PsoP1, a milk-clotting aspartic peptidase from the basidiomycete fungus Piptoporus soloniensis

The first enzyme of the basidiomycete Piptoporus soloniensis, a peptidase (PsoP1), was characterized after isolation from submerged cultures, purification by fractional precipitation, and preparative native-polyarylamide gel electrophoresis (PAGE). The native molecular mass of PsoP1 was 38 kDa with an isoelectric point of 3.9. Similar to chymosin from milk calves, PsoP1 showed a maximum milk-clotting activity (MCA) at 35-40 °C and was most stable at pH 6 and below 40 °C. The complete inhibition by pepstatin A identified this enzyme as an aspartic peptidase. Electrospray ionization-tandem MS showed an amino acid partial sequence that was more homologous to mammalian milk clotting peptidases than to the chymosin substitute from a fungal species, such as the Zygomycete Mucor miehei. According to sodium dodecyl sulfate-PAGE patterns, the peptidase cleaved κ-casein in a way similar to chymosin and hydrolyzed β-casein slowly, as it would be expected from an efficient chymosin substitute.     

Impact of cultivar, harvesting time, and seasonal variation on the content of biophenols in olive mill waste

Olive mill waste (OMW) contains substantial amounts of valuable antioxidant biophenols that can be recovered for possible applications in food, pharmaceutical, and cosmetic industries. However, the impact of cultivar, harvesting time, and seasonal variation on the phenolic composition of OMW has not yet been assessed. Total phenols, antioxidant activity, and phenol profiles of OMW extracts from five different Australian-grown cultivars (Barnea, Correggiola, Manzanillo, Mission, and Paragon) were studied at four different harvesting times in the 2004 season. The impact of seasonal variation was assessed by comparing total phenol content, antioxidant activity, and phenol profile of two cultivars (Correggiola and Mission) harvested in the 2004 and 2005 seasons. The phenol content and antioxidant activity at different harvesting times were mainly a function of the olive cultivar. Harvesting time had a quantitative effect rather than a qualitative effect on the phenol profile. Intercultivar and harvesting time variation accounted for a 2-5-fold change in the total phenol and antioxidant capacity, while levels of individual biophenols experienced up to 50-fold change. The phenol content and antioxidant capacity of OMW significantly changed between seasons with different variation patterns for different cultivars.     

Thiamine (vitamin B1) deficiency in germinating seeds of Phaseolus vulgaris L. exposed to soaking injury

Exogenous application of thiamine (vitamin B1) during imbibition improved germination and seedling development of bean (Phaseolus vulgaris L. cv. Lasso) seeds, which were exposed to soaking injury by submergence. Leaching of the vitamin into the incubation medium was not increased in submerged seeds. However, translocation of the thiamine reserves from the cotyledons to the seedling axis was reduced in response to soaking injury. After exogenous application of free thiamine to the seeds, thiaminedi-phosphate (TDP), which is the physiologically active coenzyme form of thiamine, accumulated in the seedling axis of submerged seeds, suggesting an increased demand for thiamine-dependent metabolic reactions in these tissues. Limited oxygen supply of the seeds during the soaking period induces a shift from respiration to fermentation of carbohydrates. Neither thiamine-dependent ethanolic fermentation, nor ATP production or adenylate energy charge (AEC) of the seedling axis were affected by exogenous thiamine application, suggesting that there is no limitation of thiamine-dependent reactions in the energy metabolism of the seedlings. Thus the physiological mechanisms improving germination and seedling development of submerged seeds in response to thiamine seed treatment are still not clear.     

Evaluation of sunflower (Helianthus annuus L.) single cross hybrids under heat stress condition

Sunflower is an important oilseed crop, which shows susceptibility to heat stress. In this study, 63 single cross hybrids were evaluated under heat stress condition for 2 years and compared with the two commercial hybrids. Genotype and genotype × environment (GGE) was used to differentiate single cross hybrids on the basis of multiple traits. GGE biplot showed that several single cross hybrids had higher seed yield potential than standard check. Moreover, seed yield per plant (SYP) was related to pollen viability percentage, showing that achene yield was the product of high gametophytic fertility under heat stress. Hybrids having high seed yield potential under heat stress had lower cell membrane injury. GGE biplot for SYP and its components showed that single cross hybrids were characterized into two major groups. Group I was further characterized into two sub group. Group Ia included hybrids with high 100-SW, while group Ib had the hybrids with high number of seeds per head and head diameter. Group II had the hybrids with high kernel weight and kernel to seed ratio. The hybrids could be recommended according to their potential utilization in the seed industry.     

Performance of Anaerobic Biotrickling Filter and Its Microbial Diversity for the Removal of Stripped Disinfection By-products

The objective of this research was to evaluate the biodegradation of chloroform by using biotrickling filter (BTF) and determining the dominant bacteria responsible for the degradation. The research was conducted in three phases under anaerobic condition, namely, in the presence of co-metabolite (phase I), in the presence of co-metabolite and surfactant (phase II), and in the presence of surfactant but no co-metabolite (phase III). The results showed that the presence of ethanol as a co-metabolite provided 49% removal efficiency. The equivalent elimination capacity (EC) was 0.13 g/(m³ h). The addition of Tomadol 25-7 as a surfactant in the nutrient solution increased the removal efficiency of chloroform to 64% with corresponding EC of 0.17 g/(m³ h). This research also investigated the overall microbial ecology of the BTF utilizing culture-independent gene sequencing alignment of the 16S rRNA allowing identification of isolated species. Taxonomical composition revealed the abundance of betaproteobacteria and deltaproteobacteria with species level of 97%. Azospira oryzae (formally dechlorosoma suillum), Azospira restrica, and Geobacter spp. together with other similar groups were the most valuable bacteria for the degradation of chloroform.     

Impact of 12-year field treatments with organic and inorganic fertilizers on crop productivity and mycorrhizal community structure

The effect of manure and mineral fertilization on the arbuscular mycorrhizal (AM) fungal community structure of sunflower (Helianthus annuus L.) plants was studied. Soils were collected from a field experiment treated for 12 years with equivalent nitrogen (N) doses of inorganic N, dairy manure slurry, or without N fertilization. Fresh roots of tall fescue (Festuca arundinacea Schreb.) grass collected from the field plots without N fertilization and unfumigated field soils were used as native microbial inoculum sources. Sunflower plants were sown in pots containing these soils, and three different means of manipulating the microbial community were set: unfumigated soil with fresh grass roots, fumigated soil with fresh grass roots, or fumigated soil with sterilized grass roots. Assessing the implications with respect to plant productivity and mycorrhizal community structure was investigated. Twelve AM fungal OTUs were identified from root or soil samples as different taxa of Acaulospora, Claroideoglomus, Funneliformis, Rhizophagus, and uncultured Glomus, using PCR-DGGE and sequencing of an 18S rRNA gene fragment. Sunflower plants grown in manure-fertilized soils had a distinct AMF community structure from plants either fertilized with mineral N or unfertilized, with an abundance of Rhizophagus intraradices-like (B2). The results also showed that AM inoculation increased P and N contents in inorganic N-fertilized or unfertilized plants, but not in manure-fertilized plants.     

Sulfur Alleviates Growth Inhibition and Oxidative Stress Caused by Cadmium Toxicity in Rice

A hydroponic experiment was conducted to investigate the effect of sulfur (S) on growth inhibition and oxidative stress caused by Cd²⁺ toxicity, using two rice cultivars with different grain Cd²⁺ content. Treatments consisted of factorial arrangement of three S levels (0.2, 0.4, and 0.8 mmol), two cadmium (Cd) levels (0 and 1 μ mol), and two rice cultivars (‘Bing 97252,’ a cultivar with low grain Cd²⁺ content, and ‘Xiushui 63,’ a cultivar with high grain Cd²⁺ content). The results showed that Cd²⁺ addition in the medium generally increased Cd²⁺ and malondialdehyde (MDA) content in both roots and shoots; the increases were more pronounced in ‘Xuishui 63’ than in ‘Bing 97252.’ Dramatic reductions in growth parameters, including plant height, root and shoot weight, tillers per plant, chlorophyll content, and net photosynthetic rate were found in the plants exposed to Cd stress relative to the plants without Cd²⁺ treatment. ‘Xiushui 63’ showed more sensitivity than ‘Bing 97252’ under Cd²⁺ exposure. In comparison with the lower S level (0.2 mmol), the higher S levels (0.4 and 0.6 mmol) helped alleviate Cd toxicity, characterized by a significant increase in growth parameters, and a decrease in Cd²⁺ and MDA content in both roots and shoots. In addition, superoxide dismutase (SOD) activity in the plants varied among tissues, cultivars, and Cd treatments. High Cd²⁺ and MDA content was consistently accompanied by higher SOD activity, and higher S levels caused a marked increase in glutathione content and a reduction in SOD activity, indicating a positive effect of S in alleviating oxidative stress.     

Taurine is more effective than melatonin on cytochrome P450 2E1 and some oxidative stress markers in streptozotocin-induced diabetic rats

Melatonin and taurine have alleviative effects in streptozotocin (STZ)-induced diabetic rats. Male Wistar rats were divided into nondiabetic, diabetic, diabetic melatonin supplemented and diabetic taurine supplemented groups. At the end of the study, both blood and liver were collected for determination of some oxidative stress parameters, and hepatic cytochrome P450 2E1 (CYP2E1) enzyme activity and gene expression. An increased CYP2E1 activity and expression level with a concomitant significant change in oxidative stress parameters were found in STZ-induced diabetic rats. Taurine or melatonin supplementation to the diabetic rats alleviated these experimental parameters with a more significant effect for taurine than that of melatonin. Suppression of β-hydroxybutyrate (β-HB) production by taurine can be one of the mechanisms of a reduction in CYP2E1. Taurine was effective more than melatonin in reducing CYP2E1 activity and expression; therefore antioxidants might prove beneficial in type 1 diabetes associated with manifestations of liver injury.     

棉秆切碎及压缩成型的试验研究

采用３种不同切碎方法,即锤片切碎、螺旋刀切碎和直刃刀切碎进行了棉秆切碎的性能研究。试验结果表明,直刃刀对青棉秆的一次切碎,能获得细小颗粒产量高、能耗低的满意效果。切碎物料在英国Ｓｈｉｍａｄａ螺旋压缩机上进行热压成型试验。成型燃料质量指标的测定结果表明,切碎物料含水率在８％～１２％,加热温度在１２０～１５０℃,最大粒度控制在１２．５ｍｍ以下,能获得密度为１．１３～１．２１ｇ／ｃｍ^３、低位热值为１８．３４ＭＪ／ｋｇ的优质棉秆成型燃料。     

Thiamine (vitamin B1) seed treatment enhances germination and seedling growth of bean (Phaseolus vulgaris L.) exposed to soaking injury

Seed treatment with thiamine mononitrate significantly increased germination rate of bean (Phasenius vulgaris L.) seedlings germinated at high soil moisture levels and low night-temperature (5° C). Beneficial effects of thiamine applications on germination were also observed in soil-free germination tests when the seeds were submerged in water for 1–4 d at 18°C. Optimum response was achieved at thiamine concentrations of 3 mM, applied during the first 24 h of imbibition. As an early response to the thiamine treatment, vital staining with triphenyltetrazolium chloride revealed a lower proportion of damaged tissue of the embryonic axes. In later stages of seedling development, thiamine seed treatment resulted in a lower proportion of abnormal seedlings with malformations of the shoot apex and of the primary leaves. The beneficial effect of thiamine was more pronounced when the temperature during submergence was kept at 25° C compared to 5° C. There was no response to thiamine applications when the germinating seeds were exposed to low temperature treatments only. The thiamine effect could be mimicked by raising the osmotic potential of the incubation medium, indicating a relationship to soaking injuries. The results suggest that normal seedling development may be impaired by thiamine deficiency in the embryonic tissue induced by soaking injury, whereas low temperatures are not involved as a stress-factor.