The writer already reported that large quantity of free auxin was contained in the gall tissue of the stem and leaf of
Youngia japonica infected with
Protomyces inouyei, though the determination of the hormonal substance was not yet done. This article deals with the results of experimental studies on the production of auxin and its identification regarding 3 species of
Protomyces fungi.
Four kinds of solvents such as ether, chloroform, ethyl-alcohol and water, were used for obtaining extract from the gall formed by the infection of
Protomyces inouyei. With these extracts Went's
Avena-test was carried out. The ether solvent was found to be better for the purpose of extraction than others, as shown in Table 1.
When
Protomyces was cultured in Czapek's solution or in the same one containing 1
g of L-tryptophane in substitution for KNO
3 as a N source, the filtrate from the latter medium alone was ascertained by the
Avena-test to contain a considerable quantity of auxin.
The material fungi were cultured in Czapek's medium containing tryptophane, at 26°C. for 30 days, and their filtrates were concentrated to one half. The filtrates were then extracted with trebled volume of ether, and the dry yellowish extract was obtained from
P. pachydermus and the yellowish brown one from other two fungi. The yellowish brown extract showed typical IAA color reaction with Gordon and Weber's reagent, while the yellowish one never showed the reaction. For analysing the extract one-dimentional paperchromatography was applied under the condition as described in the foot-note of Table 4. The reddish pink spots at Rf 0.54 and 0.55 developed by isopropanol-ammonia-water solvent mixture and also the spots at 0.79 and 0.80 developed by ethanol solvent were all identified as IAA, by comparing with the Rf value shown by synthetic IAA as the control. The IAA spots appeared was found to be smaller in area than other spots.
Figure 1 shows the data of analysis of these extracts by the paperchromatographic bioassay method. The results also clearly show the presence of IAA similarly to the experimental data mentioned above.
It seems to the writer that
Protomyces may produce tryptamine and indoleacetaldehyde as the intermediates in the metabolic process of tryptophane, because some similar spots to the very intermediates in respect of color reaction and Rf value were shown in the paperchromatogram.
Protomyces fungi were proved to have lower activity in IAA metabolic production from tryptophane than certain various fungi belonging to
Ustilago, Taphrina, Exobasidium, Fusarium etc. as far as the writer's experiments concerned.
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