不同贮藏年限老芒麦种胚线粒体抗氧化功能的变化规律

种子在自然贮藏过程中常常伴随着内部生理机能的恶化，线粒体作为种子内活性氧（reactive oxide species，ROS）产生的主要位点是最先遭到破坏的细胞器。为探讨不同贮藏年限对老芒麦种胚线粒体抗氧化功能的影响，本试验以室温贮藏0～4年的老芒麦种子为材料，分析比较其老化规律及种胚线粒体抗氧化特性的变化规律。结果表明：随着贮藏年限的延长，老芒麦种子发芽势、发芽率和种苗鲜重逐渐下降，死种子逐渐增多，种胚线粒体苹果酸脱氢酶（mitochondria malate dehydrogenase，MDH）、谷胱甘肽还原酶（glutathione reductase，GR）、单脱氢抗坏血酸还原酶（monodehydroascorbate reductase，MDHAR）活性逐渐下降，但在死种子中超氧化物歧化酶（superoxide dismutase，SOD）活性显著升高。此外，在贮藏过程中老芒麦种胚线粒体O₂^·-产生速率不断上升，而H₂O₂含量则呈现逐渐降低的趋势，表明线粒体中O₂^·-的积累与细胞氧化损伤密切相关。     

Effect of AIR on Inflammatory Reaction Infected by Brucella

In order to investigate the effect of AIR on inflammatory reaction infected by Brucellamelitensis (16M), the AIR domain of Tecpr1 gene of murine macrophages RAW264.7 were knocked down (I-A), overexpressed (O-A) and reversed (OA-IA). Using the chlorine fluorescein (DCFH-DA) as a probe, we detected the variation of ROS production and mitochondria distribution by confocal laser scanning microscopy. We observed the expression changes of NLRP3, ASC and Caspase-1 by qRT-PCR and the expression changes of IL-18,IL-1β and Caspase-1 in host cells by ELISA. The results showed that 16M could stimulate RAW264.7 cells to produce ROS by time-dependent pathway, and I-A group and O-A group showed more abnormal accumulation of mitochondrial. The results of qRT-PCR and ELISA suggested that it had effect on the expression levels of NLRP3, ASC,Caspase-1 and IL-18, IL-1β and Caspase-1 in cells of different groups. Those results indicated that with AIR gene deletion, the release amount of ROS changed, mitochondrial clustered abnormally, and AIR was closely related to the activation of inflammasomes and induction of inflammatory reactions.     

Betaine Alleviates Heat Stress-Induced Hepatic and Mitochondrial Oxidative Damage in Broilers

The aim of this study was to evaluate the effects of dietary betaine (BET) on growth performance, redox state, and related gene expression in broilers under heat stress (HS). A total of 144 21-day-old male broiler chickens with similar body weights were assigned randomly to three treatments with six replicates (eight chickens per replicate cage). Broilers in the control (CON) group were kept at thermoneutral (TN, 22±1°C) conditions and fed a basal diet until they were 42 days of age. Broilers in the other two groups (defined as HS and HS + BET) were exposed to HS (34±1°C, 8 h/day) and fed the basal diet without or with 1000 mg/kg BET, respectively. Rectal and cockscomb temperature of broilers was increased (P<0.05) in HS and HS + BET groups compared with the CON group, whereas there was no difference between HS and HS + BET groups. Dietary BET supplementation restored (P<0.05) average daily gain (ADG) and average daily feed intake (ADFI) of broilers and reversed (P<0.05) the increase in serum alanine transaminase (ALT) activity and malondialdehyde (MDA) content in the liver tissue of broilers under HS. The HS + BET group had higher (P<0.05) activities of superoxide dismutase (SOD) and glutathione peroxidase (GPX) in the liver tissue and mitochondria than the HS group, and the same pattern was observed for glutathione (GSH) and GSH/glutathione disulphide (GSSG) in the liver tissue. The decreased mRNA levels of GPX1 and uncoupling protein (UCP) in the liver induced by HS were restored by BET supplementation. In conclusion, dietary BET supplementation can alleviate HS-induced hepatic and mitochondrial oxidative damage of broilers by regulating mRNA expressions of GPX1 and UCP.     

紫花苜蓿种子吸胀期胚根线粒体AsA-GSH循环对低温胁迫的响应

为探究胚根线粒体抗坏血酸-谷胱甘肽(AsA-GSH)循环响应低温胁迫的抗氧化作用机制,以紫花苜蓿种子为材料,研究了不同温度(10和20℃)处理下发芽特性、不同吸胀时间(6、12和24 h)胚根线粒体AsA-GSH循环酶活性、抗氧化物以及过氧化氢(H₂O₂)含量的变化规律。结果表明,吸胀12和24 h后,10℃处理的胚根线粒体H₂O₂含量高于20℃。吸胀24 h期间,10℃处理的胚根线粒体谷胱甘肽还原酶(GR)活性均低于20℃。吸胀12和24 h后,10℃处理的胚根线粒体抗坏血酸(AsA)含量均低于20℃。10℃条件下吸胀24 h后,与20℃处理相比,胚根线粒体谷胱甘肽(GSH)含量显著降低(P<0.05)。苜蓿种子在10℃条件下吸胀萌发时,主要通过降低AsA-GSH循环中GR、单脱氢抗坏血酸还原酶(MDHAR)、过氧化物酶(POD)活性和AsA、GSH含量,使胚根线粒体内的H₂O₂积累,产生氧化损伤,继而影响种子萌发的正常进程。     

葡萄果实转色过程中线粒体呼吸代谢变化

为明确线粒体呼吸代谢与果实成熟期间颜色形成的关系,本试验探究了3种葡萄果实(夏黑、巨峰和金手指)成熟期间颜色、总花色苷含量、呼吸强度、内源乙烯产生和线粒体呼吸代谢的变化规律。结果表明,夏黑和巨峰葡萄果实的果皮颜色指数(CIRG)值和总花色苷含量随着果实成熟呈上升趋势,而金手指葡萄果实CIRG值在整个成熟期间变化不显著;葡萄果实成熟期间呼吸强度和内源乙烯均呈先上升后下降趋势,在花后63d达到峰值。同时,葡萄果实成熟期间,线粒体膜通透性转换孔(MPTP)逐渐增加,线粒体膜流动性(MMF)持续下降;线粒体琥珀酸脱氢酶(SDH)活性随着成熟进程逐渐下降,而细胞色素氧化酶(CCO)活性在花后63d开始上升。表明盛花63d之后是果实色泽快速形成阶段,可能为葡萄果实色泽形成的关键时期。线粒体呼吸代谢增强促进了果实成熟期间呼吸强度增加和内源乙烯释放,从而使夏黑和巨峰果实中花色苷大量积累,加快了金手指葡萄果实叶绿素的降解和类胡萝素的合成。本研究结果为调控南方葡萄果实着色提供了理论依据。     

氮对低氧胁迫下樱桃根系抗氧化酶活性及线粒体功能的影响

 以‘吉塞拉5号’樱桃组培苗为试材, 研究短期低氧胁迫下氮对樱桃幼苗根系抗氧化酶活性 及线粒体功能的影响。通过溶氧调节仪人为控制营养液溶氧浓度为2 mg·L ^{- 1} , NO₃^- 浓度共设4个水平:0、7.5、15.0、22.5 mmol·L ^{- 1} , 以1 /2 Hoagland营养液正常低氧处理(NO₃^- 浓度7.5 mmol·L ^{- 1} ) 为对照。结果表明: 各处理SOD、POD、CAT酶活性均呈现先升高后降低的趋势。与对照相比, 营养液加氮处理的SOD、POD、CAT酶活性升高, 膜质过氧化程度减轻, 且氮浓度越大酶活性升高幅度越大, 而营养液缺氮处理的酶活性降低, 膜质过氧化程度加重; 营养液缺氮处理樱桃根系线粒体中的H₂O₂和MDA含量、O₂^-·生成速率升高, 而营养液加氮处理可降低O₂^-·的产生及MDA含量, 提高线粒体的氧化磷酸化水平, 保护樱桃根系线粒体膜的完整性, 对线粒体呼吸功能的破坏明显减轻, 且NO₃^- 离子浓度高时效果更好, 说明适当提高营养液中的氮素水平可以缓解低氧胁迫对樱桃根系的伤害。     

Inheritance pattern of chloroplast and mitochondrial genomes in artificial hybrids of Porphyra yezoensis (Rhodophyta)

ABSTRACT:   To investigate inheritance of chloroplasts and mitochondria (organelles) in the marine red alga Porphyra yezoensis , cleaved amplified polymorphic sequence (CAPS) profiles of the two strains, TU-2 and KGJ, as parental gametophytes and 44 hybrid sporophytes (i.e. female of TU-2 and male of KGJ) were examined. A carbamyl phosphate synthase small subunit gene ( carA ) and ribosomal protein S11 gene ( rps11 )-small subunit ribosomal RNA gene ( rns ) spacer region were used as molecular markers for the chloroplast and mitochondrial DNAs, respectively. Thirty-eight of 44 (86.4%) conchocelis colonies showed maternal banding patterns, five (11.4%) colonies showed biparental bands and one (2.2%) colony showed paternal bands in CAPS analyses both with chloroplast and mitochondrial markers. These results suggest that organelles of P. yezoensis are inherited uniparentally from the female parent.     

四种菊酯类杀虫剂对枸杞蚜虫线粒体膜流动性的影响

为明确菊酯类杀虫剂对昆虫线粒体膜流动性的影响,以1,6-二苯基-1,3,5-己三烯（1,6-diphenyl-1,3,5-hexatriene,DPH）为荧光探剂,采用荧光偏振法研究了4种菊酯类杀虫剂对枸杞蚜虫Aphis sp.线粒体膜流动性的影响。结果表明,4种菊酯类杀虫剂对枸杞蚜虫线粒体膜流动性均有显著影响,其荧光偏振度显著增大,线粒体膜流动性显著减小。当药剂浓度为100 μmol/L时,对照组枸杞蚜虫线粒体膜的荧光偏振度为0.239,高效氯氰菊酯的影响最大,线粒体膜的荧光偏振度为0.315,联苯菊酯的影响最小,线粒体膜的荧光偏振度为0.287。4种菊酯类杀虫剂对线粒体膜流动性的影响都存在一定的浓度效应,当药剂浓度在0~100 μmol/L范围内,浓度越大影响越大。4种菊酯类杀虫剂在不同温度下对枸杞蚜虫线粒体膜流动性的影响不同,高效氯氰菊酯、联苯菊酯和甲氰菊酯对线粒体膜流动性的影响基本不受温度影响,而氯菊酯在高温条件下产生的影响大,在低温条件下产生的影响小,表现为明显的正温度系数。表明DPH可以作为荧光探剂研究枸杞蚜虫线粒体膜的流动性。     

两个人工选择奥利亚罗非鱼群体系统发育 及其遗传多样性分析

[目的]了解人工选择偏好对奥利亚罗非鱼遗传多样性的影响,为今后人工选育及生产提供参考依据.[方法]对两个奥利亚罗非鱼群体(那马群体和武鸣群体)各70尾个体的线粒体DNA的D-loop区序列、CoI基因和Cytb基因进行测序及系统进化研究,并选用20对微卫星标记对其遗传多样性进行分析.[结果]基于D-loop区序列两个群体共检测出32个单倍型,其中共享单倍型6个;基于CoI基因共检测出23个单倍型,其中共享单倍型16个;基于Cytb基因共检测出14个单倍型,其中共享单倍型10个.基于D-loop区序列、CoI基因和Cytb基因单倍型分别构建的系统发育进化树显示,武鸣群体和那马群体的个体交错在一起,地理差异不明显,且采用NJ法和ME法构建系统发育进化树的进化拓扑结构基本相似.20对微卫星引物均能在奥利亚罗非鱼中获得稳定有效的扩增条带,其中有18个微卫星位点呈多态性;那马、武鸣群体的平均等位基因数(Na)分别为6.5000和7.9444,平均有效等位基因数(Ne)分别是3.9857和4.7268,平均观测杂合度(Ho)分别为0.7123和0.7752,平均期望杂合度(He)分别为0.9614和0.9711、平均Nei期望杂合度分别为0.7017和0.7636,均表现为武鸣群体略高于那马群体.两个群体的遗传分化系数(Fst)在不同微卫星位点间差异明显,其变化范围为0.0173(GM241)~0.2318(UNH868),平均0.0997;从单群体近交系数(Fis)和总群体近交系数(Fit)来看,所有微卫星位点的数值均为负值.[结论]经短期人工选择的武鸣群体奥利亚罗非鱼遗传多样性较被长期人工选择的那马群体遗传多样性丰富,即短期内的不同人工选择偏好对线粒体DNA的遗传影响较小.     

Mitochondrial NAD(P)-dependent malic enzyme from herring testicular tissue: Purification,kinetic behaviour and regulatory properties

Mitochondrial NAD(P)-dependent malic enzyme [EC 1.1.1.39, L-malate: NAD⁺ oxidoreductase (decarboxylating)] was purified from herring testicular tissue to a specific activity of 26.4 μmol NADH/min/mg protein. Herring testicular tissue is one of the most abundant sources of this enzyme. The purification procedure involved differential centrifugation of mitochondria and then chromatography on DEAE-Sephacel, Red Agarose and Sephacryl S-300. This enzyme catalyzes the oxidative decarboxylation of malate in the presence of Mn²⁺ and either NAD or NADP. Under V_max conditions the ratios for the rate of NAD/NADP reduction was 1.8. A study of the reductive carboxulation reaction indicated that this enzyme reaction is reversible; at pH 7.0 the reverse reaction exhibited 22% of the activity of forward reaction. Some kinetic characteristics of the enzyme were determined. ATP was found to be a competitive inhibitor with respect to malate. Fumarate reversed ATP inhibition. Regulation of NAD(P)-dependent malic enzyme from herring testicular tissue mitochondria could respond to changing levels of mitochondrial ATP and fumarate in vivo.