Observation of the scale of patchy stomatal behavior in leaves of Quercus crispula using an Imaging-PAM chlorophyll fluorometer

Patchy stomatal closure occurs in plants with heterobaric leaves, in which vertical extensions of bundle sheath cells delimit the mesophyll and restrict the diffusion of CO(2). The scale of patchy stomatal behavior was investigated in this study. The distribution of PSII quantum yield (Φ(II)) obtained from chlorophyll fluorescence images was used to evaluate the scale of stomatal patchiness and its relationship with leaf photosynthesis in the sun leaves of 2-year-old saplings of Quercus crispula Blume. Fluorescent patches were observed only during the day with low stomatal conductance. Comparison of numerical simulation of leaf gas exchange and chlorophyll fluorescence images showed that heterogeneous distribution of electron transport rate through PSII (J) was observed following stomatal closure with a bimodal manner under both natural and saturated photosynthetic photon flux densities. Thus, fluorescence patterns can be interpreted in terms of patchy stomatal closure. The mapping of J from chlorophyll fluorescence images showed that the scale of stomatal patchiness was approximately 2.5-fold larger than that of anatomical patches (lamina areas bounded by bundle sheath extensions within lamina). Our results suggest the spatial scale of stomatal patches in Q. crispula leaves.     

A new DNA marker for sex identification in purple asparagus

Garden asparagus (Asparagus officinalis L.) is an economically-important perennial crop. This plant is dioecious, as there are both male and female individuals; male individuals are preferred over females for agricultural production. To reduce the time required for garden asparagus breeding, various male-specific DNA markers are utilized. Male-specific DNA markers, such as Asp1-T7sp and MSSTS710, are currently available for sex identification in many asparagus cultivars. In the current study, we found that these markers are not suitable for sex identification in the purple asparagus cultivar ‘Pacific Purple’, as male-specific amplification of this marker was detected in some male individuals of this cultivar but not in other males. The Asp1-T7sp marker is suitable for use in sex identification in various Asparagus species related to A. officinalis, indicating that the region around this marker is conserved among these species. Thus, we isolated a DNA fragment around this marker by inverse PCR and produced a new DNA marker, MspHd, based on this sequence. However, like Asp1-T7sp and MSSTS710, MspHd was not suitable for sex identification in the cultivar ‘Pacific Purple’. Since all ‘Pacific Purple’ males have morphologically similar male flowers with functional stamens, we produced a new male-specific marker based on the sex determination gene, MSE1/AspMYB35/AspTDF1, which is responsible for stamen development. This marker, named AspMSD, is suitable for sex identification in ‘Pacific Purple’. In addition, this marker can be utilized for sex identification in various asparagus cultivars and some related Asparagus species.     

Decomposition of soil organic carbon influenced by soil temperature and moisture in Andisol and Inceptisol paddy soils in a cold temperate region of Japan

Purpose

Understanding the effects of temperature and moisture on soil organic carbon (SOC) dynamics is crucial to predict the cycling of C in terrestrial ecosystems under a changing climate. For single rice cropping system, there are two contrasting phases of SOC decomposition in rice paddy soils: mineralization under aerobic conditions during the off-rice season and fermentation under anaerobic conditions during the growth season. This study aimed to investigate the effects of soil temperature and moisture on SOC decomposition under the aerobic and subsequently anaerobic conditions.

Materials and methods

Two Japanese paddy soils (Andisol and Inceptisol) were firstly incubated under four temperatures (±5, 5, 15, and 25°C) and two moisture levels (60 and 100% water-filled pore space (WFPS)) under aerobic conditions for 24 weeks. Then, these samples were incubated for 4 weeks at 30°C and under anaerobic conditions. Carbon dioxide (CO₂) and methane (CH₄) productions were measured during the two incubation stages to monitor the SOC decomposition dynamics. The temperature sensitivity of SOC was estimated by calculation of the Q₁₀ parameter.

Results and discussion

The total CO₂ production after the 24-week aerobic incubation was significantly higher in both soils for increasing soil temperature and moisture (P < 0.01). During the subsequent anaerobic incubation, total decomposed C (sum of CO₂ and CH₄ productions) was significantly lower in samples that had been aerobically incubated at higher temperatures (15 and 25°C). Moreover, CH₄ production was extremely low in all soil samples. Total decomposed C after the two incubation stages ranged from 256.8 to 1146.1 mg C kg^?1 in the Andisol and from 301.3 to 668.8 mg C kg^?1 in the Inceptisol. However, the ratios of total decomposed C to SOC ranged from 0.29 to 1.29% in the Andisol and from 2.21 to 4.91% in the Inceptisol.

Conclusions

Both aerobic and anaerobic decompositions of SOC in two paddy soils were significantly affected by soil temperature and moisture. Maintaining optimal soil temperature and medium moisture during the off-rice season might be an appropriate agricultural management to mitigate CH₄ emission in the following rice growth season. Although it is high in SOC content, Andisol has less biodegradable components compared to Inceptisol and this could be a probable reason for the distinct difference in temperature sensitivity of SOC decomposition between two paddy soils.

     

Determination of physical and chemical properties and degradation of archeological Japanese cypress wood from the Tohyamago area using near-infrared spectroscopy

Here, we evaluated the application of near-infrared (NIR) spectroscopy for estimating the degradation level of archeological wood samples from the Tohyamago area, the dendrochronological ages of which were also determined. The wood samples were radially cut from three logs obtained from the Tohyamago area. NIR reflectance spectra were measured from the tangential faces of air- and oven-dried wood samples using a Fourier transform NIR spectrophotometer. The second derivative spectra within the wavenumber range of 6400–5200 cm^?1, in which the effect of moisture content in wood is suspected to be insignificant, showed a characteristic behavior with age. By comparing the second derivative spectral change in our wood samples with that in wood degraded by aging, thermal treatment, fungal attack, and lightning reported in the literature, we found that the second derivative spectra of wood samples from one log was similar to those of wood degraded by hygro-thermal treatment, whereas those of wood samples from another log was similar to those of wood degraded by brown-rot fungi. The physical and chemical properties of archeological wood were well predicted using a combination of partial least square regression analysis and NIR spectroscopy.     

Selective assessment of duplex heat-treated wood by near-infrared spectroscopy with principal component and kinetic analyses

We selectively assessed the thermal and hygrothermal treatment times of duplex heat-treated samples from the softwood hinoki cypress (Chamaecyparis obtusa) and the hardwood Japanese zelkova (Zelkova serrata) using near-infrared (NIR) spectroscopy with principal component analysis (PCA) and spectral-kinetic analysis. Wood samples from each species were thermally or hygrothermally treated at 120, 130, 150, and 180 °C, and the second-derivative spectra of these samples in the 6300–5450 cm^?1 range, where moisture content has the smallest effect, were then subjected to PCA. The master curve that was calculated by kinetic analysis successfully explained changes in the first principal component (PC1) scores with thermal treatment time for all temperatures. The angles between the PC1 loadings that explained the spectral variation due to thermal and hygrothermal treatment were 79° for hinoki and 80° for zelkova. Thus, calculation of the inner product between the second-derivative spectra of duplex heat-treated wood and a loading vector that explained the spectral variation due to thermal or hygrothermal treatment allowed us to selectively assess the thermal and hygrothermal treatment times.     

Seroprevalence of bovine immunodeficiency virus and bovine leukemia virus in dairy and beef cattle in hokkaido

Serological survey of bovine immunodeficiency virus (BIV) and bovine leukemia virus (BLV) infection was conducted in dairy cattle from 10 different regions of Hokkaido, Japan. Among 390 cattle, 11.0% of cattle were BIV-seropositive and 3.3% were BLV-seropositive. Moreover, in two dairy farms, where bovine leukosis has been reported, prevalence of BIV infections were 6.4 and 9.1%, respectively. In contrast, among 150 beef cattle, 16.6% were BIV-seropositive while none was BLV-seropositive. Dual infections with BLV and BIV in dairy cattle were tested by using 107 BLV-seropositive sera, and 20 sera were found BIV-positive (18.7%). These results indicate that BIV infection was widespread in Hokkaido.     

Plasmodium coatneyi-infected erythrocytes bind to C32 amelanotic melanoma cells under static and flow conditions

The ability of Plasmodium coatneyi-infected red blood cells (IRBCs) to bind to C32 amelanotic melanoma cells was examined under static and physiologic flow conditions in vitro. Six blood samples obtained from P. coatneyi-infected Japanese macaques (Macaca fuscata) with severe manifestations of disease were used in the static adhesion assay. All blood samples constantly exhibited binding of IRBCs to C32 cells under static conditions. Immunofluorescence staining with anti-CD36 mAb revealed a positive reaction at the surface of C32 cells with the infected erythrocytes, while the reaction with C32 cells without IRBCs was negative. To further examine the specificity of the interaction between P. coatneyi-infected erythrocytes and C32 cells, we carried out the binding assay under physiological flow conditions. In flow adhesion assay, three blood samples were used. Adhesion and rolling of IRBCs on C32 cells were detected at several rates of shear stress under flow conditions. At a shear stress of 1.0 dyne/cm(2), the number of IRBCs adherent to C32 cell averaged 5 to 6, and the number of IRBCs rolling on C32 cells averaged 6 to 11. The anti-CD36 mAb OKM5 inhibited 75-100% of IRBC adhesion and rolling, while the inhibitory effect of anti-ICAM-1 mAb 84H10 varied between 20-40%. The combination of anti-CD36 and anti-ICAM-1 mAb resulted in 83-100% inhibition of rolling and 100% inhibition of adhesion. These findings suggest that CD36 is one of the principal adhesion receptors of P. coatneyi-infected erythrocytes.     

New melanin biosynthesis inhibitors and their structural similarities

Relationships between their activities as blast control agents, and their abilities to inhibit mycelial melanisation on a nutrient agar, are described for 103 substituted benzothiazol-2(3H)-ones, benzoxazol-2(3H)-ones, indolin-2-ones, quinolin-2(1H)-ones, 1,2,3,4-tetrahydroquinolin-2-ones, benzo-1,4-thiazin-3(2H)-ones and benz-1,4-oxazin-3(2H)-ones, and some corresponding thiones. Several compounds in the respective series had a high protective activity and an antimelanisation activity against the blast fungus Pyricularia oryzae; furthermore, there was a good correlation in both of these activities, indicating that these compounds belong to the group of melanin biosynthesis inhibitors. Structural similarities of these compounds can be identified as follows: (a) having a benzo-bicyclic ring system; (b) containing a nitrogen atom in one ring at a position alpha to the benzene ring system; and (c) substitution, at the ring nitrogen atom, at the peri position in the aromatic ring relative to the nitrogen atom, and at the position alpha to the nitrogen atom in the ring system, with a double bond such as in carbonyl and thiocarbonyl groups. Among the compounds that have been proposed as melanin biosynthesis inhibitors, the chemical structures of tricyclazole, pyroquilon, 4,5-dihydro-4-methyltetrazolo[1,5-a]quinazolin-5-one (PP-389), [1,2,4]-triazolo[4,3-a]quinoline and 1-methylquinolin-2(1H)-one exhibit the structural similarities described above; however, 4,5,6,7-tetrachlorophthalide, 2,3,4,5,6-pentachlorobenzyl alcohol and N-substituted-2,3,4,5-tetrachloro-6-(hydroxymethyl)benzamides do not have such similarities in their chemical structures.     

The expression of sialyl Lewis X in canine and feline mammary gland tumors

The expression of sialyl Lewis X (sLe(x)) in 93 canine and 15 feline mammary gland tumors (MGT) obtained by surgical resection at Veterinary Medical Center, the University of Tokyo was examined by immunohistochemistry. Their clinicopathological features and prognosis were also reviewed. Approximately 60% of MGT tissues showed sLe(x) positive expressions, while all normal mammary gland tissues were negative. However, its expression was not correlated with clinicopathological features and prognosis significantly. This study suggests that sLe(x) may be a tumor-associated antigen in canine and feline MGTs.     

Examination of the rat eye at the early stage of development with osmium tetroxide staining

This communication describes the benefit of osmium tetroxide (OsO4) staining on the examination of the eye during the early stage of organogenesis of rat embryos. The embryos were obtained by laparotomy on embryonic day 12 (ED 12) and were stained with OsO4 for examination of the ocular tissues with a binocular stereo-microscope, light microscope and scanning electron microscope. At the binocular stereo-microscopic level, the invaginated lens placode, lens pit and optic cup were clearly distinguished. The osmium-stained lens placode and the optic cup were light brown and dark brown in color, respectively. Light microscopic examination revealed that OsO4 postfixation could provide superior paraffin-embedded embryonic sections. Scanning electron microscopic examination revealed the lens pit as a round opening between the lateral nasal prominence and maxillary prominence. Thus, a rapid technique by which the ocular tissues of rat embryos can be examined under a binocular stereo-microscope was developed. This OsO4 staining method will provide a useful tool for research on organogenesis and ocular development.