Ultrastructural Localization of Hydrogen Peroxide in Host Leaves Treated with AK-toxin I Produced by Alternaria alternata Japanese Pear Pathotype

The rapid generation of reactive oxygen species (ROS), called the oxidative burst, is one of the earliest host responses to pathogen infection or elicitor treatments. Therefore, we looked for the induction of ROS generation in Japanese pear leaves by the host-specific toxin, AK-toxin I using a cytochemical method for detecting H₂O₂. A small amount of non-specific generation of H₂O₂ was found in the cell walls in toxin- and water-treated susceptible and resistant leaves. Thus, the generation of H₂O₂ at cell walls appears to be caused by wounding stress during sampling. Specific generation of ROS, however, was found only in the membrane fragments and extended desmotubules characteristic of modified sites of the plasma membrane in the toxin-treated susceptible leaves. In addition, generation of H₂O₂ at plasma membranes was observed with higher frequency in toxin-treated susceptible leaves. This result indicates that the H₂O₂ generation was associated with damaged sites in the plasmalemma after toxin treatment and perhaps with the formation of membrane fragments from altered portions of the invaginated plasma membrane. Received 21 September 2001/ Accepted in revised form 25 October 2001     

Lipid-rich bovine serum albumin improves the viability and hatching ability of porcine blastocysts produced in vitro

The effects of lipid-rich bovine serum albumin (LR-BSA) on the development of porcine blastocysts produced in vitro were examined. Addition of 0.5 to 5 mg/ml LR-BSA to porcine blastocyst medium (PBM) from Day 5 (Day 0 = in vitro fertilization) significantly increased the hatching rates of blastocysts on Day 7 and the total cell numbers in Day-7 blastocysts. When Day-5 blastocysts were cultured with PBM alone, PBM containing LR-BSA, recombinant human serum albumin or fatty acid-free BSA, addition of LR-BSA significantly enhanced hatching rates and the cell number in blastocysts that survived compared with other treatments. The diameter, ATP content and numbers of both inner cell mass and total cells in Day-6 and Day-7 blastocysts cultured with PBM containing LR-BSA were significantly higher than in blastocysts cultured with PBM alone, whereas LR-BSA had no effect on mitochondrial membrane potential. The mRNA levels of enzymes involved in fatty acid metabolism and β-oxidation (ACSL1, ACSL3, CPT1, CPT2 and KAT) in Day-7 blastocysts were significantly upregulated by the addition of LR-BSA. The results indicated that LR-BSA enhanced hatching ability and quality of porcine blastocysts produced in vitro, as determined by ATP content, blastocyst diameter and expression levels of the specific genes, suggesting that the stimulatory effects of LR-BSA arise from lipids bound to albumin.     

生物质资源高压连续输送的研究

建立了近中试规模的生物质资源高压连续输送装置，通过将生物质资源与水混成浆料的方法实现了生物质资源的高压连续输送、浆料的固含量可达10％（wt)以上，压力可达40MPa，最大浆料供应量为2．4L／h。     

Effects of acid extracts of tomato (Lycopersicon esculentum Mill.) and carrot (Daucus carota L.)wastes from the food industry on the growth of some crops and weeds

The effects of acid extracts of tomato ( Lycopersicon esculentum Mill.) and carrot ( Daucus carota L.) juice wastes from the food industry on the growth of five species of crops and four weeds were examined. The acid extract of the tomato juice waste promoted the shoot and root growth of tomato, Chinese cabbage, corn, and radish, but not the growth of oat seedlings. The extract did not promote the shoot growth of weeds tested, and it inhibited the root growth of the weeds that included barnyardgrass ( Echinochloa oryzicola Ohwi), southern crabgrass ( Digitaria ciliaris Koel.) and Japanese barnyard millet ( Echinochloa utilis Ohwi). The carrot juice waste was also examined as another waste from the food industry. The effect of the acid extract of carrot juice waste showed significant promoting effects on the root growth of Chinese cabbage; however, the effects on other plants were lower than that of tomato juice waste. It also did not inhibit the growth of weeds. These results suggested that the acid extract from tomato juice waste is useful as plant-growth substances because they have a promoting effect on the shoot and root growth of crops, and an inhibitory effect on the root growth of some weeds.     

Need for flagella for complete virulence of Pseudomonas syringae pv. tabaci: genetic analysis with flagella-defective mutants ?fliC and ?fliD in host tobacco plants

The flagellins purified from Pseudomonas syringae pv. tabaci induce a hypersensitive reaction in nonhost tomato cells. To investigate the role of flagella and flagellin in the compatible interaction, we generated two types of flagella-defective mutant. The fliC mutant lost the fliC gene that encodes flagellin protein, whereas the fliD mutant lost the fliD gene that encodes HAP2-capping protein. The two mutants had markedly reduced ability to cause disease symptoms in tobacco leaves. Furthermore, propagation of the mutants in tobacco leaves was less than that in wild-type pv. tabaci. Compared to the inoculation with wild-type pv. tabaci, inoculation with the two mutants did not markedly induce the expression of typical defense response-related genes such as PAL and hsr203J. Complementation of each fliC and fliD gene to the corresponding deficient mutant restored motility and virulence. These results indicate that flagella of P. syringae pv. tabaci are indispensable organelles for complete virulence on host tobacco plants.     

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.     

Comparison of homogeneity and heterogeneity of residual variance using random regression test-day models for first lactation Japanese Holstein cows

Using a large‐scale data set that included first lactation test day records from 1975 to 2000 for Japanese Holsteins, genetic parameters for milk yield were estimated by using random regression (RR) test‐day models (TDM) with heterogeneous and homogeneous residual variances. It is necessary for the RR‐TDM to include a function that explains the shape of the lactation curve. The RR‐TDM with the LW curve, which combined Wilmink's curve and a Legendre polynomial, was used for fitting the model for milk yield. In recent years, increases in residual variance have been noted for Japanese dairy cattle. Thus, three kinds of heterogeneous residual variance over the calving year were considered: H1, H2 and HG. Linear and quadratic exponential functions for the calving year were used in H1 and H2, respectively. Residual variance of HG was divided into five groups according to calving year. Homogeneous residual variance was HO. All heterogeneous residual variances increased with calving year in an almost linear fashion. Residual variance increased over the study period. However, there is no need to consider heterogeneous residual variances in genetic evaluations, because the heterogeneity of residual variance over the years did not affect the ranking of top sires and cows.     

Influence of feeding regime on timing of parturition in beef cattle and the relationship of vaginal temperature to parturition

The timing of parturition was recorded for a total of 56 beef cattle (Japanese Black × Holstein Friesian) on different dietary treatments. The rate of calving during daylight hours in cows night‐fed (18.00 hours) with a roughage diet was significantly higher than that in cows night‐fed with a high concentrate diet (79.2% vs 38.5%, P < 0.05). Subsequently, the vaginal temperature (VT) of these cows was analyzed using a cosinor method. When the feeding schedule was changed from twice daily (08.30 and 15.30 hours) to night feeding, the periodicity, the acrophase and the bathyphase, which were the parameters of the cosine curve, were unstable from the first day of night feeding until after day 6 (P < 0.05). Prior to parturition, the midline‐estimating statistic of rhythm (MESOR) and the amplitude for the cows that were fed a high‐roughage diet at night and that calved at night‐time were lower and larger, respectively, than that for the other treatments (P < 0.01). Based on these results, the time of parturition in most of the beef cows was influenced by feeding time and diet composition. Those cows that calved at night‐time in spite of night feeding had lower vaginal temperatures.