镉对玉米苗中钙调蛋白含量和Ca2+-ATPase活性的影响

试验采用营养液培养的方法,以玉米为试材,研究了不同供镉浓度（0﹑5﹑20和100 µmol/L）和处理时间（12﹑24﹑48﹑96、168 h）对植株体内钙调蛋白（CaM）含量及生物膜上的Ca²⁺-ATPase活性的影响。结果表明,植株可溶性Ca²⁺含量在镉胁迫后较不加镉处理增加,镉处理在叶和根中分别在48和24 h后达最高,然后随镉处理浓度和处理时间的增加逐步下降;同时镉诱导了植株CaM的合成,其含量随镉处理浓度和处理时间增加逐步增加,但20 µmol/L和100 µmol/L镉处理在168 h后有所下降;与不加镉处理相比,镉胁迫导致植株生物膜上的Ca²⁺-ATPase活性迅速升高,但随镉处理浓度提高和时间延长,镉胁迫植株的Ca²⁺-ATPase活性在48 h（质膜、液泡膜和内质网膜）和24 h（线粒体膜）后逐步降低。各膜上的Ca²⁺-ATPase活性依次为质膜> 液泡膜> 内质网膜> 线粒体膜,且同一微囊膜,根中的活性大于叶中。     

PEG胁迫下Ca2+和钙调素拮抗剂W7对不同品种番茄幼苗抗氧化系统的影响

以耐旱性不同的2个番茄品种为试材,采用营养液水培法,研究了钙、钙调素拮抗剂W7[N-(6-aminohexyl) -5-chloro-1-naphthalene sulfonami de]预处理对聚乙二醇模拟干旱胁迫下番茄幼苗抗氧化系统的影响.结果表明,钙调素拮抗剂W7浸种处理显著提高了番茄幼苗丙二醛(MDA)、H2O2含量和O2 ()/(*)]产生速率,抑制了超氧化物歧化酶(SOD)、过氧化物酶(POD)活性,加剧了抗坏血酸(A sA)和还原型谷胱甘肽(GSH)的破坏;而Ca 2+处理显著降低了PEG胁迫下番茄幼苗MDA 、H2O2含量和O2 ()/(*)产生速率,提高了SOD、POD活性,减轻了胁迫对AsA和GSH的破坏.相同处理条件下,耐旱性强的品种'毛粉802'较耐旱性弱的品种'皇冠'伤害程度轻,Ca 2+-CaM信号系统对缓解番茄PEG胁迫具有重要作用.     

利用 CRISPR/Cas9 技术定向编辑水稻 OsIQD1 基因

【目的】植物特异性 IQD 家族蛋白质是一类植物特有的钙调素结合靶蛋白,在植物的生物防 御和发育调节中发挥重要作用。前期酵母双杂筛选试验表明,Pik-H4 与 OsIQD1 存在互作关系,为进一步 揭示 OsIQD1 在水稻抗稻瘟病中的生物学功能,利用 CRISPR/Cas9 编辑技术对 OsIQD1 基因进行定点编辑。 【方法】通过序列比对获得水稻 OsIQD1。在 OsIQD1 第 1 外显子和第 5 外显子（DUF4005 结构域）区域设计 2 个 20 bp 的编辑靶点,将 2 个靶点核苷酸片段克隆至 pRGEB32 载体,获得 pRGEB32-OsIQD1-gRNA 载体,利用 农杆菌介导法转化水稻 Pik-H4 NIL 愈伤组织,经再生培养、潮霉素检测获得转基因阳性植株,并对 T0 代转基 因植株靶点区域序列进行 PCR 和测序,分析 osiqd1 的突变类型。【结果】转基因再生植株经过对抗潮霉素基因 HPT Ⅱ的 PCR 鉴定,获得 30 株阳性株系。对 T0 代植株靶点附近序列的测序结果表明,OsIQD1 基因被成功编 辑,两个位点的编辑效率分别为 21.67% 和 26.67%,其中,靶点 2 区域有 1 个纯合突变株系。【结论】研究结 果为进一步利用 osiqd1 突变体开展其参与钙离子 / 钙调素信号转导通路的机制研究提供了遗传材料,初步推定 OsIQD1 参与植物的基础免疫反应。     

Calmodulin is involved in the occurrence of extracellular Ca2+-dependent full-type hyperactivation in boar ejaculated spermatozoa incubated with cyclic AMP analogs

In mammals, hyperactivation is essential for sperm fertilization with oocytes in vivo. Two types of hyperactivation “full-type and nonfull-type patterns” can be observed in the spermatozoa from boars, bulls, and mice. We have a hypothesis that the full-type hyperactivation is a physiological (in vivo) pattern and are elucidating its molecular bases. The aims of this study were to detect calmodulin in boar sperm flagella by Western blotting and indirect immunofluorescence and to investigate effects of extracellular Ca²⁺ and calmodulin antagonists “W-7 and W-5 (W-5; a less potent antagonist)” on the occurrence of full-type hyperactivation in boar spermatozoa. Calmodulin was specifically detected as the 17-kDa antigen in the flagella and postacrosomal region of the heads. Full-type hyperactivation could be induced effectively in the samples incubated with 3.42 mM CaCl₂ for 120–180 min, and it was significantly reduced in the concentration-dependent manners of W-7 and W-5. Suppressing effects of W-7 on the full-type hyperactivation were stronger than those of W-5. These observations indicate that flagellar calmodulin is involved in the occurrence of extracellular Ca²⁺-dependent full-type hyperactivation in boar spermatozoa. This is the first indication of the intracellular Ca²⁺-sensing molecule which can function in the full-type hyperactivation.     

CaM与ZmCCaMK相互作用参与BR诱导的玉米叶片抗氧化防护

以玉米(Zea mays L.)为材料,通过酶联免疫法、实时荧光定量PCR技术、免疫共沉淀激酶反应以及原生质体瞬时表达体系等方法研究了在油菜素内酯(Brassinosteroids,BR)诱导的玉米叶片抗氧化防护过程中钙调素(Calmodulin,Ca M)的作用以及Ca M和Ca2+/Ca M依赖的蛋白激酶(Zm CCa MK)之间的关系。结果显示:外源BR处理显著提高了玉米叶片叶肉细胞原生质体中的Ca M含量,Ca M拮抗剂的预处理几乎完全抑制了BR诱导的抗氧化防护酶活性,表明Ca M参与了BR诱导的抗氧化防护。而且Zm CCa MK原生质体瞬时表达及瞬时沉默结果显示Zm CCa MK影响BR诱导的Ca M含量,Ca M拮抗剂的预处理也显著抑制了BR诱导的H2O2的产生、Zm CCa MK基因表达以及激酶活性。说明BR诱导产生的Ca M增强H2O2的积累,进一步诱导抗氧化防护酶活性增加,并且Ca M和Zm CCa MK之间存在互动机制。     

New taxa in Aspergillus section Usti

Based on phylogenetic analysis of sequence data, Aspergillus section Usti includes 21 species, inclucing two teleomorphic species Aspergillus heterothallicus (= Emericella heterothallica) and Fennellia monodii. Aspergillus germanicus sp. nov. was isolated from indoor air in Germany. This species has identical ITS sequences with A. insuetusCBS 119.27, but is clearly distinct from that species based on β-tubulin and calmodulin sequence data. This species is unable to grow at 37 °C, similarly to A. keveii and A. insuetus. Aspergillus carlsbadensis sp. nov. was isolated from the Carlsbad Caverns National Park in New Mexico. This taxon is related to, but distinct from a clade including A. calidoustus, A. pseudodeflectus, A. insuetus and A. keveii on all trees. This species is also unable to grow at 37 °C, and acid production was not observed on CREA. Aspergillus californicus sp. nov. is proposed for an isolate from chamise chaparral (Adenostoma fasciculatum) in California. It is related to a clade including A. subsessilis and A. kassunensis on all trees. This species grew well at 37 °C, and acid production was not observed on CREA. The strain CBS 504.65 from soil in Turkey showed to be clearly distinct from the A. deflectus ex-type strain, indicating that this isolate represents a distinct species in this section. We propose the name A. turkensis sp. nov. for this taxon. This species grew, although rather restrictedly at 37 °C, and acid production was not observed on CREA. Isolates from stored maize, South Africa, as a culture contaminant of Bipolaris sorokiniana from indoor air in Finland proved to be related to, but different from A. ustus and A. puniceus. The taxon is proposed as the new species A. pseudoustus. Although supported only by low bootstrap values, F. monodii was found to belong to section Usti based on phylogenetic analysis of either loci BLAST searches to the GenBank database also resulted in closest hits from section Usti. This species obviously does not belong to the Fennellia genus, instead it is a member of the Emericella genus. However, in accordance with the guidelines of the Amsterdam Declaration on fungal nomenclature (Hawksworth et al. 2011), and based on phylogenetic and physiological evidence, we propose the new combination Aspergillus monodii comb. nov. for this taxon. Species assigned to section Usti can be assigned to three chemical groups based on the extrolites. Aspergillus ustus, A. granulosus and A. puniceus produced ustic acid, while A. ustus and A. puniceus also produced austocystins and versicolorins. In the second chemical group, A. pseudodeflectus produced drimans in common with the other species in this group, and also several unique unknown compounds. Aspergillus calidoustus isolates produced drimans and ophiobolins in common with A. insuetus and A. keveii, but also produced austins. Aspergillus insuetus isolates also produced pergillin while A. keveii isolates produced nidulol. In the third chemical group, E. heterothallica has been reported to produce emethallicins, 5'-hydroxyaveranthin, emeheterone, emesterones, 5'-hydroxyaveranthin.     

Ca2＋信号通路对本氏烟叶位介导的核盘菌抗性的影响

组合采用药理学、分子生物学和反向遗传学等技术,分析叶位对本氏烟抗核盘菌( Sclerotinia sclerotiorum)的影响及其机制。结果表明,叶位显著影响本氏烟对核盘菌的抗性,随着叶位自上而下,该抗性逐渐增强。药理学分析结果显示,Ca2＋通道抑制剂 LaCl3和 NaVO3处理消除了本氏烟叶片对核盘菌抗性的叶位间差异。定量反转录聚合酶链反应检测结果显示,3个 Ca2＋信号通路基因 NbCNGC20、NbCA M TA3和 NbCML1在本氏烟不同叶位叶片中的表达存在显著差异,随着叶位自上而下逐渐增加。病毒诱导的基因沉默( virus‐induced gene silencing ,VIGS)分析结果表明,钙调素类似蛋白基因 NbCML1的沉默导致叶位介导的本氏烟对核盘菌抗病性的丧失。这些结果说明叶位对本氏烟抗核盘菌具有显著影响,揭示了包括 NbCML1基因在内的 Ca2＋信号通路对叶位介导的核盘菌抗性的重要调控作用。     

镉胁迫下外源钙对白菜氧化应激和NO含量的影响

为了解镉胁迫下外源钙对白菜生理响应的调控机制,通过水培实验研究了在Cd(100μmol·L-1)胁迫下Ca(5 mmol·L-1)对白菜(Brassica pekinensis)叶片钙离子和镉离子含量、ROS和MDA含量、抗氧化酶活性、还原性谷胱甘肽和内源NO含量变化的影响,并使用0.2 mmol·L-1的三氟拉嗪(TFP)研究了Ca2+-CaM信号在其中的转导功能。结果表明:(1)Cd胁迫降低了白菜叶片的Ca含量,而外源Ca增加了Ca含量并降低Cd含量,但TFP对叶片Ca、Cd含量无明显影响;(2)Cd诱导白菜叶片ROS爆发,引起质膜过氧化,并抑制了SOD活性,提高了APX、CAT和POD活性及GSH含量,增加了叶片过氧化压力,施加外源钙后降低了这种压力,而TFP的加入却降低了这种缓解作用;(3)Cd抑制了叶片NO含量,补充外源钙后提高了NO含量,而TFP降低了外源钙对NO的上调。研究可知,外源钙能降低白菜叶片Cd含量,增强Ca含量,并通过Ca2+-CaM信号途径调节抗氧化酶和GSH含量,维持叶片ROS平衡,提高叶片内源NO的表达,从而增强白菜Cd的耐性。     

Verapamil和CPZ对金冠苹果衰老生理的影响

对金冠苹果经Ca2 通道阻塞剂Verapamil,CaM拮抗剂Chloropromaize（CPZ）处理后的一些生理生化变化做了研究。结果表明,Verapamil和CPZ促进果肉软化,增加膜透性和可溶性蛋白质含量,分别提早呼吸跃变、提高呼吸速率的峰值,并且加大了过氧化物酶和超氧化物歧化酶活性的变化幅度。