Simultaneous down-regulation of DORMANCY-ASSOCIATED MADS-box6 and SOC1 during dormancy release in Japanese apricot (Prunus mume) flower buds

In temperate deciduous fruit crops such as Prunus spp., bud endodormancy is an important physiological phase affecting the timing of blooming and subsequent fruit development. Japanese apricot (Prunus mume) bears unmixed flower buds, separate from vegetative buds, that bloom slightly more than a month before vegetative bud burst. Seasonal expression of Prunus mume DORMANCY ASSOCIATED MADS-box genes (PmDAMs) has previously been analyzed only in vegetative buds, with an association between these genes and flower bud endodormancy release not yet confirmed. In this study, we performed a seasonal expression analysis of PmDAM1–6 genes in flower buds of two Japanese apricot genotypes – namely, high-chill and low-chill cultivars. The analysis revealed that PmDAM3, PmDAM5, and PmDAM6 expressions are closely associated with dormancy release in both flower and vegetative buds. In addition, a yeast two-hybrid screening demonstrated that PmDAM6 can interact in yeast with the homolog of Arabidopsis SOC1 (PmSOC1). Synchronized expression patterns were detected in PmDAM6 and PmSOC1 during dormancy release in flower buds of the two genotypes. Taken together, these results suggest that the dimer of PmDAM6 and PmSOC1 may play a role in the regulation of dormancy transition and blooming time in Japanese apricot flower buds.     

Change of fatty acid composition of the lumbar longissimus during the final stage of fattening in the Japanese Black cattle

Consideration of the shortened fattening period seems to be worthwhile for the realization of profitable beef production. In this study, change of fatty acid composition of the lumbar longissimus during the final stage of fattening was investigated in Japanese Black cattle. Each of 110 fattening animals was sampled three times: the initial two samples were taken by biopsy (25.7 months and 27.5 months on average) and the final one was from carcasses (29.9 months on average). Preliminary analysis indicated that removing muscle tissues from the constant body position of the living animals should be essential for sampling. Average monounsaturated fatty acids (MUFA) at three sampling points were 58.1%, 58.5% and 60.5%, and the differences of the third sampling with the first and second samplings were significant. Both in steers and heifers, MUFA also increased as the fattening stage proceeded, and MUFA of the heifers at all the sampling points were significantly higher than those of the steers. The increasing rate of MUFA rose from 0.21 percentage points (pp)/month at period 1 (from the first sampling to the second sampling) to 0.84 pp/month at period 2 (from the second sampling to the slaughter).     

Effects of voltage strength during electroporation on the development and quality of in vitro‐produced porcine embryos

This study was conducted to determine suitable conditions for an experimental method in which the CRISPR/Cas9 system is introduced into in vitro‐produced porcine zygotes by electroporation. In the first experiment, when putative zygotes derived from in vitro fertilization (IVF) were electroporated by either unipolar or bipolar pulses, keeping the voltage, pulse duration and pulse number fixed at 30 V/mm, 1 msec and five repeats, respectively, the rate of blastocyst formation from zygotes electroporated by bipolar pulses decreased compared to zygotes electroporated by unipolar pulses. In the second experiment, the putative zygotes were electroporated by electroporation voltages ranging from 20 V/mm–40 V/mm with five 1‐msec unipolar pulses. The rate of cleavage and blastocyst formation of zygotes electroporated at 40 V/mm was significantly lower (p < .05) than that of zygotes electroporated at less than 30 V/mm. Moreover, the apoptotic nuclei indices of blastocysts derived from zygotes electroporated by voltages greater than 30 V/mm significantly increased compared with those from zygotes electroporated by voltages less than 25 V/mm (p < .05). When zygotes were electroporated with Cas9 mRNA and single‐guide RNA (sgRNA) targeting site in the FGF10 exon 3, the proportions of blastocysts with targeted genomic sequences were 7.7% (2/26) and 3.6% (1/28) in the embryos derived from zygotes electroporated at 25 V/mm and 30 V/mm, respectively. Our results indicate that electroporation at 25 V/mm may be an acceptable condition for introducing Cas9 mRNA and sgRNA into pig IVF zygotes under which the viability of the embryos is not significantly affected.     

Tomato yellow leaf curl virus accumulates in vesicle-like structures in descending and ascending midgut epithelial cells of the vector whitefly, Bemisia tabaci, but not in those of nonvector whitefly Trialeurodes vaporariorum

Begomoviruses are transmitted by a single species of vector insect, the whitefly Bemisia tabaci, in a circulative manner. However, the mechanisms of this strict vector specificity have not been clarified. By immunoelectron microscopy, we showed that a begomovirus, Tomato yellow leaf curl virus (TYLCV), can enter midgut epithelial cells of the vector whitefly B. tabaci but not those of a nonvector whitefly, Trialeurodes vaporariorum, belonging to the same family. In midgut epithelial cells of viruliferous B. tabaci, the virus was localized in vesicle-like structures, suggesting endocytosis as an entry mechanism. These structures were also observed in midgut cells of nonviruliferous B. tabaci that had fed on healthy plants and in those of the nonvector T. vaporariorum that had fed on virus-infected plants. Vesicles containing TYLCV particles were observed most frequently in cells in the anterior part of the descending midgut, suggesting that this is the major entry site. These results clearly demonstrated that the virus-containing vector and nonvector whiteflies differ in the cellular localization of the virus and strongly suggest that a critical step in determining the vector insect specificity of begomoviruses is the entry of the viruses into midgut epithelial cells.     

Growth recovery of young hinoki (Chamaecyparis obtusa) subsequent to late weeding

The effects of late weeding treatment on the growth of young hinoki (Chamaecyparis obtusa) during the sixth growing season after planting were examined. Furthermore, the contribution toward hinoki growth recovery of crown productivity (NAR) and biomass allocation to crown (CAR) were determined. In the late weeding plot, no decline in height growth was observed subsequent to weeding, and growth in diameter at breast height (DBH) and crown projection area (CPA) began to recover subsequent to weeding; however, DBH and CPA were reduced and experienced a 1- to 1.5-year growth delay compared to values associated with the weeding plot at the end of the second year subsequent to weeding. Relative growth rate (RGR) and NAR in the late weeding plot recovered and possessed similar values to those of the weeding plot in the second year subsequent to weeding. CAR values of the late weeding plot were similar to those of the weeding plot both before and after weeding. These results suggested that the recovery of NAR rather than that of CAR was primarily responsible for the recovery of RGR. The present study demonstrated that hinoki were able to quickly acclimate to an environment dramatically altered by late weeding and recover growth rate within a short period of time. The present study also showed that delays in crown expansion associated with late weeding may have impeded subsequent matter production. Therefore, it was concluded that late weeding treatments should be employed only when the associated delays in growth are taken into account.     

The rubble-pile asteroid Itokawa as observed by Hayabusa

During the interval from September through early December 2005, the Hayabusa spacecraft was in close proximity to near-Earth asteroid 25143 Itokawa, and a variety of data were taken on its shape, mass, and surface topography as well as its mineralogic and elemental abundances. The asteroid's orthogonal axes are 535, 294, and 209 meters, the mass is 3.51 x 10(10) kilograms, and the estimated bulk density is 1.9 +/- 0.13 grams per cubic centimeter. The correspondence between the smooth areas on the surface (Muses Sea and Sagamihara) and the gravitationally low regions suggests mass movement and an effective resurfacing process by impact jolting. Itokawa is considered to be a rubble-pile body because of its low bulk density, high porosity, boulder-rich appearance, and shape. The existence of very large boulders and pillars suggests an early collisional breakup of a preexisting parent asteroid followed by a re-agglomeration into a rubble-pile object.     

B‐cell lymphoma with Mott cell differentiation in a cat

A 12‐year‐old, male castrated Domestic Shorthair cat was presented to Animal Medical Center of Gifu Univeristy with anorexia and vomiting. Physical examination revealed an enlarged left tonsil and right mandibular lymph node (approximately 2–3× the normal size), and a submucosal mass on the right side of the epiglottis (1.5 × 2.0 cm). On computed tomography images, an enlarged left tonsil, and enlarged right mandibular, right pharyngeal, and left and right cervical lymph nodes were observed. Cytologic examination of smears of tonsil and lymph nodes revealed numerous medium‐ to large‐sized neoplastic lymphoid cells, approximately half of which contained one or several light‐blue homogenous globoid cytoplasmic inclusions (5–10 μm), which stained magenta with periodic acid–Schiff (PAS) stain. Histopathologic examination of the left tonsil revealed diffuse proliferation of medium‐ to large‐sized neoplastic lymphoid cells effacing the original lymphoid architecture. Half of the cells contained one or several eosinophilic globoid cytoplasmic inclusions, which stained magenta with PAS and showed positive immunohistochemical reactions for immunoglobulin M (IgM) and λ light chain. Neoplastic lymphoid cells were also CD20⁺, Pax5⁺, and MUM1⁺, and CD3^?. Thus, the neoplastic lymphoid cells expressed a B‐cell immunophenotype, and the globoid cytoplasmic inclusions represented an aberrant IgM λ light chain accumulation, similar to Russell bodies. B‐cell lymphoma with Mott cell differentiation was diagnosed based on cytologic, histopathologic, and immunohistochemical features. This is the first report of B‐cell lymphoma with Mott cell differentiation in a cat.     

Genetic nature of bushy plant type in the variant strain induced by grafting in Capsicum annuum L.

Summary To elucidate the genetic nature of bushy plant type in the variant strains derived from grafting, testcrosses were made on the one hand between one of the variant strains, G₅S₁₇, and both Yatsubusa and Spanish Paprika, and on the other hand between the two cultivars.The bushy strain has more nodes and branches on the main stem than the two cultivars.This bushy plant character was recessive to cv. Yatsubusa used as a scion and over-recessive to cv. Spanish Paprika used as a root stock.Such graft-induced change from recessive to over-recessive as bushy plant type was quite different from other varied characters such as, fruit shape, capsaicin content and guard cell size, which were generally intermediate between the two cultivars.     

Mapping of EST-derived CAPS markers in Italian ryegrass (Lolium multiflorum Lam.)

New molecular markers derived from expressed sequence tag (EST) sequences were mapped on linkage maps of Italian ryegrass by a two-way pseudo-testcross strategy. cDNA sequences were obtained from various tissues of Italian ryegrass ( Lolium multiflorum ) and converted into cleaved amplified polymorphic sequence (CAPS) markers. Of 260 EST primer pairs that amplified a single band, 74 generated bands that showed clear polymorphisms among individuals of an F₁ mapping family. Of the 74 polymorphic marker loci, 69 were mapped on an Italian ryegrass linkage map previously constructed using amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism (RFLP), and simple sequence repeat (SSR) markers. The newly-developed EST-CAPS markers would be useful as an efficient tool to identify genetic markers and to identify candidate genes for quantitative trait loci (QTLs) associated with important traits in Italian ryegrass.