Unequal VH gene rearrangement frequency within the large VH7183 gene family is not due to recombination signal sequence variation, and mapping of the genes shows a bias of rearrangement based on chromosomal location.

Much of the nonrandom usage of V, D, and J genes in the Ab repertoire is due to different frequencies with which gene segments undergo V(D)J rearrangement. The recombination signal sequences flanking each segment are seldom identical with consensus sequences, and this natural variation in recombination signal sequence (RSS) accounts for some differences in rearrangement frequencies in vivo. Here, we have sequenced the RSS of 19 individual V(H)7183 genes, revealing that the majority have one of two closely related RSS. One group has a consensus heptamer, and the other has a nonconsensus heptamer. In vitro recombination substrate studies show that the RSS with the nonconsensus heptamer, which include the frequently rearranging 81X, rearrange less well than the RSS with the consensus heptamer. Although 81X differs from the other 7183-I genes at three positions in the spacer, this does not significantly increase its recombination potency in vitro. The rearrangement frequency of all members of the family was determined in microMT mice, and there was no correlation between the in vitro recombination potential and V(H) gene rearrangement frequency in vivo. Furthermore, genes with identical RSS rearrange at different frequencies in vivo. This demonstrates that other factors can override differences in RSS potency in vivo. We have also determined the gene order of all V(H)7183 genes in a bacterial artificial chromosome contig and show that most of the frequently rearranging genes are in the 3' half of the region. This suggests that chromosomal location plays an important role in nonrandom rearrangement of the V(H)7183 genes.     

Characterization of Novel Simian Immunodeficiency Viruses from Red-Capped Mangabeys from Nigeria (SIVrcmNG409 and -NG411)

Two novel simian immunodeficiency virus (SIV) strains from wild-caught red-capped mangabeys (Cercocebus torquatus torquatus) from Nigeria were characterized. Sequence analysis of the fully sequenced SIV strain rcmNG411 (SIVrcmNG411) and gag and pol sequence of SIVrcmNG409 revealed that they were genetically most closely related to the recently characterized SIVrcm from Gabon (SIVrcmGB1). Thus, red-capped mangabeys from distant geographic locations harbor a common lineage of SIV. SIVrcmNG411 carried a vpx gene in addition to vpr, suggesting a common evolutionary ancestor with SIVsm (from sooty mangabeys). However, SIVrcm was only marginally closer to SIVsm in that region than to any of the other lentiviruses. SIVrcm showed the highest similarity in pol with SIVdrl, isolated from a drill, a primate that is phylogenetically distinct from mangabey monkeys, and clustered with other primate lentiviruses (primarily SIVcpz [from chimpanzees] and SIVagmSab [from African green monkeys]) discordantly in different regions of the genome, suggesting a history of recombination. Despite the genetic relationship to SIVcpz in the pol gene, SIVrcmNG411 did not replicate in chimpanzee peripheral blood mononuclear cells (PBMC), although two other viruses unrelated to SIVcpz, SIVmndGB1 (from mandrills) and SIVlhoest (from L'Hoest monkeys), were able to grow in chimpanzee PBMC. The CCR5 24-bp deletion previously described in red-capped mangabeys from Gabon was also observed in Nigerian red-capped mangabeys, and SIVrcmNG411, like SIVrcmGB1, used CCR2B and STRL33 as coreceptors for virus entry. SIVrcm, SIVsm, SIVmndGB1, and all four SIVlhoest isolates but not SIVsun (from sun-tailed monkeys) replicated efficiently in human PBMC, suggesting that the ability to infect the human host can vary within one lineage.     

The synaptic behaviour of Triticum turgidum with variable doses of the Ph1 locus

Cultivated wheat Triticum turgidum is an allotetraploid (AABB) with diploid-like behaviour at metaphase I. This behaviour is mainly influenced by the action of the Ph1 locus. To study the effect of Ph1 on chromosome pairing in T. turgidum we have analysed the synaptic pattern in fully traced spread nuclei at mid- and late-zygotene and at pachytene of three different genotypes: a standard line, ph1c mutant and a duplication mutant, with zero, two and four doses of Ph1, respectively. The number of synaptonemal complex (SC) bivalents and of the different SC multivalent associations were determined in each nucleus. The mean number of lateral elements involved in SC multivalent associations (LEm) at mid-zygotene was relatively high in all lines and was similar in two and zero doses of Ph1. These means changed little with the progression of zygotene but decreased at pachytene because of the transformation of multivalents into bivalents. Multivalent correction was more efficient in the presence than in the absence of Ph1. The four doses of Ph1 genotype showed a higher number of SC bivalents at mid-zygotene, and the frequency of multivalents decreased progressively throughout zygotene and pachytene. The results suggest that the main action of the Ph1 locus on the diploidisation mechanism would be related to a process of checking for homology operating during prophase I. Received: 27 February 2000 / Accepted: 12 July 2000     

Effects of Angular Leaf Spot and Rust on Leaf Gas Exchange and Yield of Common Bean (Phaseolus Vulgaris)

Isolated and interactive effects of angular leaf spot (caused by Phaeoisariopsis griseola) and rust (caused by Uromyces appendiculatus) on leaf gas exchange and yield was studied in common bean (Phaseolus vulgaris L. cv. Carioca) plants. Gas exchange was measured on 37, 44, 51, and 58 d after planting using a portable photosynthesis system. The inoculation of plants with P. griseola (P), U. appendiculatus (U), and the combination of both pathogens (P+U) caused a significant reduction of net photosynthetic rate (P _N) and yield. The reduction of stomatal conductance (g _s), P _N, and yield was higher under P and combination of P+U than under U treatment. By effect of U, the reduction on yield was higher than the reductions on gas exchange parameters. On the treatment P+U, a reduction of 23 % in P _N and a correspondent reduction of 32 % in yield was observed. The interactive effects of the pathogens on yield could be explained in part by the decreases in g _s and in P _N of diseased bean leaves. The combined effect of both diseases on yield and gas exchange parameters suggests an antagonistic interaction.     

Expression of complement receptor type 1 (CR1) on erythrocytes of paracoccidiodomycosis patients

Complement receptor type 1 (CR1) is a membrane glycoprotein that acts as a receptor for the C3b, iC3b and C4b fragments of complement. In primates, one function of erythrocytes is to promote safe clearance of immunocomplexes (IC) from the circulation through CR1. Theoretically, in diseases characterized by high levels of circulating IC, an erythrocyte CR1 (CR1/E) deficiency may favor IC deposition in tissues or facilitate inappropriate activation of leukocytes in the circulation. Depression of the cell immune response occurs in paracoccidioidomycosis (PCM), especially in the more severe cases, and is frequently associated with high serum IC levels. In the present study we quantified the number of CR1/E in patients with the acute and chronic forms of PCM before and after treatment and correlated it with serum IC levels and CD4+ and CD8+ T cell concentration in the peripheral blood of these patients. Patients with PCM, particularly those with active disease and who had received treatment for shorter periods of time, had low numbers of CR1/E. In addition, an increase in serum IC concentration and a reduction in the CD4+/CD8+ T cell ratio were observed. After treatment there was a significant increase in mean CR1/E number and a reduction in serum IC levels. In patients with the chronic form of the disease the CD4+/CD8+ T cell ratio tended to increase after treatment and was associated with increased CR1/E levels. These results suggest that the reduction in CR1/E observed in patients is a phenomenon acquired with the disease and that CR1 could play a role in the pathogenesis of PCM.This revised version was published online in October 2005 with corrections to the Cover Date.     

Survival of Salmonella enterica in Freshwater and Sediments and Transmission by the Aquatic Midge Chironomus tentans (Chironomidae: Diptera)

Survival of a nalidixic acid-resistant strain of Salmonella enterica serovar Typhimurium mr-DT-104 in water and sediments was tested using artificially contaminated aquaria. Water samples remained culture positive for salmonella for up to 54 days. Sediment samples were culture positive up to 119 days. In addition, potential mechanisms for spreading salmonella in the environments by chironomid larvae and adults were tested. We evaluated the acquisition of mr-DT-104 by chironomids from contaminated aquatic sediments and subsequent spread to uncontaminated sediments. Larval chironomids raised in contaminated sediments became culture positive, and the bacteria were carried over to adults after emergence. Contamination of clean sediments by chironomid larvae was not demonstrated. These findings clearly suggest that mr-DT-104 serovar organisms can survive in aquatic sediments for at least several months. Uptake of salmonellae by chironomid larvae and adults suggests that they are possible vectors of mr-DT-104 in both aquatic and terrestrial environments, although the role of larval defecation in movement of bacteria to new sediments was not demonstrated.     

应用红外荧光和加尾引物法进行向日葵SSR遗传分析中的多聚PCR和多聚凝胶电泳的优化

在向日葵(Helianthus annuus L.)自交系SSR遗传分析中,为了提高SSR荧光分析效率、简化分析程序和降低研究费用,我们进行了多聚PCR和多聚凝胶电泳两项技术的优化研究。结果表明,优化多聚PCR和多聚凝胶电泳的影响因子(如逐步降低的退火温度的循环数、各个多聚位点引物浓度的平衡、PCR缓冲液浓度以及Taq DNA多聚酶的浓度等)可以收到更好的实验效果。基于以上的优化研究,系统地提出了一套优化的加尾引物法的策略。同时,提出了在多聚PCR和多聚凝胶电泳中常常遇到的技术难题的有效防止和解决的方法。     

Modelling the interactions between water and nutrient uptake and root growth

A model of three-dimensional root growth has been developed to simulate the interactions between root systems, water and nitrate in the rooting environment. This interactive behaviour was achieved by using an external-supply/internal-demand regulation system for the allocation of endogenous plant resources. Data from pot experiments on lupins heterogeneously supplied with nitrate were used to test and parameterise the model for future simulation work. The model reproduced the experimental results well (R ² = 0.98), simulating both the root proliferation and enhanced nitrate uptake responses of the lupins to differential nitrate supply. These results support the use of the supply/demand regulation system for modelling nitrate uptake by lupins. Further simulation work investigated the local uptake response of lupins when nitrate was supplied to a decreasing fraction of the root system. The model predicted that the nitrate uptake activity of lupin roots will increase as the fraction of root system with access to nitrate decreases, but is limited to an increase of around twice that of a uniformly supplied control. This work is the first example of a modelled root system responding plastically to external nutrient supply. This model will have a broad range of applications in the study of the interactions between root systems and their spatially and temporally heterogeneous environment.