Postharvest resistance in hard spring and winter wheat varieties of the northern Great Plains to the lesser grain borer (Coleoptera: Bostrichidae)

Wheat (Triticum aestivum L.) varietal resistance to the lesser grain borer, Rhyzopertha dominica (F), was evaluated in hard spring and winter wheat produced 1997, 1998 (Bozeman, Montana). We tested the hypothesis that six Montana-grown spring wheat varieties, 'Ernest', 'Scholar', 'Hi-Line', 'McNeal', 'Newana', and 'Amidon', were equally and strongly resistant to R. dominica at low moisture contents (9-10%). Mortality/Feeding damage occurred in all varieties. In most assays, Ernest had significantly greater feeding damage from R. dominica than other varieties, usually not significantly different from the susceptible control. Mean adult mortality was significantly greater in McNeal (93%) and Hi-Line (92%) than in Ernest (34%) and Montana-grown, soft white spring wheat (Penawawa), the susceptible control (36%). In 9 wk, twice as many adult progeny were produced on Ernest than on McNeal, Hi-Line, or Scholar. We also compared three Montana-grown winter wheat varieties for resistance to R. dominica attack at low moisture contents (9-10%). Significantly more mortality after 6 wk was associated with all winter wheat varieties than the susceptible control. In 'Nuwest', 'Rocky', and 'Vanguard', significantly fewer progeny were produced than in the susceptible control; these varieties appeared more resistant. 'Tiber' and 'Neeley', in contrast, appeared more susceptible than other winter wheat varieties evaluated. Susceptibility decreased significantly with a 1.2% decrease in moisture content. Percentage of total protein was positively correlated with percentage of sound kernels and negatively with total progeny (r2 = -0.69). Kernel hardness was positively correlated with percentage of sound kernels, but negatively correlated with total progeny (r2 = -0.87) and dry weight loss.     

Inhibition of amylases from different origins by albumins from the wheat kernel.

The amylase activity of water extracts from 18 insect species, from 23 marine species and from 17 different species of birds and mammals was determined quantitatively. The inhibition of amylase in these extracts by three albumin fractions from the mature wheat kernel, which had been separated according to their molecular weights (60 000, 24 000 and 12 500 D), was determined as well. The inhibition activity of the three albumin fractions toward amylases extracted from a number of cereal species or from immature and germinating wheat kernel was also tested. The extracts from insects that are destructive of wheat grain and stored wheat products showed much higher amylase activities as compared to the other insect species that do not attack wheat and wheat products. On the basis of the effectiveness with which the three albumin fractions inhibit their activities, the amylase preparations tested were divided into susceptible, partially susceptible and resistent. Susceptible amylases, inhibited by any of the three albumin fractions, were found mainly in insects that attack wheat and in marine species. Partially susceptible amylases, inhibited by only one or two of the three albumin fractions, were present in a few avain and mammalian species including man. Resistent amylases were largely distributed in cereal, avian and mammalian species as well as in insect species that do not usually attack wheat grain or wheat flour products. At no stage of development, wheat alpha-amylase was inhibited by the albumin fractions from the mature kernel. The 12 500 dalton albumin fraction was the most effective in inhibiting insect amylases, but it was inactive toward avian and mammalian amylases. The 24 000 dalton albumin fraction was the most effective in inhibiting amylases from marine avian and mammalian species and inhibited as much as 33 amylases over 66 different amylases tested. It is suggested that protein inhibitors of amylase contributed to natural selection of polyploid wheats by giving some insect resistence to such wheats, even though some insect species were able to overcome this biochemical defense toa large degree by producing higher amylase activities.     

Interactions of wheat variety, production environments, and prior insect damage on postharvest resistance to the lesser grain borer

Wheat, Triticum aestivum L., varietal resistance to the lesser grain borer, Rhyzopertha dominica (F), was evaluated in hard spring wheat produced in 2001 and 2002 (Bozeman, Moccasin, and Huntley, MT). We tested the hypothesis that seed coat factors, not the endosperm, cause feeding resistance to R. dominica and that this resistance is genetic, not affected by agronomic conditions. Using a rapid, intensive feeding bioassay (frass production), we found, with one exception, no significant difference in resistance to R. dominica, among sound kernels of hard red wheat, comparing all locations and cultural conditions (irrigated versus dryland production). The most significantly resistant samples as indicated by lowest feeding activity (measured by lowest frass production) were 'Amidon' produced at Moccasin under dryland conditions and Amidon produced at Huntley under irrigated conditions. As with previous studies done in our laboratory, sound kernels of all hard wheat varieties from these new crop year studies (2001, 2002) were attacked. When subsamples of these varieties from the same locations and cultural conditions as the previous test were first subjected to a heavy infestation of Plodia interpunctella (Hübner) and then to the same age adult R. dominica, damage was significantly greater. Each kernel chosen for this test had been equally damaged by P. interpunctella larvae, germ consumed, endosperm not damaged, but fully exposed behind germ, no other damage. This collaborative damage caused feeding damage by R. dominica to increase 2- to 7.5-fold (as measured by R. dominica frass production). Particularly notable was 'McNeal' that switched from one of the most resistant varieties to the most fed upon, when the endosperm was exposed by P. interpunctella larvae. Therefore, we confirmed that at least one factor conferring resistance in McNeal is located in the kernel pericarp.     

Treatment of adult Coleoptera with a chitin synthesis inhibitor affects mortality and development time of their progeny

A group of insect growth regulators, the chitin synthesis inhibitors (CSIs), are being more widely used as many insects have become resistant to broad-spectrum insecticides. This study investigated the mode of action of one CSI, particularly looking at the delayed effect of treating adult insects on the survival and development of their progeny. The study describes the responses of adult stored grain beetles Sitophilus oryzae (L.) (Curculionidae) and Rhyzopertha dominica (F.) (Bostrichidae) to wheat treated with chlorfluazuron. For both species, wheat treated with chlorfluazuron reduced progeny survival at a constant rate from 1 day to 8 weeks after it had been treated. There was a direct, negative concentration-time relationship between the treatment of the adults with chlorfluazuron-treated wheat and the survival and developmental rate of their progeny. Adults that were pre-exposed for one or more weeks before oviposition to wheat treated with chlorfluazuron had significantly lower progeny survival than adults that were not pre-exposed. The EC₉₅ values were 16-fold higher for R. dominica and 40-fold higher for S. oryzae from assays without pre-exposure, than with pre-exposure. Pre-exposure of R. dominica with 0.1 mg kg^-1 and of S. oryzae with 0.4 mg kg^-1 of chlorfluazuron reduced progeny survival by 95%. Higher concentrations of chlorfluazuron produced a reduction in mean population developmental time of several days. These findings have important implications for bioassays of CSIs, since pre-exposure of the adults can significantly reduce the numbers of F1 progeny. Therefore standard bioassays may seriously underestimate the efficacy of the CSI being assayed.     

Comparison of aeration and spinosad for suppressing insects in stored wheat

Field studies were conducted from July 2002 to January 2003 for evaluating the effects of controlled aeration and a commercial biological insecticide, spinosad, in suppressing insect populations in stored wheat. Six cylindrical steel bins were filled with newly harvested (2002 crop year) hard red winter wheat on 9 and 10 July 2002. Each bin contained 30.7 metric tons (1,100 bu) of wheat. Wheat in two bins was left untreated (control), whereas wheat in two bins was treated with spinosad, and in another two bins was subjected to aeration by using aeration controllers. Spinosad was applied to wheat at the time of bin filling to obtain a rate of 1 mg ([AI])/kg. Aeration controllers were set to run the fans when ambient air temperature fell below 23.9, 18.3, and 7.2 degrees C for the first, second, and third cooling cycles, respectively. We added 400 adults each of the rusty grain beetle, Cryptolestes ferrugineus (Stephens); lesser grain borer, Rhyzopertha dominica (F.); and red flour beetle, Tribolium castaneum (Herbst), to the grain at monthly intervals between July and October 2002. Insect density in the bins was estimated monthly by taking 3-kg grain samples from 21 locations within each bin by using a pneumatic grain sampler. No live T. castaneum or C. ferrugineus and very low densities of R. dominica (<0.008 adults per kilogram) were found in wheat treated with spinosad during the 6-mo sampling period. Density of C. ferrugineus and T. castaneum in aerated bins did not exceed two adults per kilogram (the Federal Grain Inspection Service standard for infested wheat), whereas R. dominica increased to 12 adults per kilogram in November 2002, which subsequently decreased to three adults per kilogram in January 2003. In the untreated (control) bins, R. dominica density increased faster than that of C. ferrugineus or T. castaneum. Density of R. dominica peaked at 58 adults per kilogram in October 2002 and decreased subsequently, whereas T. castaneum density was 10 adults per kilogram in October 2002 but increased to 78 adults per kilogram in January 2003. Density of C. ferrugineus increased steadily during the 6-mo study period and was highest (six adults per kilogram) in January 2003. This is the first report comparing the field efficacy of spinosad and aeration in managing insects in farm bins. Our results suggest that spinosad is very effective in suppressing R. dominica, C. ferrugineus, and T. castaneum populations in stored wheat.     

Effect of triterpenoid glycosides on alpha- and beta-amylase activity and total protein content in wheat seedlings

Influence of the aleanolic acid glycosides from Silphium perfoliatum L. (silphioside B, C, E and G) and their progenins on the amylase activity and total protein content in wheat seedlings was studied. Treatment of the Triticum aestivum L. seeds with 1-10 microM water solutions of mono- and diglycosides (mono- and bisdesmosines) elevated the alpha-amylase and total amylase activities in seedlings. Silphioside E containing three glucose moieties in its molecule did not change alpha-amylase activity, but it did if bis-triglycoside acetylated carbohydrate (as in silphioside C). Effects of 5-10 microM solutions of the active glycosides was comparable with that of exogenous gibberellin A3 and 6-benzylaminopurine.     

Molecular cloning and characterization of a midgut chymotrypsin-like enzyme from the lesser grain borer, Rhyzopertha dominica

A cDNA encoding a chymotrypsinogen-like protein in midguts of the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) was cloned and sequenced. The 901 bp cDNA contains an 816-nucleotide open reading frame encoding 272-amino acids. The predicted molecular mass and pI of the mature enzyme are 23.7 kDa and 4.64, respectively. The encoded protein includes amino acid sequence motifs that are conserved with 5 homologous chymotrypsinogen proteins from other insects. Features of the putative chymotrypsin-like protein from R. dominica include the serine proteinase active site (His(90), Asp(133), Ser(226)), conserved cysteine residues for disulfide bridges, the residues (Gly(220), Gly(243), Asp(252)) that determine chymotrypsin specificity, and both zymogen activation and signal peptides. A TPCK-sensitive caseinolytic protein (P6) with an estimated molecular mass of 24 kDa is present in midgut extracts of R. dominica and can be resolved by electrophoresis on 4-16% polyacrylamide gels. The molecular mass of this caseinolytic enzyme is similar to that of the chymotrypsin deduced from cDNA. Midgut extracts of R. dominica readily hydrolyzed azocasein and N-succinyl-alanine-alanine-proline-phenylalanine-p- nitroanilide (SAAPFpNA), a chymotrypsin-specific substrate. Properties of the enzymes responsible for these activities were partially characterized with respect to distribution in the gut, optimum pH, and sensitivity toward selected proteinase inhibitors. Optimal activity against both azocasein and SAAPFpNA occurs in a broad pH range from about 7 to 10. Both azocasein and SAAPFpNA activities, located primarily in the anterior midgut region, are inhibited by aprotinin, phenylmethyl sulphonylfluoride (PMSF), and soybean trypsin inhibitor (STI). TPCK (N-alpha-tosyl-L-phenylalanine chloromethyl ketone) and chymostatin inhibited more than 60% of SAAPFpNA but only about 10-20% of azocasein activity. These results provide additional evidence for the presence of serine proteinases, including chymotrypsin, in midguts of R. dominica. Arch. Insect Biochem. Physiol. 43:173-184, 2000.Published 2000 Wiley-Liss, Inc.     

Effect of temperature and commodity on insecticidal efficacy of spinosad dust against Sitophilus oryzae (Coleoptera: Curculionidae) and Rhyzopertha dominica (Coleoptera: Bostrychidae)

The insecticidal effect of spinosad dust, a formulation that contains 0.125% spinosad, was evaluated against adults of Sitophilus oryzae (L.) and Rhyzopertha dominica (F.) at three temperature levels (20, 25, and 30 degrees C) and four commodities (wheat, Triticum aestivum L.; barley, Hordeum vulgare L.; rice, Oryza sativa L.; and maize, Zea mays L.). For this purpose, quantities of the above-mentioned grains were treated with spinosad at two dose rates (20 and 50 ppm of the formulation, corresponding to 0.025 and 0.06 ppm AI, respectively), and mortality of the exposed adults in the treated grains was measured after 7 and 14 d, whereas progeny production was assessed 65 d later. Generally, for both species, mortality increased with dose, exposure interval, and temperature. For S. oryzae, adult survival and progeny production were lower on wheat than the other grains. After 14 d of exposure, mortality of S. oryzae adults on wheat treated with 50 ppm ranged between 61 and 98%, whereas in the other three commodities it did not exceed 42%. Mortality of R. dominica after 14 d on grains treated 50 ppm ranged between 91 and 100%. For this species, progeny production from exposed parental adults was low in all commodities regardless of temperature. Results indicate that spinosad dust can be used as an alternative to traditional grain protectants, but its effectiveness is highly determined by the target species, commodity, dose, and temperature.     

Effectiveness of spinosad on four classes of wheat against five stored-product insects

Spinosad is a commercial reduced-risk pesticide that is naturally derived. Spinosad's performance was evaluated on four classes of wheat (hard red winter, hard red spring, soft red winter, and durum wheats) against adults of the lesser grain borer, Rhyzopertha dominica (F.); rice weevil, Sitophilus oryzae (L.); sawtoothed grain beetle, Oryzaephilus surinamensis (L.); red flour beetle, Tribolium castaneum (Herbst); and larvae of the Indianmeal moth, Plodia interpunctella (Hübner). Beetle adults (25) or P. interpunctella eggs (50) were exposed to untreated wheat and wheat treated with spinosad at 0.1 and 1 mg (AI)/kg of grain. On all untreated wheat classes, adult beetle mortality ranged from 0 to 6%, and P. interpunctella larval mortality ranged from 10 to 19%. The effects of spinosad on R. dominica and P. interpunctella were consistent across all wheat classes. Spinosad killed all exposed R. dominica adults and significantly suppressed progeny production (84-100%) and kernel damage (66-100%) at both rates compared with untreated wheat. Spinosad was extremely effective against P. interpunctella on all wheat classes at 1 mg/kg, based on larval mortality (97.6-99.6%), suppression of egg-to-adult emergence (93-100%), and kernel damage (95-100%), relative to similar effects on untreated wheats. The effects of spinosad on S. oryzae varied among wheat classes and between spinosad rates. Spinosad was effective against S. oryzae, O. surinamensis and T. castaneun only on durum wheat at 1 mg/kg. Our results suggest spinosad to be a potential grain protectant for R. dominica and P. interpunctella management in stored wheat.     

Macroaffinity ligand-facilitated three-phase partitioning (MLFTPP) of alpha-amylases using a modified alginate

The crude extracts of alpha-amylases when mixed with alginate, tert-butyl alcohol, and ammonium sulfate resulted in an interfacial precipitate containing polymer-bound amylase. The precipitate was dissolved in 1 M maltose to recover alpha-amylase activity. The recovery of alpha-amylases were 74%, 77%, and 92% in the case of Bacillus amyloliquefaciens, wheat germ, and porcine pancreas, respectively. All purified preparations showed a single band on SDS-PAGE.     

Interaction of treatment of both adult and immature Coleoptera with a chitin synthesis inhibitor affects mortality and development time of their progeny

Chitin synthesis inhibitors, like many other insect growth regulating insecticides, do not kill adult insects but cause mortality of the immature stages. Pre-exposure of adult stored grain Coleoptera, Rhyzopertha dominica (F.) (Bostrichidae) and Sitophilus oryzae (L.) (Curculionidae) and development of their progeny in grain treated with the chitin synthesis inhibitor, chlorfluazuron, influenced the mortality and development rate of the progeny. Hatch rate of eggs from R. dominica adults that had both developed and laid on wheat treated with 0.75 mg kg–1 chlorfluazuron was reduced by almost 50% compared with untreated eggs, with an LC50 of 0.84 mg kg–1 . Eggs laid on treated wheat by R. dominica adults that had been exposed only to treated wheat for 2 weeks before oviposition showed greater reduction in hatch: 75% reduction of normal hatch rate at 0.25 mg kg–1 and almost 100% reduction at 2 mg kg–1 chlorfluazuron, with an LC50 of 0.19 mg kg–1 chlorfluazuron. X-rays were used to monitor the development and mortality of the immature stages of S. oryzae that developed within the wheat grains. Numbers of eggs laid were not affected by chlorfluazuron treatment. The combination of pre-exposure of adults and chlorfluazuron concentration had an additive effect on mortality of immature S. oryzae. Pre-exposure of adults caused most mortality in the first three weeks of development (eggs and larvae), whereas development in treated wheat caused mortality from weeks 3 to 8 (pupae and adults); higher concentrations of chlorfluazuron caused higher mortality. Development in wheat treated with 1 mg kg–1 chlorfluazuron caused 12% corrected overall mortality of progeny while pre-exposure to the same concentration and development in untreated wheat caused 29% corrected mortality. Pre-exposure combined with development in wheat treated with 1 mg kg–1 caused 30% corrected mortality. Thus, pre-exposure of adults appears to have a greater effect on mortality of S. oryzae progeny than development of immature stages in treated grain. Development on treated grain had no effect on development rate. Pre-exposure of adults did not appear to affect the rate of immature development, as assessed by X-rays, but did slow the emergence of adults, lengthening development time by about 2 days. This significant, additive effect of pre-exposure of adults on the mortality of their progeny will enhance the toxicity of chitin synthesis inhibitors such as chlorfluazuron, since most adults receive treatment when the immature stages are treated in crops either before they are harvested or in storage. Assessing the proportion of eggs that hatch from pre-exposed adults would be a simpler bioassay for CSIs.     

Isolation and partial characterization of three isoamylases of Rhyzopertha dominica F. (Coleoptera: Bostrichidae)

Three isoamylases of Rhyzopertha dominica (termed RdA70, RdA79, and RdA90 according to their relative mobility in gel electrophoresis) were isolated by ammonium sulfate fractionation and hydrophobic interaction chromatography. RdA70 and RdA79 showed an optimal pH of 7.0, whereas for RdA90 the optimal pH was 6.5. The three isoamylases remained stable at 50 °C for 1 h, but at 60 °C, all lost 50% of their activity in 20 min and were completely inactivated in 1 h. RdA70 and RdA79 were inhibited by albumin extracts from wheat samples varying widely in amylase inhibitory activity; however, RdA90 was highly resistant to inhibition. β-Mercaptoethanol up to 30 mM increased the activity of the three isoamylases by 2.5-fold. The action pattern of the three isoamylases was typical of endoamylases; however, differences were observed on the hydrolytic efficiency rates measured as V_max/K_m ratio on starch, amylopectin, and amylose. The hydrolyzing action of RdA90 on starch and amylopectin (V_max/K_m = 90.4 ± 2.3 and 78.9 ± 6.6, respectively) was less efficient than that on amylose (V_max/K_m = 214 ± 23.2). RdA79 efficiently hydrolyzed both amylopectin and amylose (V_max/K_m = 260.6 ± 12.9 and 326.5 ± 9.4, respectively). RdA70 hydrolyzed starch and amylose at similar rates (V_max/K_m = 202.9 ± 5.5 and 215.9 ± 6.2, respectively), but amylopectin was a poor substrate (V_max/K_m = 124.2 ± 7.4). The overall results suggest that RdA70 and RdA79 appear to belong to a group of saccharifying isoamylases that breaks down long fragments of oligosaccharide chains produced by the hydrolytic action of RdA90. The simultaneous action of the three isoamylases on starch, aside from the high resistance of RdA90 to wheat amylase inhibitors, might allow R. dominica to feed and reproduce successfully on the wheat kernel.     

Insecticidal effects of extracts of seven plant species on larval development, alpha-amylase activity and offspring production of Tribolium castaneum (Herbst) (Insecta: Coleoptera: Tenebrionidae)

Bioinsecticidal effects of methanol extracts from seven plant species on Tribolium castaneum were investigated. Centaurium erythraea, Peganum harmala, Ajuga iva, Aristolochia baetica, Pteridium aquilinum and Raphanus raphanistrum extracts inhibit growth of larvae. C. erythraea was the most toxic with 63% mortality 10 days after treatment, followed by P. harmala with 58%. C. erythraea and P. aquilinum reduce the emergence rate respectively of 66% and 19%. The duration of larval period was shortened by Launaea arborescens, P. aquilinum and A. iva extracts, whereas R. raphanistrum and P. harmala extracts extend the larval period when compared to the control. Extracts of C. erythraea, P. harmala, A. iva and A. baetica inhibited F1 progeny production. Larvae possess three alpha-amylase isoforms as determined by SDS-PAGE. Larvae fed on treated diet had lower alpha-amylase activity than larvae feed on untreated diet. C. erythraea and P. harmala are the most potent extracts. These plant extracts could be useful to reduce seed damage caused by this pest species.     

Interaction between Acarophenax lacunatus (Cross & Krantz) (Prostigmata: Acarophenacidae) and Anisopteromalus calandrae (Howard) (Hymenoptera: Pteromalidae) on Rhyzopertha dominica (Fabricius) (Coleoptera: Bostrichidae)

The interaction between Acarophenax lacunatus (Cross & Krantz) and Anisopteromalus calandrae (Howard) may be a promising tool for the integrated pest management of stored grain insect pests. The objective of this study was to evaluate the compatibility of these two natural enemies on Rhyzopertha dominica (Fabricius). The experimental units were petri dishes (140 x 10 mm) containing 30 g of whole wheat grains (13% water content) infested with 20 adults of R. dominica. The treatments consisted of inoculation of A. lacunatus and A. calandrae, separately and associated, in eight replicates. Three inoculations of five adult females of the natural enemies were carried out in each petri dish at five, ten and fifteen days after the infestation of R. dominica. All treatments were stored during 60 days in environmental chamber at 30 +/- 1 degrees C, 60 +/- 5% relative humidity and 24 h scotophase. The smallest numbers of physogastric females of A. lacunatus and of adults of A. calandrae were obtained when the natural enemies were in association. The use of A. calandrae alone demonstrated a low instantaneous rate of increase (r(i)) of R. dominica and a high protection of the wheat grains. The association of A. calandrae with A. lacunatus led to the lowest number of immatures of R. dominica. These results demonstrate the importance of this interaction as a tool of for the integrated management of R. dominica in stored wheat grains.     

Field evaluation of spinosad as a grain protectant for stored wheat in Australia: efficacy against Rhyzopertha dominica (F.) and fate of residues in whole wheat and milling fractions

Abstract  The potential of spinosad as a grain protectant for the lesser grain borer, Rhyzopertha dominica , was investigated in a silo-scale trial on wheat stored in Victoria, Australia. Rhyzopertha dominica is a serious pest of stored grain, and its resistance to protectants and the fumigant phosphine is becoming more common. This trial follows earlier laboratory research showing that spinosad may be a useful pest management option for this species. Wheat (300 t) from the 2005 harvest was treated with spinosad 0.96 mg/kg plus chlorpyrifos-methyl 10 mg/kg in March 2006, and samples were collected at intervals during 7.5 month storage to determine efficacy and residues in wheat and milling fractions. Chlorpyrifos-methyl is already registered in Australia for control of several other pest species, and its low potency against R. dominica was confirmed in laboratory-treated wheat. Grain moisture content was stable at about 10%, but grain temperature ranged from 29.3°C in March to 14.0°C in August. Bioassays of all treated wheat samples over 7.5 months resulted in 100% adult mortality after 2 weeks exposure and no live progeny were produced. In addition, no live grain insects were detected during outload sampling after a 9 month storage. Spinosad and chlorpyrifos-methyl residues tended to decline during storage, and residues were higher in the bran layer than in either wholemeal or white flour. This field trial confirmed that spinosad was effective as a grain protectant targeting R. dominica .     

Substrate-selective activation of histidine-modified porcine pancreatic alpha-amylase by chloride ion.

Porcine pancreatic alpha-amylase (1,4-alpha-D-glucan glucanohydrolase) [EC 3.2.1.1] has both amylase activity (hydrolysis of alpha-1,4-D-glucoside bond of starch) and maltosidase activity (hydrolysis of p-nitrophenyl-alpha-D-maltoside to p-nitrophenol and maltose). By the modification of histidine residues of porcine pancreatic alpha-amylase with diethylpyrocarbonate (DEP), both amylase and maltosidase activities were decreased in the absence of chloride ion. In the presence of chloride ion, however, maltosidase activity of the modified enzyme was increased to more than 260% of that of the native enzyme, whereas amylase activity was decreased to less than 15% of the native enzyme. Since the chloride ion binding site is part of the active site loop [Buisson et al. (1987) Food Hydrocolloids 1,399-406 and Buisson et al. (1987) EMBO J. 6, 3909-3916], the special arrangements of both catalytic and modified histidine residues induced by the chloride ion binding would enhance only the maltosidase activity of the histidine-modified enzyme.     

石菖蒲根茎甲醇提取物对谷蠹的生物活性

采用冷浸法和溶剂萃取法,研究石菖蒲(Acorus gramineus Soland)根茎甲醇提取物对谷蠹Rhizopertha dominica(Fabricius)成虫的生物活性。结果表明,石菖蒲甲醇提取物以石油醚萃取物的生物活性最强,水层次之。石油醚萃取物对谷蠹具有较高的触杀、熏蒸、种群抑制活性和防治效果,并具有一定的驱避活性。药膜法触杀处理72h后对谷蠹的LC50和LC95分别为8.30μg/cm2,51.75μg/cm2。药纸熏蒸法6.25μL/L处理120h后对谷蠹的校正死亡率为94.25%。饲料拌药法250mg/kg处理对谷蠹的防效与5mg/kg马拉硫磷效果相当,且对F1代的种群抑制效果显著高于5mg/kg马拉硫磷处理。     

Cloning and characterization of alpha-amylase from Atlantic salmon (Salmo salar L.)

Amylase has a lower activity in carnivorous fish species, particularly in Atlantic salmon. We report the first cloning of a salmonid alpha-amylase cDNA from Atlantic salmon, a major species in aquaculture. By amino acid alignment of several species, we identified a seven amino acid deletion in one of the large loops of the enzyme in relatively close proximity to the active site, that could impair substrate binding. We also found the signal peptide to be less hydrophobic compared to other species. This may affect import into ER during protein synthesis. Active site residues were shown to be conserved. Amylase mRNA expression was shown in pancreatic tissue, liver, and in the heart. Using blocked p-nitrophenyl-maltoheptaoside as a substrate, we measured a low amylase activity in Atlantic salmon intestinal content, which was about half of the activity measured in Atlantic cod, whereas activity measured in rainbow trout was fourteen times higher. Amylase activities in all three species showed similar degree of reduction in hydrolytic activity in a dose-response trial with a wheat amylase inhibitor preparation. This indicates similar specific activity per amylase molecule.     

Effect of long-term application of manure and fertilizer on biological and biochemical activities in soil during crop development stages

A field experiment was conducted to investigate the effect of six long-term (34-year) fertilizer and farmyard manure (FYM) treatments (Control, N, NP, NPK, NPK+S, NPK+FYM) and three physiological stages of wheat growth on the microbial biomass carbon (MBC), nitrogen (MBN) and dehydrogenase, mineralizable N and phosphatase activities in soil. It was found that a balanced application of NPK+FYM gave the highest values for the measured parameters and lowest at the control. Values were generally highest at tillering, followed by the flowering and dough stages. A significant positive interaction between fertilizer treatments and physiological stages of wheat growth was observed, being highest at maximum tillering due to application of NPK+FYM. Stepwise regressions have revealed that grain yield of wheat was significantly associated with mineralizable N at tillering (R(2)=0.80), MBC at flowering (R(2)=0.90) and alkaline phosphatase activity (R(2)=0.70) at dough stages of wheat growth.     

Influence of growing location and cultivar on Rhyzopertha dominica （Coleoptera： Bostrichidae） and Sitophilus oryzae （Coleoptera： Curculionidae） infestation of rough rice

Long-grain rice cultivars Cocodrie, Wells, and XP 723 grown in three locations （Hazen, MO; Essex and Newport, AR, USA）, and medium-grain rice cultivars Bengal and XP 713 grown in two locations （Jonesboro and Lodge Corner, AR, USA）, were harvested and assayed for susceptibility to Rhyzopertha dominica （F.） （Coleoptera： Bostrichidae）, the lesser grain borer, and Sitophilus oryzae （L.） （Coleoptera： Curculionidae）, the rice weevil, on rice held at 27℃, 57% and 75% relative humidity （RH）. Separate samples from the same harvest lots were also analyzed for the physical characteristics of brown rice yield, percentage whole kernels and kernel thickness. Progeny production and feeding damage of R. dominica were significantly different among long-grain cultivars within two of the three locations （P 〈 0.05）, but not for location or RH （P ≥ 0.05）, while progeny production of S. oryzae was different among cultivars, location, and RH （P 〈 0.05）. On medium-grain rice, both cultivar and location were significant for progeny production of R. dominica, but not RH, while cultivar and RH were significant for progeny production of S. oryzae, but not location. On both rice types, feeding damage of R. dominica followed the same trends and was always strongly positively correlated with progeny production （P 〈 0.05）, but for S. oryzae there were several instances in which progeny production was not correlated with feeding damage （P ≥ 0.05）. Physical characteristics of both rice types were statistically significant （P 〈 0.01） but actual numerical differences were extremely small, and were generally not correlated with progeny production of either species. Results indicate that the location in which a particular rice cultivar is grown, along with its characteristics, could affect susceptibility of the rice to R. dominica and S. oryzae.