Sensitive detection of mycoplasmalike organisms in field-collected and in vitro propagated plants of Brassica, Hydrangea and Chrysanthemum by polymerase chain reaction

A polymerase chain reaction (PCR) protocol, previously designed for amplification of a DNA fragment from aster yellows mycoplasmalike organism (MLO), was employed to investigate the detection of MLO DNA in field-collected and in vitro micropropagated plants. PCR with template DNA extracted from symptomatic, naturally-infected samples of Brassica, Chrysanthemum and Hydrangea, each yielded a DNA band corresponding to 1.0 Kbp. However, no DNA product was observed when either infected Ranunculus (with phyllody disease) or Gladiolus with (symptoms of ‘germs fins’) was used as source of template nucleic acid for PCR; further experiments indicated absence of target DNA in the case of Ranunculus and the presence of substances in Gladiolus which inhibited the PCR. The MLO-specific DNA was detected by PCR using less than 95 pg of total nucleic acid (equivalent to total nucleic acid from 1.9, ug tissue) in the case of field-collected Hydrangea and less than 11.4 pg of nucleic acid (equivalent to total nucleic acid from 19 ng of tissue) in the case of field-collected Brassica. The findings illustrate highly sensitive detection of MLOs in both field-grown and in vitro micropropagated infected plants.     

Mitochondrial malate dehydrogenase from watermelon: sequence of cDNA clones and primary structure of the higher-plant precursor protein

The isolation and sequence of a cDNA clone encoding the complete mitochondrial malate dehydrogenase (mMDH) of watermelon cotyledons is presented. Taking advantage of the polymerase chain reaction technology partial cDNA clones from the central part, the 3 part and the 5 part of the mRNA were obtained with oligonucleotides based on directly determined amino acid sequences. Subsequently, two complete cDNA clones for mMDH were synthesized with a sense primer corresponding to the nucleotide sequence of the amino terminal end of pre-mMDH and two antisense primers corresponding to the major alternative adenylation sites found in the mRNA.The amino acid residues for substrate and cofactor binding identified by X-ray crystallography for pig heart cytoplasmic MDH are conserved in the 320 amino acid long mature higher-plant mMDH. A presequence of 27 amino acids is present at the amino terminal end of the precursor protein.     

Reaction center light harvesting B875 complexes from Rhodocyclus gelatinosus: characterization and identification of quinones

Reaction center-B875 pigment-protein complexes were purified from Rhodocyclus gelatinosus. The proteic components consist of 7–8 polypeptides among which some were identified by their apparent molecular weights: the light harvesting B875 polypeptides and of 8 and 6 kDa, reaction center L (23 kDa), M (28 kDa) and H (34 kDa), cytochrome c (43 kDa). Four c-type hemes were found per reaction center. Flash-induced absorbance changes showed the presence of both Q_A and Q_B in the complex. Charge recombination times were determined to be: 1.16±0.2 (n=30) for P⁺Q_AQ_B ^- and 7–10 ms for P⁺Q_A ^- in presence of herbicides. From quinone analysis on one hand and kinetics of charge recombination on the other hand, we proposed that in the reaction center of Rhodocyclus gelatinosus Q_A is menaquinone 8 and Q_B is ubiquinone 8.     

Inhibition and labelling of isolated reaction centers from Rhodobacter sphaeroides by dicyclohexylcarbodiimide

The effect of dicyclohexylcarbodiimide (DCCD) on electron transfer in the acceptor quinone complex of reaction centers (RC) from Rhodobacter sphaeroides is reported. DCCD covalently labelled the RC over a wide concentration range. At low concentrations (<10 M) the binding was specific for the L subunit. At relatively high concentrations (>100 M) DCCD accelerated the rate of charge recombination of the P⁺Q_B ^- state, consistent with a decrease in the equilibrium constant between Q_A ^-Q_B and Q_AQ_B ^-. At similar concentrations, in the presence of cytochrome c as exogenous donor, turnover of the RC was inhibited such that only three cytochromes were oxidized in a train of flashes. Both these inhibitory effects were fully reversed by dialysis, indicating that stable covalent binding was not involved. Possible mechanisms of action are discussed in terms of the putative role of specific residues in proton transfer and protonation and release of quinol from the RC.     

急性缺氧下听觉运动反应的研究

受试者为9名男性青年。在低压舱模拟海拔3000、4000、5000和6000m,分别进行1h实验,共36人次。各高度上停留时作听觉检测,并记录了脑电和主诉。结果表明,在3000m,人的听觉运动反应基本正常;而在5000m、6000m,听觉运动反应的正确率、延时率、遗漏率和反应时等多项指标发生显著变化,工效降低。本研究为急性缺氧防护生理标准和缺氧耐力提供了工效的依据。     

多聚酶链反应技术鉴别肾综合征出血热病毒血清型的初步研究

肾综合征出血热(HFRS)为一组抗原性密切相关的布尼亚科汉坦病毒引起的急性传染病。在我国存在至少两种临床表现、动物宿主及流行特征截然不同的血清型别,即血清Ⅰ型(汉坦型)和血清Ⅱ型(汉城型)。这两型病毒间的血清学定型已有报道。近年来,除啮齿类动物外,从临床病人以及非啮齿类动物体内也分离到了HFRS病毒。同时出现两类型别毒株共存,以及从家鼠体内分离到野鼠型毒株或从野鼠体内分离到家鼠型毒株的复杂情形。为此,准确检定并鉴别不同来源毒株型别,将为深入了解其病原学、流行病学以及制定疫苗生产策略提供重要信息。     

Carbohydrates and fertilization in animals

A frequently used mechanism for sperm-egg recognition in many species involves complementary protein-carbohydrate interaction. The usual paradigm includes complex glycoconjugates in reproductive tract fluids or on the eggs which are recognized by carbohydrate-binding proteins on the sperm surface. Various glycoconjugates are utilized in the steps of sperm capacitation, sperm binding to the egg extracellular matrix and vitelline membrane and induction of the acrosome reaction. Several types of complex glycoconjugates are involved in these processes, including proteoglycans, lactosaminoglycans, sulfated fucose-containing glycoconjugates, and glycoproteins. There appear to be some structural similarities between active glycoconjugates; they are large in molecular weight and complex, and they are often sulfated, fucosylated, and attached to a protein through serine or threonine residues. In some species, the protein core of the glycoconjugates also participates in the interaction by limiting the binding of carbohydrates to sperm only of the relevant species, likely by providing the proper steric arrangement for the interaction. In other cases the protein core seems to serve more as a crosslinker of the carbohydrate moieties. This review discusses the types of glycoconjugates implicated in fertilization and the complementary lectin-like proteins found on sperm.     

Environmental and endogenous control of diapause inTrichogramma species

The 1st feature revealed typical for diapause phenomena inTrichogramma species is the interaction of environmental conditions in both parental and filial generations in induction of diapause in the latter. Lowered temperature during larval development is the ultimate factor evoking diapause in pronymphs but the norm of this thermal reaction is not fixed but varies depending on photoperiod and temperature in the previous generation due to maternal influence. Short day and in some cases high temperature in parental generation enhance tendency to diapause in the progeny. Unlike maternal influence the development to filial generation itself is not (or almost is not) governed by its own photoperiodic reaction. The 2nd typical feature revealed is the occurrence of endogenous process running in the sequence of generations and causing changes in the diapause tendency and underlying reaction norms even under constant rearing conditions.