The populations of Poa annua L. were monitored on green, fairway and rough in a golf course in Kyoto for a year to characterize the selection pressures on this species. Populations in the green were irregularly recruited during a whole growing period (October to June). They suffered from severe mortality (over 80%) caused by hand weeding in March and April. At the weeding time, the matured large plants were mainly removed by hand, because their panicles stood out in the green. However, over 50% of the green populations produced seeds before that time. Their irregular recruitments and quick maturation seem to be adaptive characteristics to mortal risks caused by hand weeding. In contrast, plants in the fairway and rough were recruited uniformly by December. Most individuals of those populations were killed by herbicides before their maturity.

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Summer-seeded sorghum (Sorghum bicolor Moench, cv. Pioneer 956) was subjected to different periods of foggage storage in the field, i.e. until December, January or March. The storage effects on the spring regrowth of the plants were analyzed mainly in relation to the tiller growth. The prolonging foggage storage in the field decreased foggage yield. On the contrary, it increased the survived percentage of emerged tiller buds (ETBs) while it scarcely affected the number of total tiller buds (TTBs) on a stubble tiller, the emerged percentage of TTBs and the regrown percentage of survived tiller buds. The increase in the survived percentage of ETBs with the prolonging storage period resulted in the increase in the regrown percentage of plant and consequently in the increase in the regrown plant yield from the next spring to heading. Sum of the foggage and regrown plant yields increased with the prolonging storage and it suggested the potentiality of summer-seeded foggage sorghum to be used as a biennial crop.

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This study was carried out to identify protein changes occurring through hardening in Lolium temulentum L. and to analyze N-terminal amino acid sequences of these hardening related proteins. The development of the freezing tolerance was mostly obtained after 2 weeks exposure to hardening treatment. Thus, the proteins extracted from seedlings of non-hard-ened and hardened for 2 weeks were analyzed by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). One newly induced protein and six proteins increased in intensity were identified in hardened seedlings, as compared to non-hardened seedlings. To get some information about these proteins, N-terminal amino acid sequences analysis were carried out using gas-phase protein sequencer and N-terminal amino acid sequences of five proteins were determined. As the result of homology search with a protein sequence data bank, several of these proteins were homologous to abscisic acid (ABA)-inducible protein of alfalfa (Medicago sativa L.), ribulosebisphoshate carboxylase (Rubisco) large-subunit binding protein beta chain in garden pea (Pisum sativum L.) and leaf fructose diphoshate aldolase in spinach (Spinacia oleracea L). The information of partial amino acid sequences would be helpful in estimating the function of hardening related proteins, and for molecular cloning.

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Seasonal changes of estimated dry matter digestibility of 6 temperate grasses were compared and relationship between the digestibility and meteorological elements was investigated. Lolium perenne, Dactylis glomerata, Festuca arundinacea, Agrostis alba, Festuca rubra and Poa pratensis were sown in 1984, and the digestibility was investigated from 1985 to 1989 under frequent cutting based on the pattern derived by an overlapping sequence of 3-weekly cuts. The digestibility was estimated from a linear regression equation showing relation between the digestibility and disappearance rate by the two-stage neutral detergent-cellulase technique. Mean values of digestibility for 5 years were high in early spring, declined gradually toward summer with or without troughs on the way, and recovered well toward late autumn thereafter. It seemed that the seasonal changes of digestibility were always affected by average air temperature for all grasses, affected by precipitation especially for the 1st and 2nd harvest years, and the stage of internode elongation also brought some effects to Dactylis and Festuca. The highest mean percentage digestibility for 5 years was 77% in Lolium, followed by 70% in Dactylis, 69% in Festuca arundinacea, 67% in Agrostis and Festuca rubra, and 56% in Poa. And the highest digestible dry matter yield was obtained in Lolium.

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In order to classify the grazing behavior of cattle statistically, grazing experiments were carried out from 1990 to 1992. Grazing bahavior of cattle was observed with video recorder. Grazing time were measured every two minutes from just before daybreak to just after sunset, and the percentages of grazing time in each two minutes (gt) were calculated. On all experimental days, although there were two active period of grazing at early morning and before sunset, the patterns of gt around midday were different day by day. Therefore, according to the difference in the pattern of gt around midday, the patterns of gt of the experimental days were classified into three types of grazing behavior as follows ; (1) a type with a active period of grazing around middday, (2) a type without a active one around midday, and (3) a intermediate type between the types of (1) and (2). Grazing behavior units (GUs), which were sets of gt within every one hours, were classified into four groups of grazing behavior A, B, C and D, according to the combination of the values of skewness and kurtosis in the distribution of gt of each GU. On the average of the three years, mean values of GT, which was a percentage of grazing time in each GU, of the groups of A, B, C and D were 79.7, 60.2, 35.4 and 10.4%, respectively, and those were 48, 36, 21 and 6 minutes, respectively, as converting into grazing time per hour.

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The induction of ruminant bloat is usually caused by the ingestion of too much clover on mixed sward. This study was to estimate grazing pattern of white clover on the clover/grass mixed sward by ruminants. Eating pattern of white clover (Trifolium repens L.) and orchardgrass (Dactylis glomerata L.) was investigated using Suffolk and Corriedale crossbred wether sheep (mean BW 38.0kg) housed in the metabolic cages. Voluntary intake of each herbage was measured with the elapse of time by fixed feeding (07 : 00 and 17 : 00) (Exp. 1) or by free feeding (Exp. 2). Furthermore, to examine grazing pattern of white clover on the pure stand, the voluntary intake in sole feeding of white clover was compared with the voluntary intake in white clover and orchardgrass feeding (Exp. 3). In fixed feeding, daily intake of white clover was larger than that of orchardgrass (white clover ; 6.44 kg/head, orchardgrass ; 1.28 kg/head, P<0.001). The intake of white clover was markedly high for first 20 minutes after feeding and decreased with the elapse of time, whereas the intake of orchardgrass was increased gradually. In free feeding, daily intake of white clover was about equal to that of orchardgrass (white clover ; 4.99 kg / head, orchardgrass ; 4.93 kg/head, P>0.1). Although intake rate (g/min) of white clover was higher than that of orchardgrass immediately after feeding, the rate decreased by 90 minutes, thereafter both herbages were eaten with a certain rate. It was suggested that, on the white clover/grass mixed sward, a large amount of white clover might be grazed by animals at a good appetite such as immediately after changing paddock or setting up the grazing, but with the elapse of the time of grazing, the intake of coexisting grass by ruminants increase. The difference was not observed between the total daily intake of herbage in white clover sole feeding (WC) and that in white clover and orchardgrass feeding (WC + OG) (WC ; 10.54 kg/head, WC + OG ; 10.10 kg/head, P>0.1) (Exp. 3), even though the digestibility of white clover is higher than that of orchardgrass. From these experiments, white clover appears to regulate the ingesting of white clover itself by ruminants under a stationary state of appetite as long term grazing. The clarification of the mechanism on the regulation is expected.

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Experiments were carried out to determine the changes of chemical composition of plant parts with advancing maturity, and efficient methods for conserving sesbania rostrata (Sesbania rostrata) as silage. The contents of crude protein (CP) of stem and whole plant decreased and cell wall (OCW) and acid detergent fiber (ADF) of whole plant mostly increased with advancing maturity. The CP content of sesbania at 74 days after seeding was about two thirds of the value observed at 60 days after seeding. The contents of OCW and ADF decreased and in vitro dry matter digestibility increased by the cellulase treatment in both silages made at 60 and 80 days after seeding. The pH value, acetic acid content and the percentage of the volatile basic nitrogen to total nitrogen (VBN/TN%) decreased and lactic acid content increased by the cellulase treatment in the silage made at 60 and 80 days after seeding. On the other hand, the wilting and crushing of the material did not have any apparent effect on fermentative quality of the silages. The silages prepared by adding both lactic acid bacteria and cellulase showed similar quality with those treated only with cellulase. From the above, it was concluded that sesbania should be harvested at an early stage of maturity from the nutritional view point and that cellulase addition was more effective than the other treatments to make high quality silage from sesbania.

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Experiments were carried out to determine the effect of mixing concentrates and corn (Zea mays L.) soilage with sesbania rostrata (Sesbania rostrata) on chemical composition and fermentative quality silages. In experiment 1, concentrates (brown rice, beet pulp, wheat bran or flaked corn) was added to sesbania at 15% on fresh matter basis. In experiment 2, sesbania was ensiled with corn soilage at the rates of 0, 25, 50, 75 and 100% on fresh matter basis. The addition of concentrates improved the silage quality by increasing lactic acid content and reducings the pH value and acetic acid content. In vitro dry matter digestibility (IVDMD) was the highest in silages mixed with wheat bran and followed by those mixed with brown rice, flaked corn and beetpulp. The contents of crude protein, organic cell wall and acid detergent fiber decreased and IVDMD increased with increasing the ratio of corn soilage to sesbania. The pH value, acetic acid content and volatile basic nitrogen as a percentage of total nitrogen (VBN/TN%) decreased and lactic acid content increased with increasing the ratio of corn soilage. From the above, it was concluded that sesbania should be mixed with concentrates or with corn soilage to make good quality silage from it.

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Urease activity was determined for cereal straws, grass and legume forages to clarify the applicability of urea treatment for these materials. Moreover, influences of preparative condition such as cutting length, dehydration and storage period on the activity were also studied using rice straw and orchardgrass. A wide variation in the activity was observed among the cultivars and preparative conditions. The highest activity was observed in rice straw, whereas legume forages had the lowest. The older and withering leaves showed usually higher activity than stem and younger leaves. Fresh materials cut under 2 mm length showed lower activity than those above 5mm, but drying did not affect on the activity. Under wet and warm environment, the activity in rice straw increased by 5 times of the original on 5 days after harvesting. It was assumed that those higher activity was induced with the vigorous growth of aerobic microbes which had strong urease activity. Those results indicated that the rate of urea hydrolysis was more accelerated when urea added into the straw preserved for a few days under wet and warm condition than into the fresh straw after harvesting.

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Alfalfa was harvested (4th cutting in 1991 and 1st to 4th cutting in 1993) and wilted overnight, then chopped into 2 cm lengths. The forage mixed with cellulases (0-1.0% of the forage) or glucose (1%), and lactic acid bacteria (0.001%) was tightly packed in 1 liter experimental silos or in nylon/polyethylene bagsilos. Both the silos were kept at 26℃ for 30 and 14 days, respectively. Each ensiling had three replications. Treatment of alfalfa from the 4th cutting in 1991 with cellulase preparation originated from Acremonium cellulolytius Y-94 (ACS 2) at 0.01% resulted in a lower pH, a higher lactic acid content, and a higher L/T value than obtained using a cellulase preparation from Trichoderma viride (CEP, 0.01%), or 1% glucose. Also, in the case of alfalfa from the 1st through to 4th cuttings of 1993, the addition of ACS 2 and lactic acid bacteria promoted the fermentation and much improved the quality of silage. These results showed that the addition of 0.01% ACS 2 and lactic acid bacteria was enough to improve the fermentation quality of alfalfa silage.

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Alfalfa (1st to 4th cutting in 1991 and 4th cutting in 1992) was harvested, wilted and chopped into 2 cm length. Then it was dried and powdered. The powdered alfalfa was washed with 70% aqueous ethanol to remove WSC (water soluble carbohydrates) and was used as substrate of enzymatic treatment. The powder was incubated with cellulase preparations produced by Acremonium cellulolyticus Y-94 (ACS 2) or Trichoderma viride (CEP) to compare the quantities of newly released sugar. Quantitative analysis of sugars were performed using the phenolsulfuric acid method for total sugar, the modification of Somogyi's method for reducing sugar, and the glucose oxidase-peroxidase-chromogen method for glucose. With alfalfa from the 1st cutting in 1991 as substrate, 2.4-fold more total sugar, 3.4-fold more reducing sugar, and 3.4-fold more glucose were released by ACS 2 as compared with CEP. A similar effect was observed with alfalfa from the 1 st through to 4th cutting, though the amount of reducing sugar released by each cellulase was dependent on the cutting time and decreased in order of cutting 4th, 1st, 3rd and 2nd. Treatment of NDF from the 4th cutting in 1992 with ACS 2 resulted in the production of various sugars which were consisted of 37.0% glucose, 19.0% galacturonic acid, 9.2% arabinose, 4.3% galactose, and 30.5% other sugars. These findings indicated that ACS 2 could hydrolyze not only cellulose but also pectin and hemicellulose.

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The present study was conducted to clarify the hardening responses in Italian ryegrass (Lolium multiflorum Lam.) cultivars under natural and artificial conditions. Eleven cultivars were sown four times at 2 weeks intervals from September 14 to October 26 under natural conditions. Freezing treatment was carried out at the 4-5 leaf stage of the seedlings. The effect of hardening, estimated by the changes of LT_<50> values, was rapidly induced at below 10℃ of daily mean air temperature after early November. To investigate the influences of hardening temperature and duration, Waseaoba (tolerant) and Minamiwase (sensitive) with contrasting freezing tolerance were raised in growth cabinets at day/night temperature of 20/15℃ with 16 h photoperiod. Raised seedlings at 3-4 leaf stage were exposed to five different temperatures with 8h photopriod in growth cabinets. The threshold temperature for hardening of Waseaoba was about 10℃, while for Minamiwase it was between 6℃ and 4℃. These cultivars were also hardened effectively after 14 days hardening at 2℃, 8 h photoperiod.

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Nitrogen (N) of animal wastes and capacity of farming land for N fertilization in Tochigi Prefecture were estimated by the agricultural census data in 1970, 1980 and 1990. As the results, 781.9 ton-N excreted from livestock was over beyond the total capacity. However, since the distribution of livestock and farming land in each municipality were not uniform, 30 municipalities had a surplus capacity. To solve the imbalance of excessive N among municipalities, the distribution plan in Tochigi Prefecture was drafted based on the accessibility determined by the road network. In this study, the accessibility was evaluated by Network Analysis of Geographic Information System. The drafted plan showed the municipalities accepting N from N-excess municipalities, distributing quantity and the shortest route for transportation.

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