Evidence for two genetic complementation groups in pyruvate carboxylase-deficient human fibroblast cell lines

We have examined genetic complementation in pyruvate carboxylase deficiency by comparing the enzyme activity in polyethylene glycol-induced heterokaryons with that in unfused mixtures of fibroblasts from three affected children. Complementation, manifested as a three- to sevenfold increase in pyruvate carboxylase activity, was observed in fusions between a biotin-responsive multiple carboxylase (pyruvate carboxylase, propionyl CoA carboxylase, and -methylcrotonyl CoA carboxylase) deficient fibroblast line and two other lines deficient only in pyruvate carboxylase activity. Kinetic analysis of complementing pyruvate carboxylase deficient lines, measured by the rate of restoration of enzyme activity as a function of time, revealed that maximum restoration was achieved within 10–24 hr after fusion. This profile is similar to those observed for fusions between the multiple carboxylase deficient line and two lines deficient in propionyl CoA carboxylase activity that are known to represent different gene mutations. Although the patients with pyruvate carboxylase deficiency had similar clinical findings, our studies indicate that pyruvate carboxylase deficiency is genetically heterogeneous, with at least two distinct, probably intergenic, complementation groups.This work was supported by an NIH research grant (AM 25675) and an A. D. Williams research grant (6-48360). B. Wolf is the recipient of an NIH Research Career Development Award (AM 00677) and is aided by a Basil O'Connor Starter Research Grant from The National Foundation-March of Dimes (5-263). G. Feldman is the recipient of an NIH predoctoral training grant (GM 07492). This article is No. 100 from the Department of Human Genetics at the Medical College of Virginia.     

Lack of correspondence between genetic and phenotypic groups amongst soil-borne streptomycetes

Correspondence between two distinct genetic traits, 16S rRNA gene sequences and repetitive element-sequence-based BOX-PCR DNA fingerprints, and antibiotic inhibition and resistance phenotypes was explored for a spatially explicit sample of Streptomyces from a prairie soil. There was no correspondence between 16S rRNA gene sequence groups and antibiotic phenotypes. However, 16S rRNA gene sequence groups differed significantly in mean inhibition zone sizes. Specific antibiotic phenotypes may reflect local selection pressures, as suggested by the significant differences in mean inhibition zone sizes against specific test isolates by Streptomyces from the same 16S rRNA gene sequence group but from different locations in soil. Significant correlations between antibiotic phenotypes and BOX-PCR fingerprints were found, but were small (r=0.19-0.22). Although genetic characterizations alone were not predictive of specific antibiotic phenotypes, 16S rRNA gene sequence analyses may identify isolates that are most or least likely to possess substantial inhibitory potential, providing insight into the broad ecological strategy for individual isolates.     

Comparison of isogenic lines provides evidence that phenotypic plasticity is under genetic control in rainbow trout Oncorhynchus mykiss

Comparison of nine isogenic lines of rainbow trout Oncorhynchus mykiss kept in the same environment showed significant genetic determinism of phenotypic plasticity assessed through body mass measurements. Ranking of lines differed between two tested environments.     

Lung growth of the turkey, Meleagris gallopavo: II. Comparison of two genetic lines

Comparisons of lung growth in two genetic lines of turkeys, one unselected and one selected for increased body mass, were used to evaluate the lung's alteration in structure with changes in body form or physiology. Seventy-two male turkeys, 36 genetically selected for early rapid growth and large pectoral musculature, and 36 unselected birds, were killed at 12 different ages to compare lung growth in the two lines. Body weights and lung volumes were determined. A three-level cascade sampling system was used to prepare lung tissue for qualitative and quantitative observation by light microscopy. Allometric equations describing growth of lung volume and lung compartments relative to body weight in two phases (tissue proliferation and equilibrated growth) were compared between lines of turkey for differences in slopes or intercepts. Means of data for 112- and 420-day-old birds were also compared. There were no qualitative histological differences observed between lungs of the two lines of turkey, yet there were morphometric differences in lung growth relative to body weight. During equilibrated lung growth, there was less rapid growth of air and blood capillary volumes and surfaces relative to body weight in the selected than in the unselected turkey. The gas-exchange compartment did not enlarge concomitant with the large increase in muscle mass of the selected turkeys, while large-vessel volume and small-airway volume grew similarly to body weight in both turkey lines. We conclude that the lung of the selected line of turkeys did not show an enlarged gas-exchange compartment relative to the greater body muscle mass, but it did show enlarged vessels and conducting airways.     

苦玄参40个株系表型性状遗传多样性分析

植物遗传多样性一直以来是种质资源研究的热点。为揭示苦玄参栽培种质的遗传多样性,该研究从苦玄参常年栽培种中根据形态学性状差异选择40个株系,以产量、药效物质含量及茎、叶、花等形态学性状为指标,进行物种变异和遗传多样性分析,并对其进行聚类分析,获取各株系遗传亲缘关系。结果表明:苦玄参苷IA和IB含量的遗传变异系数较高,分别为24.225%和17.853%;遗传多样性指数高,分别为1.920和2.075。产量遗传变异系数较低,仅为3.637%,但遗传多样性指数较高,达到1.884。其余表型性状指标遗传变异系数均较高,其中花色高达127.794%。数值型性状具有较高的遗传多样性,均大于1,但描述型指标遗传多样性指标较低,均小于1。聚类分析可将供试材料分为4个大的类群:第Ⅰ类群共有7个株系,此类群产量性状及相关指标平均数较高;第Ⅱ类群有8个株系,茎节长度最长,其余指标中等偏下;第Ⅲ类群数量最多,有20个株系,各指标均较低;第Ⅳ类群有5个株系,苦玄参苷含量和产量均较高,综合指标比较优。相关分析结果表明,苦玄参苷IA的含量与叶缘性状具有显著相关,产量性状与茎节长度、一级分枝和末级分枝数具有极显著相关,选择优良种质应注重该4个性状的选择。该研究结果表明目前苦玄参栽培种具有广泛的变异和较高的遗传多样性,为苦玄参资源的利用提供了参考依据。     

Effect of silkworm hemolymph on N-linked glycosylation in two Trichoplusia ni insect cell lines

A recombinant N-linked glycoprotein, secreted human placental alkaline phosphatase (SEAP), was produced in two Trichoplusia ni insect cell lines using the baculovirus expression vector. Silkworm hemolymph (SH) was added to TNMFH + 10% fetal bovine serum (FBS) medium to a concentration of 2.5% or 5%, and SEAP production and glycosylation in the presence of SH were compared with controls devoid of hemolymph. Growing Tn-4s cells in 5% SH-supplemented medium required progressive adaptation of the cells to SH, and adapted cells had a SEAP specific yield decreased by 2.5-fold compared with control cells not exposed to SH. Although SEAP produced in the control possessed little complex glycosylation (<1%), SEAP produced by SH-adapted cells in the presence of 5% SH possessed 8.7% sialylated structures, as well as unusual, asialylated, agalactosylated structures with a high degree of polymerization (DP). On the basis of enzymatic and mass-spectrometric analyses, we propose that these structures are glucosylated, high-mannose oligosaccharides. SEAP was also produced by Tn-4s cells without adaptation to SH when SH was added just prior to baculovirus infection, but SEAP specific yield was adversely affected (approximately fourfold reduction compared with control devoid of hemolymph), and glycosylation of SEAP produced under these conditions was characterized by large amounts of high-mannose and high-DP structures and an absence of complex structures. Similarly, Tn5B1-4 cells that were not adapted to SH had a SEAP specific yield reduced by approximately fivefold in SH-containing medium; however, these cells were able to produce 13.5% sialylated SEAP in the presence of 2.5% SH, whereas complex structures were not produced in the absence of SH. We propose that SH improves glycosylation either directly or indirectly by decreasing SEAP specific yield.     

Comparative population genetic structures and local adaptation of two mutualists

Similar patterns of dispersal and gene flow between closely associated organisms may promote local adaptation and coevolutionary processes. We compare the genetic structures of the two species of a plant genus (Roridula gorgonias and R. dentata) and their respective obligately associated hemipteran mutualists (Pameridea roridulae and P. marlothi) using allozymes. In addition, we determine whether genetic structure is related to differences in host choice by Pameridea. Allozyme variation was found to be very structured among plant populations but less so among hemipteran populations. Strong genetic structuring among hemipteran populations was only evident when large distances isolated the plant populations on which they live. Although genetic distances among plant populations were correlated with genetic distances among hemipteran populations, genetic distances of both plants and hemipterans were better correlated with geographic distance. Because Roridula and Pameridea have different scales of gene flow, adaptation at the local population level is unlikely. However, the restricted gene flow of both plants and hemipterans could enable adaptation to occur at a regional level. In choice experiments, the hemipteran (Pameridea) has a strong preference for its carnivorous host plant (Roridula) above unrelated host plants. Pameridea also prefers its host species to its closely related sister species. Specialization at the specific level is likely to reinforce cospeciation processes in this mutualism. However, Pameridea does not exhibit intraspecific preferences toward plants from their natal populations above plants from isolated, non-natal populations.     

A comparative proteomic analysis of parthenogenetic lines and amphigenetic lines of silkworm

Parthenogenetic strains of silkworm serve as an effective system for sex-control in silkworms. To determine the molecular mechanism of silkworm parthenogenesis, protein profiles from newly hatched silkworm of a parthenogenetic lines with high pigmentation rate and hatching rate were compared with amphigenetic lines using proteomics approach, including by two-dimensional electrophoresis (2-DE), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/TOF MS), and bioinformatics analysis. Several proteins were expressed differentially between the parthenogenetic and amphigenetic lines, and seven of nine interesting proteins were identified successfully using MALDI-TOF/TOF MS analysis. The identified proteins were muscular protein-20, odorant binding protein-LOC100301497, glutathione S-transferase delta, translationally controlled tumor protein homolog, cuticular protein RR-1 motif 19, beta-actin, actins, and muscle-type A₁ actins. These proteins may be associated with the regulation of growth, development, and reproductive processes of silkworm parthenogenetic lines.     

Impact of hygiene of housing conditions on performance and health of two pig genetic lines divergent for residual feed intake

Pigs selected for high performance may be more at risk of developing diseases. This study aimed to assess the health and performance of two pig lines divergently selected for residual feed intake (RFI) (low RFI (LRFI) v. high RFI (HRFI)) and housed in two contrasted hygiene conditions (poor v. good) using a 2×2 factorial design (n=40/group). The challenge period (Period 1), started on week zero (W0) when 12-week-old pigs were transferred to good or poor housing conditions. At week 6 (W6), half of the pigs in each group were slaughtered. During a recovery period (Period 2) from W6 to W13 to W14, the remaining pigs (n=20/group) were transferred in good hygiene conditions before being slaughtered. Blood was collected every three (Period 1) or 2 weeks (Period 2) to assess blood indicators of immune and inflammatory responses. Pulmonary lesions at slaughter and performance traits were evaluated. At W6, pneumonia prevalence was greater for pigs housed in poor than in good conditions (51% v. 8%, respectively, P<0.001). Irrespective of hygiene conditions, lung lesion scores were lower for LRFI pigs than for HRFI pigs (P=0.03). At W3, LRFI in poor conditions had the highest number of blood granulocytes (hygiene×line, P=0.03) and at W6, HRFI pigs in poor conditions had the greatest plasma haptoglobin concentrations (hygiene×line, P=0.02). During Period 1, growth rate and growth-to-feed ratio were less affected by poor hygiene in LRFI pigs than in HRFI pigs (hygiene×line, P=0.001 and P=0.02, respectively). Low residual feed intake pigs in poor conditions ate more than the other groups (hygiene×line, P=0.002). Irrespective of the line, fasting plasma glucose concentrations were higher in poor conditions, whereas fasting free fatty acids concentrations were lower than in good conditions. At the end of Period 2, pneumonia prevalence was similar for both housing conditions (39% v. 38%, respectively). During Period 2, plasma protein concentrations were greater for pigs previously housed in poor than in good conditions during Period 1. Immune traits, gain-to-feed ratio, BW gain and feed consumption did not differ during Period 2. Nevertheless, at W12, BW of HRFI previously housed in poor conditions was 13.4 kg lower than BW of HRFI pigs (P<0.001) previously housed in good conditions. In conclusion, health of the most feed efficient LRFI pigs was less impaired by poor hygiene conditions. This line was able to preserve its health, growth performance and its feed ingestion to a greater extent than the less efficient HRFI line.     

Description and analysis of genetic diversity between commercial bean lines (Phaseolus vulgaris L.)

The effectiveness of RFLP, DAMD-PCR, ISSR and RAPD markers in assessing polymorphism and relationships between 24 commercial lines of Phaseolus vulgaris L.was evaluated. We have used a Phaseolus-specific minisatellite sequence as a probe, which enabled 23 of the bean lines tested to be fingerprinted. Based on the sequence information obtained, primers corresponding to the bean-specific minisatellite core sequence were used in subsequent PCR amplifications. Our observations indicated that while the DAMD-PCR was sensitive in detecting genetic variation between bean species and between accessions of P. vulgaris, when used alone it may be limited in its ability to detect genetic variation among cultivated bean lines due to the low number of loci amplified. Only one out of the five ISSR primers tested was efficient in generating multiple band profiles, which was insufficient to distinguish all the different bean lines. Reproducible RAPD profiles were obtained, and these allowed us to differentiate all the genotypes tested with seven primers. We ultimately used only results from RFLP and RAPD markers to explore the genetic diversity among commercial bean lines. Both analyses led to the same clustering of the bean lines according to their geographical origins (United States or Europe). With respect to the European lines, the results obtained from RAPD data also enable the lines to be clustered according to their creators. Received: 15 January 2000 / Accepted: 21 March 2000     

Cowpea aphid performance and behaviour on two resistant cowpea lines

In a series of laboratory and climate chamber tests we compared the growth and behaviour of Aphis craccivora on one susceptible (ICV-1) and two aphid-resistant (ICV-11 and ICV-12) cowpea lines. The aphids' growth rates were much lower on the resistant cowpea lines than on the susceptible one, indicating strong antibiosis. In addition, the aphids invariably settled in higher numbers on the susceptible line than on either of the resistant. Compared to ICV-1, damaged leaves of the resistant line ICV-12 were settled upon to a higher degree than undamaged leaves, and leaf discs from the same line were even less resistant.On resistant lines individual aphids waited a significantly longer time before making their first test probe. Total probing time as well as the time preceding a decision to stay or leave was also longer.These results are discussed in relation to the possible mechanisms involved, and we also consider the effects of previous leaf feeding on the expression of resistance in the field.

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Résumé Au cours d'expériences au laboratoire et en chambres climatisées nous avons comparé la croissance et le comportement de A. craccivora sur une lignée sensible (ICV-1) et deux lignées résistantes (ICV-11 et ICV-12) de V. unguiculata. Les vitesses de croissance des pucerons ont été beaucoup plus lentes sur les lignées résistantes que sur la lignée sensible, ce qui révèle une forte antibiose. De plus, les pucerons atterrissent invariablement en plus grand nombre sur la variété sensible. Par comparaison avec ICV-1, les atterrissages sur lignée résistante ICV-12 étaient plus nombreux sur les feuilles endommagées que sur les feuilles intactes; les disques de feuilles de cette même lignée étaient encore moins résistants.Les pucerons ont séjourné individuellement un temps plus long sur les lignées résistantes avant de faire leur premier sondage. Le temps consacré aux sondages ainsi que le temps précédant de choix entre départ ou maintien sur la feuille étaient plus longs avec les lignées résistantes.Ces résultats ont été discutés en fonction des mécanismes impliqués. Nous avons aussi examiné les effets de la consommation antérieure sur les manifestations de la résistance dans la nature.

     

Comparison of two birnavirus-rhabdovirus coinfections in fish cell lines

Aquabirnaviruses, such as the infectious pancreatic necrosis virus (IPNV), Novirhabdoviruses, such as the infectious hematopoiteic necrosis virus (IHNV) and the viral hemorrhagic septicemia virus (VHSV), cause considerable losses to the salmonid industry worldwide. Coinfections of 2 viruses have been described, but the interactions between rhabdoviruses and birnaviruses have not been examined closely. Using virus titration, flow cytometry and RT-PCR assays, we compared the effect of IPNV on the replication of IHNV and VHSV in tissue culture cells. RT-PCR assays indicated that simultaneous infection of IPNV with VHSV does not affect the replication of the rhabdovirus either in the first or successive passages; the infective titers were similar in single and double infections. In contrast, coinfection of IPNV with IHNV induced a fall in infectivity, with reduced expression of IHNV viral antigens in BF-2 cells from Lepomis macrochirus and a loss of 4.5 log10 units of the infective titer after 3 successive passages. It was possible to stimulate BF-2 cells to produce significant interferon-like activity against IHNV but not against VHSV.     

Comparison of three inbred lines of mice by overall and genetic radiosensitivity

The radiosensitivities of three mouse strains (BALB/cLacY, C3H/SnY, and 101/HY) have been compared using the following parameters: survival after irradiation at a dose of 6-7 Gy, chromosome aberration frequency in bone marrow cells after irradiation at a dose of 1.5 Gy, and the change in testis weight and frequency of abnormal sperm heads (ASHs) after irradiation at doses from 0.5 to 4 Gy. Strain BALB/c is the most radiosensitive with respect to the survival and chromosome aberration frequency in the bone marrow but the most resistant with respect to the change in testis weight and the frequency of abnormal sperm heads. Strain 101/HY was the most resistant with respect to survival and chromosome aberration frequency in bone marrow after irradiation but the most radiosensitive with respect to testis damage.     

家蚕地方品种遗传多样性及其分子系统学研究

为检测家蚕(Bombyx mori)地方品种间的遗传差异,分析其遗传多样性和系统分化模式,为家蚕的品种鉴定和杂交育种亲本选择提供分子依据,本文对72个家蚕地方品种的淀粉酶基因(Bmamy2)片段进行测序,采用DNA序列分析技术探究了家蚕地方品种在核苷酸序列水平上的遗传差异及其系统分化特征.结果显示,家蚕地方品种在Bmamy2基因变异丰富,碱基突变率为5.6-8.2％,单倍型多样度为0.8294,核苷酸多样度为0.0236±0.00122,表明Bmamy2是一个具有丰富序列多样性的标记基因,能够较好地区分家蚕地方品种间的遗传多样性.系统发育分析、分子方差分析和Network分析等多种分析方法均检测到家蚕地方品种不按地理系统或生态类型聚类,各聚类种群均由来自不同地理系统或生态类型的品种构成,而来自同一地理系统或生态类型的品种则分别归属于不同的聚类种群.推测在家蚕的进化过程中并没有形成按地理系统或生态类型归类的分化格局,家蚕可能具有多种化性混合驯化起源的遗传背景和系统分化特征.     

Association of corticotropin-releasing hormone gene variation with performance and meat quality traits in commercial pig lines

The porcine corticotropin-releasing hormone(CRH) gene is a functional-positional candidate for quantitative tract loci on porcine chromosome 4 with major effects on growth and carcass composition. In addition, the central role of CRH in the neuroendocrine response to stress implicates the CRH gene as a functional candidate for meat quality. Association of a single nucleotide polymorphism (SNP) in the promoter region of the porcine CRH gene (g.233C > T) with several growth, carcass and meat quality traits was examined using more than 2000 individuals from four commercial lines: German Landrace (LR), Pietrain (Pi), German Large White x German Landrace (F1) and the German commercial fattening pig cross of Pietrain x F1 (PiF1). Significant association of the CRH SNP was found with feed conversion ratio in the PiF1 line, with carcass length in the LR line and with lean content in the F1, LR and Pi lines. Moreover, significant association with meat colour was found in the Pi and LR lines; however, the effects were in opposite directions. The presented results indicate that sequence variation in the porcine CRH gene has no major effect on growth and carcass composition in commercial pig lines, although it may significantly contribute to variation in meat quality. The g.233C>T SNP may be in incomplete linkage disequilibrium with causal mutations and/or exhibit effects in the context of DNA variation at other interacting loci.     

Testing the phenotypic gambit: phenotypic, genetic and environmental correlations of colour

Evolutionary theory is primarily concerned with genetic processes, yet empirical testing of this theory often involves data collected on phenotypes. To make this tenable, the implicit assumption is often made that phenotypic patterns are good predictors of genetic patterns; an assumption that coined the phenotypic gambit. Although this assumption has been validated for traits with high heritability, such as morphology, its generality for traits with low heritabilities, such as life-history and behavioural traits, remains controversial. Using a large-scale cross-fostering experiment, we were able to measure genetic, common environmental and phenotypic correlations between four colour traits and two skeletal traits in a wild population of passerine birds, the blue tit (Parus caeruleus). Colour traits had little heritable variation but common environment effects were found to be important; skeletal traits showed the opposite pattern. Positive correlations because of a shared natal environment were found between all traits, obscuring negative genetic correlations between some colour and skeletal traits. Consequently, phenotypic patterns were poor surrogates for genetic patterns and we suggest that this may be common if trade-offs or substantial parental effects exist. For this group of traits, the phenotypic gambit cannot be made and we suggest caution when inferring genetic patterns from phenotypic data, especially for behavioural and life-history traits.     

Comparison of two progestogens during out-of-season breeding in a commercial ewe flock

A comparison was made of the relative effectiveness of subcutaneous ear implants containing 2 mg Norgestomet or vagnial pessaries containing 60 mg medroxyprogesterone acetate (MAP) to induce estrus and conception in dry anestrous ewes. Groups of ewes were treated with one of the two progestogens for 14 d, and 500 IU pregnant mare serum gonadotropin (PMSG) was administered intramuscularly at the time of progestogen withdrawal. No significant differences in estrus induction, pregnancy rate or number of lambs born per ewe lambing were observed. Ewes treated with Norgestomet had 96% estrus, 60% pregnancy rate and 1.4 lambs per ewe lambing. Comparably, ewes treated with MAP had 94% estrus, 65% pregnancy rate and 1.7 lambs per ewe lambing. Norgestomet implants compared favorably with MAP pessaries for estrus induction and breeding of commercial, dry anestrous ewes.