Physiological action of the alcoholic extracts of sclerotia and mycelial mass formed by the culture of Scl, Lib. on grain byproducts (rice-bran, ctc.), after being evaporated to small vol. and treated by a proper process, was tested pn cock's comb, frog's pupil, frog's vessel and rabbit uterus.
It was found that the extracts like ergot's alkaloid suppressed the motor action of adrenaline on the iris muscle or blood vessel of frog, and uterus-active constituent was somewhat rich in the mycelial mass formed by a mutant Sm, rather than in mycelial mass, which contains sclerotia, formed by the original strain Ss.

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1. Anastomosis occurs between variants and between different strains of Koji-molds.
2. Anastomosis takes place often between conidia, between germ-tubes and between mature .hyphae, accompanied by vigorous nuclear division and nuclear migration through connecting tube or fusion bridge.
3. When the constant types S and F are grown together in mix-culture, they produce intermediate type X. In pedigree-culture..by successive single spore method of the X-type, they again separate into all 3 types, namely, X, S, and F.
4. The constant type or homo-type S and F or conidia are comparatively uniform in sizes and leave 1-4 nuclei.
The conidia of inconstant type or hetero-type X, which occur by mix-culture, are rich in variety of sizes and multiiiucleate.
5. By mix-culture between different strains of Koji-molds, they also give rise to an interstrainic dicaryophytic hybrids.
6. From the above facts and the findings hetherto reported ⁶⁾ it may be concluded that the natural variations of Koji-Molds (Asp. oryzae and Asp. sojae) is to greater extent responsible to the heterocaryosis. And the resultant heterocaryosis produce generally conidia of much greater diameters than those of the parent strains, a phenomenon similar to heterosis, through, which the heterocaryosis are very easily detectable.

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Extracted spruce wood meal, spruce holocellulose and rayon pulp were cooked with NH₂OH•HCl solution in ethanol (pH 2.5) in the boiling water bath during 1_??_20 hrs. Acetal linkages in these materials are hydrolyzed and the liberated carbonyl groups make oxime with the production of equivalent HCl. The curves (produced HCl-reaction time) for holocellulose and rayon pulp were linear, whereas in the case of spruce wood the rate of HCl production was very rapid in the earlier stage and the curve turned to straight line after about 8 hours. Extraporating the latter to time zero, it was concluded that 1g wood meal liberated 0.3 m-mol HCl within a few earlier hours. The present authors suppose that this 0.3 m-mol HCl per g wood must be produced by cleavage of acetal linkages between carbonyl group of lignin and carbohydrates, and therefore lignin has 1 mole carbonyl group per 960g.

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The plant tissues usually turn brown, when they are affected by pathogenic microorganisms: Many papers have been published about this problem, but no theory supported by chemical experiments has been established. We have selected the sweet potato infected with Ceratos, tomella fimbriata, so-called “black-rotted” sweet potato, to carry out our experiments.
In this study, the authors examined, from the point of the view mentioned above, the rise and fall of activities of some enzymes and polyphenols which were contained in the injured parts as well as in the sound parts in order to observe the abnormal metabolism caused by the infection. The results summerised are as follows.
The determination methods of the activities of oxidase, peroxidase, and catalase, and the quantity of polyphenols were improved so as to be suitable for our experiments.
It was ascertained that, using the uninoculated sound potato as the control, the oxidase and peroxidase were highly activated and polyphenols were noticeably accumulated in the sound part adjacent to the injured. This abnormal metabolic phenomena were observed more distinctly. in case of sufficient supply of oxygen than in case of lack of oxygen, and more in case of Vari-ety Norin No. 1 than in case of Variety Okinawa No. 100.
As to the reason why the injured part turned brown, it was ascertained that ate first the function of the protoplasm was destroyed and then the oxidation of polyphenols by activated`.oxidase and peroxidase, occurred 'irreversively, and the quinoid substances thus formed were polymerized immediately, as, soon as the cells which were working out the abnormal metabo-lisrn mentioned above were penetrated by this fungus.
The oxxidase and peroxidase of this fungus had some effects on the browning of the tissue though the enzyme activities of-this fungus were weak.
It was observed that the polyphenolic and quinoid substances might inhibit the penetration of the fungus into the cells cooperating with the bitter substances and resins contained in the injured part. The details including the inhibitory activity, of these substances will be published in Part V of this series.
Higher activities of catalase in the injured part was found to be due to the presence of cata-:las, e in the--mycellium of this fungus grown in the injured part.

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Fusarinic acid is proved to have no injurious action on the enzyme, which has no relation with metal ions on its formation, and then on -SH group.

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The authors experimented on the injurious action of the substances on rice-seeds' sprouting, which have analogous chemical structure with fusarinic acid and observed that the substance, which chelates with metal-ion in the molecule, showed evident activity, while the substance, which does not chelate, had no activity. The stronger the chelation is the more the activity. Fusarinic acid was proved to be deprived of a considerable amount of the injurious action. by esterifying its carboxyl group.

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One hundred g of, the extraction residue of spruce wood meal (with EtOH-C_??_H₃and hot water) was refluxed with the solution, which is composed of 1000 cc EtOH, 67.7g conc. HCl, 21g NH₂OH•HCl and 60 cc water, on the water bath during 20 hours. About 40% of lignin in the wood was removed in this process. When the filtrate and washings were poured into 5l H₂O, brownish yellow flocculent amorphous substance (α) was precipitated (5.7g). After (α) was filtered off, the filtrate was neutralized to pH 3.4, concentrated in vacuo at 55° and treated continuously with ether. Ether extraction after concentration left yellow syrup (β) (1.65g). From the determinations of OCH₃ and N contents of (α) and (β), it was concluded that (α) was lignoxime, having one N (i.e. one mole carbonyl group) vs. 3.5 lignin building stones, and (β) was low-molecular lignoxime, having one N vs. one lignin building stone.

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(1) The influences of subculture media on the survival times of spores were studied. The, spores of Bacillus natto were suspended in neutral phosphate buffer and heated at. 100°C, and. then subcultured by various media. The yeast-extract broth (10% bakers' yeast extracted cted by nutrient both, adjusted to pH 7.0). liver-extract broth (10% pork-liver extracted by nutrient broth), and glucose-broth (1% glucose added to nutrient broth), showed survival time-twice as long as plain nutrient broth. The addition of reducing agents, cysteine and thioglycollate to nutrient broth, have shown no increasing effect on the survival time. (Table 1)
(2) The effects of adding various carbohydrates and organic acids were examined. Fructose, mannose, galactose, sucrose, maltose, and soluble starch were found to have the increasing effect on the survival time of spores of Bacillus natto to the same extent as glucose, but other eight sugars, arabinose, xylose, lactose, trehalose, mannitol, glycerol glycogen, and Cori-ester have had no effects.
The addition of pyruvate or alpha-glycerophosphate to nufttent broth also increased the, survival time, but the addition of organic acids such as lactic, acetic, and succinic had noeffects.
It was found that in a synthetic medium⁽⁹⁾, which is composed of eighteen amino acids, nine vitamins, four purine-pyrimidine bases, and salts, the vegetative cellls of Bac. natto grew well without the addition of glucose, whereas the spores could not grow without glucose (carbohydrates). And it was also found that the kind of carbohydrates which were effective in increasing the survival time of the spores of Bacillus natto was the same with that which served, as the energy source and which enabled the spores to grow in the synthetic medium. (see Table 3 and 5). The minimum effective, concentration of glucose in increasing the survival time of the spores was between 10^-5M and 10^-6M, and the same order of glucose concentration also has limited the growth of spores of the strain in the synthetic medium (see Table 4 and 5).
(4) It should be noted that when subcultured by glucose broth, the survival time at each spore concentration was commonly about twice that of plain nutrient broth, throughout a wide range of spore concentration. The relation between spore concentration (N) and survival time (t) in the case of subculturing by glucose broth, can be expressed by the following general formula as reported in the previous paper⁽⁷⁾. logN=a+blogt
(5) The discussions on the mechanisms of spore survival time increasing activities of glucose were delivered.

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1. The biochemical properties of thirteen strains of the genus Pseudomonas isolated by the author were studied.
2. These strains were very weak in their oxidative abilities for ethanol.
3. These strains oxidize glucose to gluconic acid or to 2-keto-gluconic acid. In the case of shaking culture, glucose was completely oxidized in 48 to 72 hours. Five strains of them produced only gluconic acid and eight strains produced only 2-keto-glueonic acid from glucose and the Tatters oxidized gluconic acid to 2-keto-gluconic acid.
4. These strains did not oxidize sorbitol, mannitol or glycerol.
5. These strains did not oxidize maltose or lactose.

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Pharmacological study on the extrats of mycelial mass formed by the culture of Sclerotinia Libertiana was performed at Pharmacological, Institute of Medical Faculty, , Kyushu University, and it was found that uterus-contracting action of the extract against rabbit uterus, both in situ and isolated, is comparable to “gynergen”.
An antagonistic substance against adrenaline disappeared in a process of preparation of the extract.
Clinical application performed at Obstetrical Institute of Medical Faculty, Kyushu University, showed good results, with only small side reactions.

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The uterus-contracting constituent of Sclerotinia fungus extract was ascertained not to be alkaloid or biogenic amine.
Most part of the active fraction was precipitated by alcohol (60_??_95%), and the remaining liquid had a small effect.
The active fraction, which contains N and has a glycosidelike nature, lost the activity after acid hydrolysis, but it did not change after autolyzation of maceration juice.

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To get a pure culture of cellulose thermophiles, the agar plating method was examined using Na carboxylmethylcellulose, cellodextrin or cellobiose as the carbon source under a suitable, rH and an anaerobical condition with carbon dioxide or nitrogen gas. By this method nearly pure cultures of cellulose thermophiles were obtained, but these failed to digest cellulose and to recover their activity. The filtration method was also unsuccessful, because their sizes were not remarkably different and a homogeneous culture was not obtained owing to their short reproduction time.

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We have synthesized the following quaternary ammonium compounds. (1-a) dimethyl-octyl-benzyl ammonium chloride, mp. 74°, plates ; (1-b) dimethyl-dodecyl-benzyl ammonium chloride, mp. 45°, scales; (1-c) dimethyl-hexadecyl-benzyl ammonium chloride, mp. 55_??_6°, needles; (1-d) dimethyl-octadecyl-benzyl ammonium chloride, mp. 150_??_8°, scales; (2) dimethyl-dodecyl-methylthioethyl ammonium. chloride, mp. 59°, plates; (3) N, N'-bisdimethyl-N, N, N'-bisdodecyl-(N-ethyl-thio-N'-ethyl)-diammonium chloride, mp. 209°, scales; (4) dimethyldodecyl-phenylthi-oethyl ammonium chloride, mp. 82.5°, needles; (5) nicotine-N, N'-bisdodecyl chloride, mp. 99°, scales.
We have examined their germicidal activities using the spores of Gibberella Fujikurol as testing microorganism and compared with that of a 10^-3 solution of Uspulum (Uspulum contains 3% of methoxyethyl mercuric chloride and, therefore, the concentration of this Uspulum solution corresponds to a 3×10^-4 solution of methoxyethyl mercuric chloride). The figures in the following table indicate the dilution of synthesized compounds which shows the same physiological activity as that of a 10^-3 solution of Uspulum.
(1-a) 100 (1-c) 160.000 (2) 10.000 (4) 220.000
(1-b) 800.000 (1-d) 120.000 (3) 80.000 (5) 4.000

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