Effect of Application of Rice Straw and Cellulose on Methane Emission and Biological Nitrogen Fixation in a Subtropical Paddy Field

A field experiment was conducted to evaluate the effect of the incorporation of rice straw and cellulose on methane flux, soil-acetylene reduction activity (ARA) and rice plant growth under a subtropical climate. Rice straw and cellulose (as paper) were applied at the rates of 4 and 10 t ha^-1. Emission rates of CH₄ from the paddy field without and with straw and cellulose amendments were measured by using the closed chamber method. Stimulation of N₂-fixation by the amendments was measured as soil-ARA under anaerobic conditions. The measurements indicated that the application of 10 t ha^-1 cellulose resulted in a relatively high emission of CH₄, with an average flux of 106.7 mg CH₄ m^-2 h^-1, followed by 10 t ha^-1 straw, 51.7 mg m^-2 h^-1, compared with the control, 5.3 mg m^-2 h^-1. Application of straw and cellulose at the rate of 10 t ha^-1 to the paddy field increased the CH₄ emission 10 and 21 fold over the values of the control, respectively as estimated seasonal emissions. The soil-ARA levels in the treatments during the cultivation period were positive. The stimulation of ARA by the amendment with 10 t ha^-1 cellulose occurred at the early stage of rice growth, while the maximum ARA-peak occurred in the 10 t ha^-1 straw-amended soil at around the heading stage. Amendment with straw at 10 t ha^-1 significantly increased the total dry matter weight of rice, whereas growth inhibition was induced by cellulose incorporation. The differences in CH₄ flux, and soil-ARA among the treatments were most evident at the heading stage.     

Isolation and some properties of methane-oxidizing bacteria from a subtropical paddy field

Methane is the second most important greenhouse gas which contributes to global warming. As an important source of methane, rice paddy fields contribute an estimated lO% to the global methane emissions (IPCe 1992). Land use and agricultural practices significantly affect atmospheric methane fluxes (Bouwman 1989; Hütsch et al. 1994). Microbial oxidation of atmospheric methane in terrestrial environments is the only known net biological methane sink and the process consumes the equivalent of 1–l0% of the total global emission (Adamsen and King 1993). Methane-oxidizing bacteria (MOB, methanotrophic bacteria) are considered to be obligately or facultatively aerobic respiratory bacteria that can utilize methane as the sole source of carbon and energy for growth (Hanson et al. 1992; Roslev and King 1994). As a result, they are important regulators of atmospheric methane fluxes in nature (Mancinelli 1995). MOB have been isolated from a variety of environments including freshwater lakes, wetlands, and the open ocean (Whittenbury et al. 197Gb; Saralov et al. 1984; Holzapfel-Pschorn et al. 1985; Hanson et al. 1992; Omelchenko et al. 1993; Bowman et al. 1993a; Mancinelli 1995). However, reports on the isolation of MOB from rice paddy fields are limited. More information is needed on the ecology and taxonomy of MOB in paddy fields.     

Gonadotropin-releasing hormone and gonadotropin in goldfish and masu salmon

Reproductive activities in vertebrates are regulated by an endocrine system, consisting of the brain-pituitary-gonad axis. In teleosts, gonadotropin-releasing hormone (GnRH) in the brain stimulates gonadotropin (GTH) release in the pituitary gland, but because of lack of the portal vessel, it is not known when and how much GnRH is released for the regulation of GTH release. There are multiple molecular types of GnRH in teleosts and several distinct populations of GnRH neurons in the brain. However, we do not know which types and populations of GnRH neurons regulate reproductive activities. Here we summarize our recent studies on GnRH and GTH in masu salmon Oncorhynchus masou and goldfish Carassius auratus. Immunocytochemistry showed the location and molecular types of GnRH neurons. Salmon (sGnRH) and chicken-II GnRH (cGnRH-II) neuronal fibers were widely distributed in the brain of both masu salmon and goldfish. Only sGnRH fibers were observed in the pituitary of masu salmon, whereas both sGnRH and cGnRH-II fibers were observed in the goldfish pituitary, indicating that species specific GnRH profiles are involved in the regulation of pituitary function in teleosts. A series of experiments in masu salmon and goldfish suggest that among GnRH neuron populations GnRH neurons in the ventral telencephalon and the hypothalamus regulate GTH release, and that GnRH of the terminal nerve origin is not essential to gonadal maturation and ovulation. The biological function of other GnRH neurons remains unkown. Two GTHs appear to be characteristic of teleost; however, regulation of reproduction by these GTHs is a question that remains to be elucidated. In salmonid species, it is proposed that GTH I stimulates early gonadal development, whereas GTH II acts in later stages. When GTH expression was examined in goldfish, both GTH I and II mRNA levels in the pituitary gland showed increases in accordance with gonadal development, unlike the sequential expression of GTH subunits in salmonids. The expression of these GTH subunit mRNAs were affected by water temperature, starvation, and steroid hormones in goldfish, but in what manner these two GTHs regulate gonadal development remains to be clarified.     

Immunohistochemical localization of three GnRH systems in brain and pituitary of Japanese flounder

ABSTRACT:   To clarify the possible roles of gonadotropin-releasing hormone (GnRH) in the reproduction of Japanese flounder Paralichthys olivaceus , localization of salmon GnRH (sGnRH), chicken GnRH-II (cGnRH-II), and sea bream GnRH (sbGnRH) immunoreactive (ir) cell bodies and fibers in the brain and pituitary were examined together with follicle stimulating hormone (FSH) and luteinizing hormone (LH)-ir cells in the pituitary by immunohistochemistry. sGnRH-ir cell bodies were localized in the ventromedial part of the rostral olfactory bulb and cGnRH-II-ir cell bodies were restricted to the midbrain tegmentum, while sbGnRH-ir cell bodies were evident in the preoptic area. sGnRH-ir fibers were distributed throughout the brain, especially abundant in the forebrain. cGnRH-II-ir fibers were also scattered in many areas of the brain with abundance in the midbrain, but sbGnRH-ir fibers were observed in the preoptic–hypothalamic area and innervated the pituitary. In the pituitary, neither sGnRH-ir fibers nor cGnRH-II-ir fibers were found, but sbGnRH-ir fibers were profuse in the neurohypophysis and invaded the proximal pars distalis, targeting FSH and LH cells. These results suggest that three GnRH systems can play different physiological roles in the brain of Japanese flounder. Among them, sbGnRH is considered to be involved in reproduction by stimulating gonadotropin secretion, while sGnRH and cGnRH-II can function as a neurotransmitter and/or neuromodulator within the brain in this species.     

Response of soybean plants to two inoculation methods with arbuscular mycorrhizal fungus of Glomus sp. strain R-10 under field condition

ABSTRACT

Inoculation of arbuscular mycorrhizal (AM) fungi has a great potential to reduce input of phosphorus fertilizer. In this study, we tested the hypothesis that transplanting of pre-inoculated plants (pre-inoculation) with AM fungal inoculum Glomus sp. strain R-10 (R-10) is more effective for increasing AM fungal colonization and soybean yield than placing R-10 inoculum into field soil (direct inoculation). We cultivated pre-inoculated and direct inoculated plants with and without R-10 in the same field. On the contrary to the hypothesis, hyphal colonization was increased by direct inoculation, but decreased by pre-inoculation in an early growth stage. Shoot phosphorus concentration, shoot dry weight, and yield also showed the same trend as the hyphal colonization. These results indicated that pre-inoculation with R-10 would be less effective for increasing AM fungal colonization and yield than direct inoculation. It may be due to a colonization strategy of R-10 and short duration for establishment of seedling.     

Relationship between early watercore development and leaf photosynthesis or partitioning of photosynthates in apple

Seasonal changes in leaf photosynthetic rate and partitioning of ¹³C taken up by leaves were compared in relation to the development of early watercore using susceptible ‘Orin’ and resistant ‘Fuji’ apples (Malus domestica Borkh.). There was no difference between the cultivars in leaf photosynthetic rate from mid-July to mid-October. The extent of watercore in ‘Orin’ increased from mid-July and peaked in late-August, followed by a decrease through mid-October, while no watercore was detected during the experimental period in ‘Fuji’. Fruit fresh weight and soluble solids content (SSC) were higher in ‘Orin’ than ‘Fuji’ during the earlier stages. Leaf dry weight of ‘Orin’ was higher than that of ‘Fuji’ throughout the experimental period. Accumulation of dry matter in the fruit increased in both cultivars and ‘Orin’ kept significantly higher level than ‘Fuji’ from mid-July to mid-October. The amount of ¹³C distributed to ‘Orin’ fruit at 5 days after labeling was significantly higher than ‘Fuji’ in mid-July just before watercore started to occur. ¹³C taken up by ‘Orin’ fruit with prevalent watercore was also higher than ‘Fuji’ in late-August but the difference was not significant. In mid-October when early watercore was gradually reducing, ¹³C recovered by ‘Orin’ fruit decreased and was significantly lower than ‘Fuji’. The difference of partitioning pattern of ¹³C between the cultivars was associated with the susceptibility to early watercore as well as the changes in fruit growth and SSC. The seasonal changes in the amount of ¹³C distributed to ‘Orin’ fruit were also related to the fluctuations in watercore severity. The results suggest that active phloem transport might be the cause of early watercore in our previous work, which might be induced by the increased partitioning of assimilates to the fruit during summer in susceptible ‘Orin’ compared to resistant ‘Fuji’ and not by the higher potential of leaf photosynthesis.     

Involvement of sex steroids,luteinizing hormone and thyroid hormones in upstream and downstream swimming behavior of land-locked sockeye salmon <Emphasis Type="Italic">Oncorhynchus nerka</Emphasis>

The involvement of testosterone (T), estradiol-17β (E₂), 11-ketotestosterone (11-KT), 17,20β-dihydroxy-4-pregnene-3-one (DHP), luteinizing hormone (LH), thyroxine (T₄), and triiodothyronine (T₃) in the regulation of downstream and upstream movement (swimming behavior) was investigated in land-locked sockeye salmon Oncorhynchus nerka, using an artificial raceway. During the downstream migratory period, T implant resulted in high plasma T levels and inhibited the occurrence of downstream swimming behavior (negative rheotaxis) in yearling (1+) immature smolts. In terms of upstream behavior, 2-year-old (2+) males exhibited high plasma T and 11-KT levels, while 2+ females had elevated T and DHP levels. In 1+ immature fish, a T implant induced upstream swimming behavior (positive rheotaxis). In experiments 1 and 3, the plasma T₄ and T₃ levels of non-migrants tended to be higher than those of migrants. In contrast, no marked changes in plasma and pituitary LH were found in both downstream and upstream migrants. These results suggest that sex steroids, such as T, play significant roles in the regulation of downstream and upstream swimming behaviors in land-locked sockeye salmon.     

cDNA cloning and expression analysis of two heat shock protein genes,Hsp90 and Hsp60, from a sterile Ulva pertusa (Ulvales,Chlorophyta)

Molecular chaperones are synthesized and accumulated under a variety of unfavorable conditions in all organisms. Heat shock protein 90 (Hsp90) and Hsp60, which are classified into the major classes of molecular chaperones, play important roles in cellular stress responses. In this study, we characterized sterile Ulva pertusa Hsp90 (UpHsp90) and UpHsp60 genes which may be involved in tolerance to thermal and heavy metal stresses in this alga. The UpHsp90 cDNA consisting of 2,118 nucleotides encoded a polypeptide of 705 amino acids (AA). On the other hand, the UpHsp60 cDNA consisting of 1,722 nucleotides encoded a protein whose predicted length was 573 AA. The AA sequence alignment and phylogenetic analyses showed that the UpHsp90 and UpHsp60 proteins were more similar to cytoplasmic Hsp90s and mitochondrial Hsp60s, respectively, than to other types of the respective Hsps. Southern blot analysis indicated that the sterile U. pertusa genome had at least two cytoplasmic Hsp90-encoding genes and two mitochondrial Hsp60-encoding genes. The UpHsp90 and UpHsp60 mRNA levels were significantly affected by diurnal and temperature changes, and slightly affected by exposure to heavy metals. These results suggest that UpHsp90 and UpHsp60 genes play particularly important roles in adaptation to diurnal and temperature changes.     

Multiple vitellogenins and product yolk proteins in striped bass,Morone saxatilis: molecular characterization and processing during oocyte growth and maturation

The multiple vitellogenin (Vtg) system of striped bass, a perciform species spawning nearly neutrally buoyant eggs in freshwater, was investigated. Vitellogenin cDNA cloning, Western blotting of yolk proteins (YPs) using Vtg and YP type-specific antisera, and tandem mass spectrometry (MS/MS) of the YPs revealed the complex mechanisms of yolk formation and maturation in this species. It was discovered that striped bass possesses a tripartite Vtg system (VtgAa, VtgAb, and VtgC) in which all three forms of Vtg make a substantial contribution to the yolk. The production of Vtg-derived YPs is generally similar to that described for other perciforms. However, novel amino-terminal labeling of oocyte YPs prior to MS/MS identified multiple alternative sites for cleavage of these proteins from their parent Vtg, revealing a YP mixture far more complex than reported previously. This approach also revealed that the major YP product of each form of striped bass Vtg, lipovitellin heavy chain (LvH), undergoes limited degradation to smaller polypeptides during oocyte maturation, unlike the case in marine fishes spawning buoyant eggs in which LvHAa undergoes extensive proteolysis to osmotically active free amino acids. These differences likely reflect the lesser need for hydration of pelagic eggs spawned in freshwater. The detailed characterization of Vtgs and their proteolytic fate(s) during oocyte growth and maturation establishes striped bass as a freshwater model for investigating teleost multiple Vtg systems.