Effects of nitrogen limitation on water relations of jack pine (Pinus banksiana Lamb.) seedlings

The water relations of shoots of young jack pine (Pinus banksiana Lamb.) seedlings were examined 6 and 15 weeks after the initiation of four different dynamic nitrogen (N) treatments using a pressure-volume analysis. The N treatments produced a wide range of needle N concentrations from 12 to 32 mg g^?1 dry mass and a 10-fold difference in total dry mass at 15 weeks. Osmotic potential at full turgor did not change over the range of needle N concentrations observed. Osmotic potential at turgor-loss point, however, declined as N concentrations decreased, indicating an increased ability of N-deficient jack pine plants to maintain turgor. The increase could be attributed largely to an increase in cell wall elasticity, suggesting that elasticity changes may be a common, significant adaptation of plants to environmental stresses. Dry mass per unit saturated water almost doubled as needle N level dropped from 32 to 12 mg g^?1 and was inversely correlated to the bulk modulus of elasticity. This suggests that cell wall elasticity is determined more by the nature of its cross-linking matrix than by the total amount of cell wall material present. Developmental change was evident in the response of some water relation variables to N limitation.     

严重急性呼吸综合征冠状病毒2膜蛋白对宿主细胞pre-mRNA 3"UTR加工的影响

目的 研究严重急性呼吸综合征冠状病毒2（SARS-CoV-2）膜蛋白对宿主细胞mRNA前体（pre-mRNA）3"非翻译区（UTR）加工的影响。方法 本研究以人肺上皮细胞系A549为模型，利用瞬时转染在细胞内过表达SARS-CoV-2膜蛋白；利用RNA-Seq测序技术及生物信息学分析方法，系统性描绘宿主细胞选择性多聚腺苷酸化（alternative polyadenylation，APA）事件；Metascape数据库对发生显著APA变化的基因进行功能富集分析；RT-qPCR验证靶基因3"UTR长度变化；蛋白质免疫印迹（Western blot）检测目的蛋白表达水平。结果 SARS-CoV-2膜蛋白外源表达后宿主细胞内共813个基因发生显著APA变化。GO和KEGG分析显示，差异APA基因广泛参与有丝分裂细胞周期、调节细胞应激等生物过程，涉及病毒感染和蛋白质加工等。从中进一步筛选出AKT1基因，在IGV软件中显示3"UTR延长；RT-qPCR验证AKT1基因的3"UTR长度变化趋势；Western blot结果显示AKT1蛋白磷酸化水平增加。结论 SARS-CoV-2膜蛋白潜在影响宿主pre-mRNA的3"UTR加工，其中参与多种病毒性生物过程的AKT1基因 3"UTR延长，且其编码的蛋白质功能在细胞内被激活。     

Random Amplified Polymorphic DNA Variation among and within Selected Ixora (Rubiaceae) Populations and Mutants

Random amplified polymorphic DNA (RAPD) analysis was used tostudy variation among and within selectedIxora (Rubiaceae) populationsand mutants. Six populations of I. congesta yielded identicalbanding patterns suggesting genetic uniformity of this species.However, six populations of I. coccinea varieties (three red-flowered,two yellow-flowered and one red-flowered wild-type) exhibitedinfraspecific differences in RAPD profiles. Small and largeleaves of an atavistic mutant cultivar of I. coccinea were alsosubjected to RAPD analysis. An extra band was amplified in thelarge leaves that was absent in small leaves, suggesting thatthe phenotypic alteration in this taxon is due to genetic mutationrather than epigenetic changes. Similarly, an extra band wasdetected in the white sectors of I. Variegated compared to thegreen sectors, suggesting that the shoot apical meristems ofthis cultivar exist as a genetic chimera. DNA gel blot hybridizationwas performed to confirm the specificities of selected bands.Our study indicates that differences among individuals of variouspopulations and mutants may be detected using RAPD markers.Copyright 1999 Annals of Botany Company Ixora L., variegated variety, RAPD fingerprinting, DNA gel blot, intraspecific genetic similarity, atavistic mutant.     

Isolation of the lux genes from Photobacterium leiognathi and expression in Escherichia coli

Genes necessary for luminescence (lux genes) in the marine bacterium Photobacterium leiognathi, strain PL721, were isolated and expressed in Escherichia coli. A 15-kb fragment obtained from a partial digestion of PL721 DNA with HindIII was cloned into the plasmid pACYC184, resulting in the hybrid plasmid pSD721. When pSD721 was transformed into E. coli ED8654, the resulting transformants were luminous with no additions to the cells, indicating that it contained the structural genes coding for the alpha and beta subunits of luciferase (luxA and luxB), and for components involved in aldehyde biosynthesis. Hybridization analysis with luxA and luxB 32P probes confirmed the location of these two genes on the 15-kb insert. When pSD721 was transformed into four different strains of E. coli, luminescence expression varied widely in amount and in pattern. In some strains, luminescence developed like an autoinducible system, and at maximum induction was very bright, even with no addition of aldehyde, while in others, luminescence was 100-fold less, and no induction was seen. In no case was luminescence affected by shifts in temperature, osmolarity, or iron concentration. These results indicate that, while the complete lux regulon is apparently contained on the 15-kb cloned fragment, the regulation of the lux regulon in pSD721 is subject to host controls by E. coli, controls which vary widely among different E. coli strains.     

鸭C4BPα的克隆、表达及其与鸭疫里默氏菌的互作

为研究鸭C4结合蛋白(C4b-binding protein,C4BP)与鸭疫里默氏菌(Riemerella anatipestifer,RA)的相互作用,对鸭C4BPα进行克隆、原核表达,免疫小鼠制备多克隆抗体,并利用间接免疫荧光试验及斑点杂交试验验证C4BP与RA的相互作用。结果显示,鸭C4BPα核苷酸序列全长为1230bp,与鸡C4BPα的相似性最高(82.1%);系统进化树分析发现,鸭C4BPα与鸡C4BPα处于同一系统进化树分支上,两者遗传进化关系最近;C4BPα在大肠杆菌Escherichia coli BL21 (DE3)中能高效表达,重组蛋白以胞内可溶性形式存在;多克隆抗体效价超过1∶10000,并且可以与重组蛋白发生特异性反应;间接免疫荧光试验和斑点杂交试验结果显示RA与鸭C4BP可以发生相互作用。研究结果为进一步揭示RA的致病机制奠定了基础。     

不同方法治疗飞行人员慢性失眠症的疗效比较

目的:飞行人员中睡眠问题发生率高,影响白天工作效能,危及飞行安全,寻找安全有效治疗方法为保证飞行十分必要,本实验观察不同治疗方法在治疗飞行人员慢性失眠的临床疗效,为飞行人员慢性失眠的临床治疗提供一种安全有效的措施。方法:将我院自2007年至2013年收治的52例飞行人员中的慢性失眠患者,排除严重影响睡眠的器质性疾病,随机分为舒乐安定组、中药治疗组和耳针及生物反馈联合治疗组。分别在治疗前、治疗结束后第2天采用匹兹堡睡眠质量指数(PSQI)对患者的睡眠质量进行评价。结果:治疗结束后3组PSQI均较治疗前均有所改善,其中舒乐安定治疗组优于中成药及耳针及生物反馈联合治疗组,差别有统计学意义(P0.05),中成药物治疗组与耳针及生物反馈联合治疗组无明显统计学差异(P0.05)。但对于重度的睡眠障碍,单独采用舒眠胶囊或生物反馈治疗,治疗效果欠佳。结论:舒乐安定治疗失眠疗效确切,但由于其影响工作效能,危及飞行安全较少使用,中成药物、耳针及生物反馈治疗安全有效,病人易于接受,是较好的治疗飞行人员慢性失眠的方法。     

茂兰国家级自然保护区石松类和蕨类植物区系特征

本研究通过野外考查、标本采集和分类鉴定，以及文献资料整理，对贵州茂兰国家级自然保护区石松类和蕨类植物区系组成和分布区类型进行统计分析。结果显示：(1)茂兰国家级自然保护区共有石松类和蕨类植物32科76属237种，优势科为鳞毛蕨科(Dryopteridaceae)、蹄盖蕨科(Athyriaceae)、凤尾蕨科(Pteridaceae)、水龙骨科(Polypodiaceae)；优势属为双盖蕨属(Diplazium)、卷柏属(Selaginella)、凤尾蕨属(Pteris)。(2)分布区类型以东亚广布和热带亚洲分布为主，生态类型以阴生植物和土生植物居多。(3)与其他地区的区系比较显示，茂兰与广西花坪物种相似性系数最高，依次是贵州梵净山、四川峨眉山和云南马关县。