Pea extracellular Cu/Zn-superoxide dismutase responsive to signal molecules from a fungal pathogen

We previously reported that the release of O₂ ⁻ from isolated pea cell walls was enhanced by a 70-kDa glycoprotein elicitor but was suppressed by mucin-type glycopeptide suppressors (supprescins A and B) prepared from pycnospore germination fluid of Mycosphaerella pinodes, causal agent of Mycosphaerella blight of pea. Here, we show that superoxide dismutase (SOD) in the apoplast fluid/cell wall of pea seedlings responds to the fungal elicitor and suppressor molecules. In a pharmacological study and with internal amino acid sequencing, the apoplastic SOD in a pea cultivar Midoriusui was found to be a Cu/Zn type SOD. We cloned a full-length cDNA of the Cu/Zn-SOD and designated it as PsCu/Zn-SOD1. An increase in PsCu/Zn-SOD1 mRNA and the PsCu/Zn-SOD1 protein was induced by treatment with the elicitor more intensively than by wounding. Such induction by the elicitor or wounding, however, was inhibited by the concomitant presence of supprescins. The SOD activity of recombinant PsCu/Zn-SOD1 was regulated directly by these signal molecules in a manner similar to their effect on the SOD activity in the apoplastic fluid and in the cell wall-bound proteins. Based on these findings, we discuss a role for PsCu/Zn-SOD1 in the pea defense response. The nucleotide sequence data of PsCu/Zn-SOD1 reported are available in the DDBJ/EMBL/GenBank databases under accession number AB189165.     

Cultivar identification of rice (Oryza sativa L.) by polymerase chain reaction method and its application to processed rice products

As the cultivars of rice markedly affect eating quality, processing suitability, and price, identification or differentiation of rice cultivar is very important. We developed suitable 14 STS (sequence-tagged site) primers for PCR (polymerase chain reaction), and it became possible to differentiate 60 Japanese dominant rice cultivars from each other using template DNA extracted and purified from rice grains. A multiplex primer set was shown to be useful to effectively differentiate rice cultivars produced in various countries by PCR. A novel multiplex primer set for PCR has been developed to differentiate KoshihikariBL, which is closely related with the premium cultivar, Koshihikari, in Japan. The application of the cultivar identification method by PCR method to commercially processed rice products was investigated. We developed an enzyme treatment method, in which the gelatinized starch is decomposed by the heat-stable alpha-amylase at 80 degrees C, followed by the hydrolysis of proteins by proteinase K with sodium dodecyl sulfate and purification of extracted DNAs by phenol/chloroform/iso-amyl alcohol. It became possible to identify the material rice cultivars of the commercially processed rice products, such as cooked rice, rice cake, or rice cracker, by a PCR method using template DNA prepared by the enzyme treatment method and novel multiplex primer sets.     

Oryzacystatin-II, a cystatin from rice (Oryza sativa L. japonica), is a dimeric protein: possible involvement of the interconversion between dimer and monomer in the regulation of the reactivity of oryzacystatin-II

We examined the biochemical and structural properties of oryzacystatin-II, a phytocystatin in rice (Oryza sativa L. japonica), under heat-stress conditions. The enzyme inhibitory reactivity of oryzacystatin-II was enhanced by heating in a temperature-dependent manner and reached a maximum level by heating at 65 degrees C for 10 min. Size-exclusion chromatography showed that oryzacystatin-II forms a homodimer at ambient temperature and that the enhancement of inhibitory reactivity is due to the conversion of the dimeric to a monomeric form. The monomeric form of oryzacystatin-II reverted to the dimer during storage at 4 degrees C, suggesting that dimerization is an intrinsic property of oryzacystatin-II. The affinity of the monomer for cysteine proteinases was significantly higher than that of the dimer. This is the first paper to describe the noncovalent dimerization for a cystatin under nonstress conditions.     

PCR method for the detection and identification of cultivars of rice flours used in yeast leavened breads containing both wheat and rice flours

The identification of cereal grain sources in yeast leavened breads is challenging because of the mixtures of DNA from yeast and mixtures of related grains. DNA is decomposed during the fermentation and bread consists of not only rice but also wheat, yeast, sugar, butter, shortening etc.     

Characteristics of pregelatinized ae mutant rice flours prepared by boiling after preroasting

As ae mutant rice, such as EM10, lacks the starch branching enzyme IIb, its amylopectin contains more long-chain glucans than that of ordinary Indica and Japonica rice grains. Although boiled grains of ae rice cultivars are too hard and nonsticky for table rice, they are promising in terms of biofunctionality, such as prevention of diabetes. The present paper investigates the characterization of a novel group of four ae mutant rice cultivars (EM72, EM145, EM174, and EM189). They were subjected to the evaluation for their main chemical components, physical properties, and enzyme activities at different grain conditions (raw milled rice, roasted rice, boiled rice, and rice boiled after preroasting). These mutant rice grains are characterized by high apparent amylose, high protein and high glucose contents, high pasting temperature, high α-amylase activities, high resistant starch, and low degree of gelatinization. A novel method was developed to maintain the high resistant starch contents of gelatinized rice grains. Rice boild after preroasting showed a higher ratio of resistant starch and a lower amount of glucose than ordinary boiled rice. It became possible to produce high-quality and biofunctional pregelatinized rice flours by boiling with frozen fruits, such as tomatoes, after rice grains had been preroasted. These ae mutants were found to be suitable materials for rice/fruit or rice/vegetable products to serve as palatable, low-glucose, and high resistant starch rice products.     

Water penetration into rice grains during soaking observed by gradient echo magnetic resonance imaging

The water penetration into a rice grain during soaking was monitored by three-dimensional gradient echo magnetic resonance imaging, which has the advantage of a short measurement time and a sufficient sensitivity for low-moisture samples. The water penetration route was compared between the milled and brown rice grains of two japonica cultivars. In normal milled rice grains of Koshihikari, water first penetrated the ventral side surface and the embryo attachment site of the endosperm, then migrated along the central line and transverse cracks, and finally diffused to all parts of the endosperm. In milled rice grains of Yamadanishiki with a white core, water quickly infiltrated into the cracks or chalky parts on the dorsiventral line and then diffused to the lateral side. In brown rice grains of both cultivars water penetrated extremely slowly and did not infiltrate directly into transverse cracks or the white core due to inhibition by the pericarp and seed coat. The central part of every grain allowed more water to penetrate. The route, pattern and speed of water penetration were determined by the morphological structure, crack formation and hardness distribution associated with the packing of the starch granules in the grain.     

Assessment of the water quality of two rivers in Hanoi City and its suitability for irrigation water

The To Lich and Kim Nguu Rivers in Hanoi City are the main sources of irrigation water for suburban agricultural land and fish farm. Wastewater from the industrial plants located along these rivers has been discharged, and has degraded the water quality of the rivers. This study describes the chemical properties of water from the rivers, focusing on heavy metal pollution and the suitability of water quality for irrigation water. Water from the rivers was heavily polluted with organic matter and heavy metals such as Pb, Cu, Zn, Cr, Cd and Ni. Dissolved oxygen, chemical oxygen deman, and total suspended solids, and the concentrations of all heavy metals exceeded the Vietnamese standard for surface water quality in all investigated sites. The concentrations of some heavy metals such as Cu, Cd, Cr and Ni were above the internationally recommended WHO maximum level for irrigation water. A wide variation in the heavy metal concentration of water due to metal types is the result of wastewater discharged from different industrial sources.     

Permeation of urea through various polyurethane membranes

BACKGROUND: Controlled‐release systems using polymer membranes are very important in agriculture for labour‐saving and effective delivery of pesticides and other agents. Polymer‐coated granules are one of the most useful formulations, and a study of the factors for polymer design is necessary to achieve various release patterns. A permeation study using plain membranes was carried out in order to clarify parameters, and the results were compared with the release from polymer‐coated granules. RESULTS: The permeation coefficient of urea through a plain polyurethane membrane decreased significantly as the urethane and alkyl side chain content increased. The glass transition temperature and crosslink density of the polyurethanes hardly influenced its permeability. The release rate from polyurethane‐coated granules was also reduced by alkyl side chains. However, it was faster than that through a plain membrane because of capsule expansion by continuous water penetration and structural changes in the membrane. CONCLUSION: The release rate of urea through a polyurethane plain membrane and from polyurethane‐coated granules can be controlled by changing the chemical properties of the membrane. In addition, physical properties such as the glass transition temperature T_g or crosslink density should be considered to assess the release profile from polyurethane‐coated granules. Copyright © 2009 Society of Chemical Industry     

Morphological and photosynthetic responses of Quercus crispula seedlings to high-light conditions

We investigated morphological and photosynthetic responses of current-year seedlings of oak (Quercus crispula Blume) under high-light conditions. Quercus crispula seedlings were grown from seed in a relative photosynthetically active photon flux density (RPPFD) of 100, 10 or 2%. There was no difference in total dry mass between 100 and 10% RPPFD. At the end of the growing season, plants grown in 2% RPPFD had a lower total dry mass than those grown in 100 or 10% RPPFD. Seedlings grown in 100% RPPFD showed morphological acclimation, i.e., high root/shoot ratios and high leaf mass per area. De-epoxidation level in the xanthophyll cycle and activity of an antioxidant enzyme were highest in 100% RPPFD, but total chlorophyll concentration and photosynthetic rate were highest in 10% RPPFD. These results indicate that excess photons were generated in 100% RPPFD, leading to increased capacities for dissipation of received light energy through the xanthophyll cycle and for scavenging of reactive oxygen species through the water-water cycle. Nevertheless, a midday decrease in dark-adapted quantum yield of photosystem II (F(v)/F(m)) indicated that seedlings grown in 100% RPPFD were suffering from photoinhibition. We conclude that Q. crispula current-year seedlings have high morphological acclimation to high light but that photosynthetic efficiency cannot be maintained under high-light conditions even with a photoprotection system.