase growth. Threepurines, (xanthine, hypoxanthine and guanine) and three pyrimidines(uracil, methylcytosine and thymine) also promote growth inthe presence of vitamine B₁₂. (Received January 18, 1965; )