苹果LEAFY同源基因的cDNA克隆与表达分析

 从苹果的果台枝顶芽中克隆了两个长约1.4 kb的cDNA片段，分别定名为AFL1和AFL2。它们的碱基序列在编码区有90%的同源性，但在3’末端非编码区仅有约印％的同源性。它们表达的肽链氨基酸序列与小叶杨、大豆、矮牵牛等有70％以上的同源性，与番茄、金鱼草、拟南芥有近70%的同源性，证明它们是LFY的同源基因。RT-PCR的结果表明，生长期AFL2在萼片、子房、根、茎、叶以及果台枝顶芽中都有表达，而AFL1只在果台枝顶芽中表达；在不同发育时期的果台枝顶芽中，AFL1在顶芽停止营养生长转向生殖生长以后才清晰表达，而AFL2在生长期6~10月都有表达。因此认为苹果中的这两个LFY同源基因虽有共同的起源，但是在功能上存在差异，在花和营养组织的发育中起不同的作用。DNA印迹分析表明，在苹果属及近缘的梨属植物中LFY同源基因是多拷贝的。     

Estimating the equilibrium position by measuring growth stress in weeping branches ofPrunus spachiana Kitamura f.spachiana cv.Plenarosea

The equilibrium position of a current-year branch of the weeping Japanese cherry,Prunus spachiana Kitamura f.spachiana cv.Plenarosea, was estimated by measuring the released strains of growth stresses. In current-year branches that were supported with wires to prevent weeping as soon as the branches first budded, tensile growth stresses on the upper side were smaller than those of the control branches. Gelatinous fibers were rarely found on the upper part of the cross section of the supported branches, whereas the control branches had many gelatinous fibers on the upper part. The upright orientation of the supported branches was closer to the equilibrium position than the weeping orientation of the control branches. The equilibrium position of the branches was thought to be in the initial bud direction, above the horizontal plane; and the weeping style of branch was not the preferred angular orientation forP. spachiana.     

Intrafiber distribution of aluminum components in alum-treated handsheets

The distribution of aluminum components in cross sections of pulp fibers for alum-treated handsheets was successfully measured by scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) analysis. In this case, gas-phase osmium coating of paper samples is necessary for the samples to have improved stability to long-term irradiation by electron beams at high magnification during the SEM-EDX measurements. The EDX line-analysis method was superior to the elemental mapping image method in the quantification of aluminum components. According to the SEM-EDX measurement for handsheets prepared from a pulp suspension by adding 2% (on dry weight of pulp) aluminum sulfate, the aluminum components were distributed almost homogeneously in the cross sections of pulp fibers. Aluminum species with smaller sizes than those of A1(OH)₃ flocs could probably penetrate the pulp fibers. Thus, aluminum components present only at the pulp fiber surfaces may contribute to retention of colloidal substances in pulp suspensions. From this aspect, a large amount of aluminum components present in the inside pulp fibers may be wasted.This report was presented at the 49th annual meeting of the Japan Wood Research Society at Tokyo, April 1997     

Effects of dietary taurine levels on epidermal thickness and scale loss in red sea bream,Pagrus major

Taurine is often added to artificial fish diets to compensate for a reduction in fish meal (FM). However, the taurine content of FM‐based diets is typically lower than in diets consisting of raw fish, even in diets where FM is the only protein source. We evaluated the effects of dietary taurine in FM‐based diets on epidermal thickness and scale detachability in red sea bream Pagrus major. We compared the effect of diets containing 0% (control), 0.3% (Tau‐0.3%), 0.6% (Tau‐0.6%) and 1.0% (Tau‐1.0%) taurine. Red sea bream (average body weight, 39 g) were fed these diets for 7 weeks. Taurine supplementation had no effect on growth, feed intake, feeding efficiency, or survival. However, the epidermal thickness was higher in fish in the Tau‐0.6% and Tau‐1.0% groups than in the control and Tau‐0.3% groups. Similarly, scale loss was significantly higher in the control group than in the Tau‐0.6% and Tau‐1.0% groups. Our results suggest that supplementation with >0.6% taurine (1.0% in diet) improves skin condition.     

Variation in leaf morphology of Quercus crispula and Quercus dentata assemblages among contact zones: a method for detection of probable hybridization

I describe variation in leaf morphology of Quercus crispula and Quercus dentata assemblages within and among their contact zones. Such relationships provide a relatively fast and accurate method for detecting hybridization. Using discriminant function (DF) analysis based on leaf characteristics, I was able to differentiate between these two species in pure zones. In three of five contact zones, the DF scores showed broad, continuous distributions with three or more peaks. The trees showing intermediate leaf morphology are considered to be hybrids. In the other two contact zones, the DF scores showed distinct bimodal distributions, suggesting that Q. crispula and Q. dentata rarely hybridize in these sites. Thus, the method described in this study should be effective for detecting probable hybridization between these two species in contact zones, and the relative frequency of hybridization among contact zones. However, there was likely to be intraspecific variation of Q. crispula between geographic ranges, restricting interpretations to patterns occurring within sites or limited geographic ranges.     

Technical note: development of a duodenal cannula for sheep.

A single T-shaped duodenal cannula of silicone rubber with a gutter-type small inner flange was developed for sheep. The barrel of the cannula was 24 mm long with an internal diameter of 12 mm. A polyester surgical mesh (100 mm x 100 mm) was connected to the barrel of the cannula as an anchor. Fibrous tissues grew on the polyester mesh anchor and adhered firmly to the serosa of the intestine, thus leaving no gap and, hence, preventing any leakage of intestinal contents from the side of the fistula. The small (24 mm in diameter) and thin (3 mm in thickness) outer flange of the cannula became buried in the wool and prevented any mechanical disturbance of the cannula by the activity of the animal. The elasticity of the silicone rubber prevented distortion of the duodenum around the barrel of the cannula. No erosion of the tissue between the inner flange of the cannula and the mesh was seen in postmortem observations.     

Transition of the pregnancy rate of bisected bovine embryos after co-transfer with trophoblastic vesicles prepared from in vivo-cultured in vitro-fertilized embryos

Bisected bovine embryos were co-transferred with trophoblastic vesicles (TVs). These TVs were prepared by dissection of conceptuses that were collected by uterine flushing after culture for seven days in the uterus following transfer of embryos derived by in vitro fertilization (IVF). Pregnancy diagnoses were performed twice, between Day 26 and Day 43 and between Day 38 and Day 73 post-estrus by ultrasonography. The pregnancy rate was significantly increased at first pregnancy diagnosis when demi-embryos were transferred with TVs (66.7%, 16/24) compared with the control group (34.5%, 10/29) (P < 0.05). Three losses occurred in the co-transfer group between the first and second pregnancy diagnosis. The final pregnancy rates according to delivered offspring were 41.7% (10/24) and 27.6% (8/29), respectively. There were no statistically significant differences between the pregnant and non-pregnant groups with regard to the average diameter of the TVs measured before transfer at three points during the gestation period. The birth weight and gestation lengths of the offspring were almost the same for the co-transfer and control groups. In the co-transfer group, the genetic identities of calves from the separated embryos were not affected by the TVs, as confirmed by parental blood type testing. Delivered offspring in co-transferred groups showed normal morphology. In conclusion, the present study indicates that co-transfer of TVs prepared from conceptuses cultured in vivo following transfer of IVF embryos enhances the fertility of demi-embryos during the early stages of pregnancy, as has similarly been shown in previous research for those prepared from in vivo embryos.     

Cerebellopontine angle meningioma expanding into the sella turcica in a dog

A 10-year-old, female poodle was presented with left paresis and vestibular signs, following left enophthalmus and atrophy of the cheek. Magnetic resonance imaging revealed a mass along the base of the brain and extending from the left cerebellopontine angle to the sella turcica. The mass showed isointensity on the T1-weighted image and T2-weighted image, and was enhanced by contrast medium (Gd-DTPA). Although occipital craniotomy was carried out and the mass removed, the dog died without recovering from the anesthesia. The tumor was diagnosed pathologically as fibroblastic meningioma.     

Direct detection of dermatophytes in skin samples based on sequences of the chitin synthase 1 (CHS1) gene

For the direct detection of dermatophytes in skin scrapings and hairs from animals, a primer pair specific to the chitin synthase 1 (CHS1) gene of dermatophytes was constructed. By PCR analysis with the primer pair, dermatophyte DNA could be diagnosed directly and rapidly in clinical skin samples.     

Delayed-type hypersensitivity in sheep induced by synthetic peptides of bovine leukemia virus encapsulated in mannan-coated liposome

Four sheep were immunized with synthesized peptides, derived from bovine leukemia virus (BLV) envelope glycoprotein, encapsulated in mannan coated liposomes as adjuvant. On the seventh day after the immunization, the sheep were intradermally challenged with BLV antigen, or synthesized peptides. The areas challenged with antigen were increased skin thickness and biopsied sequentially for immunohistological examinations. Strong delayed-type hypersensitivity was induced in sheep immunized and challenged with peptides encapsulated in mannan-coated liposomes. The major phenotype of the infiltrating lymphocytes was CD5(+). The ratio of CD4(+) to CD8(+) cells was about 1:1.