The occurrence and effects of red clover necrotic mosaic virus in red clover (Trffbliumpratense)

In 1976, red clover necrotic mosaic virus (RCNMV) was identified in red clover variety trials at the Scottish Colleges of Agriculture and at the trial centres of the National Institute of Agricultural Botany (NIAB) in Northumberland, Dyfed, Devon and Cambridge. In 1977, RCNMV was also found in two commercial crops of red clover in South Wales. The only previous finding of this virus in Britain was in 1971.
In red clover leaves RCNMV causes veinal chlorosis, often followed by severe necrosis and deformation; the plants become stunted. All cultivars tested were infected either in field or glasshouse experiments and three of the four most susceptible cultivars were tetraploids. Yield losses in cv. Hungaropoly averaged 57% over three cuts. RCNMV was transmitted manually but not through seed or by aphids {Acyrthosiphon pisum and Myzus persicae) or weevils (Apion spp. and Sitona lineatus). Seedlings became infected when grown in pots containing RCNMV-infected plants or soil from infected sites, and the roots of infected test seedlings contained an Olpidium sp. which may be the vector.
White clover mosaic virus (WCMV), also common in red clover at some sites, was less damaging than RCNMV and in a glasshouse experiment decreased yield by only 22%. An unidentified seed-borne virus with spherical particles c. 33 nm in diameter was the only virus detected in clover seedlings screened for RCNMV.     

Evidence for a new allele at the esterase D (E.C.3.1.1.1) locus

A new rare allele for esterase D (ESD) is described in a family from Düsseldorf. The variant was tentatively named ESD Düs 2.     

显性E1基因背景下大豆开花和成熟期的全基因组关联分析

【目的】深入解析大豆开花期和成熟期的遗传基础,发掘控制性状的重要基因组区间,为分子标记辅助选择育种和新基因克隆提供依据。【方法】以224份携带显性E1基因的大豆种质为试验材料,在4个种植环境下调查表型数据,采用R-mrMLM软件包中的6种多位点关联分析模型对大豆开花和成熟期进行全基因组关联分析。【结果】共检测到91个开花期和83个成熟期QTNs（quantitative trait nucleotides）,其中6个开花期和10个成熟期QTNs能在多环境中检测到。这些环境稳定QTNs分布于15个区间大小在90~490 Kb的单倍型（基因组）内,并有6个基因组区间为本研究新检测到。【结论】研究表明显性E1基因背景下大豆开花和成熟期的QTN构成复杂,位于5号染色体39.52~40.01Mb等15个基因组区间是该群体中控制大豆开花期或成熟期的重要位点。     

A first genotyping assay of French cattle breeds based on a new allele of the extension gene encoding the melanocortin-1 receptor (Mc1r)

The seven transmembrane domain melanocortin-1 receptor (Mc1r) encoded by the coat color extension gene (E) plays a key role in the signaling pathway of melanin synthesis. Upon the binding of agonist (melanocortin hormone, α-MSH) or antagonist (Agouti protein) ligands, the melanosomal synthesis of eumelanin and/or phaeomelanin pigments is stimulated or inhibited, respectively. Different alleles of the extension gene were cloned from unrelated animals belonging to French cattle breeds and sequenced. The wild type E allele was mainly present in Normande cattle, the dominant E^D allele in animals with black color (i.e. Holstein), whereas the recessive e allele was identified in homozygous animals exhibiting a more or less strong red coat color (Blonde d''Aquitaine, Charolaise, Limousine and Salers). A new allele, named E¹, was found in either homozygous (E¹/E¹) or heterozygous (E¹/E) individuals in Aubrac and Gasconne breeds. This allele displayed a 4 amino acid duplication (12 nucleotides) located within the third cytoplasmic loop of the receptor, a region known to interact with G proteins. A first genotyping assay of the main French cattle breeds is described based on these four extension alleles.     

Presence of Leishmania infantum in red foxes (Vulpes vulpes) in southern Italy

Skin, lymph node (popliteal), and bone marrow samples were collected from 50 red foxes (Vulpes vulpes) from May 2004 to May 2005 in southern Italy. Samples were tested for Leishmania infantum by polymerase chain reaction (PCR). The parasite was detected by PCR from 20 of 50 (40%) fox carcasses. All 20 positive cases were PCR-positive from lymph node and bone marrow samples, whereas 17 of 20 positive cases were PCR-positive from skin samples. Infection status was not related to age or sex. This is the first report of leishmaniasis in red foxes in Italy based on PCR results, and these results reinforce the assumption that this wild canid can serve as a reservoir for Leishmania.     

Presence of an aromatase inhibitor, possibly heat shock protein 90, in dominant follicles of cattle.

In cattle, it has been suggested that follicular fluid has direct modulatory effects on follicular growth and maturation. In the first part of this study, an in vitro test using aromatase activity of follicular wall fragments as an end point was validated for cattle follicles and was used to test whether follicular fluid (from dominant or non-dominant follicles) modulates aromatase activity. Fluid from dominant follicles at a concentration of 24 or 12% (obtained during the luteal and follicular phases, respectively) significantly inhibited aromatase activity. Inhibitory activity was low or absent in fluid from non-dominant follicles. FSH-stimulated aromatase activity was also reduced by fluid from dominant follicles, but not to a greater extent than in basal conditions. Finally, charcoal-treated fluid from dominant follicles retained its inhibitory activity. In contrast, ovarian venous serum draining a dominant follicle had no activity at the three concentrations tested (6, 12 and 24%). In the second part of the study, identification of the compounds involved in this modulatory activity was attempted using SDS-PAGE. Comparison of the fluorographs from de novo synthesized proteins stored in follicular fluid (inhibitory medium) with those secreted in incubation medium (inactive medium) demonstrated that one protein (90 kDa, pI 5.8) was significantly (P < 0.05) more abundant in fluid from dominant follicles (2.0 +/- 0.09%) than in the culture medium (1.3 +/- 0.1% of the total proteins). This protein had characteristics similar to those of heat shock protein 90 (hsp 90). Therefore, in the final part of the study, the presence of hsp 90 in ovarian cells and follicular fluid was investigated using immunohistochemistry and western blot analysis. After immunohistochemistry, a positive signal was detected mainly in the granulosa cells of larger follicles and to a smaller extent in thecal cells and oocytes. Western blot analysis also demonstrated the presence of hsp 90 in follicular wall fragments and fluid. When blotting was achieved on a sample of follicular fluid resolved by two-dimensional PAGE, the spot detected had a similar location to that at 90 kDa and pI 5.8. Addition of purified hsp 90 to bovine follicles in vitro depressed aromatase activity by altering the K(m) value (and possibly the Vmax value) of the enzyme. It is proposed that hsp 90 is a functional regulator of follicular maturation through its action on aromatase.     

Association of Rho(D) polypeptides with the membrane skeleton in Rho(D)-positive human red cells

The Rho(D) antigen was recently identified as a 28,000 to 33,000 m.w. polypeptide expressed on the surface of human Rho(D)+ cells. We now show that 70 to 80% of the Rho(D) polypeptides remain firmly associated with the membrane skeleton (detergent-insoluble matrix) obtained after treatment of isolated membranes with Triton X-100. The same treatment solubilized most of the major sialoglycoprotein, glycophorin A. The membrane skeleton-bound Rho(D) polypeptides were not solubilized by procedures that dissociated spectrin, actin, and glyceraldehyde-3-phosphate dehydrogenase from the membrane. Affinity-purified 125I-labeled anti-Rho(D) antibodies bound to intact Rho(D)+ cells, Rho(D)+ membranes, and isolated membrane skeletons from Rho(D)+ cells, but not to Rho(D)- cells. The binding to Rho(D)+ cells was competitively inhibited efficiently by Rho(D)+ membranes and weakly by Rho(D)- membranes. When isolated unsealed Rho(D)+ and Rho(D)- membranes were labeled by lactoperoxidase-catalyzed iodination and solubilized in Triton X-100, Rho(D) polypeptides were immune precipitated only from Rho(D)+ membranes.     

A pseudodeficiency allele (D152N) of the human beta-glucuronidase gene.

We present evidence that a 480G-->A transition in the coding region of the beta-glucuronidase gene, which results in an aspartic-acid-to-asparagine substitution at amino acid position 152 (D152N), produces a pseudodeficiency allele (GUSBp) that leads to greatly reduced levels of beta-glucuronidase activity without apparent deleterious consequences. The 480G-->A mutation was found initially in the pseudodeficient mother of a child with mucopolysaccharidosis VII (MPSVII), but it was not on her disease-causing allele, which carried the L176F mutation. The 480G-->A change was also present in an unrelated individual with another MPSVII allele who had unusually low beta-glucuronidase activity, but whose clinical symptoms were probably unrelated to beta-glucuronidase deficiency. This individual also had an R357X mutation, probably on his second allele. We screened 100 unrelated normal individuals for the 480G-->A mutation with a PCR method and detected one carrier. Reduced beta-glucuronidase activity following transfection of COS cells with the D152N cDNA supported the causal relationship between the D152N allele and pseudodeficiency. The mutation reduced the fraction of expressed enzyme that was secreted. Pulse-chase experiments indicated that the reduced activity in COS cells was due to accelerated intracellular turnover of the D152N enzyme. They also suggested that a potential glycosylation site created by the mutation is utilized in approximately 50% of the enzyme expressed.     

Chinese white Rongchang pig does not have the dominant white allele of KIT but has the dominant black allele of MC1R

The mast/stem cell growth factor receptor (KIT) and melanocortin receptor 1 (MC1R) mutations are responsible for coat color phenotypes in domestic pigs. Rongchang is a Chinese indigenous pig breed with a white coat color phenotype. To investigate the genetic variability of the KIT and MC1R genes and their possible association with the coat color phenotype in this breed, a gene duplication and splice mutation of KIT were diagnosed in a sample of 93 unrelated Rongchang animals. The results show that Rongchang pigs have a single copy of KIT without the splice mutation at the first nucleotide of intron 17, indicating that the dominant white I allele of KIT is not responsible for their white phenotype. The KIT mRNA and MC1R coding sequences were also determined in this breed. Three putative amino acid substitutions were found in the KIT gene between Rongchang and Western white pigs, their association with the Rongchang white phenotype remains unknown. For the MC1R gene, Rongchang pigs were demonstrated to have the same dominant black allele (E(D1)) as other Chinese breeds, supporting the previous conclusion that Chinese and Western pigs have independent domestication origin. We also clarified that the Rongchang white phenotype was recessive to nonwhite color phenotypes. Our results provide a good starting point for the identification of the mutations underlying the white coat color in Rongchang pigs.     

A natural mutation in rc reverts white-rice-pericarp to red and results in a new, dominant, wild-type allele: Rc-g

The Rc locus regulates pigmentation of the rice bran layer, and selection for the rc allele (white pericarp) occurred during domestication of the crop. White bran is now ubiquitous among cultivated varieties throughout rice growing regions of the world. We identified a new allele that arose by natural mutation within the rc pseudogene of the cultivar 'Wells'. The mutation restored the reading frame of the gene, and reverted the bran layer pigmentation to red (wild-type). By sequencing the Rc locus in plants derived from red seeds, and linkage analysis in a segregating population, we were able to demonstrate that mutation within rc resulted in the new, dominant, wild-type allele Rc-g.     

Evidence for a Null allele at the Esterase D (EC 3.1.1.1) locus

Summary Electrophoretic and quantitative assays of esterase D in a Caucasian family demonstrate the inheritance of a null allele, which was observed in the heterozygous state in six individuals.     

(E160A)和(E160D)天花粉蛋白两种突变体晶体结构研究

培养了（Ｅ１６０Ａ）ＴＣＳ和（Ｅ１６０Ｄ）ＴＣＳ的单晶。在ＭＡＲＲｅｓｅａｒｃｈ面探测器系统上分别收集了０．１９３ｎｍ和０．２０ｎｍ分辨率的Ｘ射线衍射数据。数据处理用ＭＡＲＳＣＡＬＥ程序系统完成。用同晶差值Ｆｏｕｒｉｅｒ法解析了突变体的晶体结构,结构修正利用Ｘ－ＰＬＯＲ程序。修正结果,晶体学Ｒ因子分别为０．１７５,０．１７９,键长和键角的ＲＭＳ偏差分别为０．００１１ｎｍ和２．４５７°,０．００１３ｎｍ和２．６７５°。在这两个突变体的结构中均未见到Ｇｌｕ１８９侧链方向的改变。通过对（Ｅ１６０Ａ）ＴＣＳ和（Ｅ１６０Ｄ）ＴＣＳ的结构比较,说明（Ｅ１６０Ｄ）ＴＣＳ活性低于（Ｅ１６０Ａ）ＴＣＳ的原因：这可能是由于在（Ｅ１６０Ｄ）ＴＣＳ中Ｔｙｒ１１１和Ｔｙｒ７０的侧链都具有较大的运动性,使它们与腺嘌呤碱基的芳香堆垛作用减弱,从而导致活性的降低     

Inheritance of the allele Pr of the red cell acid phosphatase (EC. 3.1.3.2) in a Caucasian family

Summary The occurrence of the allele P^r showing an unexpected atypical isozyme band pattern in a Caucasian family is reported.     

Molecular characterization of the human red cell Rho(D) antigen

Human red cells of Rh blood groups -D-/-D- ('super-D'), -/- (Rhnull) and normal Rho(D)+ cells were radioactively surface-labeled using the lactoperoxidase 125I method. Polyacrylamide gel electrophoresis in the presence of SDS followed by fluorography showed a strong enrichment of a polypeptide with an apparent mol. wt. of 28,0000-33,000 in the 125I-labeled -D-/-D- membranes. This polypeptide was specifically immune precipitated with anti-Rho(D) antiserum. Treatment of intact cells with trypsin or Pronase did not digest the protein. The Rho polypeptide migrated identically on polyacrylamide gel electrophoresis under reducing and non-reducing conditions. It was not phosphorylated after in vitro incubation of red cells with 32P. When whole labeled membranes were solubilized in neutral detergent and applied to lectin-Sepharose columns the Rho(D) polypeptide adsorbed to Ricinus communis lectin but not to wheat germ lectin or Lens culinaris lectin. The purified molecule did not adsorb to R. communis lectin-Sepharose. Treatment of the Rho(D) antigen with endo-N-acetyl glucosaminidase H, endo-beta-galactosidase or mild alkali did not lower its apparent mol. wt.     

Compensatory extension of gestation length with advance of conception in red deer (Cervus elaphus)

Calving date in many mammals is matched to the time of greatest food availability. Out of season calving results in heavy penalties in terms of own and offspring survival or body condition. This study examined whether gestation length is affected by advancing fertilisation. Thirty-six red deer hinds (of the Iberian and Scottish subspecies) were subjected to a synchronisation treatment of oestrus, ovulation, and artificial insemination on three dates, with remaining non-pregnant females mated with an intact male in a last group. Gestation was longer the more the fertilisation was advanced; gestation lasted 241.5+/-1.3 days (d) in the first group, 237.4+/-1.2 d in the second, 235.1+/-1.3 d in the third, and 231.2+/-1.6 d in the last. Mean gestation lasted 234.2+/-0.7 d. Hinds gained less weight during gestation the more the fertilisation was advanced. The difference was due at least in part to net body weight of the hind after calving compared to that at mating, and calves did not differ in birth weight. As early born calves suffer greater mortality in the field, this enlargening of gestation might be a compensatory response of the hinds to match calving with food availability. Under natural conditions, similar small modifications of gestation length may help hinds to overcome short-term adverse conditions for calving. Because calf mortality is correlated with birth weight, hinds may have kept calf birth weight constant at the expense of greater body weight loss.     

Temporal dissection of K-ras(G12D) mutant in vitro and in vivo using a regulatable K-ras(G12D) mouse allele

Animal models which allow the temporal regulation of gene activities are valuable for dissecting gene function in tumorigenesis. Here we have constructed a conditional inducible estrogen receptor-K-ras(G12D) (ER-K-ras(G12D)) knock-in mice allele that allows us to temporally switch on or off the activity of K-ras oncogenic mutant through tamoxifen administration. In vitro studies using mice embryonic fibroblast (MEF) showed that a dose of tamoxifen at 0.05 μM works optimally for activation of ER-K-ras(G12D) independent of the gender status. Furthermore, tamoxifen-inducible activation of K-ras(G12D) promotes cell proliferation, anchor-independent growth, transformation as well as invasion, potentially via activation of downstream MAPK pathway and cell cycle progression. Continuous activation of K-ras(G12D) in vivo by tamoxifen treatment is sufficient to drive the neoplastic transformation of normal lung epithelial cells in mice. Tamoxifen withdrawal after the tumor formation results in apoptosis and tumor regression in mouse lungs. Taken together, these data have convincingly demonstrated that K-ras mutant is essential for neoplastic transformation and this animal model may provide an ideal platform for further detailed characterization of the role of K-ras oncogenic mutant during different stages of lung tumorigenesis.     

Difference in apolipoprotein E type 4 allele (APOE epsilon 4) among dentate and edentulous subjects

Objectives: To evaluate the frequency of apolipoprotein (APOE) alleles and determine whether APOE type 4 allele (?4) was associated with edentulousness even when certain factors were controlled. Background: The APOE are important in lipid homeostasis, and APOE ?4 has been found in many diseases and to have a negative impact on longevity. Tooth loss is more common in ill aged subjects with low income and education. Materials and methods: In a population‐based study involving 1860 subjects between 35 and 85 years 1321 dentate (mean age = 54; 54% women, 46% men) and 539 edentulous (mean age = 72; 62% women, 38% men) subjects were studied. Logistic regression was performed with dentate/edentulous as dependent variables and years of education, socio‐economic status, social network, stress level, handicap from birth, 23 various diseases and APOE ?4 as covariates. Thereafter, APOE ?4 frequencies were studied in 342 dentate and 336 edentulous subjects 50–85 years of age. The subjects were matched with regard to age, gender, years of education, living condition, stress level, handicap from birth and 23 various diseases. Results: APOE allele frequency in the total group was ?2 = 7.8%, ?3 = 76.4% and ?4 = 15.8%. Age, living condition, years of education and APOE ?4 were significant covariates in edentulous subjects (p ≤ 0.001). APOE ?4 in the matched groups revealed significant differences between the dentate group and the edentulous group (χ² = 5.68; p = 0.017). There was no group effect (F_(29,648) = 0.849; p < 0.696; Wilks’ lambda = 0.963). In the dentate group, the frequencies of APOE were: ?2 = 8.8%, ?3 = 77.9% and ?4 = 13.3%. Corresponding frequencies of APOE in the edentulous group were: ?2 = 6.6%, ?3 = 75.4% and ?4 = 18.0%. Conclusion: Despite matching both groups with regard to different background factors, the edentulous group had a higher frequency of APOE ?4 than the dentate group. Thus, genetic factors might contribute to greater risk in developing complex oral diseases leading to tooth loss or just be an indication that the subjects in our study carrying APOE ?4 are more fragile.