Characterization of ethylene biosynthesis and its regulation during fruit ripening in kiwifruit,Actinidia chinensis ‘Sanuki Gold’

Ethylene biosynthesis in kiwifruit, Actinidia chinensis ‘Sanuki Gold’ was characterized using propylene, an ethylene analog, and 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception. In fruit harvested between a young stage (66 days after pollination) (DAP) and an early commercial harvesting stage (143 DAP), 2 days of exposure to propylene were sufficient to initiate ethylene biosynthesis while in fruit harvested at commercial harvesting stage (154 DAP), 4 days of propylene treatment were required. This observation suggests that response of ethylene biosynthesis to propylene treatment in kiwifruit declined with fruit maturity. Propylene treatment resulted in up-regulated expression of AC-ACO1, AC-ACO2, AC-SAM1 and AC-SAM2, prior to the induction of AC-ACS1 and ethylene production, confirming that AC-ACS1 is the rate limiting step in ethylene biosynthesis in kiwifruit. Treatment of fruit with more than 5 μL L^?1 of 1-MCP after the induction of ethylene production subsequently suppressed ethylene production and expression of ethylene biosynthesis genes. Treatment of fruit with 1-MCP at harvest followed with propylene treatment delayed the induction of ethylene production and AC-ACS1 expression for 5 days. These observations suggest that in ripening kiwifruit, ethylene biosynthesis is regulated by positive feedback mechanism and that 1-MCP treatment at harvest effectively delays ethylene production by 5 days.     

Immunohistochemical analysis of pseudorabies virus spread through neurons innervating the eyeball

Pseudorabies virus (PRV) was inoculated intraocularly into BALB/c mice, and the distribution pattern of cells positive for several neurotransmitters and viral antigens in the eyeball, trigeminal nerve ganglia, and superior cervical ganglia was examined immunohistochemically to clarify the neural route of the virus spread. In the eyeball, substance P (SP)- and calcitonin gene-related peptide (CGRP)-positive cells were detected in the ipsilateral iris and ciliary body, neuropeptide tyrosine (NPY)-positive cells were detected in the choloid membrane, and tyrosine hydroxylase (TH)-positive cells were detected in the ipsilateral inner nuclear layer of the retina; all these cells contained viral antigens. In the superior cervical ganglia, viral antigen-positive cells containing TH or NPY were found at bilateral sites. In the trigeminal nerve ganglia, viral antigen-positive cells containing SP or CGRP were found at bilateral sites. These findings indicated that the SP- and CGRP-positive ganglion cells of the trigeminal nerve ganglia innervating the iris or ciliary body, and the NPY-positive ganglion cells of the superior cervical ganglia innervating the iris, ciliary body, and choroid membrane served as the route for the virus spread. These findings also suggested that dopaminergic neurons, such as the TH-positive retinal cells and TH-positive ganglion cells of the superior cervical ganglia, served as the route for virus spread.     

Langerhans cells within the follicular epithelium and the intradermal sweat duct in equine insect hypersensitivity "Kasen"

Histopathologic and electron microscopic observations were given on Langerhans cells (LCs) within the follicular epithelium (FE) and intradermal sweat duct (ISD) of equine "Kasen". By light microscopy, LCs were present in the greatest numbers within the FE and ISD than within the epidermal layer and the normal skin, with an occasional formation of several aggregated foci. By electron microscopy, LCs within the FE and ISD widely extended their dendritic processes between the keratinocytes and contained Birbeck granules (Bgs), mitochondria, rough endoplasmic reticula and ribosomes in the cytoplasm. Numerous Type 2 LCs, with a number of Bgs and endocytosis, and Type 3 LCs, with multivesicular bodies and endosomes of various sizes, were recognized within the FE and ISD, although inactive Type 1 LCs, with a narrow and lucid cytoplasm, were rarely seen. LCs observed within the FE and ISD in the "Kasen" skin lesions might express the particular stage corresponded to recognize, intake and process the antigens which permeate them.     

Ultrastructural study of Langerhans cells in equine insect hypersensitivity "Kasen"

Ultrastructural features of Langerhans cells (LCs) of equine "Kasen" were studied. Electron microscopic observation revealed that LCs were dendritic and had irregular nuclear membranes. A number of Birbeck granules (Bgs) of various types were observed in the cytoplasm of LCs. In LCs in the upper stratum spinous, many Bgs were observed (Type 2 LC). LCs in the epidermo-dermal junction (EDJ) had a few Bgs, vesicles (multivesicular bodies) and highly electron-dense granular endosomes in the cytoplasm (Type 3 LC). Inactive LCs were also observed between the keratinocytes (Type 1 LC). Various types of LCs observed in the skin lesions of equine "Kasen" were interpreted as representing those that recognize, intake and process antigens.     

Pathogenesis-related gene expression in rice is correlated with developmentally controlled Xa21-mediated resistance against Xanthomonas oryzae pv. oryzae

Disease resistance mediated by the resistance gene Xa21 is developmentally controlled in rice. We examined the relationship between Pathogenesis Related (PR) defense gene expression and Xa21-mediated developmental disease resistance induced by Xanthomonas oryzae pv. oryzae (Xoo). OsPR1a, OsPR1b, and OsPR1c genes were cloned and their induction was analyzed, in addition to the OsPR10a gene, at the juvenile and adult stages in response to a wildtype Xoo strain that induces a resistance response (incompatible interaction) and an isogenic mutant Xoo strain that does not (compatible interaction). We found that the adult stage leaves are more competent to express these OsPR1 genes and that the Xa21 locus is required for the highest levels of induction.     

Changes in carbohydrase activities during vegetative growth and development of fruit-bodies of Hypsizygus marmoreus grown in sawdust-based culture

Carbohydraseproduction of Hypsizygus marmoreus cultured on sawdust rice bran medium by bottle cultivation was investigated to elucidate the carbohydrase utilized as the growth substrate for the fruit-body formation of this fungus. Among the extracellular enzymes assayed, xylanase showed the highest activity. This activity greatly increased during the end of vegetative mycelial growth and fruit-body formation. Among the cellulases. CM-cellulase showed higher activity than avicelase. The activities of -1,3-glucanase, amylase, and chitinase were low. Among the intracellular enzymes, both xylanase and amylase showed higher activity levels than the other carbohydrases. In contrast, -1,3-glucanase, avicelase, and chitinase activities in mycelia were considerably lower. These results suggest that xylanase, CM-cellulase, and amylase play an important role in mycelial maturation and fruit-body growth of H. marmoreus.     

Screening of extracts of Japanese woods for melanin biosynthesis inhibition

The effects of 120 methanol extracts prepared from bark and heartwood of 69 types of Japanese wood on the melanin production of B16 melanoma cells were examined. The melanin content of B16 melanoma cells was determined spectrophotometrically at 405nm. The extracts were also examined for their effects on cell viability. We found that the methanol extracts of Fagus crenata (buna, wood, 100μg/ml), Sapium sebiferum (Nankinhaze, wood, bark, 10μg/ml), and Zelkova serrata (keyaki, wood, 10μg/ml) greatly inhibited the melanin production of B16 melanoma cells without significant cytotoxicity. However, these extracts did not inhibit tyrosinase activity at the concentration of 100μg/ml. These findings indicate that the depigmenting mechanism of these extracts involves the suppression of some pigmenting signals in stimulating melanogenesis rather than the inhibition of tyrosinase activity. Part of this study was presented at the 53rd Annual Meeting of the Japan Wood Research Society, Fukuoka, Japan, March 2003     

Leaf water relations in <Emphasis Type="Italic">Pinus densiflora</Emphasis> Sieb. et Zucc. on different soil moisture conditions

To elucidate the differences in the leaf water relations of Pinus densiflora Sieb. et Zucc. growing in different soil moisture conditions, we examined the pressure-volume curve and the diurnal changes in the stomatal conductance, the transpiration rate, and the leaf water potential. The leaf water relations were compared using field-grown 40-year-old pine trees growing on the upper and lower parts of a slope. We also compared the leaf water relations of potted 4-year-old saplings growing at pF 4.2 and pF 1.8 soil moisture levels for almost 1 year. The values of the ratio of symplasmic water at turgor loss point to symplasmic water at saturated point (V_p/V_o) and bulk modulus of elasticity () of both the adult trees on the upper part of the slope and the potted saplings growing on pF 4.2 soil moisture were higher than those values of both the adult trees on the lower part of the slope and the potted saplings growing on pF 1.8 soil moisture, respectively. The field-grown adult tree and the potted saplings growing under long-term water stress tended to reduce their stomatal conductance in response to the acute soil drying. It is suggested that P. densiflora growing under long-term water stress rapidly closed its stomata in response to soil drying and avoided losing water, and could also rapidly absorb water with reducing water loss because of the decrease in the leaf pressure potential derived from the high values.     

Comparison of stand dynamics after dieback caused by pine wilt disease among pine forests with different management regimes in western Japan

In pine forests damaged by pine wilt disease, in western Japan, the effect of protection regimes of pine trees on the stand dynamics were examined in the following four stands: (1) lightly damaged stand (age 30–40 years) with no procedure in operation for protecting pine trees; (2) severely damaged stand (age 30–40 years) with no procedure in place for protecting pine trees; (3) severely damaged stand (age 50 years) with a selective cutting of infected trees; (4) severely damaged stand (age 30–40 years) with a selective cutting of infected trees. All the stands had been abandoned before the pine wilt disease damage. The understory structure of the severely damaged stand with no protection procedure was similar to that of the lightly damaged stand. Frequent invasion by tree species and acceleration in the growth of understory trees occurred after the dieback in the selective cutting stand. These results suggest that a deficiency in the canopy layer caused by the dieback resulted in low disturbance intensity in the early stages after the dieback, but the selective cutting increased the intensity by the reduction in the understory as well as the canopy layer. The intensity of the disturbance in the selective cutting stands was larger in the younger stand because it had a higher density of selectively cut pine trees. The different stand structure of pine forests occurred after the dieback because the intensity of the disturbance varied as a result of the selective cutting operation and the stand age.