Application of a Molecular Method for the Identification of a Gigaspora margarita Isolate Released in a Field

A molecular technique for the identification of the Gigaspora margarita isolate CK based on the detection of a DNA sequence of 235 bp as its diagnostic marker was evaluated to investigate the survival and establishment of introduced arbuscular mycorrizal fungi (AMF) in a field ecosystem. In March 2001, roots and rhizosphere soil of Eragrostis curvula and Miscanthus sinensis were collected from the Mizunashi River at Mt. Fugendake (Nagasaki Prefecture, Japan), where plant seeds and AMF including G. margarita CK had been introduced for reforestation after the occurrence of repeated pyroclastic flows. We detected the marker sequence from DNA preparations of E. curvula roots and Gigaspora spores in the rhizosphere. This clearly showed that the isolate occurred at both hyphal and sporal stages. It was shown that the isolate survived and developed a life cycle in the revegetation area for 4 years. It was confirmed that the method was effective for tracing the isolate in samples collected from field ecosystems.     

Distribution and characterization of enzymes causing starch degradation in rice (Oryza sativa cv. koshihikari)

The thermal dependency and stability of enzymes producing reducing sugar (RS) were examined in bran, the exterior 13% part (outer endosperm), and the remaining inner endosperm of rice grains. RS-producing enzymes in the inner endosperm showed a higher optimum temperature than those in other parts of the rice grain. Diethylaminoethyl-Sephacel chromatography of crude extracts revealed two peaks of RS-producing activity with different optimum temperatures (60 and 37 degrees C) in all three parts. alpha-Glucosidase (EC 3.2.1.20) and alpha-amylase (EC 3.2.1.1) isoform G were thought to be major components of the RS-producing activities with high and low optimum temperatures, respectively. The peak with a high optimum temperature was a more abundant component in the inner endosperm, compared with other parts of the rice grain. Thus, different parts of rice were found to have distinct enzyme sets having different thermal dependency and to be involved in starch degradation to various sugars.     

Production and analysis of reciprocal hybrids between <Emphasis Type="Italic">Asparagus officinalis</Emphasis> L. and <Emphasis Type="Italic">A. schoberioides</Emphasis> Kunth

The genus Asparagus (Asparagaceae) encompasses 100–300 species, including the important vegetable crop, A. officinalis (garden asparagus). Previous attempts to hybridize garden asparagus with A. densiflorus (Kunth) Jessop, with the aim of introducing disease resistance, were unsuccessful because of the failure of endosperm development. In the present study, reciprocal interspecific hybrids between garden asparagus and A. schoberioides Kunth were generated by hand pollination. The F1 hybrids were analyzed by using both morphological and molecular techniques. This is the first report describing the production of an interspecific hybrid between garden asparagus at the diploid level (2n = 2x = 20) and its diploid (2n = 2x = 20) wild relative, A. schoberioides. Morphological characteristics of candidate hybrids were a mixture of those found in the parents, and cytological and RFLP analyses confirmed that morphologically intermediate plants were indeed diploid hybrids of those two species. In other words, post-zygotic isolation is not complete between phylogenetically distinct these two species. Our results suggest that other Asparagus species may be capable of hybridizing with A. officinalis and that introducing wild characters through interspecific hybridization may offer advantages for breeding for novel traits. Takuro Ito and Toshinori Ochiai authors are contributed equally to this work     

Structural identification of new curcumin dimers and their contribution to the antioxidant mechanism of curcumin

As a part of the research project on the elucidation of the chain-breaking antioxidant mechanism of natural phenolics against the oxidation of food components, curcumin, a main turmeric pigment, was investigated. A relatively high concentration of curcumin gave three dimers as radical termination products in addition to the coupling products with curcumin and the lipid hydroperoxide. The structural analysis of these dimers and quantitative analysis of their production rates revealed that radical-radical termination mainly occurred at the 2-position of curcumin. The contribution of the pathway for production of these dimers to the antioxidant mechanism of curcumin was estimated from the concentration-dependent data of the antioxidant activity and formation rates of these termination products. The A-A termination (dimer formation) was estimated to contribute at least about 40% of the entire antioxidant process against ethyl linoleate oxidation.     

Determination of carbamate pesticide residues in vegetables and fruits by liquid chromatography-atmospheric pressure photoionization-mass spectrometry and atmospheric pressure chemical ionization-mass spectrometry

A liquid chromatography-atmospheric pressure photoionization (APPI)-mass spectrometry method was developed for the determination of 22 carbamates including their metabolites in vegetables and fruits. For the optimization of APPI, several APPI ion source parameters were examined. As a result, many carbamates with APPI using the optimized parameter gave simple mass spectra, and a strong signal corresponding to [M + H](+) was observed except for aldicarb. However, some carbamate metabolites gave ammonium adduct ions [M + NH(4)](+) as base peak ions. The mean recovery of each carbamate from grape and onion samples spiked at 5 ng/g was 81.7-105.7%, with relative standard deviations of 3.3-5.9%. Furthermore, matrix constituents did not significantly influence the ionization efficiency. The limit of detection (S/N = 3) in grape and onion was in the range of 0.33-3.33 ng/g. For the robustness of this method, this system has been used to analyze 50 samples, and the intensities for all carbamates were found to be unaffected by the contamination of the APPI source by sample matrix constituents. This result indicates that the method is reliable.     

Antioxidative and anti-glycation activity of garcinol from Garcinia indica fruit rind

Garcinol, a polyisoprenylated benzophenone derivative, was purified from Garcinia indica fruit rind, and its antioxidative activity, chelating activity, free radical scavenging activity, and anti-glycation activity were studied. Garcinol exhibited moderate antioxidative activity in the micellar linoleic acid peroxidation system and also exhibited chelating activity at almost the same level as citrate. It also showed nearly 3 times greater DPPH (1, 1-diphenyl-2-picrylhydrazyl) free radical scavenging activity than DL-alpha-tocopherol by weight in aqueous ethanol solution. In a phenazine methosulfate/NADH-nitroblue tetrazolium system, garcinol exhibited superoxide anion scavenging activity and suppressed protein glycation in a bovine serum albumin/fructose system. Thus, garcinol might be beneficial as a potent antioxidant and a glycation inhibitor under specified conditions.     

Structure and bioactivity of thiosulfinates resulting from suppression of lachrymatory factor synthase in onion

In normal onion (Allium cepa), trans-S-1-propenyl-L-cysteine sulfoxide is transformed via 1-propenesulfenic acid into propanethial S-oxide, a lachrymatory factor, through successive reactions catalyzed by alliinase and lachrymatory factor synthase (LFS). A recent report showed that suppression of the LFS activity caused a dramatic increase in thiosulfinates previously reported as "zwiebelane isomers". After purification by recycle high-performance liquid chromatography and subsequent analyses, we established the planar structure of the putative "zwiebelane isomers" as S-3,4-dimethyl-5-hydroxythiolane-2-yl 1-propenethiosulfinate, in which two of the three molecules of 1-propenesulfenic acid involved in the formation gave the thiolane backbone, and the third molecule gave the thiosulfinate structure. Of at least three stereoisomers observed, one in the (2'R,3'R,4'R,5'R)-configuration was collected as an isolated fraction, and the other isomers were collected as a combined fraction because spontaneous tautomerization prevented further purification. Both fractions showed inhibitory activities against cyclooxygenase-1 and α-glucosidase in vitro.     

Reduction of deoxynivalenol contaminating corn silage by short-term ultraviolet irradiation: a pilot study

We evaluated the effects of short-term (up to 60 min) irradiation of corn silage with ultraviolet (UV) light (intensity: 1.5 mW/cm(2) at 254 nm UV-C wavelength), along with constant stirring of the silage, on the concentration of deoxynivalenol (DON), a major feed-contaminating mycotoxin, and those of α-tocopherol (vitamin E) and β-carotene (pro-vitamin A). The initial DON concentration in artificially contaminated silage was set at approximately 60 μg/g dry silage weight. After irradiation, the level of DON was decreased significantly (P<0.05) by approximately 13 μg/g (22%) on average at 30 min, and by 12 μg/g (21%) at 60 min. However, the concentrations of the vitamins remained relatively unaffected. Although further improvement is needed, short-term UV irradiation seems a promising on-farm method for reducing the level of DON in feedstuffs.     

Reproductive losses associated with porcine circovirus type 2 in a Japanese herd of seronegative sows

From April to July 2009, there was a high rate of reproductive losses in a 30-sow, farrow-to-finish swine herd in Yamagata Prefecture, Japan. Histopathological examinations of heart tissue from stillborn and preweaning piglets showed nonsuppurative, necrotizing lesions. Immunohistochemical staining and polymerase chain reaction analysis of myocardial lesions revealed the presence of porcine circovirus type 2 (PCV2) antigens and DNA in these tissues. Indirect immunofluorescence also showed that the PCV2 antibody positive rate in the sows was higher in May 2009 than in December 2008. The results of this study suggest that PCV2 spread to this farm and caused a high rate of reproductive losses.