Reproduction of a Robinia pseudoacacia population in a coastal Pinus thunbergii windbreak along the Kujukurihama Coast, Japan

We investigated reproduction of the clonal tree Robinia pseudoacacia in a Pinus thunbergii windbreak. Microsatellite polymorphism analysis revealed that asexual reproduction through horizontal root elongation and ramet formation was the primary mode of reproduction in this population. Germination experiments indicated that the inability of established genets to produce viable seeds inhibited sexual reproduction. The boundaries between genet areas were relatively distinct, indicating that ramets within genets spatially excluded one another. Analyses of the number of annual rings and thickness of excavated horizontal roots provided new insight regarding photosynthate transfer through horizontal roots. The number of annual rings and thickness were highest for horizontal roots near the base of ramets and decreased along portions far from ramets. This result indicates that the new horizontal root is supplied with photosynthate from the mother ramet for a few years only, and that resource translocation between neighboring ramets within the same clone ceases after several years. Hypotheses about horizontal root expansion and cessation were considered to explain the exclusive distribution pattern of the horizontal root between genets.     

Algicidal activity and identification of an algicidal substance produced by marine<Emphasis Type="Italic"> Pseudomonas</Emphasis> sp. C55a-2

A substance produced extracellularly by a marine bacterium, Pseudomonas C55a-2 strain, and possessing algal-killing activity against a diatom, Chaetoceros ceratosporum, was isolated from the culture supernatant of the bacterium in order to identify its chemical structure. The algicidal substance extracted with ethyl acetate from the culture supernatant was purified by using Sep-Pak treatment, reverse phase column chromatography with Sephadex LH-20, and high performance liquid chromatography (HPLC) with Mightysil RP-18 GP. The purified substance was identified as 2,3-indolinedione (isatin) (molecular weight 147) based on hydrogen-nuclear magnetic resonance (H-NMR) and gas chromatograph-mass spectrometry (GC-MS) analysis. Among artificial synthetic compounds examined for algicidal activity against Chaetoceros cells on the double-layer plates, 2,3-indolinedione (isatin) appeared the most effective and indoline showed relatively less activity than isatin.     

Water quality and phytoplankton stability in response to application frequency of bioaugmentation agent in shrimp ponds

Splitting the recommended weekly dosage of bioaugmentation agent and applied at varying frequencies were tested to determine its effects on water quality and phytoplankton stability in tiger shrimp (Penaeus monodon) grow-out ponds. The supplier's recommended weekly dosage was followed but at varying application frequencies namely: once per week, twice per week, and four times per week for the duration of 120 culture days. A treatment with no bioaugmentation was added as control. Results showed that ammonia and particulate organic matter levels were significantly lower and plankton die-off was greatly minimized if the product was applied four times a week application compared with twice a week and weekly applications. Significant reduction in the number of luminous Vibrio species was observed in ponds with more frequent applications than those with less. For shrimp, survival and production were significantly higher in ponds with twice and four times per week applications compared to the weekly application and the control (P < 0.05). Results of this study indicate that splitting the weekly dosage of bioaugmentation agent and applying it four times per week is highly beneficial and ideal for shrimp culture.     

Effect of synchronization of donor cells in early G1‐phase using shake‐off method on developmental potential of somatic cell nuclear transfer embryos in cattle

In this study, we compared the developmental ability of somatic cell nuclear transfer (SCNT) embryos reconstructed with three bovine somatic cells that had been synchronized in G0‐phase (G0‐SCNT group) or early G1‐phase (eG1‐SCNT group). Furthermore, we investigated the production efficiency of cloned offspring for NT embryos derived from these donor cells. The G0‐phase and eG1‐phase cells were synchronized, respectively, using serum starvation and antimitotic reagent treatment combined with shaking of the plate containing the cells (shake‐off method). The fusion rate in the G0‐SCNT groups (64.2 ± 1.8%) was significantly higher than that of eG1‐SCNT groups (39.2 ± 1.9%) (P < 0.05), but the developmental rates to the blastocyst stage of SCNT embryos per fused oocytes were similar for all groups. The overall production efficiency of the clone offspring in eG1‐SCNT groups (12.7%) per recipient cow was higher than that in G0‐SCNT groups (3%) (P < 0.05). The mean birth weight of cloned calves and the average calving score in the G0‐SCNT groups (48.1 ± 3.4 kg and 3.3 ± 0.3, respectively) was significantly higher (P < 0.05) than those of eG1‐SCNT groups (37.2 ± 2.1 kg and 2.3 ± 0.2, respectively). Results of this study indicate that synchronization of donor cells in eG1‐phase using the shake‐off method improved the overall production efficiency of the clone offspring per transferred embryo.     

Genetic Structure of Populations of the Pinewood Nematode Bursaphelenchus xylophilus, the Pathogen of Pine Wilt Disease, Between and Within Pine Forests

ABSTRACT We analyzed the genetic structure of Bursaphelenchus xylophilus populations within individual trees (subpopulations) in three distant pine forests (Tanashi, Tsukuba, and Chiba in Japan) based on the polymorphism of four microsatellite (SSR) markers. Most of the nematodes from subpopulations in Tanashi showed the same genotype over 2 years, indicating that nematodes of that genotype dominated there for years. In contrast, 16 and 15 genotypes were identified in nematode populations from Tsukuba and Chiba, respectively. Despite the high genetic diversity within the Tsukuba and Chiba populations, extremely low genetic diversity was observed within the subpopulations. The genetic difference between the Tsukuba and Chiba populations was significantly smaller than that between Tanashi and either Tsukuba or Chiba. Observed heterozygosity was significantly less than expected based on Hardy-Weinberg equilibrium. These findings are best explained by a founder effect, geographic isolation between populations, explosive nematode multiplication from a small number within individual trees, and the Wahlund effect.     

Organic-inorganic hybrid zeolites containing organic frameworks

Organic-inorganic hybrid zeolite materials containing an organic framework have been synthesized by partially superseding a lattice oxygen atom by a methylene group. The use of methylene-bridged organosilane as a silicon source gives zeolite materials containing an organic group as lattice (ZOL), with several zeolitic phases such as the MFI and the LTA structures. Through various characterization methods, the presence of methylene frameworks (Si-CH2-Si) that replace siloxane bonds (Si-O-Si) has been verified, although the Si-C bonds are partly cleaved to give rise to terminal methyl groups. The shape-selective lipophilicity of a ZOL material indicates that it is not a physical mixture of conventional zeolite and amorphous organic-containing material but contains a genuine organic-inorganic hybrid zeolite.     

Molecular characterization of South and East Asian melon, Cucumis melo L., and the origin of Group Conomon var. makuwa and var. conomon revealed by RAPD analysis

The genetic diversity and relationship among South and East Asian melon Cucumis melo L. were studied by using RAPD analysis of 69 accessions of melon from India, Myanmar, China, Korea, and Japan. The genetic diversity was large in India, and quite small in Group Conomon var. makuwa and var. conomon from East Asia, clearly indicating a decrease in genetic variation from India toward the east. Cluster analysis based on genetic distance classified 17 groups of accessions into two major clusters: cluster I comprising 12 groups of accessions from India and Myanmar and cluster II that included five groups of accessions of Group Conomon var. makuwa and var. conomon from East Asia. Cluster I was further divided into three subclusters, of which subclusters Ib and Ic included small- and large-seed type populations, respectively. Therefore, this division was based on their seed size, not cultivation area. The large-seed type from east India was differently included in the subcluster of small-seed type (Ib). A total of 122 plants of 69 accessions were classified into three major clusters and subclusters: clusters I and II comprised melon accessions mostly from India and Myanmar, and cluster III comprised Group Conomon var. makuwa and var. conomon from East Asia. The frequency of large- and small-seed types was different between clusters I and II, also indicating genetic differentiation between large- and small-seed types. One plant of the small-seed type from east India was differently included in cluster III, and two plants from east India were classified into subcluster IV. These results clearly showed that South Asian melon is genetically differentiated by their seed size, and that small-seed type melon in east India is closely related to Group Conomon var. makuwa and var. conomon.     

Bacteriolytic activity and growth of marine isolates of labyrinthulids on dead bacterial cells

ABSTRACT:   Thirteen strains of labyrinthulids were isolated from the coastal area of Kagoshima Bay, Kagoshima Prefecture, Japan and from Batan Bay, Panay Island, the Philippines by using diatom double-layer agar plates. The cells of labyrinthulid isolates grew axenically on the dead bacterial cells and extract plus egg yolk agar medium (NSBEY agar). Three representative isolates were demonstrated to belong to the labyrinthulid phylogenetic group (LPG) based on 18S  rDNA sequence analysis. As bacteriolytic activity of labyrinthulid isolates was examined, it was found that they could lyze only the dead cells of gram-negative bacteria and not those of gram-positive bacteria during incubation in both agar and liquid media. The optimum temperature range for bacteriolysis was 25–31°C. Orange carotenoid pigment was accumulated during the stationary growth phase of strain 00-Bat-05, Philippine isolate, cultured in an L-shaped tube containing a bacterial dead cell suspension. Concomitant rapid cell movement of developing zoospores was observed.     

Aluminum tolerance and aluminum-induced deposition of callose and lignin in the root tips of Melaleuca and Eucalyptus species

We exposed the roots of nine Myrtaceae species (Melaleuca bracteata F. Muell., Melaleuca cajuputi Powell, Melaleuca glomerata F. Muell., Melaleuca leucadendra [L.] L., Melaleuca quinquenervia [Cav.] S.T. Blake, Melaleuca viridiflora Sol. ex Gaertner, Eucalyptus camaldulensis Dehnh., Eucalyptus deglupta Bl., and Eucalyptus grandis W. Hill ex Maiden) to 1 mM Al in a nutrient solution for either 24 h or 20 days to evaluate their sensitivity to excess Al and to determine whether callose and lignin formation can be used as interspecific indicators of Al sensitivity. Inhibition of root elongation by Al varied among the species. Melaleuca leucadendra, M. cajuputi, E. grandis, M. quinquenervia, and E. deglupta were tolerant to 1 mM Al, whereas M. viridiflora, E. camaldulensis, M. glomerata, and M. bracteata were sensitive to 1 mM Al. We found that Al induced callose formation in the root tips of each species, but lignin was formed only in the root tips of the most sensitive species, M. bracteata. Root elongation at 1 mM Al was negatively correlated with Al-induced callose formation but not with the Al-induced lignin formation in the root tips. These results suggest that Al-induced callose formation, rather than lignin formation, can be used as an interspecific indicator of Al sensitivity.