New types of major anthocyanins detected in Japanese garden iris and its wild forms

The anthocyanins of 130 cultivars, 13 lines and 3 wild forms of Iris ensata were analyzed by HPLC, and these plants were classified into 16 types of major anthocyanins. Among these types, 8 types such as petunidin 3RGac5G – delphinidin 3RGac5G, delphinidin 3RGac5G – petunidin 3RGac5G, cyanidin 3RGac5G – peonidin 3RGac5G, delphinidin 3RG – delphinidin 3RGac, petunidin 3RG5G – malvidin 3RG5G, malvidin 3RG5G – peonidin 3RG5G, peonidin 3RG5G – cyanidin 3RG5G and peonidin 3RG – cyanidin 3RG were obtained as new types. In these new types, peonidin 3RG – cyanidin 3RG and peonidin 3RG5G – cyanidin 3RG5G types were noteworthy because cyanidin 3RG and cyanidin 3RG5G are useful for the breeding of red flowers in I. ensata. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of silviculture treatments in a hurricane-damaged forest on carbon storage and emissions in central Hokkaido,Japan

Hurricanes cause abrupt carbon reduction in forests, but silviculture treatment can be an effective means of quickly regenerating and restoring hurricane-damaged sites. This study assessed how silviculture treatments affect carbon balance after hurricane damage in central Hokkaido, Japan. We examined carbon storage in trees and underground vegetation as well as carbon emissions from silviculture operations in 25-year-old stands, where scarification and plantation occurred just after hurricane damage. The amount of carbon stored varied according to silviculture treatment. Among three scarification treatments, a scarified depth of 0 cm (understory vegetation removal) led to the largest amount of carbon stored (64.7 t·ha-1 C). Among four plantation treatments, the largest amount of carbon was stored in a Larix hybrid (L. gmelinii var. japonica × L. kaempferi) plantation (80.3 t·ha-1 C). The plantation of Abies sachalinensis was not successful at accumulating carbon (40.5·ha-1 C). The amount of carbon emitted from silviculture operations was 0.05-0.14 t·ha-1 C, and it marginally affected the net carbon balance of the silviculture project. Results indicate that silviculture treatments should beperformed in an appropriate way to effectively recover the ability of carbon sequestration in hurricane-damaged forests.     

Positional syntenic cloning and functional characterization of the mammalian circadian mutation tau

The tau mutation is a semidominant autosomal allele that dramatically shortens period length of circadian rhythms in Syrian hamsters. We report the molecular identification of the tau locus using genetically directed representational difference analysis to define a region of conserved synteny in hamsters with both the mouse and human genomes. The tau locus is encoded by casein kinase I epsilon (CKIepsilon), a homolog of the Drosophila circadian gene double-time. In vitro expression and functional studies of wild-type and tau mutant CKIepsilon enzyme reveal that the mutant enzyme has a markedly reduced maximal velocity and autophosphorylation state. In addition, in vitro CKIepsilon can interact with mammalian PERIOD proteins, and the mutant enzyme is deficient in its ability to phosphorylate PERIOD. We conclude that tau is an allele of hamster CKIepsilon and propose a mechanism by which the mutation leads to the observed aberrant circadian phenotype in mutant animals.     

Stimulation of axillary shoot formation of cuttings of Hylocereus trigonus (Cactaceae) by pre-soaking in benzyladenine solution

Either before or after curing their cut surfaces for 5 days, 7 cm- and 15-cm-long decapitated Hylocereus trigonus cuttings were treated by soaking their apical or basal ends in benzyladenine (BA) solution. They were then planted and grown in a greenhouse.For the 7 cm-long cuttings, BA (25–100 mg l^?1) applied to the apical ends for 24 h increased the ratio of cuttings with sprouted buds to 64–100%, the number of sprouted buds to 1.9–3.1 and of shoots to 1.6–2.8, and the shoot length to 35–60 mm, compared to the water control which showed 13%, 1.0, 1.0 and 12.5 mm, respectively. Soaking the basal part had only a small effect.Naphthylacetic acid (NAA) applied to the basal ends of cuttings immediately after cutting increased the number of sprouted buds and shoots by inducing early rooting. The number and length of BA-induced axillary shoots in the longer cuttings was greater than those in the shorter ones.In the 15-cm-long cuttings, increasing the soaking time from 5 min to 24 h resulted in a greater promotive effect of BA on shoot formation. BA applied before curing showed the same effect as that given after curing but caused necrosis of the tissue just under the cut surface. Enlarging the area soaked in BA solution from 5 cm to 10 cm decreased the number of sprouting buds and shoots.     

Design of hammerhead ribozymes that cleave murine Sry mRNA in vitro and in vivo

As the first step in investigating the possiblity of applying ribozyme technology to artificial control of the sex ratios at birth in farm animals, where the demand for females exceeds that for males, we designed a hammerhead ribozyme (HHRz) and 2 tRNA(val)-hammerhead ribozyme complexes (tRNARz3 and tRNARz4), and examined their effects upon murine Sry mRNA in vitro and in cells. We demonstrated that HHRz and tRNARz3 could effectively cleave the target Sry mRNA in vitro. For the purpose of experiments in vivo, HHRz was cloned into the highly efficient pUC-CAGGS mammalian expression vector (pCAG/HHRz), and the tRNA ribozyme complexes were cloned into the pol III promoter-driven pPUR-KE vector (pPUR/tRNARz3 and pPUR/tRNARz4); the ribozyme vectors were co-transfected with the target vector (pCAG/Sry). A suppressive action (up to approx. 60%) was confirmed for pCAG/HHRz and pPUR/tRNARz3 upon the transiently expressed exogenously introduced Sry in M15 cultured cells.     

Effects of AI protocol and time interval from onset of estrus to AI on conception rate in Japanese Black cows

Two different artificial insemination (AI) protocols were investigated to determine the effect of the time interval from the onset of estrus (as detected by a pedometer) to the AI on the conception rate in Japanese Black cows. Seventy‐three cows were divided into two groups that received AI either after the induction of synchronized ovulation (synchronized AI group; n = 26) or at the time of observed estrus (control AI group; n = 47). Each group was further divided into two subgroups, which were artificially inseminated either 0–12 h (early AI group; n = 21) or 12–24 h (late AI group; n = 52) after the onset of estrus. There was no significant difference in the conception rate between the synchronized AI and control AI groups. The AI protocol and the detection of estrus using a threshold of counted steps (as measured by a pedometer) in this study could be effective for planned reproduction management without the observation of standing estrus in Japanese Black cows.     

Influence of Equine Conformation on Rider Oscillation and Evaluation of Horses for Therapeutic Riding

To obtain basic knowledge about selecting horses for therapeutic riding, the influence of equine conformation on rider oscillation and relationships between these factors and the evaluation on horses as the therapeutic riding were studied. Thirty-five riding horses were used. Equine conformation was estimated by 24 indices. Rider oscillation was measured by an accelerometer fixed at the rider’s waist. The spatial position of the oscillation was estimated by a double integration of the acceleration. Horses were evaluated for therapeutic riding by a Riding for the Disabled Association instructor as a rider. Evaluations were on a scale of 1 to 5, with 5 being the highest score for 27 items. Horses were classified into 4 groups: the short and narrow (SN), short and wide (SW), tall and narrow (TN), and tall and wide (TW). The frequencies of rider oscillation both at walk and trot were higher (P<0.01), and the vertical (P<0.01) and longitudinal (P<0.05) amplitudes at trot were smaller, on short horses than on tall horses. The vertical amplitude at walk was smaller (P<0.05) and the lateral amplitude at trot was larger (P<0.01) on wide horses than on narrow horses. Short horses could be used for the rider who requires side walkers. Wide horses could be used for relieving muscular tension and for the rider who could not maintain good balance on the horse. Short and wide horses should be suitable for therapeutic riding.     

Mean retention time of digesta in the different segments of the equine hindgut

Six Thoroughbred horses were used to determine the mean retention time (MRT) of digesta in the different segments of the hindgut. The horses were fed timothy hay or silage in equal amounts (1.6% bodyweight [BW]/day) every 3 h/day. Hay or silage labeled with the rare earth elements cerium, dysprosium, neodymium, ytterbium, lanthanum, samarium and praseodymium, were fed to the horses 36, 30, 24, 18, 12, 6 and 3 h before slaughter, respectively. Just after slaughter, digesta samples in the different segments were collected and determined for the contents of the rare earth elements. Using these data, a method to calculate the MRT in the different segments was proposed. There were no significant differences between diets in the MRT of each segment. The averaged MRT of hay and silage in the cecum, right ventral colon, left ventral colon, left dorsal colon, right dorsal colon and small colon was 2.9, 3.1, 5.9, 1.0, 4.0, and 4.0 h, respectively. The dry matter weight of the digesta was related to the MRT in the right ventral colon (r = 0.94, P = 0.005), left ventral colon (r = 0.87, P = 0.03), left dorsal colon (r = 0.98, P < 0.001) and right dorsal colon (r = 0.95, P = 0.004), but it was not related to the MRT in the cecum and small colon.     

Identification of quantitative trait loci for powdery mildew resistance in highly resistant cucumber (Cucumis sativus L.) using ddRAD-seq analysis

Powdery mildew, caused by Podosphaera xanthii (syn. Sphaerotheca fuliginea ex Fr. Poll.), is one of the most economically important foliar diseases in cucumber (Cucumis sativus L.). Cucumber parental line ‘Kyuri Chukanbohon Nou 5 Go’, developed from weedy cucumber line CS-PMR1, is highly resistant to powdery mildew and is promising breeding material. We performed quantitative trait locus (QTL) analysis using double-digest restriction-site-associated DNA sequencing (ddRAD-Seq) in a population from a cross between ‘Kyuri Chukanbohon Nou 5 Go’ and the Japanese native cultivar ‘Kaga-aonaga-fushinari’, which is susceptible to powdery mildew. The resistance of the population and its parents was evaluated using leaf disc assays and image analysis. We detected one major QTL on Chr. 5 that was effective at both 20°C and 25°C and one minor QTL on Chr. 1 effective at 20°C. We detected two additional QTLs in subpopulation: one on Chr. 3 effective at 20°C and one on Chr. 5 effective at both 20°C and 25°C in a position different from the major QTL. The resistance alleles at all four QTLs were contributed by ‘Kyuri Chukanbohon Nou 5 Go’. The results of this study can be used to develop practical DNA markers tightly linked to genes for powdery mildew resistance.