苹果属无融合生殖SERKs基因家族的克隆和表达分析

本研究利用同源克隆法以苹果属平邑甜茶和杂种后代叶片为试验材料,克隆得到4个SERK基因家族片段,并分析了SERKs家族基因在平邑甜茶和杂种后代不同器官和组织中的表达情况。研究结果表明:分离获得的4个SERK基因家族片段的氨基酸序列与模式植物拟南芥、烟草等植物的同源性均在84%以上,同时与其他物种的氨基酸序列进行Blast比对都具有较高的同源性;RT-PCR定量分析结果表明SERK1、SERK2、SERK3、SERK4基因主要在生殖器官中表达。因此,推测SERKs基因在平邑甜茶和杂种后代生殖发育过程中起着重要的调控作用。本研究结果为进一步揭示无融合生殖的发生机理奠定了基础。     

Hybridizing <Emphasis Type="Italic">Brassica rapa</Emphasis> with wild crucifers <Emphasis Type="Italic">Diplotaxis erucoides</Emphasis> and <Emphasis Type="Italic">Brassica maurorum</Emphasis>

Two wide hybrids, Diplotaxis erucoides (2n = 14) × Brassica rapa (2n = 20) and B. maurorum (2n = 20) × B. rapa, were developed using the sequential ovary–ovule culture. Reciprocal crosses failed, possibly as a consequence of strong unilateral incompatibility. The F ₁ hybrids in each combination were completely male sterile and morphologically intermediate to the respective parents. DNA marker polymorphism and chromosome counts confirmed their hybrid nature. High frequency of bivalents in the F ₁ and the presence of trivalents/quadrivalents in the derived amphiploids suggested genomic duplications and homoeology of the parental genomes. Up to three homoeologous pairs between the D. erucoides (D^eD^e) and B. rapa (AA) genomes, and one between B. maurorum (B^mB^m) and B. rapa genomes were observed. Successful synthesis of the F ₁ hybrids and amphiploids of B. rapa with D. erucoides and B. maurorum, and allosyndetic chromosome pairing are expected to permit introgressions of desirable loci into the cultivated Brassica germplasm, especially for resistance to Alternaria brassicae and Albugo candida.     

Water use efficiency in a soybean field: influence of plant water stress

Measurements were made in 1980 over a fully-developed soybean (Glycine max (L.) Merrill) canopy at Mead, Nebraska to determine how crop water status influences photosynthesis, evapotranspiration and water use efficiency. Water use efficiency was calculated in terms of the CO₂—water flux ratio (CWFR). Micrometeorological techniques were used to measure the exchange rates of CO₂ and water vapor above the crop canopy. Crop water status was evaluated by reference to volumetric soil moisture (θ_v), stomatal resistance (r_s), and leaf water potential (ψ) measurements.Stomatal resistance (r_s) was independent of ψ when the latter was greater than ?1.1 MPa. r_s increased sharply as ψ dropped below this threshold. Canopy CO₂ exchange (F_c) decreased logarithmically with increasing r_s under strong irradiance. Although F_c was found to be strongly correlated with r_s, the influence of low values of ψ and of high air temperature cannot be discounted since these factors affect the enzymatic reactions associated with photosynthesis. Stomatal closure also reduced evapotranspiration and influenced the partitioning of net radiation.Under strong irradiance the CO₂ water flux ratio (CWFR) decreased with increasing stomatal resistance. This observation is at variance with predictions of certain early ‘resistance’ models, but substantiates predictions of some recent models in which leaf energy balance considerations are incorporated.     

Nodule-specific expression of Rhizobium meliloti symbiotic promoters P1 and P2 in chickpea-Rhizobium sp. symbiosis

Developmentally specific expression of Rhizobium spp. genes involved in symbiotic N₂ fixation is known to operate through cascade regulation of various nif and fix operons. Fusion constructs of lacZ under symbiotic promoters P1 (for nifHDK operon) and P2 (for fixABCX operon) of Rhizobium meliloti were mobilized into Rhizobium spp. (Cicer) strains Rcd301 and RCR13. The assays for -galactosidase activity to monitor the expression of lacZ under these promoters was performed in host backgrounds of Escherichia coli, R. meliloti, and Rhizobium spp. (Cicer). The enzyme assays indicated significant levels of expression from P1 and P2 promoters in chickpea rhizobia, specifically in symbiotic cells from nodules. However, as in R. meliloti, these promoters did not induce strong expression in free-living cells of Rhizobium spp. (Cicer). This indicates functional homology of R. meliloti promoters in rhizobium spp. (Cicer). Functional cross-reactivity of trans regulatory factors like NtrA, NtrC, and NifA between these rhizobia seems evident from the nodule-specific expression of P1 and P2 cis elements.     

Changes in colostrum of Murrah buffaloes after calving

Colostrum samples were collected from 8 Murrah buffaloes on days 1, 2, 3, 4 and 5 after calving. Levels of IgG averaged 54.0 mg/ml at calving, then decreased significantly (P < 0.01). IgA and IgM on day 1 were 3.22 mg/ml and 5.22 mg/ml, respectively; both decreased during the first five days after calving. Values of IgA and IgM were higher than those reported in cows. SCC values, which were high at calving (500 000 per ml), reduced significantly (P < 0.01) on day 2, then decreased slightly until day 5 (180 000 per ml). At calving, macrophages were the most prominent cells in buffalo colostrum, followed by lymphocytes and neutrophils. Phagocytic activity was 23% at calving and reduced significantly (P < 0.01) to 14% on day 5. Phagocytic index was highest in the first colostrum, and then decreased non-significantly.     

Prevalence and risk factors of Coxiella burnetii seropositivity in Danish beef and dairy cattle at slaughter adjusted for test uncertainty

Antibodies to Coxiella burnetii have been found in the Danish dairy cattle population with high levels of herd and within herd seroprevalences. However, the prevalence of antibodies to C. burnetii in Danish beef cattle remains unknown. The objectives of this study were to (1) estimate the prevalence and (2) identify risk factors associated with C. burnetii seropositivity in Danish beef and dairy cattle based on sampling at slaughter.     

Comparison of three enzyme-linked immunosorbant assays with the indirect immunofluorescent antibody test for the diagnosis of canine infection with Ehrlichia canis

The aim of this study was to compare three different enzyme-linked immunosorbant assays (recombinant major antigenic protein 2 (rMAP2)-ELISA, the Immunocomb (Biogal, Israel) and the Snap 3Dx assay (IDEXX Laboratories Inc., USA)) with the indirect immunofluorescent antibody test in detecting anti-Ehrlichia canis immunoglobulin-G (IgG) antibodies. Samples tested were collected from dogs suspected to be naturally infected with E. canis and from experimentally infected dogs.When qualitative results (positive/negative) were compared, there was an overall agreement of 81% (54/67) between the indirect immunofluorescence antibody (IFA) test and the rMAP2-ELISA. An overall agreement of 94% (63/67) was found between the IFA test and the Immunocomb, and an overall agreement of 91% (61/67) was found between the IFA test and the Snap 3Dx assay. In 50 of 67 (74.6%) samples tested, complete agreement in the qualitative results was found in all four tests. Sixteen of 17 samples with disagreement in the qualitative results were found to have IFA titers of 1:320 or less. The sensitivities and specificities of the tests were found to be 0.71 and 0.85 for the rMAP2-ELISA, 0.86 and 0.98 for the Immunocomb, and 0.71 and 1.00 for the Snap 3Dx assay.The tests performed in this study were found to be highly specific in detecting E. canis antibodies. Their sensitivity was found to be low with sera having IFA titers of < or =1:320, while high with sera having titers greater than 1:320. Repeating the serological tests 1-2 weeks after the first antibody assay may overcome the sensitivity problem with titers of < or =1:320.     

Characterization of bioactive recombinant woodchuck interleukin-2 amplified by RLM-RACE and produced in eukaryotic expression system

Woodchucks (Marmota monax) infected with woodchuck hepatitis virus (WHV) represent a highly valuable laboratory model of hepatitis B virus (HBV) infection, in which molecular, immunological and pathological events occurring in infected humans are adequately reflected. To advance studies on T cell immune responses and propagation of hepadnavirus in T lymphocytes in this animal model, we determined the complete sequence of woodchuck interleukin-2 (wIL-2) cDNA by utilizing RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE) reaction. The wIL-2 sequence revealed a single open reading frame encoding for the predicted precursor protein comprised of a signal peptide and a 134 amino acid-long mature protein. The mature wIL-2 protein produced in the Escherichia coli expression system, designated as ec-rwIL-2, was found to be immunogenic but not biologically active. In contrast, precursor wIL-2 protein cloned into baculovirus transfer vector and expressed in Sf9 cells, designated as bac-rwIL-2, demonstrated functional competence. Further, bac-rwIL-2 was able to stimulate proliferation and to induce multiple daughter cell generations in woodchuck T cells, as well as facilitated the survival of standard IL-2-dependent mouse CTLL-2 cells in culture. Western blot analysis of bac-rwIL-2 using antibodies generated against ec-rwIL-2 revealed a single protein band of 15.5kDa. The availability of biologically active recombinant wIL-2 should facilitate ex vivo studies on functional competence of woodchuck T lymphocytes derived from different stages of hepadnaviral hepatitis and assist in recognizing their contribution to the pathogenesis of liver injury in the woodchuck model of hepatitis B.