A New Plant Breeding Technique Using ALSV Vectors to Shorten the Breeding Periods of Fruit Trees

Fruit trees have a long juvenile phase. For example, the juvenile phase of apple lasts for 6–12 years and is a serious constraint for creating new varieties by breeding based on crossing and selection. In this chapter, we report a novel technology using the apple latent spherical virus (ALSV) vector to accelerate flowering time and life cycle in apple and pear seedlings. Inoculation of apple and pear cotyledons immediately after germination with ALSV-AtFT/MdTFL1 concurrently expressing Arabidopsis FLOWERING LOCUS T (AtFT) gene and suppressing apple TERMINAL FLOWER 1-1 (MdTFL1-1) gene can shorten the period from seeding to flowering to 1.5–3 months after germination and generation times in order to obtain next-generation seeds in 1 year or less. Most next-generation seedlings obtained from ALSV vector–infected plants were free of the virus. We also developed a method for eliminating ALSV vectors from infected apple and pear plants by only high-temperature treatment. A method combining the promotion of flowering in apple and pear by ALSV vector with an ALSV elimination technique is expected to see future application as a new plant breeding technique that can significantly shorten the breeding periods of apple and pear

Anticancer and antimetastatic effects of cordycepin, an active component of Cordyceps sinensis

AbstractCordyceps sinensis, a fungus that parasitizes on the larva of Lepidoptera, has been used as a valued traditional Chinese medicine. We investigated the effects of water extracts of Cordyceps sinensis (WECS), and particularly focused on its anticancer and antimetastatic actions. Based on in vitro studies, we report that WECS showed an anticancer action, and this action was antagonized by an adenosine A3 receptor antagonist. Moreover, this anticancer action of WECS was promoted by an adenosine deaminase inhibitor. These results suggest that one of the components of WECS with an anticancer action might be an adenosine or its derivatives. Therefore, we focused on cordycepin (3′-deoxyadenosine) as one of the active ingredients of WECS. According to our experiments, cordycepin showed an anticancer effect through the stimulation of adenosine A3 receptor, followed by glycogen synthase kinase (GSK)-3β activation and cyclin D1 suppression. Cordycepin also showed an antimetastatic action through inhibiting platelet aggregation induced by cancer cells and suppressing the invasiveness of cancer cells via inhibiting the activity of matrix metalloproteinase (MMP)-2 and MMP-9, and accelerating the secretion of tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 from cancer cells. In conclusion, cordycepin, an active component of WECS, might be a candidate anticancer and antimetastatic agent

Efficient virus-induced gene silencing in apple, pear and Japanese pear using Apple latent spherical virus vectors

<p>Abstract</p> <p>Background</p> <p>Virus-induced gene silencing (VIGS) is an effective technology for the analysis of gene functions in plants. Though there are many reports on virus vectors for VIGS in plants, no VIGS vectors available for <it>Rosaceae </it>fruit trees were reported so far. We present an effective VIGS system in apple, pear, and Japanese pear using <it>Apple latent spherical virus </it>(ALSV) vectors.</p> <p>Results</p> <p>Inoculation of ALSV vectors carrying a partial sequence of endogenous genes from apple [ribulose-1, 5-bisphosphate carboxylase small subunit (<it>rbcS</it>), alpha subunit of chloroplast chaperonin (<it>CPN60a</it>), elongation factor 1 alpha (<it>EF-1a</it>), or actin] to the cotyledons of seeds by a particle bombardment induced highly uniform knock-down phenotypes of each gene on the true leaves of seedlings from 2~3 weeks after inoculation. These silencing phenotypes continued for several months. Northern blot and RT-PCR analyses of leaves infected with ALSV containing a fragment of <it>rbcS </it>gene showed that the levels of <it>rbcS</it>-mRNA drastically decreased in the infected apple and pear leaves, and, in reverse, <it>rbcS-</it>siRNAs were generated in the infected leaves. In addition, some of apple seedlings inoculated with ALSV vector carrying a partial sequence of a <it>TERMINAL FLOWER 1 </it>gene of apple (<it>MdTFL1</it>) showed precocious flowering which is expected as a knock-down phenotype of the silencing of <it>MdTFL1 </it>gene.</p> <p>Conclusions</p> <p>The ALSV-based VIGS system developed have provides a valuable new addition to the tool box for functional genomics in apple, pear, and Japanese pear.</p

ALSV Vector Substantially Shortens Generation Time of Horticultural Plants

Flowering of plants is tightly regulated by both plant maturity and seasons in the year. Now that the Flowering LocusT (FT) gene has been revealed to encode the flowering hormone florigen, researchers are seeking to regulate and modify flowering behaviours by using florigen as a genetic tool. In place of transgenic approaches, Apple latent spherical virus (ALSV) vector was successful in promoting flowering of both model plants (Arabidopsis and tobacco), and fruit trees (e.g. apple, pear, and loquat), vegetables (e.g. tomato and cucumber), legumes (e.g. soybean), and ornamental flowers (e.g. petunia, Japanese gentian and Eustoma). In so doing, FT was expressed and/or TFL1 was suppressed by the ALSV vector. ALSV is a latent (non-pathogenic) virus isolated from an apple tree. After induction of flowering and seed production in crops, ALSV is not transferred to most of the next-generation seedlings, or it can be artificially removed from the infected plant by incubation at high temperature. Thus, the generation times of horticultural plants are approximately halved, and the generation time of apple plants is substantially shortened to within one year. Hence, ALSV technology is expected to be useful as a part of New Plant Breeding Techniques (NPBT) for agricultural application

閉塞型睡眠時無呼吸症候群患者における早朝の高分子量フォンウィルブランド因子減少は無呼吸の重症度を反映する

Plasma von Willebrand factor (VWF), produced in and released from vascular endothelial cells by various stimuli including hypoxia, induces platelet aggregation under high shear stress and plays dual pivotal roles in haemostasis and thrombosis within arterioles, which are regulated by the size of vWF multimers (VWFMs). Patients with obstructive sleep apnoea (OSA) have increased risk of thrombotic cardiovascular events, but the pathogenesis is unclear. We examined the relationship between VWF and OSA by measuring VWF antigen (VWF:Ag), VWFMs, VWF collagen binding activity (VWF:CB) and a disintegrin-like, metalloproteinase, and thrombospiondin type 1 motifs 13. A total of 58 OSA patients were enrolled. Blood samples were collected before sleep, after sleep, and after one night of nasal continuous positive airway pressure therapy. Based on VWFM analysis, OSA patients were classified into three groups; consistently normal VWFMs (group 1, n=29), increased high molecular weight (HMW)-VWFMs at 06:00 h (group 2, n=18), and decreased or absent HMW-VWFMs at 06:00 h (group 3, n=11). Patients in group 3 had significantly worse apnoea/hypopnoea index; VWF:CB followed a similar pattern. We observed a significant decrease in platelet count between 21:00 h and 06:00 h in OSA patients, potentially associated with reduced larger VWFMs together with decreased VWF:Ag levels. Severe OSA may contribute to an arterial pro-thrombotic state.博士（医学）・乙第1294号・平成24年5月28日Copyright © 2012 by the European Respiratory Societ

Detectability of the Warm/Hot Intergalactic Medium Through Emission Lines of OVII and OVIII

Most of cosmic baryons predicted by the big-bang nucleosynthesis has evaded the direct detection. Recent numerical simulations indicate that approximately 30 to 50 percent of the total baryons in the present universe is supposed to take a form of warm/hot intergalactic medium (WHIM) whose X-ray continuum emission is very weak. To identify those missing baryons, we consider in detail the detectability of WHIM directly through emission lines of OVII (561, 568, 574, 665eV) and OVIII (653eV). For this purpose, we create mock spectra of the emission lines of WHIM using a light-cone output of the cosmological hydrodynamic simulations. Since the predicted fluxes are generally below the current detection limit, the unambiguous detection requires a dedicated X-ray satellite mission that we also discuss in detail. We find that our proposed mission is sensitive to the WHIM with gas temperature $T=10^{6-7}$K and overdensity $\delta=10-100$ up to a redshift of 0.3 without being significantly contaminated by the cosmic X-ray background and the Galactic emissions. Thus such a mission provides a unique and important tool to identify a large fraction of otherwise elusive baryons in the universe.Comment: 22 pages, 10 figures. To appear in PASJ, v55, Oct. 25 (2003) issu

Arachidonic Acid Drives Postnatal Neurogenesis and Elicits a Beneficial Effect on Prepulse Inhibition, a Biological Trait of Psychiatric Illnesses

Prepulse inhibition (PPI) is a compelling endophenotype (biological markers) for mental disorders including schizophrenia. In a previous study, we identified Fabp7, a fatty acid binding protein 7 as one of the genes controlling PPI in mice and showed that this gene was associated with schizophrenia. We also demonstrated that disrupting Fabp7 dampened hippocampal neurogenesis. In this study, we examined a link between neurogenesis and PPI using different animal models and exploring the possibility of postnatal manipulation of neurogenesis affecting PPI, since gene-deficient mice show biological disturbances from prenatal stages. In parallel, we tested the potential for dietary polyunsaturated fatty acids (PUFAs), arachidonic acid (ARA) and/or docosahexaenoic acid (DHA), to promote neurogenesis and improve PPI. PUFAs are ligands for Fabp members and are abundantly expressed in neural stem/progenitor cells in the hippocampus. Our results are: (1) an independent model animal, Pax6 (+/−) rats, exhibited PPI deficits along with impaired postnatal neurogenesis; (2) methylazoxymethanol acetate (an anti-proliferative drug) elicited decreased neurogenesis even in postnatal period, and PPI defects in young adult rats (10 weeks) when the drug was given at the juvenile stage (4–5 weeks); (3) administering ARA for 4 weeks after birth promoted neurogenesis in wild type rats; (4) raising Pax6 (+/−) pups on an ARA-containing diet enhanced neurogenesis and partially improved PPI in adult animals. These results suggest the potential benefit of ARA in ameliorating PPI deficits relevant to psychiatric disorders and suggest that the effect may be correlated with augmented postnatal neurogenesis