甘蓝型油菜BnMYC2基因克隆及对植物抗虫胁迫的研究

为了研究髓细胞组织增生蛋白(Myelocytomatosis protein,MYC2)基因与甘蓝型油菜抗虫性的关系,该研究从甘蓝型油菜恢复系R18中克隆获得BnMYC2基因的cDNA序列,并进行了基因表达及转化模式植物拟南芥的抗虫分析。生物信息学分析表明,BnMYC2基因gDNA不具有内含子结构,cDNA完整开放阅读框(ORF)为1 833 bp,编码610个氨基酸,推测BnMYC2基因位于甘蓝型油菜C6染色体上,具有bHLH型转录因子的保守结构域HLH。基因同源性分析结果表明,BnMYC2与拟南芥、甘蓝、萝卜的亲缘关系最近,氨基酸的相似度都在98%以上。组织特异性表达分析显示,BnMYC2在甘蓝型油菜的根、茎、叶、花、授粉后27 d的果荚和授粉后35 d的果荚中均有表达,且授粉后27 d的果荚中的表达水平最高。抗虫胁迫实验的结果显示,过表达BnMYC2拟南芥的抗虫基因VSP2的表达水平显著高于对照,推测甘蓝型油菜BnMYC2基因可能作为重要的调节因子参与虫害胁迫响应;MeJA诱导6 h后,VSP2基因在不同基因型拟南芥的表达量都明显增加,与诱导前相比差异显著,表明BnMYC2正向调...     

甘蓝型油菜每角果粒数全基因组关联分析

为挖掘甘蓝型油菜每角果粒数显著关联单核苷酸多态性（SNP）位点及相关候选基因。本研究以300份甘蓝型油菜自交系为试验材料,对甘蓝油菜每角果粒数进行一年两地表型考察,并结合该群体前期开发的201 817个SNPs标记,采用一般线性模型（GLM）和混合线性模型（MLM）进行全基因组关联分析（GWAS）,此外,对性状显著关联SNP位点两侧100 kb区域内相关候选基因进行功能预测。300份甘蓝型油菜每角果粒数在两地均表现出广泛的表型变异,筛选出2份每角果粒数较多的油菜种质资源。基于GLM模型检测到39个与油菜每角果粒数显著关联SNPs,采用MLM分析发现,两地共检测到的3个每角果粒数显著关联SNPs位点均在GLM检测到。8个位点附近找到CIK,ERF022和EDE1等19个拟南芥已报道角果籽粒发育相关的同源基因。研究结果有助于解析甘蓝型油菜每角果粒数的遗传基础,为研究每角果粒数的调控机制、指导每角果粒数的遗传改良奠定基础。     

人工合成甘蓝型油菜特长角变异系的选育

用中国西藏东部采集的1个白菜型油菜地方品种与云南甘蓝类蔬菜白花芥蓝远缘杂交,人工合成了甘蓝型油菜新材料,从杂种后代中通过系统选育,获得了1个特长角变异系,其主花序中部角果平均长度20cm左右,果身长16～18cm,果喙长3cm左右.从中获得的极端最长角果达31.5cm,果身长26.1cm,果喙长5.4cm.这是迄今芸苔属植物中很少见到的长角果油菜材料.该材料的平均角果长度大约为普通甘蓝型油菜的3倍左右,遗传已基本稳定,定名为川农特长角.本文报导其选育经过和主要特征特性,并对其育种和研究利用价值作了简要分析.     

角果开裂方式

角果开裂方式角果是由二心皮组成的雌蕊发育而成。由二心皮腹缝线处向中央生出了隔膜即假隔膜，将于房隔成二室，胎座仍在腹缝线着生，种子就附生在假隔膜边缘两侧的胎座上。果实成熟后，十字花科中除了萝卜、群心菜等植物的角果不开裂外，大多数十字花科植物如大白菜、薄...     

RNA干涉AtSUS3影响拟南芥SUS家族表达模式及角果成熟

蔗糖合成酶（SuSy）是植物蔗糖代谢的关键酶,在植物生长发育过程中起着重要作用.为研究拟南芥中SUS3的功能,构建RNAi-SUS3干涉载体,通过农杆菌介导的真空渗透法转化拟南芥.筛选获得纯系转基因植株后,对AtSUS家族进行表达分析,利用环境扫描电子显微镜观察转基因植株表型,并对转基因拟南芥角果进行木质素组织化学染色以及透射电子显微镜检测.结果表明,RNA干涉技术能够抑制AtSUS3的表达,正常培养条件下该基因沉默后对拟南芥的表型没有显著影响,但可引起角果中AtSUS1,AtSUS2和AtSUS4表达代偿性增加,使转基因植株角果内果皮层细胞次生细胞壁增厚,木质化程度加深,同时果瓣厚度也有增加趋势.结果提示,转基因拟南芥角果的发育较野生型植株更为优先,AtSUS3基因沉默可能有利于角果的成熟.     

油菜矮秆突变WRKY转录因子cDNA克隆及表达分析

以甘蓝型油菜为材料,利用已建立的抑制性消减文库(SSH),采用RACE技术克隆到1个植物WRKY转录因子相关基因,命名为BnD11,其cDNA全长1034 bp,含有810 bp的完整开放阅读框,编码269个氨基酸。该基因编码的氨基酸序列与拟南芥WRKY40氨基酸序列相似性为79%,与拟南芥中编码WRKY-DNA结合蛋白40基因的氨基酸序列相似性达78%,与其它多种植物的WRKY转录因子的氨基酸序列也有较高的相似性。半定量RT-PCR对BnD11进行组织特异性表达分析显示:在正常生长条件下,BnD11在野生型和矮秆油菜的各个组织中均有表达,但在矮秆突变的根、茎、茎尖的相对表达量明显高于野生型。研究表明,BnD11功能区段具有很高的保守性,可能参与了油菜的茎秆发育。     

甘蓝型油菜水苏糖合成酶基因克隆及表达分析

以甘蓝型油菜品种‘德油5号’为试材,采用RT-PCR和RACE方法首次从油菜中获得水苏糖合成酶基因cDNA全长序列,命名为BnSTS,GenBank登录号为HQ285880.1。该cDNA全长3 210bp,包含一个长为2 619bp的开放阅读框,编码873个氨基酸。通过构建原核表达载体,成功表达出预期蛋白。同源性比对结果显示,该基因编码蛋白与拟南芥STS相似性最高为82.08%。荧光定量PCR结果显示,基因表达水平与油菜种子发育过程中水苏糖含量变化具有相关性,与种子脱水耐性的获得具有时间一致性。该基因可能在种子脱水耐性形成机制中具有一定的作用。     

春性甘蓝型油菜BnFY34基因的克隆及序列分析

春性甘蓝型油菜杂交种由于发育期较长,极大地限制了该油菜在高寒地区的推广。为了从春性特早熟甘蓝型油菜中筛选出苗期和蕾期差异表达的基因,该研究以春性特早熟甘蓝型油菜为材料,利用cDNA-AFLP技术筛选出1个春性特早熟甘蓝型油菜苗期特异表达的基因片段,并采用RACE技术成功分离克隆了该基因,命名为BnFY34。通过对该基因的测序和生物信息学分析,发现该基因序列大小为455 bp,包含完整的开放阅读框(ORF),编码一个由71个氨基酸残基组成的蛋白,推测的蛋白质分子量为8.04 kD,理论等电点为10.25,其三级结构中含有3个α螺旋,不含β折叠。同时,将BnFY34基因与甘蓝型油菜基因组序列进行比对,结果显示BnFY34基因位于C4染色体上,由3个外显子和2个内含子组成。另外,通过对该基因与其他植物同源基因的亲缘关系进行了分析,结果表明BnFY34基因与芸薹属植物属于同一亚族,并且与甘蓝型油菜中已报到的未诠释基因XM_013837282.1的相似度达100%,其功能有待进一步研究。该研究结果对春油菜产量和品质的早熟转基因育种具有重要意义。     

甘蓝型油菜的BR响应及BnBZL2基因的功能分析

对甘蓝型油菜(Brassica napus L.)与拟南芥(Arabidopsis thaliana L.)中保守的油菜素甾醇(Brassinosteroids,BR)信号相关基因进行对比分析,并以甘蓝型油菜品种‘沪油15’为材料,对BR信号通路相关同源基因进行了组织表达分析。结果显示,BR合成基因与信号组分在花和幼嫩种子中表达量更高;低浓度BR处理可以促进幼苗根的生长,高浓度BR处理则起抑制作用; BR合成抑制剂(Brassinozole,BRZ)处理可抑制黑暗条件下幼苗下胚轴的伸长; BR处理可以降低BR合成基因的表达水平,而BRZ处理则相反,表明甘蓝型油菜中BR信号增加能反馈抑制BR的合成。烟草(Nicotiana tabacum L.)瞬时表达实验结果发现,与拟南芥BZR1基因同源的甘蓝型油菜BnBZL2编码蛋白定位在细胞质和细胞核中,BR处理可增加BnBZL2的核定位。蛋白质免疫印迹检测结果显示,BR处理可增加去磷酸化BnBZL2的比例。本研究进一步模拟了拟南芥bzr1-1D功能获得性突变体对BnBZL2蛋白进行点突变(BnBZL2*),并构建载体转化拟南芥,黑暗条件下转基因植株幼苗对BRZ处理不敏感,提示BnBZL2*可提高转基因植株的BR信号水平。本研究结果表明甘蓝型油菜中存在与拟南芥相似且保守的BR信号通路和调控机制。     

荠菜CRABS CLAW的克隆与拟南芥遗传转化

根据GenBank收录的CRC基因cDNA序列设计引物,以短角果荠菜(Capsella bursa-pastoris)为材料,通过RT-PCR扩增出与拟南芥CRC基因同源的全长cDNA,进行测序、比对及同源性分析.结果显示,克隆的该基因cDNA序列与报道的拟南芥CRC序列一致性达到93%,可以推断其为荠菜CRC基因的cDNA(cbCRC).以Ti质粒pWM101为载体,构建了由CaMV35S启动子调控的cbCRC基因植物表达载体pWM101-cbCRC, 采用根癌农杆菌滴注柱头法转化拟南芥,获得了转cbCRC基因的拟南芥植株.转基因拟南芥心皮果荚形态大小发生了一定的变化,说明荠菜cbCRC基因在拟南芥中的表达对拟南芥心皮形态和大小都产生了一定影响,但其并没有使拟南芥表现出荠菜短角果的形态.     

白蚁与动物及微生物的共生类型及其机制

综述了白蚁螱客的主要种类、共生关系及相关机制的研究进展。白蚁螱客中,已报道的动物种类达170种。在与动物的共生关系中存在偏利共生（宾主共栖和异种共栖）、互利共生和无关共生三种;在与微生物的共生关系中,存在与内生菌（原生动物、细菌、真菌和放线菌）和外生菌（蚁巢伞菌等）间的互利关系。指出了白蚁与螱客研究中存在的问题,给出了解决方案,并提出了今后可能的研究热点或方向,为白蚁的综合利用（如纤维素酶）及今后研究物种间的协同进化提供了基础资料。     

New amino-dithiaphospholanes and phosphoramidodithioites and their rhodium and iridium complexes

New sulfur derivatives of phosphoramidite ligands were synthesized and the impact of the sulfur unit on the spectroscopic properties of their rhodium and iridium complexes was investigated. The new ligands Bn₂NPSCH₂CH₂S^a(P-S^a) (Bn = benzyl, 4), Bn₂NPSCHCHS^a(CH₂)₃C^aH₂(P-S^a)(C^a-S^a) (6) and Bn₂NP(4-XC₆H₄OMe)₂ (X = S, 7a; X = O, 7b) were converted to the rhodium and iridium complexes trans-[Rh(CO)Cl(L)₂] (L = 4, 6, 7), [RhCl(COD)(L)] (L = 4, 6, 7), [IrCl(COD)(7a)] and [IrCl₂Cp∗(6)]. For comparison, some phosphoramidite complexes of these formulations also were synthesized. The new metal complexes were spectroscopically analyzed. For the carbonyl complexes, the ν_CO IR stretching frequencies were lower than for the corresponding phosphite and phosphoramidite ligands. The ¹J_PRh coupling constants for the rhodium complexes with the new ligands were also smaller than for the respective phosphoramidite and phosphite complexes. Finally, the ¹J_PSe coupling constants of the selenides of the new ligands were lower than those of the phosphoramidite ligands but higher than for PPh₃. The spectroscopic data reveal that the new thio ligands 4, 6 and 7a are more electron donating than phosphites and phosphoramidites but less electron donating than PPh₃.     

Astrocytes and Synaptosomes Transport and Metabolize Lactate and Acetate Differently

Astrocytes transport the monocarboxylate acetate, but synaptosomes do not. The reason for this is unknown, because both preparations express monocarboxylate transporters (MCT). The transport and metabolism of lactate, another monocarboxylate, was examined in these two preparations, and the results were compared to those for acetate. Lactate transport is more rapid in astrocytes than in synaptosomes, but of lower affinity (Kms of 17 and 4 mM, respectively). Lactate (0.2 mM) is metabolized to CO2 more rapidly in synaptosomes than in astrocytes (rates of 0.37 and 0.07 nmol x mg protein(-1) x min(-1), respectively). The reason for this is unclear, but cellular differences in lactate dehydrogenase isotype expression may be involved. Acetate is metabolized to CO2 more rapidly in astrocytes than in synaptosomes (rates of 0.43 and 0.02 nmol x mg protein(-1) x min(-1), respectively). This is likely due to cellular differences in the expression of monocarboxylate transporter subtypes.     

Rapporteur report: Basics and technology and metrology and standards

The first and second sessions of the Workshop focussed on the basics of ultrasound and infrasound, their applications in both industry and medicine, and metrology and protection standards for ultrasound applications.     

Relative and combined effects of ethanol and protein deficiency on strontium and barium bone content and fecal and urinary excretion

To elucidate accumulation of minerals in human iliac arteries with aging, the content of minerals was analyzed by inductively coupled plasma atomic emission spectrometry. Bilateral common, internal, and external iliac arteries of 16 men and 8 women, ranging ages from 65 to 93 yr, were examined. It was found that an extremely high accumulation of calcium and phosphorus occurred in the common iliac artery at old age, being higher than that of the internal and external iliac arteries. It should be noted that the accumulation of calcium and phosphorus is the highest in the common iliac artery among the human arteries examined to date. Regarding sexual differences, the content of calcium and phosphorus in the common and internal iliac arteries was higher in women than in men, whereas their content in the external iliac artery was lower in women than in men.     

Prostaglandin and thromboxane production by human and guinea-pig macrophages and leucocytes

The ability of partially purified human and guinea-pig haematogenous cell populations, when cultured in vitro, to metabolise arachidonic acid (AA) has been studied. Supernatants from 24 hour cell culture have been subjected to analysis for products of AA metabolism by gas chromatography with electron-capture detection.The cell types studied were human peripheral blood monocytes (both glass adherent and non-adherent), neutrophils, eosinophils and leukemic leucocytes; thoracic duct lymphocytes and lung alveolar macrophages. From the guinea-pig, induced and non-induced macrophage or neutrophil enriched peritoneal exudate populations, lymph node cells, peritoneal eosinophils and peripheral blood platelets were examined. Supernatants were assayed for the presence of PGE₂, PGD₂, PGF_2α, TXB₂ and 6-keto-PGF_1α. In all types studied PGE₂ and TXB₂ were the major products formed. The identification of PGE₂ and TXB₂ was confirmed by GC/MS with multiple ion monitoring.The results have been compared with other reports and their possible significance discussed in relation to the proposed role of prostaglandins as mediators and modulators in immunopathology.     

Chemokines and cytokines: axis and allies in asthma and allergy

Allergic asthma can be precipitated by many factors. For the atopic person, fungus, pollen, dust mites, cockroach antigens, and diesel exhaust are all agents that may trigger an allergic attack. Cytokines and chemokines are integral mediators of fungal asthma. From the earliest time points, they recruit and activate the cells required for the clearance of fungus as well as being critical factors involved in the immunopathology of this disease. In the final analysis, it is clear that these mediators can act to the benefit or the detriment of the host.     

Oxidation and nitration of ribonuclease and lysozyme by peroxynitrite and myeloperoxidase

In spite of the many studies on protein modifications by reactive species, knowledge about the products resulting from the oxidation of protein-aromatic residues, including protein-derived radicals and their stable products, remains limited. Here, we compared the oxidative modifications promoted by peroxynitrite and myeloperoxidase/hydrogen peroxide/nitrite in two model proteins, ribonuclease (6Tyr) and lysozyme (3Tyr/6Trp). The formation of protein-derived radicals and products was higher at pH 5.4 and 7.4 for myeloperoxidase and peroxynitrite, respectively. The main product was 3-nitro-Tyr for both proteins and oxidants. Lysozyme rendered similar yields of nitro-Trp, particularly when oxidized by peroxynitrite. Hydroxylated and dimerized products of Trp and Tyr were also produced, but in lower yields. Localization of the main modified residues indicates that peroxynitrite decomposes to radicals within the proteins behaving less specifically than myeloperoxidase. Nitrogen dioxide is emphasized as an important protein modifier.