线粒体ATP合酶基因组成及生化机制研究进展

线粒体ATP合酶是能量合成过程中的一个关键酶。该酶由水溶性的F1和跨膜的脂溶性F02个亚基组成,其中F1亚基由核基因编码,F0亚基由线粒体ATP6和ATP8基因以及核基因共同编码。该酶利用线粒体内膜两侧质子梯差形成的势能,通过结合改变机理旋转合成ATP,达到了很高的能量转化效率。因此,研究其基因组成及作用机制对分析和改良与动物能量代谢密切相关的重要经济性状及其基因效应具有重要价值。本文对线粒体ATP合成酶的基因组成及其作用机制进行了阐述。     

4种菊酯类杀虫剂对亚洲小车蝗的毒力和对ATPase活性的抑制

化学防治是草原蝗虫防治的主要应急措施,亚洲小车蝗是内蒙古草原的优势种。采用点滴法测定了4种菊酯类杀虫剂联苯菊酯、高效氯氰菊酯、高效氯氟氰菊酯和甲氰菊酯对亚洲小车蝗3龄蝗蝻的毒力。结果表明,联苯菊酯对亚洲小车蝗3龄蝗蝻的毒力最强,LD50值为0.004 8 μg/头,高效氯氟氰菊酯、高效氯氰菊酯和甲氰菊酯的LD50值分别为：0.022 7、0.012 2和0.005 1 μg/头。同时明确了4种菊酯类杀虫剂对亚洲小车蝗3龄蝗蝻离体ATPase活性都有一定程度的抑制作用。当药剂浓度为1×10-4mol/L时,高效氯氟氰菊酯、高效氯氰菊酯、联苯菊酯和甲氰菊酯对Na+-K+-ATPase的抑制分别为：57.62%、55.34%、45.70%和20.94%,对Ca2+-Mg2+-ATPase的抑制分别为：58.30%、58.05%、41.80%和19.35%。在浓度范围为1×10-4~1×10-8 mol/L,4种杀虫剂对亚洲小车蝗ATPase活性的抑制存在明显的剂量效应关系。     

烟草ATP合酶F_0部分4个亚基基因转录本编辑位点分析

RNA编辑是高等植物线粒体基因转录后表达调控的一种重要方式。为探究ATP合酶F_0部分的4个亚基基因与植物雄性不育性的关系,本研究以3个烟草雄性不育系(MS中烟90、MS云烟85和MS K326)及其同型保持系为供试材料,比较分析atp6、atp9、orf25和orf B线粒体基因转录本的编辑位点。结果表明,orf25和orf B基因转录本在不育系和保持系中发生的编辑位点是一致的。atp6基因在不育系中未发生编辑,在保持系中共有6处发生了RNA编辑。与保持系相比,atp9基因在不育系中除8处共同的C→T编辑外,还缺少2个C→T的特异编辑位点,其中1个导致氨基酸类型的改变。推测不育胞质因缺少特异的线粒体基因转录本编辑而导致烟草的细胞质雄性不育。     

Decline in leaf growth under salt stress is due to an inhibition of H+‐pumping activity and increase in apoplastic pH of maize leaves

In this study, salt‐induced changes in the growth rate of maize (Zea mays L.) were investigated during the first phase of salt stress. Leaf growth was reduced in the presence of 100 mM NaCl, and effects were more pronounced for the salt‐sensitive cv. Pioneer 3906 in comparison to the hybrid SR03. While hydrolytic activity of plasma membrane remained unaffected, H⁺‐pumping activity was reduced by 47% in Pioneer 3906, but was unchanged in SR03. Changes in apoplastic pH were detected by ratiometric fluorescence microscopy using the fluorescent dye fluorescein isothiocyanate‐dextran (50 mM). Pioneer 3906 responded with an increase of 0.2 pH units in contrast to SR03 for which no apoplastic alkalization was found. With respect to the hypothesis that the apoplastic pH is influenced by salinity, it is suggested that salt resistance is partly achieved due to efficient H⁺‐ATPase proton pumping, which results in cell‐wall acidification and loosening.     

Effects of Cold Acclimation on Several Enzyme Activities in Euonymus radicans ‘Emorald & Gold’and Its Relation to Semi-lethal Temperature

The changes in activities of superoxide dismutase (SOD), peroxidase (POD) and ATPase in the leaves of Euonymus radicans were studied when seedlings were cold-acclimated (at 4 ℃) for 1 week, 2 weeks, 3 weeks and then treated for 1 d under low temperature stress (at 5 ℃). The semi-lethal temperatures of acclimated and unacclimated seedlings were also investigated. The results indicated that the activities of the three enzymes in the leaves of the seedlings treated at 4 ℃ for 1, 2 and 3 weeks were all higher than those of unacclimated seedings (treated at 22 ℃ as controls). The activities of SOD and POD increased continuously with the prolongation of the time of cold acclimation, but stepped up to summits then down to the levels of the controls. The activities of SOD culminated at the first week, and the activities of POD at the second week. When acclimated and unacclimated seedlings were both treated at 5 ℃ for 1 d, the activities of the three enzymes in the leaves of acclimated seedlings were a little lower than those before stress, but higher than those of the controls. Moreover, the decrease rate of enzyme activities was greatly lower than that of the controls. The results showed that cold acclimation could enhance the stability of the three enzymes in the leaves of seedlings under low temperature stress; the semi-lethal temperature was 19.1 ℃ when the seedlings were treated at 4 ℃ for 3 weeks, but it was 5.4 ℃ when the seedlings were treated at 22 ℃. The decline of the semi-lethal temperature caused by the adaptive changes of enzyme activities was one of the foundations of enhancing the cold tolerance.     

Dietary potassium requirement of fingerling Indian major carp,Labeo rohita (Hamilton), based on growth parameters,gill Na+‐K+ ATPase activity,tissue mineralization and antioxidant activities

To quantify the dietary potassium requirement of fingerling Labeo rohita (6.2 ± 0.12 cm; 1.98 ± 0.06 g), seven purified experimental diets (350 g/kg crude protein and 16.72 kJ/g gross energy) with graded levels of potassium (0.32, 1.35, 2.41, 3.46, 6.48, 9.47 and 12.39 g/kg diet) were fed to triplicate groups of fishes at 08:00, 12:00 and 16:00 hr to apparent satiation for 8 weeks. Live weight gain (LWG; 671.46%), specific growth rate (3.65%/day), protein efficiency ratio (2.16), protein gain (PG; 2.41 g/fish) and feed conversion ratio (1.32) were found to be best in fish fed diet containing 3.46 g/kg potassium. Gill Na⁺‐K⁺ ATPase activity was also highest in fish fed diet with 3.46 g/kg potassium. Potassium content of whole‐body, vertebrae and scales increased significantly with the increase in dietary potassium level up to 6.48 g/kg. Significant changes were also noted in serum malondialdehyde content, superoxide dismutase, catalase, glutathione peroxidase and alkaline phosphatase activity. Based on the maximum live weight and protein gain observed in the present study, the inclusion of 3.55 g/kg potassium is recommended for developing potassium‐balanced commercial feeds for intensive culture of fingerling L. rohita.     

Changes in net proton release by roots of intact maize plants after local nutrient supply in a split root system

The effect of local nutrient supply to maize roots (Zea mays L. cv. Blizzard) on net proton release was studied using the split root technique (SRNS, SRCa) to compare plants that were cultivated with their roots completely in either nutrient solution (NS) or 0.1 mM CaSO₄ (Ca). Roots in NS released more protons than roots in Ca. This higher net proton release was associated with significantly higher ATP concentrations in the root tissue. Higher net proton release and ATP concentrations were also observed after a 4 h lag phase when 20 μM abscisic acid were exogenously applied to roots in 0.1 mM CaSO₄. It is suggested that higher metabolic activity in roots supplied with nutrients increased ATP concentrations and thus the substrate supply of the plasma membrane H⁺ ATPase. When only half of the root system was supplied with nutrient solution with the other half bathed in 0.1 mM CaSO₄, the roots in the SRNS compartment released significantly higher amounts of protons relative to the NS control plants. Conversely, roots in the SRCa compartment showed net proton uptake in contrast to the roots of control plants in 0.1 mM CaSO₄ which significantly acidified the root medium. These differences in proton release by roots in the split root system and control roots could not be explained in terms of differences in ATP concentrations. It is therefore suggested that an internal signal may lead to a modification of the plasma membrane H⁺ ATPase as shown earlier during plant adaptation to low pH in the root medium.     

根际pH对水稻细胞膜质子泵基因表达的影响

 通过水培试验研究了不同根际pH对水稻(日本晴)细胞膜质子泵基因表达的影响。以栽培在pH 5.5的营养液中的水稻作为对照,然后分别转移到pH 3.5和pH 7.5的营养液中培养。研究发现,质子泵基因OsPMA1、 OsPMA2、OsPMA3、OsPMA7在根系和叶片中均有表达;当营养液pH为3.5或7.5时,上述质子泵基因的表达总体上均比对照（pH 55）降低。但植物地上部的质子泵表达不受根际pH的影响。而且,pH越低,水稻生物量与体内的氮含量越低。其可能的原因是,水稻在pH骤然降为3.5的营养液中没有及时适应高浓度的H+胁迫,导致根系细胞膜质子泵受抑制,进而影响水稻对养分的吸收。而pH 7.5与植物细胞质pH相当,质子泵无需过量表达即可用于养分吸收。说明根际pH直接影响到根系细胞膜质子泵在转录水平上的表达,并间接影响根系对养分的吸收和植物的生长。     

长牡蛎闭壳肌肌纤维的组织学特性

分别利用石蜡切片苏木精-伊红染色法、冷冻切片烟酰胺腺嘌呤二核苷酸四唑氧化还原酶(NADH-TR)组化染色法、肌球蛋白ATP酶染色法对长牡蛎闭壳肌平滑肌和横纹肌进行了组织学特性研究,并比较平滑肌和横纹肌肌纤维组成的差异。石蜡切片结果显示,闭壳肌中平滑肌和横纹肌肌纤维形态结构不同,肌纤维直径、肌纤维密度和横截面积存在显著差异,其中平滑肌肌纤维直径为(5.97±0.631)μm,肌纤维密度为(25 485.65±3917.807)根/mm~2,肌纤维横截面积为(26.25±5.365)μm~2;横纹肌肌纤维直径为(7.41±0.847)μm,肌纤维密度为(16 908.25±3917.807)根/mm~2,横截面积为(40.45±10.083)μm~2。NADH-TR染色结果显示,平滑肌中富含NADH,肌纤维染色较深,呈现深蓝紫色,主要是I型肌纤维,为氧化代谢的慢速收缩型肌纤维;横纹肌染色较浅或者基本不着色,主要是II型肌纤维,为糖酵解代谢的快速收缩肌纤维。ATPase酶染色结果显示,横纹肌ATPase活性强,染色较深,呈现黑色,主要是II型肌纤维;平滑肌中ATPase活性弱,着色浅或基本不着色,主要是I型肌纤维。ATPase酶染色结果与NADH-TR染色结果一致。研究阐明了长牡蛎闭壳肌中平滑肌和横纹肌肌纤维的组织学特性及收缩、代谢特性差异,为深入研究长牡蛎肌纤维生物学特性提供了重要的基础资料,为提高我国贝类肉质品质提供理论依据。     

间甲酚对瓯江彩鲤早期发育总ATP酶活性的影响

在水温(20±2)℃条件下,设置7 mg/L、13 mg/L、19 mg/L的间甲酚(m-cresol)浓度梯度,研究了瓯江彩鲤(Cyprinus carpio var.color)早期发育过程中总ATP酶活性变化及间甲酚对总ATP酶影响。结果表明,瓯江彩鲤总ATP酶活性在成熟卵和受精卵中无显著差异(P0.05);各浓度组总ATP酶活性表现为良好的剂量-效应和发育时期-效应关系;总体上看,瓯江彩鲤早期发育从原肠胚期开始,总ATP酶活性随着发育时间的延长和暴露浓度的增加而降低。