• 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.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.26 no.1
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• pp.1-31
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• 1981

Experiments were conducted to investigate rice varietal response to low water and air temperatures at different growth stages from 1975 to 1980 in a phytotron in Suweon and in a cold water nursery in Chooncheon. Germination ability, seedling growth, sterility of laspikelets, panicle exertion, discoloration of leaves, and delay of heading of recently developed indica/japonica cross(I/J), japonica, and indica varieties at low air temperature or cold water were compared to those at normal temperature or natural conditions. The results are summarized as follows: 1. Practically acceptable germination rate of 70% was obtained in 10 days after initiation of germination test at 15\circ_C for japonica varieties, but 15 days for IxJ varieties. Varietal differences in germination ability at suboptimal temperature was greatest at 16\circ_C for 6 days. 2. Cold injury of rice seedlings was most severe at the 3.0-and 3.5-leaf stage and it was reduced as growth stage advanced. A significant positive correlation was observed between cold injury at 3-leaf stage and 6-leaf stage. 3. At day/night temperatures of 15/10\circ_C seedlings of both japonica and I/J varieties were dead in 42 days. At 20/15\circ_C japonica varieties produced tillers actively, but tillering of I/J varieties was retarded a little. At 25/15\circ_C, both japonica and I/J varieties produced tillers most actively. Increase in plant height was proportional to the increase in all varieties. 4. In I/J varieties the number of differentiated panicle rachis branches and spikelets was reduced at a day-night temperature of 20-15\circ_C compared to 25-20 or 30-25\circ_C, but not in japonica varieties although panicle exertion was retarded at 20-15\circ_C. The number of spikelets was not correlated with the number of primary rachis branches, but positively correlated with that of secondary rachis branches. 5. Heading of rice varieties treated with 15\circ_C air temperature at meiotic stage was delayed compared to that at tillering stage by 1-3 days and heading was delayed as duration of low temperature treatment increased. 6. At cold water treatment of 17\circ_C from tillering to heading stage, heading of japonica, I/J, and cold tolerant indica varieties was delayed 2-6, 3-9, and 4-5 days, respectively, Growth stage sensitive to delay of heading delay at water treatment were tillering stage, meiotic stage, and booting tage in that order, delay of heading was greater in indica corssed japonica(Suweon 264), japonica(Suweon 235), and cold tolerant indica(Lengkwang) varieties in that order. Delay of heading due to cold water treatment was positively correlated with culm length reduction and spikelet sterility. 7. Elongation of culms and exertion of panicles of rice varieties treated with low air temperature 17\circ_C. Culm length reduction rate of tall varieties was lower than that of short statured varieties at low temperature. Panicle exertion was most severaly retarded with low temperature treatment at heading stage. Generally, retardation of panicle exertion of 1/1 varieties was more severe than that of japonica varieties at low temperature. There was a positive correlation between panicle exertion and culm length at low temperature. 8. The number of panicles was increased with cold water treatment at tillering stage, but reduced at meiotic stage. As time of cold water treatment was conducted at earlier growth stage, culm length was shorter and panicle exertion poorer. 9. Sterility of all rice varieties was negligible at 17\circ_C for three days but 30.3-85.2% of strility was observed for nine-day treatment at 17\circ_C. Among the tested varieties, sterility of Suweon 264 and Milyang 42 was highest and that of Suweon 290 and Suweon 287 was lowest. The most sensitive growth stage to low temperature induced sterility was from 15 to 5 days before heading. There was positive correlation between sterility of rice plants treated with low temperature at meiotic and heading stage. 10. Percentage of spikelet sterility was greatest at cold water treatment at meiotic stage (auricle distance -15～-10cm) and it was higher in 1/1 (Suweon 264, Joseng tongil), japonica (Nongbaek, Towada), and cold tolerance indica(Lengkwang) varieties in the order. Level of cold water and position of young-ear affected on the sterility of varieties at meiotic stage; percentage of spikelet sterility of variety, Lengkwang, of which young-ear was located above the cold water level was high, but that of short statured variety, Suweon 264, of which young-ear was located in the cold water was lower. 11. Percentage of ripened grains was not reducted at 15\circ_C air temperature for three days at full heading stage in all varieties. However, at six-day low temperature treatment Suweon 287, Suweon 264 showed percentage of ripended grains lower than 60%, but at nine-day low temperature treatment all varieties showed percentage of ripened grains lower than 60%. Low temperature treatment of 17\circ_C from 10 days after heading for 20 days did not affect on the ripening of all varieties. 12. Uptake of nitrogen, phosphorous, potassium, calcium, and magnesium in whole plants was higher at average air temperature of 25\circ_C, but concentration of the elements was lower compared to those at 19\circ_C. However, both total uptake and concentration of manganese were higher at 19\circ_C compared to 25\circ_C. 13. Higher application of nitrogen, phosphorus, silicate, and compost increased yield of rice due to increased number of panicles and spike let fertility in cold water irrigated paddy.