Tolerance of bitter compounds by an herbivore,Cavia porcellus

Many plant defensive chemicals are bitter to humans. Because of this taste characteristic, and because bitter compounds are often toxic, such substances, and the plants that contain them, are regarded as generally unpalatable to wildlife. These assumptions may be unwarranted. To test the hypothesis that herbivores are indifferent to bitter tastants, we investigated the responsiveness of guinea pigs (Cavia porcellus) to denatonium benzoate, denatonium saccharide, limonene,l-phenylalanine, naringin, quebracho, quinine, Ro-Pel (a commercial animal repellent containing denatonium saccharide) and sucrose octaacetate. Only quinine and sucrose octaacetate slightly but significantly reduced feeding (P<0.05). Our findings are inconsistent with the notion that herbivores generally avoid what humans describe as bitter tastes.Post doctoral fellow at the Monell Chemical Senses Center at the time of the experiment.     

Gustatory Responsiveness to Polycose in Four Species of Nonhuman Primates

The taste responsiveness of six squirrel monkeys, five pigtail macaques, four olive baboons, and four spider monkeys to polycose, a starch-derived polysaccharide, was assessed in two-bottle preference tests of brief duration (2 min). In experiment 1, the monkeys were given the choice between tap water and defined concentrations of polycose dissolved in tap water. In experiment 2, the animals were given the choice between polycose and sucrose, fructose, glucose, lactose, and maltose presented in equimolar concentrations of 100 and 200 mM, respectively. The animals were found to prefer concentrations of polycose as low as 10 mM (pigtail macaques), 30 mM (olive baboons and spider monkeys), and 60 mM (squirrel monkeys) over tap water. Relative taste preferences were stable across the concentrations tested and indicate an order of relative effectiveness (sucrose > polycose maltose) in squirrel monkeys, spider monkeys, and olive baboons that is similar to the order of relative sweetness in humans. Pigtail macaques, however, displayed an order of relative effectiveness (maltose > polycose sucrose) that differs markedly from that found in the other primate species tested and is similar to relative taste preferences found in rodents such as rats. Both the high sensitivity of the pigtail macaques to polycose and their vivid predilection for this polysaccharide and its disaccharide constituent maltose suggest that Macaca nemestrina, unlike other primates, but like rodents, may have specialized taste receptors for starch.     

Sour-Taste Tolerance in Four Species of Nonhuman Primates

The taste of most fruits is characterized by a mixture of sensations termed sweet and sour by humans, and the food selection behavior of primates suggests that they may use the relative salience of sweetness and sourness to assess palatability of potential food items. Therefore, taste responses of six squirrel monkeys, five pigtail macaques, four olive baboons, and four spider monkeys to sweet-sour taste mixtures were assessed in two-bottle preference tests of brief duration (2 min). Monkeys were given the choice between a reference solution of 50 mM sucrose and mixtures containing 10, 30, or 50 mM citric acid plus 10, 20, 50, 100, 200, 400, 800, or 1000 mM sucrose. We found that the four species differed markedly in their acceptance of physiological concentrations of sour-tasting citric acid. Whereas olive baboons showed the highest degree of sour-taste tolerance and actually preferred most of the sweet-sour taste mixtures over sweet-tasting reference solutions, squirrel monkeys showed the lowest degree of sour-taste tolerance and rejected most of the sweet-sour taste mixtures even when they contained considerably more sucrose than the reference solutions. Additional tests demonstrated that the preference for sweet-sour taste mixtures was not based on masking effects. Rather, the animals perceived both the sweetness and the sourness of the taste mixtures and made a trade-off between the attractive and aversive properties of the two taste qualities. The results of this study suggest that the proximate reason for the marked differences in acceptance of sweet-sour taste mixtures are differences among species in the hedonic evaluation of the sour taste of citric acid. Possible ultimate reasons, which do not necessarily exclude, but may complement each other, include evolutionary adaptation to dietary specialization, avoidance of competition pressure, and phylogenetic relatedness.     

Taste Responsiveness to Food-Associated Acids in the Squirrel Monkey (Saimiri sciureus)

The taste responsiveness of six adult squirrel monkeys to five food-associated acids was assessed in two-bottle preference tests of brief duration (5 min). In experiment 1 the monkeys were given the choice between tap water and defined concentrations of citric acid, ascorbic acid, malic acid, acetic acid, or tannic acid dissolved in tap water. In experiment 2 the animals were given the choice between a 50 mM sucrose solution and defined concentrations of the same acids dissolved in a 50 mM sucrose solution. With both procedures Saimiri sciureus significantly discriminated concentrations as low as 10 mM ascorbic acid and acetic acid, 5 mM citric acid and malic acid, and 0.2 mM tannic acid from the alternative stimulus. The results showed: (1) that squirrel monkeys respond to the same range of acid concentrations as other nonhuman primate species tested so far, (2) that Saimiri sciureus detects food-associated acids at concentrations well below those present in most fruits, and (3) that the responsiveness of this New World primate to acidic tastants was largely unaffected by the addition of a sweet-tasting substance. The results support the assumption that squirrel monkeys may use sourness and/or astringency of food-associated acids as a criterion for food selection.     

Taste Preferences for Five Food-Associated Sugars in the Squirrel Monkey (Saimiri sciureus)

The taste preferences of six adult squirrel monkeys for sucrose, fructose, glucose, lactose, and maltose were assessed in two-solution choice tests of brief duration (5 min). In experiment 1 the monkeys were given the choice between all binary combinations of the saccharides presented in equimolar concentrations of 50, 100, 200, and 400 mM, respectively. Preferences for individual sugars were stable across the concentrations tested and indicate an order of relative effectiveness (sucrose > fructose > glucose ≥ maltose ≥ lactose), which is identical to the order of sensitivity found in this species and similar to findings on relative sweetness in man. In experiment 2 the squirrel monkeys were given the choice between a standard sucrose solution of 50 mM and ascending series of concentrations of the other saccharides in order to determine concentrations that were consumed in equal amounts compared to the standard. Solutions of 100 mM fructose, 300 mM glucose, and 200 mM lactose and maltose were found to be equally effective to the 50 mM sucrose, whereas other concentrations led to marked preferences for one of the alternatives. The finding of concentrations of equal effectiveness are consistent with findings on concentration-dependent loss of discrimination ability in man and support the hypothesis that for squirrel monkeys sucrose may indeed be indiscriminate from fructose, glucose, lactose and maltose when concentrations are suitably adjusted. The results suggest that squirrel monkeys and man share important features of sweet-taste perception.     

The Frequency of Occurrence of Acyclic Monoterpene Alcohols in the Chemical Environment does not Determine Olfactory Sensitivity in Nonhuman Primates

Using a conditioning paradigm, the olfactory sensitivity of five spider monkeys, three squirrel monkeys, and three pigtail macaques for six acyclic monoterpene alcohols that differ markedly in their frequency of occurrence in plant odors was assessed. The results showed that: (1) all three primate species have a well-developed olfactory sensitivity for acyclic monoterpene alcohols; (2) squirrel monkeys are significantly more sensitive for members of this class of odorants than the other two species and are able to detect all six odorants at concentrations below 0.1 ppm; and (3) there is a lack of positive correlations between olfactory sensitivity and the abundance of the acyclic monoterpene alcohols in flower odors and etheric oils. The results lend support to the growing body of evidence that suggests between-species comparisons of the number of functional olfactory receptor genes or of neuroanatomical features are poor predictors of olfactory performance. The findings do not support the hypothesis that olfactory sensitivity for members of a chemical class may be related to the frequency of occurrence of such odorants in a species' chemical environment.     

Role of cucurbitacin C in resistance to spider mite (Tetranychus urticae) in cucumber (Cucumis sativus L.)

Cucurbitacins are bitter triterpenoid compounds that are toxic to most organisms and occur widely in wild and cultivated Cucurbitaceae. The only cucurbitacin identified in Cucumis sativus is cucurbitacin C. The bitter taste of cucumber has been correlated with resistance to the spider mite Tetranychus urticae, but a quantitative relationship has not been established. We determined the spider mite resistance and cucurbitacin C content in the dihaploid progeny derived from the F₁ generation of a cross between a bitter, spider-mite-resistant cucumber line and a bitter-free, spider-mite-susceptible line. The ratio of the number of bitter to bitter-free dihaploids conformed to the expected 1:1 ratio, based on a monogenic segregation pattern. Genetic analysis ascribed 69% of the variance of the difference in spider mite survival rate to the bitterness locus. Within the group of bitter dihaploids, cucurbitacin C content was significantly correlated with spider mite resistance. Thus, a quantitative relationship between cucurbitacin C content and spider mite resistance could be established.     

Serum α-lipoprotein responses to variations in dietary cholesterol,protein and carbohydrate in different non-human primate species

Serum α-lipoprotein responses to variations in dietary cholesterol, protein, and carbohydrate were studied in different nonhuman primate species. Chimpanzee, rhesus, green, patas, squirrel and spider monkeys all showed significant interspecies differences in serum total cholesterol responses to 1.84 mg/kcal exogenous cholesterol. Dietary cholesterol significantly increased the α-lipoprotein cholesterol in all species except rhesus and chimpanzee. Among these species, there was no relationship between the basal serum lipoprotein profile and subsequent lipoprotein responses to dietary cholesterol. Although the level of dietary protein at 6%, 12%, and 37% of calories had no appreciable main effect on serum total cholesterol in spider monkeys, very low protein diet (6% of calories) produced a significant elevation in α-lipoprotein cholesterol. Serum α-lipoprotein responses to exogenous cholesterol (1.84 mg/kcal) was highest for the very low protein diet and lowest for low protein diet (12% of calories). Diets with high sucrose (76.5% of calories) and low saturated fat (12.5% of calories) containing no added cholesterol were tested in squirrel and spider monkeys and produced a consistent serum total cholesterol response; the α-lipoprotein response was significantly higher in squirrel monkeys than in spider monkeys. The above findings have implications in experimentally induced and comparative atherogenesis. Presented at the Lipoprotein Symposium AOCS meeting, St. Louis, Missouri, May 1978.     

Taste preferences of the common vampire bat (Desmodus rotundus)

Taste preference tests, with simultaneous presentation of treated and untreated food, were administered to 24 common vampire bats (Desmodus rotundus). The bats received brief exposures to four different stimuli representing sweet, salty, sour, and bitter tastes, each at four different concentrations. Despite a strong location bias, the bats significantly (P < 0.01) avoided the highest concentrations of the salty, sour, and bitter tastes. Consumption of the sweet stimulus at all concentrations was similar to that of the untreated standard. Vampires evidently can discriminate based on taste, although their ability is apparently poorly developed when compared with some euryphagous species such as the rat. Hence, taste is probably not a factor in host selection by the vampire.This research was conducted, in part, with funds provided to the U.S. Fish and Wildlife Service by the U.S. Agency for International Development under PASA RA (ID) 1–67.     

Comparisons of Contact Chemoreception and Food Acceptance by Larvae of Polyphagous Helicoverpa armigera and Oligophagous Bombyx mori

We compared food choice and the initial response to deterrent treated diet between fifth instars of Helicoverpa armigera, a polyphagous generalist pest, and Bombyx mori, an oligophagous specialist beneficial. Bombyx mori was more behaviorally sensitive to salicin than to caffeine. The relative sensitivities were reversed for H. armigera, which was tolerant to the highest levels of salicin found in natural sources but sensitive to caffeine. A single gustatory receptor neuron (GRN) in the medial styloconic sensillum of B. mori was highly sensitive to salicin and caffeine. The styloconic sensilla of H. armigera did not respond consistently to either of the bitter compounds. Phagostimulants also were tested. Myo-inositol and sucrose were detected specifically by two GRNs located in B. mori lateral styloconic sensillum, whereas, in H. armigera, sucrose was sensed by a GRN in the lateral sensillum, and myo-inositol by a GRN in the medial sensillum. Myo-inositol responsiveness in both species occurred at or below 10^?3 mM, which is far below the naturally occurring concentration of 1 mM in plants. Larval responses to specific plant secondary compounds appear to have complex determinants that may include host range, metabolic capacity, and gustatory repertoire.     

Taste receptors for pyrrolizidine alkaloids in a monophagous caterpillar

Caterpillars of Utetheisa ornatrix are monophagous on species of Crotalaria from which they obtain pyrrolizidine alkaloids (PAs) for defense and which the males convert to a pheromone. We show that a taste receptor neuron in each of the lateral and medial galeal styloconic sensilla responds to PAs of three different types. Monocrotaline, commonly present in Crotalaria species, is the most strongly stimulating with thresholds of response below 10^–11 and 10^–9 M in the two sensilla. These are among the lowest known taste thresholds in any insect and are similar to the thresholds for PAs in a polyphagous arctiid caterpillar that also sequesters PAs and uses them as the source of male pheromone. The receptors also respond to heliotrine, a type of PA that is probably never encountered by the insects. Monocrotaline and senecionine N-oxide are shown in behavioral assays to be phagostimulants. The data show that there is no tight link between taste receptor sensitivity to specific PAs and hostplant selection in these caterpillars. Perhaps the adults are primarily responsible for hostplant selection.     

Common Variants of <Emphasis Type="Italic">ABCB4</Emphasis> and <Emphasis Type="Italic">ABCB11</Emphasis> and Plasma Lipid Levels: A Study in Sib Pairs with Gallstones,and Controls

Most epidemiological surveys have confirmed the association of low HDL-cholesterol and high triglyceride levels with cholesterol gallstones. Our objective was to analyze the relationship between plasma lipid levels and common polymorphisms of ABCB11 (encoding the bile salt export pump, BSEP) and ABCB4 (encoding the phospholipid transporter into bile, MDR3) genes. Plasma lipids were measured in 108 index patients of sib pairs with gallstones and in 260 controls. Using PCR-based assays with 5′-nuclease and fluorescence detection (TaqMan), the ABCB11 coding SNP p.A444V and four haplotype-tagging SNPs covering the ABCB4 gene (c.504C > T, c.711T > A, p.R652G, rs31653 in intron 26) were genotyped. Plasma lipids were compared in carriers of the common versus rare allele of these polymorphisms using Student’s t test and Pearson’s correlation. BMI and triglyceride levels were higher and HDL-cholesterol levels were lower in affected siblings than in controls. Among cases, triglyceride and cholesterol levels were higher in carriers of the common versus rare (hetero/homozygous carriers) allele of the SNPs p.A444V of ABCB11 and C.504C > T of ABCB4. HDL-cholesterol was lower in carriers of the common allele of rs31653. In controls, significant differences of cholesterol and HDL-cholesterol levels were found in carriers of ABCB4 polymorphisms. Our results do not support the hypothesis of a link between ABCB4 and ABCB11 polymorphisms, lithogenic dyslipidemia, and gallstone risk.     

Use of surface response methodology to optimize culture conditions for iturin A production by Bacillus subtilis in solid-state fermentation

Response surface methodology (RSM) was employed to optimize the cultivation conditions of Bacillus subtilis S3 for the enhancement of iturin A, a lipopeptide antibiotic used as biological pesticide, production in solid-state fermentation (SSF). The statistic experimental model predicted a maximum iturin production of 11.435 mg/g-wet solid material. Verification of the calculated maximum was done with experiments that were performed in the culture media representing the optimum combination found, and the iturin A production of 11.447 mg/g-wet solid material (average of three repeats) was obtained when B. subtilis S3 was cultivated at 25 °C for 5 days in solid fermentation containing high gluten flour 10 g and rice bran 50 g in addition to glucose 1.15%, KH₂PO₄ 1.27 mM, MgSO₄ 5.08 mM, peanut oil 1.01%, inoculum 19.49% and water content 44.97%. The iturin A production by B. subtilis S3 was increased significantly by 23%, from 9.26 mg/g-wet solid material to 11.447 mg/g-wet solid material when the strain was cultivated in the optimal medium developed by surface response methodology, as compared to medium conventionally developed by one-factor-at-a-time. The yield of iturin A (11.447 mg/g-wet solid material, with 45% moisture content) produced by B. subtilis S3 reported in this study is the highest reported to date for B. subtilis species in SSF. In addition, the use of rice bran as a substrate in solid-state fermentation for iturin A production by B. subtilis is unique.     

Platinum(VI) dihydroxo tetramesitylporphyrin; characterization and chemical reactivity

Tetramesitylporphyrin platinum(VI) dihydroxo complex, TMPPt(OH)₂ · 2 benzoate was synthesized by addition of two equivalent of meta-chloroperbenzoic acid molecules to the tetramesitylporphyrin platinum(II). This complex was characterized by X-ray diffraction and spectroscopic methods, and is capable to convert two molecules of triphenylphosphine to triphenylphosphine oxide.     

Identification of host-plant chemicals stimulating oviposition by swallowtail butterfly,Papilio protenor

The ovipositional response of a Rutaceae-feeding papilionid butterfly,Papilio protenor, toCitrus host plants was evoked by the synergistic action ofl-(–)-stachydrine,d-(–)-quinic acid, (–)-synephrine, andl-(–)-proline that characterize the chemical compositions of the leaves and epicarp ofCitrus plants (C. natsudaidai andC. unshiu). The stimulatory activity of their mixture was enhanced by the addition of flavanone glycosides, naringin and hesperidin, which coexist in these plants and have previously been demonstrated to serve as oviposition stimulants. However, sugars such as sucrose, glucose, and inositols, which abound in plant tissues, exerted no effect on egg-laying by the females. On the other hand, chlorogenic acid present in the leaves of another host plant,Fagara ailanthoides, was found to act as an excellent synergist. However, there existed significant qualitative dissimilarities between the chemical compositions of the leaves ofC. unshiu andF. ailanthoides. This strongly suggests thatP. protenor is likely to utilize different categories of compounds as chemical cues in recognizing each plant as a host.     

Within-plant variation in concentrations of amino acids,sugar, and sinigrin in phloem sap of black mustard,Brassica nigra (L.) Koch (Cruciferae)

Although within-plant variation in the nutrient and allelochemical composition of phloem sap has been invoked to explain patterns of host use by phloem-feeding insects, little is known about within-plant variation in phloem chemistry. Here I describe a new technique in which I use the green peach aphid,Myzus persicae Sulz., to investigate within-plant variation in the concentrations of chemicals in the phloem sap of black mustard,Brassica nigra (L.) Koch (Cruciferae). Relationships between the concentrations of chemicals in aphid diets and honeydew were established using honeydew from aphids fed on artificial diets with known concentrations of amino acids, sucrose, and sinigrin. These relationships were applied to honeydew from aphids fed on different aged leaves of black mustard to estimate the concentrations of the chemicals in phloem sap. Sinigrin concentration was estimated to be high (>10 mM) in phloem sap in young leaves, calling into question the prevailing opinion that phloem sap contains only low concentrations of allelochemicals. High concentrations may function as defenses against sap-feeding herbivores. Within-plant variation in phloem sap composition was high: (1) young leaves had high concentrations of nutrients (216 mM amino acids, 26% sugar) and sinigrin (>10 mM); (2) mature and presenescent leaves had lower concentrations of nutrients (77–83 mM amino acids, 19–20% sugar) and low concentrations of sinigrin (1–2 mM); and (3) senescing leaves had high concentrations of nutrients (199 mM amino acids, 25% sugar) and low concentrations of sinigrin (3 mM).     

Ethanol and (−)-α-Pinene: Attractant Kairomones for Bark and Ambrosia Beetles in the Southeastern US

In 2002–2004, we examined the flight responses of 49 species of native and exotic bark and ambrosia beetles (Coleoptera: Scolytidae and Platypodidae) to traps baited with ethanol and/or (−)-α-pinene in the southeastern US. Eight field trials were conducted in mature pine stands in Alabama, Florida, Georgia, North Carolina, and South Carolina. Funnel traps baited with ethanol lures (release rate, about 0.6 g/day at 25–28°C) were attractive to ten species of ambrosia beetles (Ambrosiodmus tachygraphus, Anisandrus sayi, Dryoxylon onoharaensum, Monarthrum mali, Xyleborinus saxesenii, Xyleborus affinis, Xyleborus ferrugineus, Xylosandrus compactus, Xylosandrus crassiusculus, and Xylosandrus germanus) and two species of bark beetles (Cryptocarenus heveae and Hypothenemus sp.). Traps baited with (−)-α-pinene lures (release rate, 2–6 g/day at 25–28°C) were attractive to five bark beetle species (Dendroctonus terebrans, Hylastes porculus, Hylastes salebrosus, Hylastes tenuis, and Ips grandicollis) and one platypodid ambrosia beetle species (Myoplatypus flavicornis). Ethanol enhanced responses of some species (Xyleborus pubescens, H. porculus, H. salebrosus, H. tenuis, and Pityophthorus cariniceps) to traps baited with (−)-α-pinene in some locations. (−)-α-Pinene interrupted the response of some ambrosia beetle species to traps baited with ethanol, but only the response of D. onoharaensum was interrupted consistently at most locations. Of 23 species of ambrosia beetles captured in our field trials, nine were exotic and accounted for 70–97% of total catches of ambrosia beetles. Our results provide support for the continued use of separate traps baited with ethanol alone and ethanol with (−)-α-pinene to detect and monitor common bark and ambrosia beetles from the southeastern region of the US.     

Sensory Thresholds of Selected Phenolic Constituents from Thyme and their Antioxidant Potential in Sunflower Oil

The odor detection thresholds of carvacrol (5-isopropyl-2-methyl-phenol), thymol (2-isopropyl-5-methyl-phenol) and p-cymene 2,3-diol (2,3-dihydroxy-4-isopropyl-1-methyl-benzene) in sunflower oil, determined by the three-alternative, forced-choice procedure, were 30.97, 124 and 794.33 mg kg⁻¹, respectively. Sunflower oil containing 13, 70, or 335 mg kg⁻¹ of carvacrol, thymol or p-cymene 2,3-diol, respectively, was judged to be similar (P < 0.01) in taste and odor to its antioxidant-free counterpart. The rate constant of sunflower oil oxidation, measured from the increase in peroxide value during storage at 25 °C, was 9.2 × 10⁻⁹ mol kg⁻¹ s⁻¹ while the rate constants were 9.3 × 10⁻⁹, 9.8 × 10⁻⁹, and 4.3 × 10⁻⁹ mol kg⁻¹ s⁻¹ in the presence of 13 mg kg⁻¹ carvacrol, 70 mg kg⁻¹ thymol, and 335 mg kg⁻¹ p-cymene 2,3-diol, respectively. At a level of 335 mg kg⁻¹, p-cymene 2,3-diol did not impart flavor taints and effected a 46.7% reduction in the rate of oxidation of sunflower oil. These findings indicate that the diphenolic p-cymene 2,3-diol could potentially replace synthetic antioxidants and is a valuable addition to the antioxidants used by the food industry in its quest to meet consumer demands for synthetic-additives-free and ‘natural’ foods.     

Differences in host use efficiency of larvae of a generalist moth, Operophtera brumata on three chemically divergent Salix species

The food selection, growth, and fecundity of insect herbivores are largely dictated by the chemical composition and nutritive values of plant foliage. We studied the host-use efficiency of larvae of the generalist moth, Operophtera brumata (Lepidoptera: Geometridae) on three chemically divergent but nutritively similar willows (Salix spp.). The 4th instars were able to use the salicylate-free leaves of S. phylicifolia efficiently. Growth was slightly reduced on S. pentandra, which contained a moderate level of acetylated salicylates. The high concentration of salicylates found in the leaves of S. myrsinifolia seemed to provide efficient protection against non-specialized O. brumata. We also studied assimilation of nutrients and degradation of salicylates and other secondary compounds in the digestive tract of O. brumata larvae. Neither the assimilation of nitrogen nor of carbon were affected by secondary chemicals of ingested food. Salicylates were shown to be degraded to salicin and catechol, while further degradation of salicin to saligenin was rather slow. In an artificial diet experiment, we showed that two degradation products of salicylates, catechol and saligenin markedly reduced the growth of the larvae. Neither salicin nor chlorogenic acid affected larval growth. We conclude that salicylates reduced the growth of the generalist winter moth mainly by feeding deterrence caused by 6-hydroxy-2-cyclohexenone and catechol. Compared to the deleterious effects of salicylates the effects of other secondary compounds were minor.     

Possible Mechanism of Inhibition of 6-Methoxy-Benzoxazolin-2(3H)-One on Germination of Cress (Lepidium sativum L.)

6-Methoxy-benzoxazolin-2(3H)-one (MBOA) inhibited the germination of cress (Lepidium sativum L.) seeds at concentrations greater than 0.03 mM. Inhibition was overcome by sucrose, suggesting that MBOA may inhibit sugar metabolism in cress seeds. Induction of α-amylase activity in seeds was also inhibited by MBOA at concentrations greater than 0.03 mM. Inhibition of both germination and induction of α-amylase activity increased with increasing concentrations of MBOA, and the extent of germination correlated positively with the activity of α-amylase in the seeds. MBOA added to a reaction mixture for α-amylase assay did not affect enzyme activity, indicating that MBOA does not inhibit in vitro α-amylase activity. Cress seeds germinated approximately 16 hr after incubation, and inhibition of α-amylase by MBOA occurred within 6 hr after incubation. These results suggest that MBOA may inhibit the germination of cress seeds by inhibiting the induction of α-amylase activity, because α-amylase plays a key role in the conversion of reserve carbohydrate into soluble sugars, a prerequisite for seed germination.