Accumulation of gamma-aminobutyrate (GABA) caused by heat-drying and expression of related genes in immature vegetable soybean (edamame)

We studied the effects of drying of immature seeds of vegetable soybean (Glycine max L. Merrill) on the accumulation of gamma-aminobutyrate (GABA) in the seeds. GABA accumulated after heat-drying, with the maximum at 40°C. The GABA content (447.5 mg/100 g DW) increased to more than 5 times the value in untreated seeds (79.6 mg/100 g DW). In contrast, the glutamate content decreased rapidly to 1/3 the level in the untreated seeds. The GABA content increased early in the heat-drying treatment: after 30 min, it had increased to 1.5 times the value in the untreated seeds. GABA did not accumulate in the vacuum-drying treatment. Among genes related to the GABA shunt, the gene for glutamate decarboxylase (EC 4.1.1.15), which catalyzes the decarboxylation of glutamate to produce GABA, showed relatively high expression, decreasing to only 70% of the value in untreated seeds even after 4 h of treatment. In contrast, expression of the genes for two catabolic mitochondrial enzymes, GABA transaminase (GABA-T; EC 2.6.1.19) and succinate semialdehyde dehydrogenase (SSADH; EC 1.2.1.16), decreased rapidly during heat-drying. These results suggest that the accumulated GABA was not metabolized rapidly by GABA-T and SSADH and therefore remained at high levels.     

Effect of enzymes of microbial origin on in vitro digestibilities of dry matter and crude protein in soybean meal

The effects of supplementation with cellulase, xylanase, pectinase, hemicellulase, glucanase, phytase and protease of microbial origin on digestibility of crude protein (CP) and dry matter (DM) of soybean meal in vitro were examined in the present study. Changes in viscosity during in vitro digestion were also examined as an index of carbohydrate digestibility. Hemicellulase and all enzyme combinations without protease showed significant improvement of CP digestibility. Cellulase, xylanase, phytase and glucanase tended to improve CP digestibility. Dry matter digestibility was improved significantly by pectinase and all enzyme combinations. Cellulase and phytase tended to improve DM digestibility. Crude protein and DM digestibilities were the highest when all the enzymes except protease were added. Protease, cellulase, pectinase, xylanase, glucanase and hemicellulase significantly reduced viscosity, while phytase had no effect on viscosity. Viscosity was increased unexpectedly when all the enzymes were added. These results strongly suggest that a combination of carbohydrases improves the CP and DM digestibility of soybean meal while protease inhibits the actions of these enzymes. The present study also suggests that viscosity is not always a good index of digestibility. Moreover, excluding the protease from commercial crude enzyme preparations may improve digestibility.     

Perchloric acid treatment and use of chromogenic substrate in the Limulus test: Application to veterinary diagnosis

The Limulus test was carried out for endotoxin determination using a synthetic chromogenic substrate after perchloric acid(PCA) treatment to remove inhibiting substances in the blood.PCA treatment completely removed non-specific amidolytic activity and other factors which interfered with the Limulus test. The recovery of added endotoxin was about 100% irrespective of animal species.Endotoxin levels in PCA-treated blood plasma of healthy domestic animals were invariably below 1 pg ml^?1.Many of the cows with gangrenous mastitis, from which Gram-negative rods were isolated, showed high endotoxin levels ranging from 22.8 to 138.0 pg ml^?1. Endotoxin was also detected in the blood of cows affected with salmonellosis, colibacillosis, Haemophilus somnus infection and urethritis associated with Proteus.     

Number of nitrate- and nitrite-reducing Selenomonas ruminantium in the rumen, and possible factors affecting its growth

The cell number of Selenomonas ruminantium (S. ruminantium) that reduces nitrate and nitrite in the rumen was usually 8–10% of the total number of S. ruminantium (an order of 10⁶/mL). The percentage was not affected by the roughage/concentrate ratio or nitrate content of the diet in 2 weeks. However, feeding a high‐nitrate diet for 12 weeks increased the percentage. The percentage of lactate‐using S. ruminantium, such as the ssp. lactilytica, was less than 1% of the total number of S. ruminantium. No S. ruminantium was found that used formate as an electron donor for nitrate and nitrite reduction. Lactate and H₂ appeared to be important for nitrate and nitrite reduction by S. ruminantium. Nitrate reduction by S. ruminantium was enhanced by the coexistence of amylolytic bacteria in a medium containing starch, and as a result, nitrite accumulation increased. Coexistence of cellulolytic bacteria facilitated the growth of S. ruminantium in a medium containing cellulose, and consequently increased nitrite reduction. In order to suppress nitrite accumulation in the rumen, it may be important to enhance fiber digestion.     

Mixed feed containing dextran improves milk production of holstein dairy cows

Total 37 Holstein daily cows (body weight: 631.76 +/- 18.45 kg, age: 5.47 +/- 1.94 years, parturition: 3.71 +/- 1.76 times) which became pregnant and gave birth to calves in the same season and lactated continuously were selected for this study. They were randomly divided into two groups: Group A-control, Group B-fed with 30 g/head/day of mixed feed containing supplemental dextran for one year from October 2001. After supplementation of the mixed feed, milk yields and components (fat, protein and solid non-fat) of Group B were compared with those of Group A in the 8th, 10th and 11th months (May, July and August of 2002). Milk yields of Group B were greater than the yields of Group A. In particular, there was a significant difference (p<0.001) between these groups in the July and August values. Milk components of Group B slightly differed from those of Group A before the supplementation, but after the supplementation, concentrations and total amounts of fat, protein and solid non-fat significantly increased more in Group B than in Group A. Thus mixed feed containing dextran can increase the milk production of Holstein dairy cows in the hot season.     

Characteristics of water balance in a rainfed paddy field in Northeast Thailand

Rice productivity in rainfed paddy fields varies with seasonal changes of water availability in which the conditions of flooding are affected by the water balance. Hydrometeorological measurements were performed in a rainfed paddy field in Northeast Thailand from July 2004 to December 2006 to analyze the water balance. As a result of our measurements, climatologically conditions were classified as semi-humid with an annual precipitation of 1,100 mm/year and annual potential evaporation of 1,660 mm/year in both the year. The surface layer of the paddy soil was clayey and the hydraulic conductivity was very low, so groundwater levels remained below the soil surface even under flooded conditions during the rainy season. Seasonal changes in the amount of soil water were very small, comprising only less than 16% of the total precipitation during the rainy season. Consequently, an effective precipitation of less than 180 mm was enough to establish standing water in the rainfed paddy field. Shinkichi Goto, Tsuneo Kuwagata and Pisarn Konghakote contributed equally to the paper.     

Contrast harmonic imaging of canine hepatic tumors

Six adult healthy Beagles were used to investigate the hepatic perfusion dynamics of Levovist, a contrast agent used in contrast harmonic imaging (CHI). In addition, 8 dogs with hepatocellular carcinoma (HCC) and 2 dogs with metastatic hepatic hemangiosarcoma (HSA) were used to characterize both the CHI findings with Levovist. In the Beagles, the start of intravenously injected Levovist into the aorta between the cranial mesenteric and renal arteries and the portal vein at the hepatic hilum were 5.47 +/- 1.52 sec and 16.03 +/- 3.39 sec, respectively. As a characteristic CHI finding in the 8 dogs with HCC, the early arterial phase showed a fine network of blood flow enhanced at the surrounding region and within the tumor in all the 8 dogs (100%), and the post vascular phase demonstrated a defect in the whole tumor and an enhancement of the surrounding hepatic tissues in 7 dogs (87.5%). In the 2 dogs with HSA, characteristic finding in which the early arterial and late vascular phases showed a rim contrast enhancement pattern, and the post vascular phase revealed that the whole tumor lacked contrast enhancement and the surrounding hepatic tissues was clearly enhanced. In dogs, the start of the early arterial and late vascular phases, and the characterizations of the CHI findings in HCC and HSA were suggested to be similar to those in humans. Therefore, CHI is thought to be useful for the diagnosis of HCC and metastatic hepatic HSA in dogs as well as in humans.     

A novel frameshift mutant allele, <Emphasis Type="Italic">fzp</Emphasis>-<Emphasis Type="Italic">10</Emphasis>, affecting the panicle architecture of rice

The rice FRIZZY PANICLE (FZP) locus on chromosome 7, in which an ERF and acidic domain are present, is concerned with the regulation of spikelet meristem identity and the determination of panicle architecture. Many fzp mutants drastically alter panicle morphology with higher-order rachis-branches developing successively instead of the development of floral organs in these mutants. A new mutant showing the same fzp phenotype was induced by γ-ray irradiation of seeds of a rice cultivar “Gimbozu”. Examination of this fzp-like mutant for its allelism to a known allele of fzp-1, nucleotide sequence, and panicle and agronomic characteristics clearly indicated that the allele of this fzp-like mutant is located in FZP. Because there is a previously identified allele called fzp-9, we designated this new allele as fzp-10. fzp-10 has a single nucleotide (cytosine) deletion between the ERF domain and the acidic domain, which results in a frameshift mutation and a premature stop codon, thereby altering the C-terminus of the encoded protein. The degree of higher-order branching in the panicles was significantly reduced in the fzp-10 mutant compared with that of fzp-1. Moreover, the fzp-10 mutant showed highly depressed culm and panicle lengths and panicle number, as well as delayed heading dates compared with its wild type and also with fzp-1. fzp-10 has several new characteristics, its altered nucleotide position and severity of phenotype alteration, and, therefore, could be a new gene resource to examine the function of FZP and the determination of rice panicle architecture.     

Distorted genetic segregation of the transposon mPing at the long arm of chromosome 12 in rice

A class II transposable element, mPing exists in the rice genome ubiquitously and can transpose even in ordinary cultivation conditions. A copy of mPing was identified at the long arm of chromosome 12. In reciprocal backcrossed F₁s between a heterozygote and a homozygote without mPing, the male gametes with this mPing from heterozygotes were transmitted to the next generation at a lower frequency than those without mPing, resulting in distorted genetic segregation in self-fertilized progenies, as well as in F₁s after backcrossing. Pollens with mPing tended to germinate on stigma less than those without mPing. These results, however, could not explain the lower transmission of male gametes with mPing. In addition, no excision of mPing was observed in a homozygote. Thus, it was suggested that male gametes with mPing were eliminated partly from pollination to fertilization by negative competition against male gametes without mPing. Less formation of microspores with mPing in meiosis could also be a cause for the distorted segregation, although this could not be examined. At least two ORFs, whose functions have not been identified, are located near this mPing. It is plausible that either of these ORFs or both are necessary for the normal functioning of male gametes.