Functional Glycosylation of Dystroglycan Is Crucial for Thymocyte Development in the Mouse

Background

Alpha-dystroglycan (α-DG) is a cell surface receptor providing a molecular link between the extracellular matrix (ECM) and the actin-based cytoskeleton. During its biosynthesis, α-DG undergoes specific and unusual O-glycosylation crucial for its function as a high-affinity cellular receptor for ECM proteins.

Methodology/Principal Findings

We report that expression of functionally glycosylated α-DG during thymic development is tightly regulated in developing T cells and largely confined to CD4⁻CD8⁻ double negative (DN) thymocytes. Ablation of DG in T cells had no effect on proliferation, migration or effector function but did reduce the size of the thymus due to a significant loss in absolute numbers of thymocytes. While numbers of DN thymocytes appeared normal, a marked reduction in CD4⁺CD8⁺ double positive (DP) thymocytes occurred. In the periphery mature naïve T cells deficient in DG showed both normal proliferation in response to allogeneic cells and normal migration, effector and memory T cell function when tested in acute infection of mice with either lymphocytic choriomeningitis virus (LCMV) or influenza virus.

Conclusions/Significance

Our study demonstrates that DG function is modulated by glycosylation during T cell development in vivo and that DG is essential for normal development and differentiation of T cells.     

Micronutrient Deficiencies and Plasmodium vivax Malaria among Children in the Brazilian Amazon

Background

There is a growing body of evidence linking micronutrient deficiencies and malaria incidence arising mostly from P. falciparum endemic areas. We assessed the impact of micronutrient deficiencies on malaria incidence and vice versa in the Brazilian state of Amazonas.

Methodology/Principal Findings

We evaluated children <10 years old living in rural communities in the state of Amazonas, Brazil, from May 2010 to May 2011. All children were assessed for sociodemographic, anthropometric and laboratory parameters, including vitamin A, beta-carotene, zinc and iron serum levels at the beginning of the study (May 2010) and one year later (May 2011). Children were followed in between using passive surveillance for detection of symptomatic malaria. Those living in the study area at the completion of the observation period were reassessed for micronutrient levels. Univariate Cox-proportional Hazards models were used to assess whether micronutrient deficiencies had an impact on time to first P. vivax malaria episode. We included 95 children median age 4.8 years (interquartile range [IQR]: 2.3–6.6), mostly males (60.0%) and with high maternal illiteracy (72.6%). Vitamin A deficiencies were found in 36% of children, beta-carotene deficiency in 63%, zinc deficiency in 61% and iron deficiency in 51%. Most children (80%) had at least one intestinal parasite. During follow-up, 16 cases of vivax malaria were diagnosed amongst 13 individuals. Micronutrient deficiencies were not associated with increased malaria incidence: vitamin A deficiency [Hazard ratio (HR): 1.51; P-value: 0.45]; beta-carotene [HR: 0.47; P-value: 0.19]; zinc [HR: 1.41; P-value: 0.57] and iron [HR: 2.31; P-value: 0.16]). Upon reevaluation, children with al least one episode of malaria did not present significant changes in micronutrient levels.

Conclusion

Micronutrient serum levels were not associated with a higher malaria incidence nor the malaria episode influenced micronutrient levels. Future studies targeting larger populations to assess micronutrients levels in P. vivax endemic areas are warranted in order to validate these results.     

氮肥和6-BA对花后受渍冬小麦抗渍性的调控效应

渍水是黄淮南部麦区和长江中下游麦区冬小麦生育后期的主要气象灾害因子,试验以小麦品种‘烟农19’为材料,采用裂区设计研究氮肥和6-BA对花后受渍冬小麦抗渍性的调控效应。结果显示:(1)花后渍害显著降低冬小麦根系活力、总吸收面积、活跃吸收面积、旗叶叶绿素含量、旗叶净光合速率,极显著降低穗粒数、千粒重和籽粒产量,而对穗数无显著影响。(2)受渍后叶面喷施氮肥+6-BA、氮肥、6-BA均能显著提高冬小麦花后抗渍性,其根系活力分别提高18.7%、14.6%、12.0%,根系总吸收面积分别提高31.9%、22.1%、19.6%,根系活跃吸收面积分别提高32.5%、21.7%、18.1%,叶绿素含量分别提高16.2%、9.4%、10.3%,净光合速率分别提高32.6%、23.3%、23.3%,穗粒数分别提高13.8%、9.3%、8.5%,千粒重分别提高17.3%、11.5%、10.5%,籽粒产量分别提高35.9%、23.5%、20.9%。研究表明,氮肥和6-BA正交互作用显著,即花后同时喷施氮肥和6-BA缓解小麦渍害的效果更佳。     

刈割对冬小麦再生积温需求及其籽粒产量和品质的影响

对黄土高原塬区冬小麦‘陇育216’于分蘖期、拔节期及孕穗期进行刈割处理,以不刈割为对照,测定冬小麦再生生长发育的积温需求及籽粒产量及其品质,探究刈割利用时间对冬小麦再生生长积温需求及产量构成的影响。结果表明:(1)分蘖中期(6个分蘖)前刈割利用,小麦再生各阶段的有效积温需求较对照无显著差异,在收获小麦青干草0.9～1.5t/hm2的同时,能保证籽粒、秸秆产量及其品质较对照均无显著下降;分蘖期后期(9个分蘖)及其后刈割利用,拔节至开花及成熟期的有效积温需求显著减少,尽管较分蘖期可多收获60%的青干草,但籽粒、秸秆产量较对照均显著降低。(2)通径分析发现,刈割主要通过降低再生植株高度及减少单位面积穗数而导致冬小麦籽粒减产。研究表明,为维持冬小麦再生生长节律、籽粒产量及品质形成的稳定,冬小麦刈割利用的时期应不迟于分蘖中期(6个分蘖)。     

The Development of Waterlogging Damage in Young Wheat Plants in Anaerobic Solution Cultures

Anaerobic (anoxic) solution cultures were used to investigatethe effect of a restricted oxygen supply to roots on the developmentof symptoms of waterlogging damage in young wheat plants, especiallyeffects on growth and nutrient uptake by the shoots. Anaerobicconditions produced by bubbling solutions with oxygen-free nitrogengas caused premature senescence of the lower leaves, slowedshoot fresh weight accumulation, and arrested the growth ofthe seminal roots. However the shoot dry weight initially increasedabove that of the aerobic controls. Nutrient accumulation bythe shoot was severely inhibited by anoxia, the uptake of nitrate,phosphate, and potassium being more affected than that of calciumand magnesium. The calculated concentrations in the xylem streamof all these ions (except nitrate) were equal to, or less than,those in the external solution, suggesting that the slow butcontinuous accumulation of nutrients in the shoot could haveoccurred passively by the mass flow of solution across damagedroots in response to transpiration. Aerenchymatous nodal rootsextended into the anoxic solutions to a maximum length of 12cm but there were few produced, and the size of the root systemremained small and may have limited shoot growth. Inclusionof carbon dioxide (10 kPa partial pressure) in the nitrogengas stream had little additional effect on plants to that causedby anoxia alone. All the responses of wheat to the anaerobic solutions were similarto those observed previously in waterlogged soil, indicatingthat many of the early symptoms of waterlogging damage to wheatcan be caused simply by the direct effects of inadequate oxygensupply to the roots. The results are discussed in relation tocurrent views of the mechanisms contributing to waterloggingdamage to plants.     

Desmocollin 3-mediated Binding Is Crucial for Keratinocyte Cohesion and Is Impaired in Pemphigus

Desmocollin (Dsc) 1–3 and desmoglein (Dsg) 1–4, transmembrane proteins of the cadherin family, form the adhesive core of desmosomes. Here we provide evidence that Dsc3 homo- and heterophilic trans-interaction is crucial for epidermal integrity. Single molecule atomic force microscopy (AFM) revealed homophilic trans-interaction of Dsc3. Dsc3 displayed heterophilic interaction with Dsg1 but not with Dsg3. A monoclonal antibody targeted against the extracellular domain reduced homophilic and heterophilic binding as measured by AFM, caused intraepidermal blistering in a model of human skin, and a loss of intercellular adhesion in cultured keratinocytes. Because autoantibodies against Dsg1 are associated with skin blistering in pemphigus, we characterized the role of Dsc3 binding for pemphigus pathogenesis. In contrast to AFM experiments, laser tweezer trapping revealed that pemphigus autoantibodies reduced binding of Dsc3-coated beads to the keratinocyte cell surface. These data indicate that loss of heterophilic Dsc3/Dsg1 binding may contribute to pemphigus skin blistering.Desmogleins (Dsg)² and desmocollins (Dsc) are members of the Ca²⁺-dependent cadherin family of adhesion molecules that extend with their outer domains into the extracellular core of desmosomes. Desmosomal cadherins include four Dsg (Dsg1–4) and three Dsc3 isoforms (Dsc1–3) (1, 2). Desmosomal cadherins share a common domain organization with five N-terminally located extracellular subdomains (EC1–5). The membrane-distal EC1 domain is thought to contain the adhesive interface necessary for trans-interaction as could be concluded from structural analysis and blocking studies using peptides and antibodies (3–5). By establishing trans- and cis-interacting adhesive complexes, desmosomal cadherins participate in providing mechanical strength to stratified epithelia (6). In human epidermis Dsg1 and Dsc1 expression decreases from the outermost granular layer toward deeper layers, whereas Dsg3 and Dsc3 are primarily found in the basal layer and display an inverse expression gradient (7, 8). In contrast to classical cadherins present in adherens junctions that primarily undergo homophilic trans-interaction, desmosomal cadherins are generally believed to mediate both homo- and heterophilic binding (9). Recently, an important role of Dsc3 for integrity of murine epidermis was demonstrated in animals with conditional epidermal Dsc3 deficiency that suffered from severe intraepidermal blister formation (10) comparable with the phenotype of the autoimmune bullous skin disease pemphigus vulgaris (PV) (11). PV is associated with antibodies (Abs) against Dsg3, in part combined with Abs targeting Dsg1, whereas Dsg1 Abs alone are associated with pemphigus foliaceus (PF). However, PV and PF sera usually do not contain autoantibodies targeting Dsc3 (12). In view of the apparently important role of Dsc3 in epidermal adhesion, we addressed whether Dsg1 and Dsg3 might heterophilically interact with Dsc3 and whether Abs in pemphigus might interfere with such type of interaction.     

Aromatic L-Amino Acid Decarboxylase (AADC) Is Crucial for Brain Development and Motor Functions

Aromatic L-amino acid decarboxylase (AADC) deficiency is a rare pediatric neuro-metabolic disease in children. Due to the lack of an animal model, its pathogenetic mechanism is poorly understood. To study the role of AADC in brain development, a zebrafish model of AADC deficiency was generated. We identified an aadc gene homolog, dopa decarboxylase (ddc), in the zebrafish genome. Whole-mount in situ hybridization analysis showed that the ddc gene is expressed in the epiphysis, locus caeruleus, diencephalic catecholaminergic clusters, and raphe nuclei of 36-h post-fertilization (hpf) zebrafish embryos. Inhibition of Ddc by AADC inhibitor NSD-1015 or anti-sense morpholino oligonucleotides (MO) reduced brain volume and body length. We observed increased brain cell apoptosis and loss of dipencephalic catecholaminergic cluster neurons in ddc morphants (ddc MO-injected embryos). Seizure-like activity was also detected in ddc morphants in a dose-dependent manner. ddc morphants had less sensitive touch response and impaired swimming activity that could be rescued by injection of ddc plasmids. In addition, eye movement was also significantly impaired in ddc morphants. Collectively, loss of Ddc appears to result in similar phenotypes as that of ADCC deficiency, thus zebrafish could be a good model for investigating pathogenetic mechanisms of AADC deficiency in children.     

The Development of Eyespot on Seedling Leaf Sheaths in Winter Wheat and Winter Barley Crops Inoculated with W-type or R-type Isolates of Pseudocercosporella herpotrichoides

In 1986–88 the development of eyespot lesions in winter wheat or winter barley differed plots inoculated with W-type isolates of Pseudocercosporella herpotrichoides and plots inoculated with R-type isolates. In the spring of 1986, after a cold winter, the incidence (%shoots infected) and severity (number of leaf sheaths penetrated) of eyespot lesions in wheat before GS 30/31 were greater in plots inoculated with R-type isolates than in those inoculated with W-type isolates. In 1987, after amild winter, eyespot incidence and severity in both wheat and barley were initially greater in W-type plots than in R-type plots. However, by GS 30/31 or 1987. In 1988, when the crop was October-sown, eyespot incidence and severity were greater in W-type than in R-type plots at GS 30/31. Differences in eyespot incidence and severity between W-type and R-type plots were smaller in barley than in wheat. Both the incidence and severity of eyespot were greater in early-sown than in late-sown plots. Seed rate, had little effect on the rate of lesion development in 1987, but in 1988 the rate of penetration was less at the low seed rate for both wheat and barley.     

氮肥后移对花后受渍冬小麦灌浆特性及产量构成的影响

2010～2012年连续2个生长季,以小麦品种‘烟农19’为材料,大田试验按基肥∶拔节肥∶孕穗肥比例设为氮肥处理N1(10∶0∶0)、N2(7∶3∶0)、N3(5∶5∶0)、N4(3∶5∶2),采用多项式、Logistic方程拟合不同氮肥运筹对花后受渍冬小麦灌浆期籽粒体积与干重变化过程,研究不同氮肥运筹对花后受渍冬小麦灌浆特性及产量构成的影响.结果表明:(1)两年度间各指标变化趋势基本一致,故各指标结果按其两年度平均值计算.(2)花后渍水使冬小麦籽粒体积减小11.8％,灌浆速率降低15.4％,灌浆历期缩短9.2％,千粒重降低20.9％,穗粒数减少7.0％,籽粒产量降低26.6％,而对穗数无显著影响.相对于氮肥全部作为基肥处理N1,氮肥后移处理N2、N3、N1的小麦籽粒体积分别增加10.2％、15.0％、19.4％,灌浆速率分别提高8.5％、16.0％、18.2％,灌浆历期分别延长5.0％、9.4％、11.5％,千粒重分别增加11.9％、20.2％ 、25.6％,穗粒数分别增加7.9％、12.3％、14.6％,产量分别提高18.1％ 、29.6％、37.6％,而穗数分别降低2.2％、3.8％、4.3％.(3)各灌浆参数与冬小麦籽粒产量的关联度表明,花后渍水逆境对灌浆速率的影响大于对灌浆历期的影响,氮肥后移主要通过提高小麦中后期灌浆速率缓解渍水逆境胁迫伤害.研究认为,在花后土壤受渍冬小麦生产区,生产中可获得最高小麦产量的最佳氮肥运筹模式为N4(3∶5∶2)处理.     

Chondroitin Sulfate Is a Crucial Determinant for Skeletal Muscle Development/Regeneration and Improvement of Muscular Dystrophies

Skeletal muscle formation and regeneration require myoblast fusion to form multinucleated myotubes or myofibers, yet their molecular regulation remains incompletely understood. We show here that the levels of extra- and/or pericellular chondroitin sulfate (CS) chains in differentiating C2C12 myoblast culture are dramatically diminished at the stage of extensive syncytial myotube formation. Forced down-regulation of CS, but not of hyaluronan, levels enhanced myogenic differentiation in vitro. This characteristic CS reduction seems to occur through a cell-autonomous mechanism that involves HYAL1, a known catabolic enzyme for hyaluronan and CS. In vivo injection of a bacterial CS-degrading enzyme boosted myofiber regeneration in a mouse cardiotoxin-induced injury model and ameliorated dystrophic pathology in mdx muscles. Our data suggest that the control of CS abundance is a promising new therapeutic approach for the treatment of skeletal muscle injury and progressive muscular dystrophies.     

Oligomerization Is Crucial for the Stability and Function of Heme Oxygenase-1 in the Endoplasmic Reticulum

Heme oxygenase-1 (HO-1), a stress-inducible enzyme anchored in the endoplasmic reticulum (ER) by a single transmembrane segment (TMS) located at the C terminus, interacts with NADPH cytochrome P450 reductase and biliverdin reductase to catalyze heme degradation to biliverdin and its metabolite, bilirubin. Previous studies suggested that HO-1 functions as a monomer. Using chemical cross-linking, co-immunoprecipitation, and fluorescence resonance energy transfer (FRET) experiments, here we showed that HO-1 forms dimers/oligomers in the ER. However, oligomerization was not observed with a truncated HO-1 lacking the C-terminal TMS (amino acids 266–285), which exhibited cytosolic and nuclear localization, indicating that the TMS is essential for the self-assembly of HO-1 in the ER. To identify the interface involved in the TMS-TMS interaction, residue Trp-270, predicted by molecular modeling as a potential interfacial residue of TMS α-helices, was mutated, and the effects on protein subcellular localization and activity assessed. The results showed that the W270A mutant was present exclusively in the ER and formed oligomers with similar activity to those of the wild type HO-1. Interestingly, the W270N mutant was localized not only in the ER, but also in the cytosol and nucleus, suggesting it is susceptible to proteolytic cleavage. Moreover, the microsomal HO activity of the W270N mutant was significantly lower than that of the wild type. The W270N mutation appears to interfere with the oligomeric state, as revealed by a lower FRET efficiency. Collectively, these data suggest that oligomerization, driven by TMS-TMS interactions, is crucial for the stabilization and function of HO-1 in the ER.Heme oxygenase (HO)³ catalyzes the NADPH cytochrome P450 reductase-dependent oxidative degradation of cellular heme to biliverdin, carbon monoxide (CO), and free iron (1, 2). Biliverdin is subsequently converted to bilirubin by biliverdin reductase in the cytosol. Two HO isoforms have been identified in mammalian systems. HO-1 is a 288 amino acid protein and is expressed at high amounts in a variety of pathological conditions associated with cellular stress. There is compelling evidence that HO-1 induction represents an important cytoprotective defense mechanism against oxidative insults by virtue of the anti-oxidant properties of the bilirubin and the anti-inflammatory effect of the CO produced (2). HO-1 is anchored in the endoplasmic reticulum (ER) through a single transmembrane segment (TMS) located at the C terminus, while the rest of the molecule is cytoplasmic (3). HO-1 is sensitive to proteolytic cleavage (4), and it was recently shown that HO-1 can be proteolytically cleaved from the ER and translocated to the nucleus under certain stress conditions (5). Although the catalytic site in the cytoplasmic domain remains intact, the activity of soluble HO-1 is drastically reduced (5), indicating that ER localization is important for its full enzymatic function.Self-assembly to form dimers and higher oligomers is a common phenomenon in many membrane proteins (6, 7). Numerous studies have revealed that interactions between TMSs play an important role in the structure and function of many membrane proteins. Examples include receptors, enzymes, neurotransmitter transporters, and ion channels, in which oligomerization is crucial for their proper cellular localization and function (8). HO-1 does not contain any cysteine residues and has therefore been assumed to function as a monomer (1). To determine whether HO-1 forms oligomers in native membranes, in the present study, we performed chemical cross-linking, co-immunoprecipitation, and FRET analysis using fluorescent protein tags fused to the N terminus of HO-1. The results showed that HO-1 formed dimers/oligomers in the ER and that the TMS provided the interface for the protein-protein interactions. Interference with the TMS-TMS interaction resulted in destabilization of HO-1 and a reduction in enzymatic function.     

Phosphorylation of the Activation Loop Tyrosine 823 in c-Kit Is Crucial for Cell Survival and Proliferation

The receptor tyrosine kinase c-Kit, also known as the stem cell factor receptor, plays a key role in several developmental processes. Activating mutations in c-Kit lead to alteration of these cellular processes and have been implicated in many human cancers such as gastrointestinal stromal tumors, acute myeloid leukemia, testicular seminomas and mastocytosis. Regulation of the catalytic activity of several kinases is known to be governed by phosphorylation of tyrosine residues in the activation loop of the kinase domain. However, in the case of c-Kit phosphorylation of Tyr-823 has been demonstrated to be a late event that is not required for kinase activation. However, because phosphorylation of Tyr-823 is a ligand-activated event, we sought to investigate the functional consequences of Tyr-823 phosphorylation. By using a tyrosine-to-phenylalanine mutant of tyrosine 823, we investigated the impact of Tyr-823 on c-Kit signaling. We demonstrate here that Tyr-823 is crucial for cell survival and proliferation and that mutation of Tyr-823 to phenylalanine leads to decreased sustained phosphorylation and ubiquitination of c-Kit as compared with the wild-type receptor. Furthermore, the mutated receptor was, upon ligand-stimulation, quickly internalized and degraded. Phosphorylation of the E3 ubiquitin ligase Cbl was transient, followed by a substantial reduction in phosphorylation of downstream signaling molecules such as Akt, Erk, p38, Shc, and Gab2. Thus, we propose that activation loop tyrosine 823 is crucial for activation of both the MAPK and PI3K pathways and that its disruption leads to a destabilization of the c-Kit receptor and decreased survival of cells.     

The Number of Catalytic Elements Is Crucial for the Emergence of Metabolic Cores

Background

Different studies show evidence that several unicellular organisms display a cellular metabolic structure characterized by a set of enzymes which are always in an active state (metabolic core), while the rest of the molecular catalytic reactions exhibit on-off changing states. This self-organized enzymatic configuration seems to be an intrinsic characteristic of metabolism, common to all living cellular organisms. In a recent analysis performed with dissipative metabolic networks (DMNs) we have shown that this global functional structure emerges in metabolic networks with a relatively high number of catalytic elements, under particular conditions of enzymatic covalent regulatory activity.

Methodology/Principal Findings

Here, to investigate the mechanism behind the emergence of this supramolecular organization of enzymes, we have performed extensive DMNs simulations (around 15,210,000 networks) taking into account the proportion of the allosterically regulated enzymes and covalent enzymes present in the networks, the variation in the number of substrate fluxes and regulatory signals per catalytic element, as well as the random selection of the catalytic elements that receive substrate fluxes from the exterior. The numerical approximations obtained show that the percentages of DMNs with metabolic cores grow with the number of catalytic elements, converging to 100% for all cases.

Conclusions/Significance

The results show evidence that the fundamental factor for the spontaneous emergence of this global self-organized enzymatic structure is the number of catalytic elements in the metabolic networks. Our analysis corroborates and expands on our previous studies illustrating a crucial property of the global structure of the cellular metabolism. These results also offer important insights into the mechanisms which ensure the robustness and stability of living cells.     

The Clostridium perfringens Germinant Receptor Protein GerKC Is Located in the Spore Inner Membrane and Is Crucial for Spore Germination

The Gram-positive, anaerobic, spore-forming bacterium Clostridium perfringens causes a variety of diseases in both humans and animals, and spore germination is thought to be the first stage of C. perfringens infection. Previous studies have indicated that the germinant receptor (GR) proteins encoded by the bicistronic gerKA-gerKC operon as well as the proteins encoded by the gerKB and gerAA genes are required for normal germination of C. perfringens spores. We now report the individual role of these GR proteins by analyzing the germination of strains carrying mutations in gerKA, gerKC, or both gerKB and gerAA. Western blot analysis was also used to determine the location and numbers of GerKC proteins in spores. Conclusions from this work include the following: (i) gerKC mutant spores germinate extremely poorly with KCl, l-asparagine, a mixture of asparagine and KCl, or NaP_i; (ii) gerKC spores germinate significantly more slowly than wild-type and other GR mutant spores with a 1:1 chelate of Ca²⁺ and dipicolinic acid and very slightly more slowly with dodecylamine; (iii) the germination defects in gerKC spores are largely restored by expressing the wild-type gerKA-gerKC operon in trans; (iv) GerKC is required for the spores'' viability, almost certainly because of the gerKC spores'' poor germination; and (v) GerKC is located in the spores'' inner membrane, with ∼250 molecules/spore. Collectively, these results indicate that GerKC is the main GR protein required for nutrient and nonnutrient germination of spores of C. perfringens food-poisoning isolates.     

The Influence of Leaf Removal upon the Development of the Grain of Winter Wheat

The paper describes the effects on grain development of theremoval of half the flag leaf, the entire flag leaf and allthe leaves from selected main shoots of a field crop of wheat,cultivar Maris Ranger, 7 days after anthesis. None of the treatmentsreduced grain weights in the 14 days following defoliation.Even in the next 14 days only the more severe defoliations reducedgrain weight, with the most severe treatment causing the greatestreduction. Ultimately the rate of grain growth in plants onwhich only a half of the flag leaf had been removed also fellbehind that of the controls. Final grain yields were in theorder, control plants, > plants with half the flag leaf removed,> the entire flag leaf removed, > all the leaves removed.The significance of these results in interpreting the factorslimiting grain growth in wheat is discussed.     

白菜型冬油菜不育系LRCMS花器生理生化特性及其雄蕊发育特征研究

以白菜型冬油菜‘陇油8号’ב陇油9号’杂交后代中选育出的不育系LRCMS,以及代表性甘蓝型油菜不育系(J39A、J68A、Pol10A)和它们的保持系为材料,研究LRCMS在盛花期花蕾和花中POD、SOD、CAT活性和可溶性蛋白含量、过氧化物酶(POD)同工酶谱变化特征及其雄蕊败育过程。结果显示:(1)所有参试材料花朵中POD活性普遍高于花蕾。(2)不育材料POD活性高于保持材料,甘蓝型不育材料POD活性较保持材料增加幅度大于LRCMS与‘陇油9号’,LRCMS花蕾与花的POD活性分别较保持系材料‘陇油9号’高出15.0%和9.0%;不育材料花的CAT活性高于保持系材料89%~293%,而其花蕾的CAT活性低于保持系材料25%~43%,LRCMS与其它不育材料间无显著差异;不育系材料花SOD活性低于保持系材料15%~25%,LRCMS与其它不育材料无显著差异,其花蕾SOD活性高于保持材料6%~112%,但LRCMS增加幅度低于其它不育材料;保持系材料花蕾与花中蛋白质含量为0.014~0.020mg/g,不育系材料花蕾与花中蛋白质含量为0.012~0.016mg/g,所有不育材料蛋白质含量均低于保持材料。(3)LRCMS的POD同工酶谱在花中较对应的可育系‘陇油9号’和其它甘蓝型雄性不育多1条迁移率(Rf)为0.623的条带。(4)LRCMS雄蕊败育起始于造孢细胞期,造孢细胞粘连,无核仁,绒毡层未发育,为无花粉囊型不育。研究表明,LRCMS与其他类型油菜不育系有着不同的遗传背景,是一个新的白菜型冬油菜不育类型。