• Korean journal of applied entomology
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• v.13 no.3 s.20
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• pp.105-133
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• 1974

The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.

• Magazine of the Korean Society of Agricultural Engineers
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• v.15 no.3
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• pp.3059-3088
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• 1973

The Object of research was laid on the dry paddy field which had a low level of underground water, rather than on a paddy field with a high level of underground water. In the treatment of the clay paddy field before transplanting we employed 3 kinds of methods; deep plowing, development of cracks by drying the surface of the field under which pipe drain was built. This study was to find which one, among these three methods, is the most effective to let roots extend to deep zone and increase the yield of rice and at the same time, for trafficability of large scale machinery which will be introduced to the harvest, in the light of the earth bearing capacity in relation with underground drainage. In the treatments of plots, 1) the kyong plot was plowed 39 days before transplanting and dried, 2) the kyun plot was plowed again 2days before transplanting after plowing 39 days before transplanting, leveling field surface in the saturation with water and developing the cracks by drying, 3) the kyunam plot was plowed again 2 days before transplanting after setting the drainage pipe and at the same time plowing 39 days before transplanting, leveling field surface in the saturation with water and developing the cracks by drying. Also each plot above had three different levels of soil depth, respectively; that is 15cm, 25cm, 35cm. The kyong plot with 15cm-depth was he control. The results obtained were as follows; 1. The kyunam plot showed a remarkably lager amount of water consumption by better underground drainage than the kyong and the kyun plot, and the kyong plot indicated a greater amount of water consumption than the kyun plot. Therefore the amount of available rainfall was decreased in the order of kyunam>kyong>kyun. The net duty of water decreased in the order of kyunam>kyong>kyun and its showed about 105cm in depth at the kyunam plot, about 70cm in depth at the kyong plot and about 45cm in depth at kyun plot, regardless of soil depth. 2. According to the tendency that the weight of the total root was effected by the maximum depth of the crack, it seemed that the root development was more affected by the depth of the crack than by only the crack itself. The weight of the total roots tended to increase as the depth of the crack got deeper and deeper, and the weight of the total roots was increased in the order of kyun<kyunam<kyong. 3. In the growing of the plant height, the difference did not appear at the beginning of growing(peak period of tillering) of any plot, But for the mid period of growing(ending period of tillering) to the period of young panicle formation, the deeper the depth of plot is, the more the growing goes down. On the contrary at the late period of growing, growth was more vigorous in the plot with deep depth than in the plot with shallow depth. Since the midperiod of growing, in the light of experimental treatment, the kyun plot was not better in growing than the other two plots and no remarkable defference was shown between the kyunam and the kyong plot, but the kyunam plot had the tendency of superiority in growing plant height. 4. As the depth of plot went deeper, the decreasing tendency was shown in the number of tillers through a whole period of growingi. When the above results were observed concering each plot of experimental treatment, the kyun plot was always smaller in the number of tiilers than the kyunam and the kvong plot, and the kyong plot was slightly larger than the kyunam plot in the number of tillers. 5. When each plot of the different experimental treatments was compared with the control plot(15-kyong), yield(weight of grains) was increased by 17% for the 35-kyong plot, by 10% for the 35-kyunam and yields for the other plots were less or nomore than the control plot. On the whole, as the depth of plot went deeper, yields for plots was increased in the order of kyong>kyunam>kyun. 1% of significance between the levels of depths and 5% of significance between the treatments were shown. 6. The depth of consumptive water which was more effective on the weight of grains is that of the last half period. When the depth of consumptive water was increased at the range of less than 2.7cm/day in the 15cm plot, 3.0cm/day in the 25cm plot and 3.3cm/day in the 35cm plot, the weight of grains was increased, and at the same time the weight of grains was increased as the depth of plot went deeper. The deeper plots was of advantage to the productivity at the same depth of consumptive water. 7. The increase in the weight of grains in propertion to the weighte of root showed a tendency to increase depending on the depth of plot at each plot of the same weight of roots. The weight of roots and grains together increasezd in the order of kyun>kyunam>kyong, considering each treatment of experimental plot. The weight of grains was in relation to the minimum water content ratio during the midperiod of surface drainage and the average earth temperature was mainly affected by the minimum water content ratio because it was relatively increased in proportion to the water content ratio(at less than 40%) 8. The weight ratio of straw to grain showed an increasing tendency at the plot of shallow depth and had a relation of an inversely exponental function to the weight of roots. At the same depth of plot except the 15cm plot, the weight ratio of straw to grain was increased in proportion to the depth of consumptive water. The weight of grains was increased as the depth of consumptive water was increased to some extent, but at the same time the weight of ratio of straw to grain was increased. 9. At a certain texture of soils the increase in the amount of the cracks depends on meteorological conditions, especially increase in amounts of pan evaporation. So if it rains during the progressing of field drying the cracks largely decrease. The amount of cracks of clay soil had relation of inversely exponental function to the water content ratio(at more than 25%). The maximum depth of crack kept generally a constant value at less than 30% of water content ratio. 10. The cone index showed the tendency that it was propertional to the amount of cracks within a certain limit but more or less inversely proportional over a certain limit. The water content ratio at the limit may be about 25%. 11. The increase in the cone index with the progressing of time after final surface drainage showed the tendency that it was proportional to the depth of consumptive water at the last half of growing period. Based on the same depth of if the cone index in the kyunam plot was much larger than in the other two plots and that in the kyong plot was much smaller than in the kyun plott, as long as the depth of plot was deeper, especially in the 35-kyong plot. 12. In the light of a situation where water content ratio of soil decreased and the cone index increased after final surface drainage the porogress of the field dryness was much more rapid in the kyunam plot than in the kyong plot and the kyun plot, especially slowest in the kyong plot. In the plot with deeper zone the progress was much slower. The progress requiring the value of the cone index, $2.5kg/cm^2$, that working machinary can move easily on the field changed with the time of final surface drainage and the amount of rainfall, but without nay rain it required, in the kyunam plot, about 44mm in total amount of pan evaporation and more than 50mm in the other two plots. Therefore the drying in the kyunam plot was generally more rapid in the kyunam plot was generally more rapid over 2days than in the kyun plot, and especially may be more rapid over 5days than in the 35-kyong plot.