Biological evaluation of neopeptins isolated from a Streptomyces strain

BACKGROUND: Microbial secondary metabolites are a rich source of antifungal agents and have merit as alternatives to synthetic fungicides. To develop disease control agents against powdery mildew, the lipopeptide antibiotic neopeptins were identified from the culture broth of a Streptomyces sp., and in vivo control efficacy of the compounds was evaluated on cucumber plants under glasshouse conditions. RESULTS: The Streptomyces sp. KNF2047 antagonistic against powdery mildew development in cucumber plants was isolated from a soil sample. Antifungal compounds were purified from the culture broth and identified as neopeptin A and B. In vitro microtitre assays revealed the inhibitory activities of the compounds in the range 128-512 microg mL(-1) against the mycelial growth of Alternaria mali, Botrytis cinerea, Cladosporium cucumerinum, Colletotrichum lagenarium, Didimella bryoniae and Magnaporthe grisea. Although neither compound showed remarkable in vitro antifungal activity against other plant pathogenic fungi, a mixture of neopeptins (484 mg of neopeptin A and 290 mg of neopeptin B per gram of partially purified powder) showed potent protective and curative activity against cucumber powdery mildew in vivo. The disease control activity of the neopeptins at a concentration of 2.4 mg L(-1) was 92.1%, which was similar to that of the commercial fungicide fenarimol (89.3% at 63 mg L(-1)) and that of the commercial biocontrol agent Actinovate (67.4% at 2 x 10(7) cfu L(-1)). CONCLUSION: Neopeptin mixtures isolated from Streptomyces sp. KNF2047 showed potent disease control activity against powdery mildew development on cucumber plants. .     

Isolation and antifungal activity of lignans from Myristica fragrans against various plant pathogenic fungi

BACKGROUND: In a search for plant extracts with potent in vivo antifungal activity against various plant diseases, we found that treatment with a methanol extract of Myristica fragrans Houttyn (nutmeg) seeds reduced the development of various plant diseases. The objectives of the present study were to isolate and determine antifungal substances from My. fragrans and to evaluate their antifungal activities. RESULTS: Three antifungal lignans were isolated from the methanol extract of My. fragrans seeds and identified as erythro-austrobailignan-6 (EA6), meso-dihydroguaiaretic acid (MDA) and nectandrin-B (NB). In vitro antimicrobial activity of the three lignans varied according to compound and target species. Alternaria alternata, Colletotrichum coccodes, C. gloeosporioides, Magnaporthe grisea, Agrobacterium tumefaciens, Acidovorax konjaci and Burkholderia glumae were relatively sensitive to the three lignans. In vivo, all three compounds effectively suppressed the development of rice blast and wheat leaf rust. In addition, EA6 and NB were highly active against the development of barley powdery mildew and tomato late blight, respectively. Both MDA and NB also moderately inhibited the development of rice sheath blight. CONCLUSION: This is the first study to demonstrate the in vitro and in vivo antifungal activities of the three lignans from My. fragrans against plant pathogenic fungi.     

Novel life stage targets against plum curculio, Conotrachelus nenuphar (Herbst), in apple integrated pest management

In this study the authors employed the plant-insect-chemistry (PIC) triad to investigate two novel life stage targets against the plum curculio (PC), Conotrachelus nenuphar (Herbst), in apple integrated pest management (IPM). Laboratory treated apple bioassays were used to determine if the insect growth regulator (IGR) insecticides novaluron and tebufenozide have physiological effects on PC larvae following adult exposure. Curative activity bioassays were conducted for IGR, neonicotinoid, oxidiazine and organophosphate insecticides on PC larvae post-infestation, and fruit penetration profiles of insecticides were developed. The results revealed that novaluron exhibits activity on PC larvae via vertical transmission following exposure of mated females to treated substrate. Surface treatments of azinphos-methyl, thiacloprid and thiamethoxam to preinfested fruit resulted in significant reductions in larval emergence. For all compounds, 50% or more of the total recovered active ingredient was from apple skin, and for azinphos-methyl, indoxacarb and thiamethoxam it was greater than 80%. For azinphos-methyl, novaluron, methoxyfenozide and thiacloprid, however, active ingredient was recovered from every section of the apple, from skin to core. Implications for twenty-first century IPM are discussed.     

Photoinduced generation of 2,3-butanedione from riboflavin

The volatile compound formation from riboflavin solution of a phosphate buffer (0.1 M, pH 6.5) under light for 15 h was studied by SPME-GC and SPME-GC/MS analysis. Only one major compound in the riboflavin solution was formed and increased as the light exposure time increased. The light-exposed riboflavin solution had a buttery odor. The compound of riboflavin solution under light was analyzed by gas chromatography and olfactometry. The major volatile compound eluted from the gas chromatograph had a buttery odor. The buttery odor compound was positively identified as 2,3-butanedione by a combination of gas chromatographic retention time, mass spectrum, and odor evaluation of authentic 2,3-butanedione. The addition of sodium azide, a singlet oxygen quencher, to riboflavin solution minimized the formation of the buttery odor compound. Singlet oxygen was involved in the formation of the buttery odor. The 2,3-butanedione was produced from the reaction between riboflavin and singlet oxygen. Singlet oxygen was formed from triplet oxygen by riboflavin photosensitization mechanism. This is the first reported oxidation reaction between riboflavin and singlet or triplet in food and biological systems.     

Evaluation of enhanced hygroscopicity, bifidogenicity, and anticariogenicity of enzymatically synthesized beta-galactosyl-trehalose oligosaccharides

beta-Galactosyl-trehalose oligosaccharides (beta-GTOs) were enzymatically prepared as a mixture of 6-beta-galactosyl-trehalose (1) and 4-beta-galactosyl-trehalose (2) with a 9:1 ratio (w/w). The beta-GTO mixture showed a highly enhanced hygroscopicity as compared to those of trehalose and other sugars used. At 72 h of incubation under 90% relative humidity and room temperature, it had a large increase in weight due to its moisture absorption, which was five times larger than that of trehalose, 1.9 times larger than that of sucrose, and 1.5 times larger than that of maltotriose. It was very effective in the growth promotion of Bifidobacteria, such as Bifidobacterium longum and Bifidobacterium bifidum, which was better than the growth promotion in the cases of trehalose and galactooligosaccharide. It also showed a highly anticariogenic property; it had only 10% cell proliferation of Streptococcus sobrinus for that of the sucrose control and 60% inhibition of insoluble glucan synthesis. Its effectiveness of inhibition was two and 1.5 times better than that of trehalose and one and two times than xylitol, respectively, against cell growth and glucan synthesis. Conclusively, the functionality of the beta-GTO in terms of hygroscopicity, bifidogenicity, and anticariogenicity was considerably improved as compared to that of trehalose. It is thus suggested that the beta-GTO might be applied as an effective humectant and prebiotic substitute with enhanced noncariogenicity in food applications.     

Simple purification (desalting) procedure to facilitate structural analysis of an alkali-solubilized/neutralized starch solution by intermediate-pressure size-exclusion chromatography

A technique was established to remove impurities (e.g., salts) from starch dissolved in strong alkali and neutralized with acid to accommodate starch structural analysis via intermediate-pressure size-exclusion chromatography (IPSEC). Starch (corn and wheat) subjected to an alkaline-microwave dissolution scheme (35 s microwave heating in a mixture of 6 M urea and 1 M KOH) was either treated with ion-exchange resin or passed through a desalting column to remove salt/urea contaminants. Control (untreated) starch solution analyzed by IPSEC displayed a significant interfering peak (attributable to salt/urea), which coeluted with the starch amylose peak. The interfering peak was most efficiently eliminated by first passing the starch solution through a desalting column, which process effectively removed impurities (e.g., salts/urea) without appearing to adversely impact the starch structural analysis. This simple technique coupled with the rapid alkaline-microwave starch dissolution procedure greatly expedites structural investigation of starch by facilitating analysis by IPSEC.     

Hypoglycemic effects of a phenolic acid fraction of rice bran and ferulic acid in C57BL/KsJ-db/db mice

Rice bran contains many phenolic acids, the most abundant of which is the antioxidant, ferulic acid (FA). We evaluated the hypoglycemic effects of a phenolic acid fraction (the ethyl acetate fraction, EAE) of rice bran and of FA in C57BL/KsJ db/db mice. Type 2 diabetic mice were allocated to a control group, an EAE group, or an FA group. Animals were fed a modified AIN-76 diet, and EAE or FA was administered orally for 17 days. There was no significant difference in body weight gain between groups. Administration of EAE and FA significantly decreased blood glucose levels and increased plasma insulin levels. EAE or FA groups had significantly elevated hepatic glycogen synthesis and glucokinase activity compared with the control group. Plasma total cholesterol and low density lipoprotein (LDL) cholesterol concentrations were significantly decreased by EAE and FA administration. These findings suggest that EAE and FA may be beneficial for treatment of type 2 diabetes because they regulate blood glucose levels by elevating glucokinase activity and production of glycogen in the liver.     

Structural identification of anthocyanins and analysis of concentrations during growth and flowering in buckwheat (Fagopyrum esculentum Moench) petals

The anthocyanin profiles and variety/breeding-line differences of anthocyanin concentrations in petals of common buckwheat flowers have been studied. Four anthocyanins, cyanidin 3-O-glucoside, cyanidin 3-O-rutinoside, cyanidin 3-O-rhamnoside, and cyanidin 3-O-galactosyl-rhamnoside were isolated from the petals of common buckwheat (Fagopyrum esculentum Moench), separated using high performance liquid chromatography and identified using reversed-phase liquid chromatography-electrospray ionization-tandem mass spectrometry techniques. In every variety/breeding line tested, cyanidin 3-O-rutinoside was detected as the major anthocyanin and the next is cyanidin 3-O-glucoside whereas cyanidin 3-O-rhamnoside and cyanidin 3-O-galactosyl-rhamnoside were trace or not detectable in white and pink flowered buckwheat. Of all the varieties/breeding lines tested, Gan-Chao, a Chinese variety, contained the highest amount of anthocyanins. The largest part of cyanidin moiety was presented as a proanthocyanidin form (PAs-Cy). Anthocyanins and PAs-Cy in petals were increased along with increase of flower development stages. Therefore, fully developed petals of red flowered buckwheat, especially Gan-Chao, are promising as a new anthocyanin-rich material for food processing.