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.     

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。     

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.     

褪黑素对内毒素血症山羊肝细胞线粒体呼吸功能的影响

将48只健康山羊随机分为4组：对照组（A组）、内毒素（LPS）组（B组）、内毒素＋褪黑素（LPS＋MT）组（C组）、褪黑素（MT）组（D组）,在处理后第3 h和6 h提取肝细胞线粒体,采用Clark氧电极技术测定线粒体呼吸控制率（RCR）、磷氧比值（P/O）、氧化磷酸化效率（OPR）的变化,探讨了MT对LPS诱导的山羊内毒素血症机体线粒体呼吸功能的影响。结果显示,内毒素组山羊在第3 h和6 h的RCR、P/O、OPR显著低于对照组（P〈0.05）;应用褪黑素后,即内毒素＋褪黑素组第3 h的肝细胞线粒体RCR、P/O、OPR明显升高（P〈0.05）;褪黑素组第3 h的RCR明显高于对照组（P〈0.05）,而褪黑素组第3 h的P/O、OPR与对照组差异不显著（P〉0.05）。证实,山羊内毒素血症时线粒体的呼吸功能明显下降,褪黑素对肝细胞线粒体呼吸功能具有一定的保护作用。     

短高温对小麦花器官及旗叶的超微结构变化的直接影响

在春小麦花粉母细胞期和单核期进行短期高温杀雄处理后，用透射电子显微镜观察小麦花器官和旗叶的超微结构。结果表明：短期高温直接使小孢子和绒毡层细胞中线粒体等双层细胞器严重破坏，并且使绒毛层细胞膨胀与小孢子粘连。而胚囊的线粒体和旗叶片中的 纯粒体及叶绿体结构变化不明显。     

Mitochondrial NAD(P)-malic enzyme from herring skeletal muscle

Mitochondrial NAD(P)-dependent malic enzyme [EC 1.1.1.39, L-malate: NAD⁺ oxidoreductase (decarboxylating)] was purified from herring skeletal muscle to a specific activity of 8.2 mol/min/mg. The purification procedure involved chromatography on DEAE-cellulose, Red Agarose and a Sephacryl S-300 with a final recovery of 38% of enzyme activity. This enzyme catalyzes the oxidative decarboxylation of malate in the presence of either NAD or NADP in the presence of Mn²⁺. Some kinetic characteristics of this enzyme were determined. The pH optimum of activity is 7.0. ATP was shown to be a competitive inhibitor with malate. The inhibition by ATP displayed hyperbolic competitive kinetics with a K_i (ATP) of 0.28 mM in the presence of NAD and 0.75 mM in the presence of NADP. Fumarate reversed ATP inhibition.In vivo, regulation of NAD(P)-dependent malic enzyme might respond to changing levels of mitochondrial ATP and fumarate with the enzyme undergoing kinetic activation by an increase in the concentration of mitochondrial fumarate which could reverse enzyme inhibition by ATP.     

Programmed Cell Death Induced by (?)-8,9-Dehydroneopeltolide in Human Promyelocytic Leukemia HL-60 Cells under Energy Stress Conditions

(+)-Neopeltolide is a marine macrolide natural product that exhibits potent antiproliferative activity against several human cancer cell lines. Previous study has established that this natural product primarily targets the complex III of the mitochondrial electron transport chain. However, the biochemical mode-of-actions of neopeltolide have not been investigated in detail. Here we report that (−)-8,9-dehydroneopeltolide (8,9-DNP), a more accessible synthetic analogue, shows potent cytotoxicity against human promyelocytic leukemia HL-60 cells preferentially under energy stress conditions. Nuclear morphology analysis, as well as DNA ladder assay, indicated that 8,9-DNP induced significant nuclear condensation/fragmentation and DNA fragmentation, and these events could be suppressed by preincubating the cells with a pan-caspase inhibitor, N-benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (zVAD). Immunoblot analysis demonstrated the release of cytochrome c from the mitochondria and the cleavage of full-length caspase-3 and poly(ADP-ribose) polymerase (PARP). These results indicated that 8,9-DNP induced caspase-dependent apoptotic programmed cell death under energy stress conditions. It was also found that 8,9-DNP induced non-apoptotic cell death in the presence/absence of zVAD under energy stress conditions. Immunoblot analysis showed the intracytosolic release of apoptosis-inducing factor (AIF), although it did not further translocate to the nucleus. It appears most likely that, in the presence of zVAD, 8,9-DNP triggered necrotic cell death as a result of severe intracellular ATP depletion.     

线粒体分子标记技术在水产养殖行业的应用

分子标记技术的使用在分子层面上对水产动物进行深度剖析,推动了水产行业的发展和创新.对线粒体分子标记技术的种类、构造和它们在水产行业中各个方面的应用方式进行了综述,探讨了线粒体分子标记技术在起源演化、亲缘和亲权关系、遗传距离和遗传多样性等方面的研究,并比较了不同线粒体分子标记技术的区别,讨论了该分子标记技术在未来水产行业中的发展前景.     

采用Extracellular Flux Analyzer方法测定植物线粒体能量代谢

线粒体是植物进行呼吸和能量代谢的主要场所,但目前关于线粒体能量代谢的测定方法较为繁琐,亟需一种快速有效测定线粒体能量代谢的手段。本研究采用Extracellular Flux Analyzer(Seahorse Bioscience,America)方法对玉兰(Magnolia denudata)花部器官线粒体代谢情况进行测定,并对测试技术进行改进。通过直接测定氧气消耗速率,进而检测组织细胞内的线粒体呼吸速率和能量代谢。采用该方法对同一组织样品进行多次测量,测量结果稳定,表明该方法是一种可靠的测定线粒体呼吸和能量代谢的方法。并且,通过加入不同的呼吸抑制剂来鉴定细胞的呼吸途径,可快速获取有效数据。此外,还可直接对组织内线粒体的呼吸代谢进行测定,简便性、实时性均优于其他线粒体代谢测定方法,是一种快速有效,且可以进行实时动态观测的试验方法。     

热胁迫对大豆花荚离层细胞HSP70基因表达、能量供应及花荚脱落率的影响

实验运用Northern blot检测和生理生化分析等技术研究热胁迫对大豆(Glycine max)品种花荚离层细胞的HSP70基因表达、能量供应及花荚脱落的影响。结果显示，热胁迫能够提高所有6个供试大豆品种花荚离层细胞的HSP70基因表达水平、线粒体及其蛋白含量以及细胞色素氧化酶表达量，而不同程度地降低了所有供试品种的花荚脱落率。结果说明，大豆花荚离层细胞HSP70基因的高效表达可能是通过有效地改善花荚离层细胞的能量供应，从而达到抑制花荚脱落的目的。关键词: 大豆；热胁迫；HSP70基因；线粒体；细胞色素氧化酶；花荚脱落率