Immunohistochemical localization of enzymes related to lignin biosynthesis in the primary xylem of hybrid aspen

We have investigated the spatial regulation of the accumulation of enzymes involved in the biosynthesis of shikimate and lignin during differentiation of primary xylem from the apical meristem via procambium in hybrid aspen (Populus sieboldii x Populus grandidentata). Immuohistochemical staining revealed that, in the top part of shoots, lignification began in a single or just a few adjacent vessel elements and subsequently spread to neighboring cells. The spatial localization of 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase (DAHPS), which is one of the key enzymes in the shikimate pathway, was tightly correlated with the cell-specific deposition of lignin in the primary xylem. We also found that the spatial localization of enzymes in the general phenylpropanoid pathway and in the lignin-specific pathway was closely associated with the cell-specific deposition of lignin and the accumulation of DAHPS. Our data suggest that enzymes that act in the shikimate, general phenylpropanoid, and lignin-specific pathways are initially produced and function coordinately in a single or a few adjacent elements at the start of primary xylem development.     

The effect of bark decortication for hiwada production on xylem and phloem formation in Chamaecyparis obtusa

Of all plant materials used to cover the roofs of traditional Japanese buildings, Japanese cypress (Chamaecyparis obtusa) bark, hiwada, has the longest service life and has been used from ancient times. However, wood and bark properties after hiwada harvest have not been evaluated in detail. We studied whether decortication for hiwada production in winter affected xylem and phloem formation. Decorticated trees still preserved all inner bark and part of the outer bark, and both decorticated and control trees had similar annual ring structures at all stem heights in the xylem and phloem. In both xylem and inner bark, no significant difference in ring width at any stem height was found between annual rings before and after decortication. Thus, this study revealed that the decortication of bark for hiwada production does not affect the formation of xylem and the inner and outer bark if decortication is carried out by highly skilled workers in winter.     

Hybrid aspen with a transgene for fungal manganese peroxidase is a potential contributor to phytoremediation of the environment contaminated with bisphenol A

To assess the possible utility of a fungal gene for manganese-dependent peroxidase (MnP) produced by a transgenic plant in phytoremediation, we transformed hybrid aspen with a chimeric gene for MnP. Our gene construct allowed expression of the gene for MnP in plants and relatively high MnP activity was detected in the hydroponic medium in which roots of plants that expressed the transgene had been cultured. Some of our transgenic plants were able to remove bisphenol A from the medium more efficiently than wild-type plants. Our results demonstrate that, without any modification of the coding sequence, a chimeric gene for fungal MnP can be expressed in a woody plant, with secretion of active MnP from roots into the rhizosphere. Our strategy suggests new options using woody plants for phytoremediation.     

Manipulation of fish germ cell: visualization, cryopreservation and transplantation

Germ-cell transplantation has many applications in biology and animal husbandry, including investigating the complex processes of germ-cell development and differentiation, producing transgenic animals by genetically modifying germline cells, and creating broodstock systems in which a target species can be produced from a surrogate parent. The germ-cell transplantation technique was initially established in chickens using primordial germ cells (PGCs), and was subsequently extended to mice using spermatogonial stem cells. Recently, we developed the first germ-cell transplantation system in lower vertebrates using fish PGCs and spermatogonia. During mammalian germ-cell transplantation, donor spermatogonial stem cells are introduced into the seminiferous tubules of the recipient testes. By contrast, in the fish germ-cell transplantation system, donor cells are microinjected into the peritoneal cavities of newly hatched embryos; this allows the donor germ cells to migrate towards, and subsequently colonize, the recipient genital ridges. The recipient embryos have immature immune systems, so the donor germ cells can survive and even differentiate into mature gametes in their allogeneic gonads, ultimately leading to the production of normal offspring. In addition, implanted spermatogonia can successfully differentiate into sperm and eggs, respectively, in male and female recipients. The results of transplantation studies in fish are improving our understanding of the development of germ-cell systems during vertebrate evolution.     

Characterization of Erysipelothrix rhusiopathiae strains isolated from recent swine erysipelas outbreaks in Japan

The objective of the present study was to characterize Erysipelothrix sp. strains from recent erysipelas outbreaks in Japan. Eighty-three (100%) strains were identified as E. rhusiopathiae, based on serotyping and spaA PCR. Fifty (60.3%), 5 (6.0%), and 28 (33.7%) strains were isolated from animals with acute, subacute and chronic outbreaks, respectively, of which 79 (95.2%), 1 (1.2%), and 3 (3.6%) belonged to serotypes 1a, 2a, and untypeable, respectively. Fifteen strains (including 3, 2, and 10 from acute, subacute, and chronic cases, respectively) were sensitive to acriflavine, and showed high levels of virulence in mice; of which strains from acute cases, and from subacute and chronic cases killed 100%, and 80 to 100% mice, respectively at challenge doses of 10(2) CFU per mouse. Based on sequence analysis of a 432-bp hypervariable region in spaA gene, 83 strains could be divided into 3 groups: (i) group 1 (3 strains of serotype 1a) had Ala-195 and Ile-203; (ii) group 2 (76 strains of serotype 1a and 3 of untypeable) had Asp-195 and Met-203; and (iii) group 3 (one strain of serotype 2a) had Asn-195 and Ile-203. The results of the present study suggest that the serotype 1a strains belonging to the group 2 might be widespread in pig populations in Japan.     

Pathological Findings in a Case of Equine T-Cell Lymphoma Associated with Ataxia

A 15-year-old Dutch Warmblood gelding suddenly developed incoordination and hindlimb stumbling. The horse had a history of eyelid lymphoma. Necropsy revealed yellow-white or dark reddish-brown masses adhering to the outer surface of the spinal dura mater from the first cervical vertebra to the seventh thoracic vertebra. The spinal cord close to the first cervical vertebra and the seventh thoracic vertebra was markedly compressed by the masses filling the epidural space. The masses were also observed in the larynx, eyelids, and adipose-rich tissues, including the joints and orbits. They appeared similar in shape. The mandibular, retropharyngeal, axillary, superficial inguinal, deep inguinal, and lateral iliac lymph nodes were solid and enlarged. Histologically, the masses were composed of small or medium-sized lymphocyte-like tumor cells, but atypical cells and mitotic figures were rare. There were moderate infiltrations of macrophages and multinucleated giant cells, which were occasionally ingesting the surrounding tumor cells. Immunohistochemically, the tumor cells were classified as T-cell-derived cells. Throughout the spinal cord, enlargement or loss of nerve axons, dilation of periaxonal spaces, and macrophage infiltration into periaxonal spaces were observed, mainly in the ventral funiculus. Spinal cord compression by the tumor mass was suggested as a cause of the locomotive dysfunction. This is the first report of equine lymphoma with ataxia located from the proximal cervical to middle thoracic dura mater and in joint cavities.     

Use of bean husk as an easily digestible fiber source for activating the fibrolytic rumen bacterium Fibrobacter succinogenes and rice straw digestion

A series of in sacco and in vitro studies were carried out to evaluate bean husks for activation of fibrolytic rumen bacteria and rice straw digestion. First, lablab bean husk, chickpea husk and rice straw were suspended in the rumen of sheep to analyze the bacterial consortium developed on each fiber source. Known members of fiber‐associating bacteria were found on both lablab bean husk and rice straw, but some of these bacteria were lacking on chickpea husk. Second, a pure culture study was carried out using six strains of Fibrobacter succinogenes. Both husks stimulated the growth of all tested strains, including a strain that did not grow on rice straw. The strain OS128 that showed the highest growth on rice straw displayed even higher growth on lablab bean husk without a time lag. Finally, two‐step incubations were carried out to determine whether prior incubation of rumen fluid with husks stimulates subsequent rice straw digestion. Higher digestibility of rice straw was recorded in the second‐round incubation following the first incubation with bean husks. These results suggest that the tested bean husks improve the digestion of rice straw by activating fibrolytic F. succinogenes and other associated bacteria.     

Race distribution of Phytophthora sojae on soybean in Hyogo, Japan

Since 1987, Phytophthora root and stem rot of soybean [Glycine max (L.) Merr. cv. Tanbakuro], caused by Phytophthora sojae Kaufman and Gerdemann, has been increasing in the Sasayama, Nishiwaki, and Kasai regions in Hyogo, the most famous soybean (cv. Tanbakuro)-producing areas in Japan. In 2002 to 2004, 51 isolates (one from each field) of P. sojae were recovered from 51 fields in Hyogo. These isolates were tested for virulence on six Japanese differential soybean cultivars used for race determination in Japan, and three additional ones containing four Rps genes used in Indiana, USA. Race E was the most prevalent from 2002 to 2004, followed by races A, C, D, and four new races (proposed as races K, L, M, and N). Interestingly, none of the new races had high virulence on the Japanese differential cultivars, compared with other races in each area. One (race N) was avirulent on all six soybean differentials. There was a difference in race distribution on each of three individual areas; race E seemed to be a major component of the P. sojae population in Sasayama, whereas race A and the new race M were the most prevalent in Nishiwaki and Kasai, respectively. Rps6 (cv. Altona) and Rps1a + Rps7 (cv. Harosoy 63) were infected by 90.2% and 33.3% of all isolates, respectively. However, Rps1d (cv. PI103091) was not susceptible to any of the 51 isolates, nor was cv. Gedenshirazu-1. These two soybean cultivars were considered to be potential sources of resistance to breed new resistant cultivars with the desirable characteristics of cv. Tanbakuro for this region.