• 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.11 no.4
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• pp.1775-1782
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• 1969

The results of the study on the consumptine use of irrigated water in paddy fields during the growing season of rice plants are summarized as follows. 1. Transpiration and evaporation from water surface. 1) Amount of transpiration of rice plant increases gradually after transplantation and suddenly increases in the head swelling period and reaches the peak between the end of the head swelling poriod and early period of heading and flowering. (the sixth period for early maturing variety, the seventh period for medium or late maturing varieties), then it decreases gradually after that, for early, medium and late maturing varieties. 2) In the transpiration of rice plants there is hardly any difference among varieties up to the fifth period, but the early maturing variety is the most vigorous in the sixth period, and the late maturing variety is more vigorous than others continuously after the seventh period. 3) The amount of transpiration of the sixth period for early maturing variety of the seventh period for medium and late maturing variety in which transpiration is the most vigorous, is 15% or 16% of the total amount of transpiration through all periods. 4) Transpiration of rice plants must be determined by using transpiration intensity as the standard coefficient of computation of amount of transpiration, because it originates in the physiological action.(Table 7) 5) Transpiration ratio of rice plants is approximately 450 to 480 6) Equations which are able to compute amount of transpiration of each variety up th the heading-flowering peried, in which the amount of transpiration of rice plants is the maximum in this study are as follows: Early maturing variety ; Y=0.658+1.088X Medium maturing variety ; Y=0.780+1.050X Late maturing variety ; Y=0.646+1.091X Y=amount of transpiration ; X=number of period. 7) As we know from figure 1 and 2, correlation between the amount evaporation from water surface in paddy fields and amount of transpiration shows high negative. 8) It is possible to calculate the amount of evaporation from the water surface in the paddy field for varieties used in this study on the base of ratio of it to amount of evaporation by atmometer(Table 11) and Table 10. Also the amount of evaporation from the water surface in the paddy field is to be computed by the following equations until the period in which it is the minimum quantity the sixth period for early maturing variety and the seventh period for medium or late maturing varieties. Early maturing variety ; Y=4.67-0.58X Medium maturing variety ; Y=4.70-0.59X Late maturing variety ; Y=4.71-0.59X Y=amount of evaporation from water surface in the paddy field X=number of period. 9) Changes in the amount of evapo-transpiration of each growing period have the same tendency as transpiration, and the maximum quantity of early maturing variety is in the sixth period and medium or late maturing varieties are in the seventh period. 10) The amount of evapo-transpiration can be calculated on the base of the evapo-transpiration intensity (Table 14) and Tablet 12, for varieties used in this study. Also, it is possible to compute it according to the following equations with in the period of maximum quantity. Early maturing variety ; Y=5.36+0.503X Medium maturing variety ; Y=5.41+0.456X Late maturing variety ; Y=5.80+0.494X Y=amount of evapo-transpiration. X=number of period. 11) Ratios of the total amount of evapo-transpiration to the total amount of evaporation by atmometer through all growing periods, are 1.23 for early maturing variety, 1.25 for medium maturing variety, 1.27 for late maturing variety, respectively. 12) Only air temperature shows high correlation in relation between amount of evapo-transpiration and climatic conditions from the viewpoint of Korean climatic conditions through all growing periods of rice plants. 2. Amount of percolation 1) The amount of percolation for computation of planning water requirment ought to depend on water holding dates. 3. Available rainfall 1) The available rainfall and its coefficient of each period during the growing season of paddy fields are shown in Table 8. 2) The ratio (available coefficient) of available rainfall to the amount of rainfall during the growing season of paddy fields seems to be from 65% to 75% as the standard in Korea. 3) Available rainfall during the growing season of paddy fields in the common year is estimated to be about 550 millimeters. 4. Effects to be influenced upon percolation by transpiration of rice plants. 1) The stronger absorbtive action is, the more the amount of percolation decreases, because absorbtive action of rice plant roots influence upon percolation(Table 21, Table 22) 2) In case of planting of rice plants, there are several entirely different changes in the amount of percolation in the forenoon, at night and in the afternoon during the growing season, that is, is the morning and at night, the amount of percolation increases gradually after transplantation to the peak in the end of July or the early part of August (wast or soil temperature is the highest), and it decreases gradually after that, neverthless, in the afternoon, it decreases gradually after transplantation to be at the minimum in the middle of August, and it increases gradually after that. 3) In spite of the increasing amount of transpiration, the amount of daytime percolation decreases gadually after transplantation and appears to suddenly decrease about head swelling dates or heading-flowering period, but it begins to increase suddenly at the end of August again. 4) Changs of amount of percolation during all growing periods show some variable phenomena, that is, amount of percolation decreases after the end of July, and it increases in end August again, also it decreases after that once more. This phenomena may be influenced complexly from water or soil temperature(night time and forenoon) as absorbtive action of rice plant roots. 5) Correlation between the amount of daytime percolation and the amount of transpiration shows high negative, amount of night percolation is influenced by water or soil temperature, but there is little no influence by transpiration. It is estimated that the amount of a daily percolation is more influenced by of other causes than transpiration. 6) Correlation between the amount of night percoe, lation and water or soil temp tureshows high positive, but there is not any correlation between the amount of forenoon percolation or afternoon percolation and water of soil temperature. 7) There is high positive correlation which is r=+0.8382 between the amount of daily percolation of planting pot of rice plant and amount and amount of daily percolation of non-planting pot. 8) The total amount of percolation through all growin. periods of rice plants may be influenced more from specific permeability of soil, water of soil temperature, and otheres than transpiration of rice plants.