Sulphur isotope characteristics of two north Bohemian forest catchments

Sulphate concentrations and ³⁴S ratios were monitored in bulk precipitation, spruce throughfall, and soil water (depth of 30 and 90 cm) at ervená jáma (CER) and Naetín (NAC), two severely polluted sites in the Czech Republic, between December 1992 and September 1994. Throughfall [SO₄ ^2–], up to 80 mg L^–1 in winter and as low as 7 mg L^–1 in summer, was higher than [SO₄ ^2–] in bulk precipitation (annual average 6 mg L^–1). There was a distinct seasonaity in S isotope abundances, with lower ³⁴S_BULK in summer (+4 per mil CER, + 6 per mil NAC) and lower ³⁴S_TF in winter (+3 per mil CER, +4 per mil NAC). Wintertime ³⁴S_BULK was around +8 per mil at CER and +10 per mil at NAC, summertime ³⁴S_TF was close to +7 per mil at both sites. For only a 1- month period in spring, bulk precipitation S became isotopically lighter than throughfall S. Bulk precipitation data from CER were in good agreement with those from the nearby monitoring station Lesná (LES), typically differing by less than 10 mg L^–1 and 2 per mil in [SO₄ ^2–] and ³⁴S, respectively. Suction lysimeters (soil depth of 30 and 90 cm) yielded higher sulphate concentrations and lower ³⁴S ratios compared to both bulk and throughfall precipitation. Little seasonality was observed in [SO₄ ^2–] at 30 cm (around 40 mg L^–1); at 90 cm [SO₄ ^2–] was higher in winter (70 mg L^–1) than in summer (45 mg L^–1). ³⁴S at 90 cm was <+5 per=" mil=" in=" 1993=" and=" up=" to=" +7.5=" in=" 1994,=" lower=" in=" the=" first=" year=" and=" higher=" in=" the=" second=" year=" compared=" to=" the=" depth=" of=" 30=" cm.=" sulphur=" fluxes=" at=" cer=" and=" nac=" are=" characterized=" by=" distinct=" isotope=" compositions=" and=" can=" therefore=" be=" used=" to=" trace=" s=" pathways=" and=" transformations=" in=" the=" forest=">     

Malting enzyme activities, grain protein variation and yield potentials in the displaced genetic resources of barley landraces of Finland

Thirty-two unimproved landraces of barley ( Hordeum vulgare L.) based on single plant selections in SE Finland were studied. Some of the lines still had ancient features, e.g., rachis brittleness in the tip of the spike. The lines showed 17 different and 16 unique hordein patterns and three lines showed more than one pattern, suggesting heterozygosity, and six different residual grain protein patterns. The three enzyme activities (-amylase, -amylase and -glucanase) of grains germinated aseptically for 120 h were determined. The average activity levels were high compared with a standard of five global barleys and with those determined previously in wild barley (H. vulgare ssp. spontaneum (Koch) Archers et Graebn.) grown in Finland. The ssp. spontaneum sample of 257 accessions showed significantly (P<0.001) less variation in -amylase and significantly (P<0.001) lower mean activity of all three enzymes. The high variation of these chemotypes indicates great potential variation of possible use by breeders has been lost by the disappearance and displacement of local barleys with commercial cultivars since 1950 in Finland before which barley cultivation and adaptation to the local environment had occurred over more than 3000 years. Selection for currently preferable plant characteristics in the descendants of the cross of HA52 (a landrace selection) × Adorra discriminated the hordein pattern of HA52 not being directly selected. The best landraces outyield the standard cultivars especially when there was no lodging. Top yield and small grains appeared to be associated characteristics under the environmental selection pressure, conflicting with the man-made regulations of the EU.     

The use of nitrogen-15 natural abundance and nitrogen yield of non-nodulating isolines to estimate nitrogen fixation by soybeans (Glycine max L.) across three elevations

Summary Dissimilarities in soil N uptake between N₂-fixing and reference non-N₂-fixing plants can lead to inaccurate N₂ fixation estimates by N difference and ¹⁵N enrichment methods. The natural ¹⁵N abundance ( ¹⁵N) method relies on a stabilized soil ¹⁵N pool and may provide reliable estimates of N₂ fixation. Estimates based on the ¹⁵N and differences in N yield of nodulating and non-nodulating isolines of soybean were compared in this study. Five soybeans from maturity groups 00, IV, VI, and VIII and their respective non-nodulating isolines were grown at three elevations differing in ambient temperature and soil N availability. Despite large differences in phenological development and N yield between the non-nodulating isolines, the ¹⁵N values measured on seeds were relatively constant within a site. The ¹⁵N method consistently produced lower N₂ fixation estimates than the N difference method, but only in three of the 15 observations did they differ significantly. The average crop N derived from N₂ fixation across sites and maturity groups was 81% by N difference compared to 71% by ¹⁵N. The magnitude of difference between the two methods increased with increasing proportions of N derived from N₂ fixation. These differences between the two methods were not related to differences in total N across sites or genotypes. The low N₂ fixation estimates based on ¹⁵N might indicate that the nodulating isolines had assimilated more soil N than the non-nodulating ones. A lower variance indicated that the estimates by N difference using non-nodulating isolines were more precise than those by ¹⁵N. Since the differences between the estimates were large only at high N₂ fixation levels (low soil N availability), either method may be used in most situations when a non-nodulating isoline is used as the reference plant. The ¹⁵N method may have a comparative advantage over N difference and ¹⁵N enrichment methods in the absence of a suitable non-N₂-fixing reference plant such as a non-nodulating isoline.     

Abiotic controls on the functional structure of soil food webs

Summary The hypothesis that the trophic structure of soil food webs changes as a result of the abiotic environment was examined by reviewing studies of soil biota. In dry soils with a water potential below –1.5 MPa, most bacteria, protozoans, and many species of nematodes are not active. These taxa persist in the soil in a state of anhydrobiosis. Because soil fungi grow at soil water potentials of –6.0 to –8.0 MPa, soil food webs in dry environments appear to be fungal-based and fungal grazers in dry environments appear to be predominantly fungiphagous mites. There is indirect evidence that some species of fungiphagous mites remain inactive in dry soils in a state of cryptobiosis. In habitats where there is insufficient vegetative cover to shade and modify the soil surface, the functional soil food web consists of fungi and a few taxa of soil acari for extended periods of time.     

Variation and genomic polymorphism of lectin-related proteins in Lima bean (Phaseolus lunatus L.) seeds

Variation of the lectin and the two lectin-related proteins, AIL (-amylase inhibitor-like) and ARL (arcelin-like) was examined in wild and cultivated accessions of Lima bean (Phaseolus lunatus L.) using electrophoresis of total seed proteins, immunoblot and RFLP analysis of lectin-related genes. Results confirm that divergence of the two major Lima bean gene pools, Andean and Mesoamerican, also apply to this protein family. All three members of the family are present in both gene pools, with differences in size, abundance and composition between gene pools, giving the possibility to distinguish Andean from Mesoamerican lectin pattern types. Both patterns show some variants, such as lack of lectin or its presence as an abundant protein. The observed variation reflects, at least in part, into genomic polymorphism. The presence of arcelin- and -amylase-related proteins in Lima bean could represent a tool to increase our knowledge in the evolution of the lectin family in Phaseolus species.     

Gymnanthemum coloratum (Willd.) H. Rob. et B. Kahn (Compositae-Vernonieae): the correct name for a cultivated bitterleaf of Africa

Gymnanthemum coloratum (Willd.) H. Rob. et B. Kahn is shown to be the correct name for the species to which the name G. chapelieri (Drake) H Rob. was erroneously applied in the new Mansfeld     

Activity,origins and location of cellulases in a silt loam soil

Summary Cellulase activity in a silt loam soil was assayed and characterised using a microcrystalline cellulose substrate (Avicel). Activity was maximal between pH 5.3 and pH 6.0. A 64% loss in activity was observed on air-drying the soil. However, the residual activity was stable to storage at 40°C for 7 days and was resistant to the action of added protease. The component endoglucanase and -D-glucosidase activities in field-moist and air-dried soil were also assayed. The proportion of the soil microbial population able to utilise cellulose was investigated and the persistence of two free (soluble) cellulase preparations of microbial origin was determined following their addition to soil. A rapid decline in the endoglucanase activity of a Streptomyces sp. cellulase preparation was observed while 30% of the original activity of a Trichoderma viride cellulase preparation could still be detected after 20 days. From the data obtained in this study it appears that the major portion of the -D-glucosidase activity is bound to and protected by the soil colloids. By contrast, the major portion of the exo- and endoglucanase activity appears to be free in the soil solution, attached to the outer surfaces of cellulolytic microorganisms or associated in enzyme substrate complexes. The low residual activity measured in air-dried soil may owe its stability to an association with soil colloids or with recalcitrant cellulosic material present in soil.     

Extraction of humic-β-glucosidase fractions from soil

Humic compounds with -glucosidase activity were extracted from soil using tetrasodium pyrophosphate as extracting solution. Mixture of soil samples with 0.01 M pyrophosphate in a ratio of 1:5 (w/v), adjustment to pH 7.0–7.3, extraction for 18 h with reciprocating agitation, and bacteriological filtration after centrifugation were the optimum conditions for extraction of the -glucosidase complexes. Otherwise, experimental conditions for extraction indicated that the concentration and pH of pyrophosphate were the factors with the most influence on enzymatic extraction yields. The results indicated that the -glucosidase was extracellular and associated with soil humates.     

Differentiation and geographical distribution of β-amylase isozyme in barley

This study reports on the investigation of the polymorphism and geographical distribution of -amylase isozymes by isoelectric-focusing (IEF) analysis in samples of world barley. The isozyme pattern of high thermostability type A and low thermostability type C varieties was restricted in isozyme type II. However, the isozyme pattern of the middle thermostability type B varieties was polymorphic. The -amylase isozyme of 1,554 lines of the middle thermostability group was investigated next. Besides types I and II, new isozyme types III and IV were observed. Most lines were grouped into type II in all growing areas, except for Europe where type I and type II were even in number. Types III and IV were observed in only one line in India and Afghanistan, respectively. In Northern Europe, especially in Denmark, type I was major. It was concluded that a mutant IEF isozyme type I originated from the type II prototype of the thermostability B group in Northern Europe.     

Nitrite production by Nitrosomonas europaea and Nitrosospira sp. AV in soils at different solution concentrations of ammonium

Although it remains unclear why NH₃-oxidizing bacteria (AOB) of the genus Nitrosospira dominate soil environments, and why Nitrosomonas spp. are less common, virtually no studies have compared their behavior in soil. In this study, the NH₃ oxidation rates of Nitrosomonas europaea (ATCC 19718) and Nitrosospira sp. AV were compared in three differently textured soils containing a range of extractable contents (2-11 μg soil). Soils were adjusted to pH 7.0-7.4 with CaCO₃ and sterilized with γ-radiation. Cell suspensions of each bacterium were inoculated into the soils to bring them to two-third of water-holding capacity and cell densities ∼2.5×10⁶ g⁻¹ soil. In virtually all cases, rates of production for both N. europaea and Nitrosospira sp. AV were linear over 48 h, and represented between 13 and 75%, respectively, of the maximum rates achieved in soil-free bacterial suspensions. Soil solution concentrations that supported these rates ranged between 0.2 and 1.5 mM. Addition of 21-36 μg soil raised soil solution levels to 1.8-2.5 mM and stimulated production to a greater extent in N. europaea (3.3-6.6-fold) than in Nitrosospira sp. AV (1-2.1-fold). Maximum rates of production were obtained by raising soil solution levels to 3-4 mM with a supplement of ∼80-90 μg soil. K_s values in soil for Nitrosospira sp. AV and N. europaea were estimated as 0.14 and 1.9 mM , respectively, and estimates of V_max were about 3.5-times higher for N. europaea (0.007 pmol h⁻¹ cell⁻¹) than for Nitrosospira sp. AV (0.002 pmol h⁻¹ cell⁻¹). The cell density of N. europaea increased in sterile Steiwer soil independent of supplemental . In the case of treatments receiving supplemental , growth yields of N. europaea calculated from either produced or consumed were similar to those reported in literature (3.5×10⁶-6×10⁶ cells μmol⁻¹). A higher growth yield was measured in the case of zero added (2.7×10⁷ cells μmol⁻¹), indicating that use of organic carbon compounds might have occurred and resulted in some energy sparing. Our results suggest that Nitrosospira spp. with a K_s similar to Nitrosospira sp. AV may have an advantage for survival in soil environments where soil solution levels are less than 1 mM. However, it is apparent that AOB like N. europaea are poised to take advantages of modest increases in extractable that raise soil solution levels to about 2.0-2.5 mM.     

Reducing NH3, N2O and {\text{NO}}_3^ - –N losses from a pasture soil with urease or nitrification inhibitors and elemental S-amended nitrogenous fertilizers

A 3-month field experiment comparing nitrogen (N) losses from and the agronomic efficiency of various N fertilizers was conducted on a sandy loam (Typic Hapludand) soil at Ruakura AgResearch farm, Hamilton, New Zealand during October to December 2003. Three replicates of seven treatments: urea, urea + the urease inhibitor N-(n-butyl) thiophosphoric triamide (trade name Agrotain), urea + Agrotain + elemental sulphur (S), urea + double inhibitor [DI; i.e., Agrotain + dicyandiamide (DCD)], diammonium phosphate (DAP), DAP + S, each applied at 150 kg N ha⁻¹, and control (no N). After fertilizer application, soil ammonium () and nitrate () concentrations (7.5-cm soil depth), ammonia (NH₃) volatilization, nitrate () leaching, nitrous oxide (N₂O) emission, pasture dry matter, and N uptake were monitored at different timings. Urea applied with Agrotain or Agrotain + S delayed urea hydrolysis and released soil at a slower rate than urea alone or urea + DI. Urea applied with DI increased NH₃ volatilization by 29% over urea alone, while urea + Agrotain and urea + Agrotain + S reduced NH₃ volatilization by 45 and 48%, respectively. Ammonia volatilization losses from DAP were lower than those from urea with or without inhibitors. Total reduction in leaching losses for urea + DI and urea + Agrotain compared to urea alone were 89% and 47%, respectively. Application of S with urea + Agrotain reduced leaching losses by an additional 6%. Nitrous oxide emissions were higher from the DAP and urea alone treatments. Urea applied with DI and urea + Agrotain reduced N₂O emissions by 37 and 5%, respectively, over urea alone. Compared to urea alone, total pasture production increased by 20, 17, and 15% for urea + Agrotain + S, urea + Agrotain, and urea + DI treatments, respectively, representing 86, 71, and 64% increases in N response efficiency. Total N uptake in urea + Agrotain, urea + Agrotain + S, and urea + DI increased by 29, 22, and 20%, respectively, compared to urea alone. These results suggest that the combination of both urease and nitrification inhibitors may have the most potential to reduce N losses and improve pasture production in intensively grazed systems.     

Zymogram patterns of α-amylase isozymes inEthiopian tetraploid wheat landraces: insight into their evolutionaryhistory and evidence for gene flow

The variation for -amylaseisozymes was analyzed in 71 tetraploid wheat (AABB genome)landraces from Ethiopia, including accessions of Triticumdicoccon Shrank, T.turgidum L., T.durum Desf., T.pyramidale Percival and T.aethiopicum Jakubz., by thin-layerpolyacrylamide gel isoelectric focusing. Four zymogram phenotypeswere obtained from all the materials studied. Except inT. dicoccon, however, thestandard pattern of the durum wheat variety, LD222, was predominant.T. dicoccon showed twozymogram types that differed for the absence or consistently weakactivity of band 18(-Amy-B1)of the malt type. Band 1(-Amy-B3)of the malt type was fixed in T.dicoccon and present in only 10% of thefree-threshing (FT) types. Generally, the resultsindicated that the variation for-amylase isozymes in cultivatedtetraploid wheats, including landraces from secondary centers, is lowpossibly due to the founder effect or as a result of selection. Thedata were useful for inferences about the evolutionary history ofEthiopian wheat landraces. It is speculated thatT. dicoccon was the firstwheat to arrive in the Ethiopian highlands ca.5000 years ago. However, it is not known whether the present day FTEthiopian tetraploid wheat landraces are direct descendants ofT. dicoccon, or whetherthey were introduced independently. The clear differences in-amylase zymogram patterns favor the latterhypothesis. However, the presence of band1 in some of the FT types indicated the occurrence of geneflow between the FT types and T.dicoccon, which may also explain thequantitative nature of spike threshability in these landraces.Possible implications for durum wheat breeding arediscussed.     

Distribution of fertiliser N among fixed ammonium fractions as affected by moisture and fertiliser source and rate

The relevance of fixed in certain soils and its categorisation has made it necessary to re-examine N behaviour. A replicate factorial experiment was designed to investigate the influence of soil type, soil moisture and fertiliser source and rate on fixation dynamics with particular attention to the distribution between weakly and strongly fixed pools. Fixation of was <20% of added N for all soils except River Estate. The percentage of added N present as fixed was greater for the low application rate. Soil moisture did not significantly influence weakly fixed . However, the dry soil treatment showed greater fertiliser ¹⁵N present as strongly fixed . Fertiliser ¹⁵N present as weakly and strongly fixed decreased and increased, respectively, at the second sampling, indicating movement between the pools. The importance of the weakly fixed fraction as a transitory pool between strongly fixed and available was observed.     

Nitrate uptake by Eriophorum vaginatum controls N2O production in a restored peatland

The clear dependence of N₂O production through denitrification on available nitrate in soil has been shown in many studies. Since N availability similarly limits the growth of plants, the resource competition with vegetation limits the activity of denitrifying microbes and may consequently moderate the N₂O emissions from peatlands. We used uptake by Eriophorum vaginatum L. as a vegetation competition factor for microbes. The species was selected for the experiment because it has high nutrient use efficiency in low-nutrient conditions and high nutrient uptake efficiency in luxuriant nutrient conditions. We measured gaseous N flux as N₂O (end product of denitrifier activity) in a restored peatland in central Finland with acetylene inhibition technique over a growing season from sample plots with varying addition levels and E. vaginatum cover. The resource competition effects were analysed with a model that used exponential decay dependence of N₂O flux on the leaf area of E. vaginatum, and saturating response of N₂O flux to addition level. The model explained the variation in N₂O fluxes well (R²=0.86). The model simulation showed that the increasing nutrient uptake of E. vaginatum decreased the N₂O fluxes exponentially. Simultaneously, denitrification appeared to saturate even in conditions with high availability of and low level of competition by vegetation. Thus, E. vaginatum is an effective competitor for in sedge-dominated peatlands that controls the availability of for denitrification, and consequently moderates the N₂O emissions from peatlands.     

Prolonged elevated atmospheric CO2 does not affect decomposition of plant material

Prolonged elevated atmospheric CO₂ might alter decomposition. In a 90-day incubation study, we determined the long-term (9 years) impact of elevated CO₂ on N mineralization of Lolium perenne and Trifolium repens plant material grown at ambient and elevated CO₂ and low- and high-¹⁵N fertilizer additions. No significant differences were observed in recovery rates between any of the treatments, except an N addition effect was observed for L. perenne (0.4 versus 0.5% day⁻¹ in high versus low N). The results suggest that elevated CO₂ did not change plant N mineralization in any of the soils, because of a surplus of available N in the fertilized and leguminous systems, and because of insignificant plant responses to elevated CO₂ in the low soil N availability systems.     

Characterisation of white clover populations collected from the Caucasus and high altitude regions of eastern Turkey

White clover populations collected from the Caucasus region and eastern Turkey were compared with seven white clover cultivars in a moist, lowland, highly fertile environment. The Caucasus collection came from 15 sites below 1000 m and 19 sites above 1000 m; all seven sites in Turkey were above 1000 m. The populations were generally small leaved and except for those from low altitude sites in Dagestan and Azerbaijan populations were acyanogenic. None of the collected populations were significantly higher yielding than Grassland Huia in any season. However, several populations from the Caucasus were higher yielding than Kent Wild White and similar to that of Grassland Tahora. Survival was poor with only four populations having more than 50% of plants surviving after 18 months in this environment. High altitude Caucasus populations were earlier flowering than low altitude populations, an adaptation to allow adequate seed set.There was a negative relationship between altitude of population origin and proportion of cyanogenic plants; and between altitude and both leaf size and canopy height. High incidence of no leaf mark (vv) was associated with populations from colder rather than warmer environments. Populations from sites with low rainfall were larger leaved and more upright than populations from wetter sites. Populations from severely grazed sites were less upright and less cyanogenic than populations from infrequently or nil defoliated sites.Deceased     

Forest floor gross and net nitrogen mineralization in three forest types in Quebec, Canada

The effect of high nitrogen (N) depositions on forest ecosystems is an important concern in North America and may lead to N saturation of forest ecosystems and contribute to soils and surface water acidification. In this study, nitrogen dynamics in the FH layers of a sugar maple (SM), a balsam fir (BF) and a black spruce (BS) forest was characterized using a short term ¹⁵N isotopic pool dilutions approach and mid-term FH material incubation both in situ and in the laboratory. The short term dilutions approach indicated that the mean residence times of and in the FH material of the three sites were low (<1 d). The amount of inorganic nitrogen () recycled annually within the exchangeable forest floor reservoir was between one and two orders of magnitude larger than the annual atmospheric N deposition found at each of the sites. The BS site was clearly distinct than the two other forest types in that net N mineralization was negligible, even in absence of root uptake, suggesting that soil microorganisms were severely N limited. While net nitrification was not observed within the FH material of the BF site, did accumulate in the FH of the SM despite a low pH of 3.72 presumably because of heterotrophic nitrification or as a result of acid-tolerant autotrophic nitrification. The difference in N dynamics between the sites were most probably caused by dominant tree species. Transformation rates of inorganic N were higher in SM, followed by BF and BS stands. Given that the potential to mineralize inorganic N matches with a superimposed N atmospheric deposition gradient in Québec, the sugar maple forest is more likely to be affected by N saturation than coniferous forests.     

Agricultural potential of Mediterranean grain and forage legumes: 2) Anti-nutritional factor concentrations in the genus Vicia

As part of an investigation into the agricultural potential of under-exploited Mediterranean Viceae, anti-nutritional factor concentrations were measured in the seed of 7 species. These included: V. sativa(subsp. amphicarpa(L.) Batt., subsp. macrocarpa(Moris) Arcang, subsp. nigra(L.) Ehrh., subsp. sativa), V. ervilia L., V. faba L. (var. faba, var. minor Peterm.), V. narbonensis L. (var. aegyptiaca Kornhuber ex Asch. et Schweinf., var. affinis Kornhuber ex Asch. et Schweinf., var. jordanica H. Schaef., var. narbonensis, var. salmonea(Mout.) H. Schaef.), and relatives, including V. johannis Tamamsch., (var. johannis, var. procumbens H. Schaef.), V. hyaeniscyamus Mout., and V. serratifolia Jacq. Both quantitative and qualitative differences in seed chemistry were observed. Condensed tannins, phenolics and proteinase inhibitors were found in varying concentrations in all taxa. The highest concentrations of phenolic secondary metabolites (0.8–1.4% leucocyanidin equivalents, 9.5–16.4 mg/g phenolics) were found in V. faba, the V. narbonensis relatives and V. sativa subsp. macrocarpa. These taxa also tended to contain low levels of proteinase inhibitors (0.25-0.55 mg trypsin/mg protein), whereas the opposite was the case for V. ervilia(2.1 mg t/mg p). V. narbonensis was intermediate both in terms of tannin and proteinase inhibitor levels. In contrast, the non-protein amino acids tended to be more specific in their distribution. Concentrations of canavanine varied from 0.04-0.11% in the seed of V. ervilia, and may constrain the end-use of the grain, given that pigs are sensitive to concentrations as low as 0.08% in their diet. In V. sativa variation of -glutamyl--cyanoalanine within and between subspecies (0.41–1.36%) is not sufficient to make the grain palatable to monogastrics, since > 0.075% inclusion in the diet causes mortality in chicks. In V. narbonensis, there was no significant variation between most varieties for -glutamyl-S-ethenyl cysteine (GEC), a non-protein amino acid peptide which reduces grain palatability. However, lower concentrations were found in V. n. var. jordanica and its near relatives, V. johannis, V. hyaeniscyamus and V. serratifolia(1.3-1.6%, compared with 1.9+/–0.02% in all remaining V. narbonesis sub-species). Anti-nutritional factor concentrations were used to visualize taxonomic relationships using hierarchical cluster analysis. The V. narbonensis varieties were closely aligned with their near relatives, V. johannis, V. hyaeniscyamus and V. serratifolia. In contrast V. faba was positioned closer to the V. sativa subspecies than either to V. narbonesis or its near relatives. V. ervilia was the most distantly related species. Across all taxa proteinase inhibitors and condensed tannins or total phenols were negatively correlated (r=– 0.72, P < 0.009). This may be a mechanism which minimizes wasteful resource allocation to chemical defense, because the 2 anti-nutritional factors are functionally substitutable, since they both reduce the availability of protein in plant material. Thus taxa investing in high concentrations of proteinase inhibitors do not duplicate chemical defense mechanisms by simultaneously producing large amounts of tannins and phenolics.