Chloroplasts require glutathione reductase to balance reactive oxygen species and maintain efficient photosynthesis

Thiol‐based redox‐regulation is vital for coordinating chloroplast functions depending on illumination and has been throroughly investigated for thioredoxin‐dependent processes. In parallel, glutathione reductase (GR) maintains a highly reduced glutathione pool, enabling glutathione‐mediated redox buffering. Yet, how the redox cascades of the thioredoxin and glutathione redox machineries integrate metabolic regulation and detoxification of reactive oxygen species remains largely unresolved because null mutants of plastid/mitochondrial GR are embryo‐lethal in Arabidopsis thaliana. To investigate whether maintaining a highly reducing stromal glutathione redox potential (E_GSH) via GR is necessary for functional photosynthesis and plant growth, we created knockout lines of the homologous enzyme in the model moss Physcomitrella patens. In these viable mutant lines, we found decreasing photosynthetic performance and plant growth with increasing light intensities, whereas ascorbate and zeaxanthin/antheraxanthin levels were elevated. By in vivo monitoring stromal E_GSH dynamics, we show that stromal E_GSH is highly reducing in wild‐type and clearly responsive to light, whereas an absence of GR leads to a partial glutathione oxidation, which is not rescued by light. By metabolic labelling, we reveal changing protein abundances in the GR knockout plants, pinpointing the adjustment of chloroplast proteostasis and the induction of plastid protein repair and degradation machineries. Our results indicate that the plastid thioredoxin system is not a functional backup for the plastid glutathione redox systems, whereas GR plays a critical role in maintaining efficient photosynthesis.     

定点突变提高解淀粉芽孢杆菌JP-21脲酶应用特性

氨基甲酸乙酯(Ethylcarbamate,EC)是一种存在于发酵食品和酒精饮料中的可致癌物,过量摄入可能会影响人体健康。酶法降解是减少发酵食品中氨基甲酸乙酯及其前体尿素含量的有效方法之一。脲酶具有氨基甲酸乙酯水解酶和尿素酶两种活性,因此在减少发酵食品中氨基甲酸乙酯及其前体尿素方面具有良好的应用前景。目前脲酶降解发酵酒精饮料中氨基甲酸乙酯面临的主要问题是脲酶对氨基甲酸乙酯的催化活性及亲和力较低,因而其降解效果不理想。文中成功在大肠杆菌Escherichia coli中表达了来源于解淀粉芽孢杆菌Bacillus amyloliquefaciens JP-21的脲酶,表达水平为尿素酶3 291.74 U/L,氨基甲酸乙酯水解酶227.26 U/L。通过模拟脲酶中催化亚基UreC与氨基甲酸乙酯对接的结构,确定了M326和M374这两个影响酶与底物结合的位点。采用点饱和突变获得了3株氨基甲酸乙酯水解酶活性提高的突变体M374A、M374T和M326V,以EC为底物时的Km分别为101.84mmol/L、129.49 mmol/L和121.67 mmol/L,比野生型分别降低了37.47%–50.82%。突变体可以降解黄酒中97%的尿素,M374T对黄酒中EC的降解效果最好,可将黄酒中EC从513.90μg/L降至393.57μg/L,降解率是野生脲酶的1.97倍。研究结果对今后改造脲酶催化特性和改善其应用特性具有重要意义,可为开发减控发酵食品中的微生物代谢氨(胺)类危害物策略提供参考。     

‘培矮64S’与其eui突变体‘长选3S’穗颈节间蛋白质组差异分析

为从蛋白质表达水平了解长穗颈温敏核不育水稻穗颈节间伸长机理,该研究以长穗颈(EUI)温敏核不育水稻‘长选3S’为材料,温敏核不育水稻‘培矮64S’为对照,采用固相pH梯度双向凝胶电泳和质谱分析方法,对2个水稻材料抽穗前2 d的穗颈节间蛋白质进行分离,并进行差异蛋白质组学的比较研究。结果表明:(1)获得了分辨率和重复性较好的双向凝胶电泳图谱。(2)对40个差异蛋白质点进行MALDI-TOF-TOF-MS肽质谱指纹图谱分析,成功鉴定其中27个差异蛋白质点;与‘培矮64S’相比,‘长选3S’中有17个上调表达和10个下调表达的蛋白质。(3)差异蛋白质按照其功能可分为6类,其中主要是与细胞代谢相关蛋白,其次是与细胞壁重建相关蛋白;并且这些差异蛋白质可能与‘长选3S’抽穗期穗颈节间剧烈伸长生长有关,尤其是细胞壁重建相关蛋白与细胞的伸长密切相关。(4)实时荧光定量PCR对随机挑选的蛋白点2、7、8、24、35和36所对应的基因在两个材料最上节间的表达结果显示,‘长选3S’的2(Os10g08550)、7(Os12g42876)、8(Os01g55830)基因的表达量较‘培矮64S’明显下调,而24(Os06g48760)、35(Os05g25850)、36(Os07g42300)基因的表达量较‘培矮64S’显著上调,表明q-PCR的结果与蛋白凝胶图分析结果一致。研究认为,水稻eui基因可能是通过调节抽穗期穗颈节间这些蛋白质的表达,从而促进穗颈节间细胞分裂,尤其是细胞的伸长生长。     

典型草原大针茅种群空间格局及对长期过度放牧的响应

种群空间格局是生态学研究的基本问题之一。典型草原带由于过度放牧退化严重, 原生群落罕见, 探讨原生群落的种群空间格局具有重要生态学意义。大针茅(Stipa grandis)草原是典型草原区广泛分布的主要群落类型, 1979年围封的大针茅样地, 是目前保存完整的大针茅草原原生群落。本文选择大针茅草原原生群落和长期过度放牧群落, 应用O-Ring函数结合不同零假设模型分析了大针茅种群的空间格局。结果表明: 在原生群落中大针茅种群在小尺度范围内呈均匀分布, 而在长期过度放牧群落中则表现为聚集分布。这说明在大针茅草原原生群落中竞争是主要的相互作用, 而在长期过度放牧群落中正相互作用居主导, 验证了胁迫梯度假说; 同时证明长期过度放牧改变了种群空间格局。     

Heterogeneous genetic invasions of three insecticide resistance mutations in Indo‐Pacific populations of Aedes aegypti (L.)

Nations throughout the Indo‐Pacific region use pyrethroid insecticides to control Aedes aegypti, the mosquito vector of dengue, often without knowledge of pyrethroid resistance status of the pest or origin of resistance. Two mutations (V1016G + F1534C) in the sodium channel gene (Vssc) of Ae. aegypti modify ion channel function and cause target‐site resistance to pyrethroid insecticides, with a third mutation (S989P) having a potential additive effect. Of 27 possible genotypes involving these mutations, some allelic combinations are never seen whereas others predominate. Here, five allelic combinations common in Ae. aegypti from the Indo‐Pacific region are described and their geographical distributions investigated using genome‐wide SNP markers. We tested the hypothesis that resistance allele combinations evolved de novo in populations versus the alternative that dispersal of Ae. aegypti between populations facilitated genetic invasions of allele combinations. We used latent factor mixed‐models to detect SNPs throughout the genome that showed structuring in line with resistance allele combinations and compared variation at SNPs within the Vssc gene with genome‐wide variation. Mixed‐models detected an array of SNPs linked to resistance allele combinations, all located within or in close proximity to the Vssc gene. Variation at SNPs within the Vssc gene was structured by resistance profile, whereas genome‐wide SNPs were structured by population. These results demonstrate that alleles near to resistance mutations have been transferred between populations via linked selection. This indicates that genetic invasions have contributed to the widespread occurrence of Vssc allele combinations in Ae. aegypti in the Indo‐Pacific region, pointing to undocumented mosquito invasions between countries.     

生境梯度影响下的天然红松种群空间格局与种内关联

为理清生境梯度下天然红松种群空间分布规律,以黑龙江省凉水国家级自然保护区内自坡底至坡顶的四种生境(生境Ⅰ,谷地平坡潮湿生境;生境Ⅱ,坡下缓坡潮湿生境;生境Ⅲ,坡上斜坡半湿润生境;生境Ⅳ,坡顶陡坡半干旱生境)类型中建群种红松种群为研究对象。分析不同林层(主林层、亚林层、演替层、更新层)红松种群的数量特征,并基于O-ring函数,在重复采样条件下评价了红松种群空间分布格局。结果表明:(1)下坡位生境(生境Ⅰ、Ⅱ)红松更新层种群数量显著大于上坡位(生境Ⅲ、Ⅳ)(P0.05),演替层中差异不显著;自坡底至坡顶的生境梯度上,主林层和亚林层中红松株数比例增加,红松种群年龄结构结构由倒J型向J型转变。(2)下坡位原始红松林更新层和演替层中红松种群聚集规模与聚集强度均高于上坡位,坡顶(生境Ⅳ)原始红松林更新层中红松种群聚集规模与聚集强度高于坡上(生境Ⅲ)。(3)四类生境的原始红松林中主林层与更新层中红松种群总体表现为空间独立,在个别尺度上表现出微弱的空间正关联或负关联;在0—5 m尺度上,下坡位红松林原始林内演替层与主林层红松种群表现为空间负关联,而上坡位生境林分对应林层间空间关系则为空间正关联;四类生境中,亚林层与主林层中红松种群在全部研究尺度上均表现为空间独立。可以认为,造成上坡位生境的原始红松林中幼树、幼苗聚集规模与强度弱于下坡位的原因是上坡位更新与演替层中种群数量较少;而随坡位上升,更现层、演替层与主林层中红松种群间的空间关系由空间负关联向空间正关联转变是生物与非生物因子共同作用的结果。研究结果为阔叶红松林的经营管理与生态修复提供了理论依据。     

2种调查方法对四川黑竹沟国家级自然保护区3种雉类种群密度调查的比较

种群数量是物种的重要生态学基础资料,合适的密度调查方法是数量估算的基础。2016年4-5月,采用广泛应用于鸡形目Galliformes鸟类种群密度调查的样线法和样点法,调查了四川黑竹沟国家级自然保护区3种鸡形目鸟类(白腹锦鸡Chrysolophus amherstiae、红腹角雉Tragopan temminckii和血雉Ithaginis cruentus)的种群密度。样线法和样点法估算的雄体密度分别是:白腹锦鸡1.20只/km^2和(6.31±0.98)只/km^2,红腹角雉5.41只/km^2和(0.39±0.17)只/km^2,血雉3.01只/km^2和(5.97±2.70)只/km^2。除红腹角雉外,样点法估算的白腹锦鸡、血雉种群密度均大于样线法。建议针对不同鸡形目鸟类采用不同的调查方法,并尽量扩大样本数量,从而提高调查结果的准确性。     

Impacto del ejercicio físico en variables relacionadas con el bienestar emocional y funcional en adultos mayores

Background and aimThe benefits of the physical exercise in aging, and specially in frailty, have been associated with reduced risk of mortality, chronic disease, and cognitive and functional impairments. Multi-component training, which combines strength, endurance, balance, and gait training, represents the most beneficial kind of physical exercise in older adults.MethodsGiven the effectiveness of the multi-component training, a physical exercise program «Actívate» (based on the methodology Vivifrail), with the focus on «active aging», was conducted in the present study. Forty-nine older adults over 60 years participated in this program.ResultsThe physical exercise intervention led to a reduction in diastolic blood pressure, pain threshold and sleep disturbances (e. g. hypersomnia) (t ≥ 2.72, p < 0.01), as well as an increase of walking speed (t = 7.84, p ≤ 0.001). Further, quality of life factors (GENCAT scale), like emotional well-being, personal development, physical well-being, self-determination, and social inclusion, were greater after intervention (t ≥ ?2.06, p < 0.05).ConclusionsThese findings underline the benefits of multi-component training in functionality of older adults, and further, provide relevant aspects about the modulation of pain perception, sleep disturbances, social factors and physical and emotional well-being. Physical exercise programs such as «Actívate» should be promoted, in order to encourage healthy lifestyle habits, in the older adults’ population.     

雏鸡源致病性粪肠球菌的分离鉴定及其耐药性分析

【背景】粪肠球菌(Enterococcus faecalis)是人和动物肠道正常菌群之一,也是一种条件性致病菌。近年来,粪肠球菌引起人和动物感染的报道越来越多。【目的】探明引起某养鸡场雏鸡发病死亡的病原及其致病性和有效治疗药物。【方法】结合临床症状和病理剖检,开展病原菌分离、生理生化特性检测和16S rRNA基因序列分析、致病性试验、耐药分析和对患病鸡群的药物治疗。【结果】患病鸡有昏睡、瘫痪或共济失调等临床症状;肝、脾肿大,肝脏发黄、少量出血点、质脆易碎,肠道粘膜增厚、出血,脑轻微水肿;从肝脏组织分离得到一株革兰氏阳性球菌,经纯化培养后命名为CJ517;依据该菌株形态特征、生理生化特性和16S rRNA基因序列分析鉴定为粪肠球菌;致病性试验显示CJ517菌株能致死小鼠,致死率为66.67%;该菌对头孢噻肟、磷霉素、丁胺卡那等药物敏感,对多西环素、卡那霉素、新霉素、氟苯尼考等药物耐药;经用敏感药物和提高免疫力结合治疗后,鸡群病情得到控制。【结论】研究结果可为临床诊断和治疗动物粪肠球菌感染提供参考。     

组织工程细胞片的构建

作为一种无支架的组织工程技术,组织工程细胞片不仅可以避免支架材料带来的不利影响,而且可通过组装进一步形成更为复杂的三维功能化组织,因而在生物医学领域备受关注。细胞片的构建主要是基于敏感性材料所构建的培养基底,通过改变温度、酶、光、离子、氧化还原pH、糖等刺激因素,调节基底对细胞的粘附行为使细胞发生自然脱附,从而获取细胞片。近年来,随着研究的深入进行,特别是各种新型的敏感性培养基底的不断发展,各种简单高效的细胞片构建技术不断涌现,得到的各种具有优良性能的细胞片极大地扩展了其应用的广度和深度。文中对组织工程细胞片的各种构建方法进行了阐述,对其存在的问题及发展前景进行了分析和展望。