Mottle necrosis of sweet potato caused by <Emphasis Type="Italic">Pythium scleroteichum</Emphasis> in Japan and varietal difference in susceptibility to the disease

Severe mottle necrosis (Shirogusare-byo in Japanese) was found on mature tubers of sweet potato (Ipomoea batatas) in Ibaraki Prefecture in October 2004. The causal organism was identified as Pythium scleroteichum hitherto unknown in Japan. Sweet potato cultivar Purple Sweet Lord was more susceptible than cultivars Beniazuma, Benimasari, Koukei-14, and Tamayutaka to the pathogen at 25°C, while this difference in the susceptibility was not clear at 15°C.     

Genetic Variability in Phosphorus Acquisition and Utilization Efficiency from Sparingly Soluble P-Sources by Brassica Cultivars under P-Stress Environment

Plants display an array of classical strategies to maximize phosphate (Pi) acquisition from sparingly soluble P sources. Acclimation to Pi-stress via elegant Pi-starvation induced (PSI) adjustments would reduce our current overreliance on expensive, polluting and non-renewable Pi-fertilizers. Nevertheless, differences in the ability of various species to solubilize sparingly soluble P-sources have been often evidenced; inter-cultivar variations are scarcely documented. Brassica is known as an effective, non-mycorrhizal user of sparingly soluble P-sources. Various growth parameters and biomass accumulation by genetically diverse Brassica cultivars were determined in four experiments using hydroponics and quartz sand culture media. Role of PSI root mediated pH changes, organic anions (OAs) exudation and altered root architecture in mobilization and acquisition of sparingly soluble P-forms [Jordan rock-P (RP) at 2 g l⁻¹ and Ca₃(PO₄)₂ (TCP) at 0.2 g l⁻¹ respectively] was investigated. Cultivars showed considerable genetic variations in biomass accumulation, various growth parameters and root–shoot ratio. Concentration and total uptake of P, specific absorption rate of P, P-transport rate and P-utilization efficiency (PUE) were also significantly (P < 0.001) different for various cultivars and their dry matter was significantly correlated with P-uptake [r = 0.94** (significant at 1% level)]. P-tolerant cultivars showed substantial decrease in solution media pH because of H⁺ efflux and exuded more carboxylates than low P-sensitive cultivars under P-starvation. P-uptake by cultivars increased linearly with decreasing pH. The amount and types of OAs exuded from the roots of P-starved plants differed from those of plants grown under P-sufficient environment. In split pot study, with TCP and RP supplied spatially separated from other nutrients, efficient cultivars were still able to mobilize RP and TCP more efficiently than inefficient cultivars. In rhizobox study, the elongation rates of primary roots decreased but the elongation rates of the branched zones of primary roots and the length of lateral roots increased under P-starvation. Tested cultivars showed genetic diversity in accessing, mobilization, acquisition and utilization of Pi from sparingly soluble P forms. An arrange marriage of plant traits can explain cultivar’s access to different forms of sparingly soluble P, and in addition to altered lateral root topology and enhanced P-uptake and PUE, enhanced H⁺ efflux and OAs exudation are key factors in Pi scavenging from extra cellular sparingly soluble P-forms.     

Developmental changes in pituitary–thyroid axis, and formation of gonads in leptocephali and glass eels of Anguilla spp.

SUMMARY: Pituitary, thyroid gland and gonads in leptocephali of Japanese eel Anguilla japonica (19.8–32.6 mm in total length), A. obscura (45.0 mm), and A. bicolor pacifica (49.5 mm) and those in glass eels of the Japanese eel were histologically and immunohistochemically examined in order to observe the developmental changes of these endocrine organs in the Anguillidae. The pituitary, consisting of adenohypophysis and neurohypophysis in Japanese eel leptocephali over 22.5 mm, did not contain thyroid stimulating hormone (TSH) immunoreactive cells. Such cells were, however, detectable in the more developed pituitaries of leptocephali of A. obscura and A. bicolor pacifica and in those of glass eels. Conversely, thyroxine (T₄)-immunoreactive thyroid follicles could be detected in all specimens, both leptocephalic and glass eel. Only in glass eels, gonads were found in the body cavity, and these gonads harbored one or two primordial germ cells (PGC) per cross-section. Our results indicate that thyroid hormones (TH) production started prior to TSH production, and that TSH and TH are both secreted during the metamorphosis from leptocephalus to glass eel. Therefore, it is plausible that the TSH–TH axis is involved in the metamorphosis from leptocephalus to glass eel, but not in the early growth from preleptocephalus to leptocephalus.     

The effect of strain, growth stage, and cultivating condition of Ganoderma lucidum on 5α-reductase inhibition

The inhibitory effects of 102 methanol extracts of 40 mycelia, 9 culture fluids, and 53 fruiting bodies of 40 strains of Ganoderma lucidum on 5-reductase were investigated. The methanol extract of the fruiting body of each strain was found to show the strongest 5-reductase inhibitory activity among the extracts tested.     

The therapeutic effects of SET/I2PP2A inhibitors on canine melanoma

Canine melanoma is one of the most important diseases in small animal medicine. Protein phosphatase 2A (PP2A), a well conserved serine/threonine phosphatase, plays a critical role as a tumor suppressor. SET/I2PP2A is an endogenous inhibitor for PP2A, which directly binds to PP2A and suppresses its phosphatase activity. Elevated SET protein levels have been reported to exacerbate human tumor progression. The role of SET in canine melanoma, however, has not been understood. Here, we investigated the potential therapeutic role for SET inhibitors in canine melanoma. The expression of SET protein was observed in 6 canine melanoma cell lines. We used CMeC-1 cells (primary origin) and CMeC-2 cells (metastatic origin) to generate cell lines stably expressing SET-targeting shRNAs. Knockdown of SET expression in CMeC-2, but not in CMeC-1, leads to decreased cell proliferation, invasion and colony formation. Phosphorylation level of p70 S6 kinase was decreased by SET knockdown in CMeC-2, suggesting the involvement of mTOR (mammalian target of rapamycin)/p70 S6 kinase signaling. The SET inhibitors, OP449 and FTY720, more effectively killed CMeC-2 than CMeC-1. We observed PP2A activation in CMeC-2 treated with OP449 and FTY720. These results demonstrated the potential therapeutic application of SET inhibitors for canine melanoma.     

Adventitious root formation of Japanese cedar (Cryptomeria japonica D. Don) cuttings is stimulated by soaking basal portion of cuttings in warmed water while cooling their apical portion

To improve the propagation of Japanese cedar (Cryptomeria japonica D. Don), we investigated the effects of apical and basal temperatures during a water soaking treatment on the adventitious root formation of 70-mm long shoot tip cuttings which have an apical bud. The basal portion of the cuttings was soaked for 28 days in water with temperatures ranging from 10 to 35 °C, at an air temperature of 5 or 10 °C. Control cuttings were soaked in water at 25 °C, with an air temperature of 25 °C. Treated cuttings were then planted in vermiculite rooting medium and grown at an air temperature of 25 °C for 35 days. Adventitious roots initiated earlier and developed more in the cuttings treated with apical temperatures of 5 or 10 °C and basal temperatures of 20–30 °C than in the control cuttings. The rooting percentage was greatest (93 %) in the cuttings treated with a 10/25 °C apical/basal temperature, whereas few control cuttings rooted (13 %). This suggests that the temperature gradient created by warming the basal portion of the cuttings while cooling their apical end stimulates adventitious root formation. When we tested seasonal variation of rootability at 10/25 °C, the rooting percentage increased from early autumn to winter, and decreased from winter to summer. The soluble sugar contents did not directly affect the formation of adventitious roots in the present study.     

8S globulin of mungbean [Vigna radiata (L.) Wilczek]: cloning and characterization of its cDNA isoforms, expression in Escherichia coli, purification, and crystallization of the major recombinant 8S isoform

Three isoforms of the cDNA of the major 8S globulin of mungbean, 8Salpha, 8Salpha', and 8Sbeta, were isolated, cloned, and characterized. The cDNA sequences of 8Salpha, 8Salpha', and 8Sbeta had open reading frames of 1362, 1359 or 1362, and 1359 bp, respectively, which code for 454, 453 or 454, and 453 amino acids corresponding to molecular weights of 51 973, 51 627 or 51 758, and 51 779, respectively. Homology in terms of cDNA and amino acid sequences was 91-92% between 8Salpha and 8Salpha', 87% between 8Salpha and 8Sbeta, and 86-88% between 8Salpha' and 8Sbeta. The signal peptide was found to be 1-25, 1-24 or 25, and 1-23 for 8Salpha, 8Salpha', and 8Sbeta, respectively, using the signalP website (Nielsen, H.; Engelbrecht, J.; Brunak, S.; von Heijne, G. Protein Eng. 1997, 10, 1-6). The propeptide was determined to be IVHREN. A single site for glycosylation (N-X-S/T) was observed about 90 amino acids from the C terminus. Homology between mungbean 8S isoforms and other 7-8S proteins ranged from 45 to 68% within members of the legume family and 29 to 34% for crops of different species. The major isoform 8Salpha was expressed in Escherichia coli and purified by successive ammonium sulfate fractionation, hydrophobic interaction, and Mono Q column chromatography. The recombinant 8Salpha, but not the native form, was successfully crystallized producing rhombohedral crystals.     

Nitrogen-fixing activity in decomposing litter of three tree species at a watershed in eastern Japan

Nitrogen fixation during litter decomposition was studied for 34 months using litterbags containing newly fallen litter of coniferous species Cryptomeria japonica and Pinus densiflora and that of deciduous species Quercus serrata. Litterbags were set in contact with the forest floor in a deciduous broad-leaved forest near the top of a slope and in a C. japonica stand at the middle of the slope at a watershed in eastern Japan. Nitrogen-fixing activity, estimated by acetylene reduction after 16 and 19 months of incubation, was 62.65–3.86 nmoles C₂H₄ h⁻¹ g⁻¹ DW in Cryptomeria litter, but only 1.07–0.09 in Pinus and 0.72–0.04 in Quercus. The rate of N increase in decomposing litter was highest in Cryptomeria. Fungal biomass in decomposing litter, estimated by ergosterol content, increased during the initial 16 months of incubation in Cryptomeria and Quercus, and during the initial 19 months of incubation in Pinus. The litter decomposition rate was highest in Cryptomeria among the three species, due to increased N content and fungal biomass in Cryptomeria litter. Thus, N increase in decomposing Cryptomeria litter affects the subsequent N dynamics and decomposition pattern.     

Mungbean [Vigna radiata (L.) Wilczek] globulins: purification and characterization

Vicilin type (8S) and basic 7S globulins and legumin type (11S) globulins were isolated from mungbean [Vigna radiata (L.) Wilczek]. The native molecular weights of the different globulin types were 360000 for legumin, 200000 for vicilin, and 135000 for basic 7S. Some of the 8S globulin apparently complexed and coeluted with the 11S on gel filtration. On SDS-PAGE, 11S was composed of two bands of 40000 and 24000, 8S was composed of 60000, 48000, 32000, and 26000 bands, and basic 7S was composed of 28000 and 16000 bands. The percent composition of total globulins was estimated to be as follow: 8S, 89%; basic 7S, 3.4%; and 11S, 7.6%. The basic 7S and 11S but not the 8S globulins were found to have disulfide bonds. The presence of carbohydrates by conjugated peroxidase reaction was observed in all bands of 8S, the acidic polypeptide of basic 7S, and its complex but not in 11S. The 28000 basic 7S band and its 42000 complex and the first three major bands of 8S cross-reacted with antibodies to all types of soybean conglycinin subunits (alpha, alpha', and beta), whereas the fourth band cross-reacted only with the anti-beta subunit. None of the mungbean globulins cross-reacted with anti-soybean glycinin. Basic 7S was found to be easily extracted with 0.15 M NaCl, 11S was extracted with 0.35 M NaCl,and 8S was extracted over a wide range of NaCl concentrations. The N-terminal sequences of the different subunits/fragments of the globulins were determined and found to have strong homology with storage proteins of other legumes and crops.