INHIBITION OF NITRIFICATION BY CLIMAX ECOSYSTEMS. III. INHIBITORS OTHER THAN TANNINS

We obtained considerable evidence in earlier work that inhibition of nitrification begins during old-field succession and increases to a maximum in the climax (Rice and Pancholy, 1972, 1973). Moreover, we found that tannins and tannin derivatives appear to be important inhibitors of nitrification. In the present project, other potential phenolic inhibitors of nitrification were identified in acetone extracts of entire plants of most herbaceous species and leaves of tree species important in an intermediate stage of succession and the climax in three vegetation types in Oklahoma. Attempts were made also to identify potential inhibitors in acetone extracts of soil from the top 15 cm of the oak-pine climax. Seventeen potential inhibitors were identified from the eleven important species of plants surveyed. These were mostly phenolic acids and flavonoids, but one coumarin compound, scopolin, was found in high amounts in several species. The potential inhibitors were most common in green tops or green leaves, but roots, dead tops (of previous year), and dead leaves had high amounts of some compounds. Caffeic and ferulic acids were prominent in dead leaves or dead tops, and one flavonoid, myricetin, occurred in sizeable amounts in dead tops of Sorghastrum nutans. The aglycones of most of the compounds were tested against nitrification in soil suspensions, and all completely inhibited oxidation of NH⁺₄ to NO⁻₂ by Nitrosomonas at concentrations as low as 10⁻⁶ to 10⁻⁸ M. Oxidation of NO⁻₂ to NO⁻₃ by Nitrobacter, however, was affected much less severely by these inhibitors. The greater resistance of Nitrobacter is not significant biologically because inhibition of the first step carried out by Nitrosomonas effectively inhibits the entire process of nitrification. The 3-glucoside of quercetin, isoquercitrin, inhibited the activity of Nitrosomonas completely at the same concentration as quercetin. We found a compound in large quantities in the oak-pine climax soil which appeared in all tests to be a flavonoid aglycone, but we were never able to identify it to our satisfaction. This substance was extremely inhibitory to germination and seedling growth of ‘Crimson Giant’ radish seeds. These have hard seed coats and germinate very rapidly so most inhibitors do not affect their germination at all. It is likely that some, if not all, of the nitrification inhibitors identified may be important in inhibition of nitrification in the later stages of succession and in the climax along with the tannins.     

INHIBITION OF NITRIFICATION BY CLIMAX ECOSYSTEMS. II. ADDITIONAL EVIDENCE AND POSSIBLE ROLE OF TANNINS

The same nine plots were used in this study as in our previous study on inhibition of nitrification (Rice and Pancholy, 1972). These consisted of three stands representing two stages of old field succession and the climax in each of three vegetation types in Oklahoma: tall grass prairie, post oak-blackjack oak forest, and oak-pine forest. Soil samples were analyzed three times during the growing season of 1972 for exchangeable ammonium nitrogen, nitrate, and numbers of Nitrosomonas and Nitrobacter. Results were similar to those obtained during the entire year of 1971. The amount of ammonium nitrogen was lowest in the first successional stage, intermediate in the intermediate successional stage, and highest in the climax. The amount of nitrate was highest in the first successional stage, intermediate in the intermediate successional stage, and lowest in the climax. The numbers of nitrifiers were highest in the first successional stage usually and decreased to a very low number in the climax. These data furnish additional evidence that the nitrifiers are inhibited in the climax so that ammonium nitrogen is not oxidized to nitrate as readily in the climax as in the successional stages. This would aid in the conservation of nitrogen and energy in the climax ecosystem. Some inhibition of nitrification occurred in the intermediate stage of succession also. Previous studies of tannins indicated that these are inhibitory to nitrification, so all important plant species in the intermediate successional stage and the climax were analyzed for total tannin content. A method for extracting and quantifying condensed tannins from soils was developed and the amounts of tannins were determined in each 15-cm level down to 60 cm in the same two plots in each vegetation type. Gallic and ellagic acids, which result from the digestion of hydrolyzable tannins in oak species, were also extracted and quantified in the climax oak-pine forest. All the important herbaceous species, including the grasses, were found to have considerable amounts of condensed tannins. The highest amounts of tannins occurred in the oaks and pine, however. Condensed tannins, hydrolyzable tannins, ellagic acid, gallic acid, digallic acid, and commercial tannic acid (hydrolyzable tannin), in very small concentrations, were all found to completely inhibit nitrification by Nitrosomonas in soil suspensions for 3 weeks, the duration of the tests. Slightly larger concentrations were required to inhibit nitrification by Nitrobacter under similar conditions. The concentrations of tannins, gallic acid, and ellagic acid found in the soil of the research plots were several times higher than the minimum concentrations necessary to completely inhibit nitrification. The inhibition of nitrification was always greater in the climax stand than in the intermediate successional stage in each vegetation type, and the concentration of tannins in the top 15 cm of soil was always higher in the climax stand than in the intermediate successional stage. Moreover, the amounts of tannins calculated to be added to each plot each year are much less than the amounts found in the soil, indicating that the tannins accumulate over a period of time. Thus, it appears that the tannins and tannin derivatives may play a continuous and rather prominent role in the inhibition of nitrification by vegetation.     

IN VITRO ALLELOPATHIC INHIBITION OF NITRIFICATION BY BALSAM POPLAR AND BALSAM FIR

A soil percolation apparatus was used to study the effect of plant extracts on nitrification. Foliar leachates, leaf extracts, and bud extracts of balsam fir (Abies balsamea) and balsam poplar (Populus balsamifera) were added to the ammonium percolation solution. Color tests and quantitative measurement of residual ammonium and formed nitrate were applied. Foliar leachates of balsam fir strongly inhibited nitrification; leachates of balsam poplar were slightly less inhibitory. Balsam fir needle and balsam poplar dormant bud extracts (5% w/v) completely prevented the oxidation of ammonium. The same estracts (2% w/v) still prevented the oxidation of ammonium to nitrate to a large extent. Once again balsam fir extracts inhibited the process more strongly than balsam poplar. No evidence was obtained for the presence of a specific inhibitor of nitrifier microorganisms.     

蛋白酶抑制剂对小鼠胚泡着床的作用

本文采用子宫内注射和胚胎与子宫上皮细胞在体外共培养的方法,研究几种蛋白酶抑制对小鼠胚胎着床的影响。实验结果显示,蛋白酶抑制剂ＳＴＩ,ＥＡＣＡ和ＮＰＧＢ均能显著抑制胚泡在子宫内着床;ＳＴＩ和ＥＡＣＡ能有效地抑制胚泡在子宫上皮细胞单层上的扩展,而ＮＰＧＢ对胚泡在子宫上皮细胞单层上的粘附和扩展均有显著的抑制作用。     

THE INHIBITION OF CYPRIDINA LUMINESCENCE BY LIGHT

The luminescence of Cypridina luciferin-luciferase solution is inhibited by illumination from a carbon arc of 15,000 foot candles in between 1 and 2 seconds. The blue to violet rays are the effective ones, the limits lying somewhere around 4,600 Å. u. to 3,800 Å. u. The luciferin, not the luciferase, is the substance affected by the light. The effect is partially reversible in the dark. The chemiluminescences obtained by oxidizing phosphorus, lophin, and chlorphenylmagnesium bromide are not inhibited by light under the above conditions.     

类胡萝卜素对亚油酸甲酯氧化的抑制作用

研究溶液中类胡萝卜素对2 ,2’ -偶氮二 (2 ,4短杠二甲基戊腈 ) (AMVN)引发的亚油酸甲酯氧化的抑制作用及色素的消耗变化。表明 β-胡萝卜素 ,叶黄素及胭脂树橙都按依赖于剂量的方式抑制亚油酸甲酯氢过氧化物的形成。41.7×10 -6mol/L的叶黄素及玉米黄质的抑制活性相近 ,高于β-胡萝卜素与胭脂树橙的活性 ,后两者活性近似。在83.3×10 -6mol/L浓度下鸡油菌黄质活性高于叶黄素 ,后者又高于 β-胡萝卜素与胭脂树橙 ,最后两者仍近似。结果表明下列五种类胡萝卜素对亚油酸甲酯氧化的抑制能力为 :鸡油菌黄质>玉米黄质≈叶黄素> β-胡萝卜素≈胭脂树橙。类胡萝卜素在抑制脂质氧化过程中 ,本身逐渐损失消耗 ,4h后在试验所用浓度下最高剩余量不超过20 %。其中鸡油菌黄质与胭脂树橙的消耗速度慢于 β-胡萝卜素与玉米黄质。     

INHIBITION BY BACTERIA OF PSEUDOPLASMODIUM FORMATION OF DICTYOSTELIUM DISCOIDEUM

A mutant of Dictyostelium discoideum was isolated, the development of which was normal at 18.5°C but stopped at the aggregate stage when cultured with bacterial associate at 26°C. Probably the bacteria release a factor inhibitory to pseudoplasmodium formation, the inactivation of the inhibitor being temperature-sensitive in this mutant. This bacterial factor is different from the one reported by W eber and R aper (16), judging from the synergism shown by these two types of mutants.
The temperature-shift experiment gave the result that the inhibited developmental step was at about the time of initiation of tip formation. However, once a tip was formed under the permissive conditions, the re-formation of a new tip after removal of the original was not inhibited under the nonpermissive conditions. These results indicate that the bacterial inhibitor acts just before tip formation.