小麦突变体返白系返白阶段叶绿素代谢的变化

小麦 ( Triticum aestivum)返白系在返白阶段 Chl、胡萝卜素 ( Caro)含量均下降 ,但 Caro/ Chl的比值大于对照 ,表明叶片白化不是因 Caro减少引起的。Chl下降的同时 ,Chla和 Chlb均下降 ,表明该突变体属阶段性缺总 Chl型。返白初期 Chlase活性增高 ,返白中期活性下降 ,表明 Chl降解不是造成叶片失绿的主要原因 ;Chl合成中间物 δ-氨基酮戊酸 ( ALA)、胆色素原 ( PBG)积累 ,尿卟啉 ( Uro )、原卟啉 ( Proto )、镁 -卟啉 ( Mg- Proto )、原叶绿素酸 Pchl减少 ,特别是 Uro 在返白中期含量最低 ,复绿初期却急剧积累 ,表明叶绿素合成受阻于尿卟啉原 ( Urogen )的形成上。     

油菜叶绿素b减少突变体Cr3529叶绿素生物合成的研究

利用吸收光谱和荧光光谱法测定了油菜叶绿素b减少突变体Cr3529子叶叶绿素生物合成途径中几种主要前体物质的含量.结果显示:突变体子叶中叶绿素生物合成第一个限速步骤的前体物质δ-氨基乙酰丙酸(ALA)含量与野生型油菜大致相同,饲喂ALA后的突变体及野生型油菜子叶中ALA含量均显著增加,但二者无显著差异;胆色素原含量在突变体中也未降低,而尿卟啉原Ⅲ含量仅为野生型的一半,粪卟啉原Ⅲ、原卟啉Ⅸ、镁原卟啉Ⅸ和原植基叶绿素的含量都明显低于野生型.结果证明,Cr3529突变体中叶绿素生物合成受阻于由胆色素原形成尿卟啉原Ⅲ的步骤,其叶绿素合成缺陷的机制和前体物质的累积与其它叶绿素b减少突变体明显不同.     

海水胁迫对菠菜叶绿素代谢的影响

以耐海水品种‘荷兰3号’和海水敏感品种‘圆叶菠菜’为试验材料,采用营养液栽培,研究海水胁迫对菠菜叶绿素代谢的影响。结果表明,海水胁迫下,2个菠菜品种叶片的叶绿素a(Chl a)、叶绿素b(Chl b)和总叶绿素含量以及叶绿素合成前体———原叶绿素酸(Pchl)、镁原卟啉IX(Mg-proto IX)、原卟啉IX(Proto IX)和尿卟啉原III(UroIII)含量均明显降低,而胆色素原(PBG)和δ-氨基酮戊酸(ALA)积累,‘圆叶菠菜’的变化幅度大于‘荷兰3号’。海水胁迫下,‘荷兰3号’叶片的叶绿素酶(Chlase)活性无显著变化,胆色素原脱氨酶(PBGD)和尿卟啉原III合酶(UROS)活性在胁迫第3天显著下降,而‘圆叶菠菜’Chlase活性显著上升,PBGD和UROS活性显著下降。研究发现,在海水胁迫条件下,菠菜叶片的叶绿素合成代谢受阻,受阻位点位于PBG→UroIII的转化过程,其中‘圆叶菠菜’的受阻程度大于‘荷兰3号’;耐海水品种‘荷兰3号’叶片叶绿素含量降低主要由叶绿素合成代谢受阻引起,而海水敏感品种‘圆叶菠菜’叶绿素含量的降低则是由叶绿素合成受阻和叶绿素降解共同作用的结果。     

宽叶吊兰叶绿素生物合成的昼夜节律变化

在被子植物中,从谷氨酰-tRNA到叶绿素的生物合成是由许多酶催化的级联反应,其中间代谢产物具有较强的光反应活性和细胞毒性,因此这一过程在细胞内受到严格的调控。本研究通过检测宽叶吊兰叶片叶绿素生物合成途径的14种中间产物含量随昼夜节律的变化,探讨昼夜节律对宽叶吊兰叶绿素生物合成的影响。结果表明,中间产物ALA(δ-氨基乙酰丙酸)、PBG(胆色素原)、ProtoⅨ(原卟啉Ⅸ)、Heme(血红素)、Mg-ProtoⅨ(镁原卟啉Ⅸ)、Chlide a(叶绿素酸酯a)、Chlide b(叶绿素酸酯b)、Chl a(叶绿素a)、Chl b(叶绿素b)受光诱导,而UrogenⅢ(尿卟啉Ⅲ)、CoprogenⅢ(粪卟啉Ⅲ)和Pchlide(原叶绿素酸脂)受黑暗诱导,尤其是Pchlide在黑暗中的积累量显著增加;Mpe(镁原卟啉Ⅸ单甲酯)和Mpde(镁原卟啉Ⅸ二酯)具有2个积累峰值,分别出现在中午12∶00和夜间24∶00。说明叶绿素生物合成受昼夜节律的调控,但其中间代谢产物含量的变化规律与昼夜节律并不完全一致。     

叶面喷施亚精胺对盐碱胁迫下番茄幼苗生长及其叶绿素合成前体含量的影响

以盐碱敏感型番茄品种‘中杂9号’幼苗为试验材料,研究叶面喷施0.25mmol·L-1亚精胺(Spd)对75mmol·L-1盐碱溶液胁迫下番茄幼苗生长、净光合速率及叶绿素合成前体物质含量的影响,探讨Spd在缓解番茄盐碱胁迫伤害的生理机制。结果显示:(1)盐碱胁迫下番茄叶片叶绿素合成前体物质原卟啉Ⅸ(ProtoⅨ)、镁-原卟啉Ⅸ(Mg-protoⅨ)、原叶绿素酸(Pchl)含量均显著降低,而δ-氨基酮戊酸(ALA)、胆色素原(PBG)、尿卟啉原Ⅲ(UroⅢ)显著积累,UroⅢ到ProtoⅨ的转化受阻,引起叶片叶绿素a(Chl a)、叶绿素b(Chl b)和总叶绿素(Chl)含量显著降低,以及幼苗叶片净光合速率(Pn)、叶面积、叶片相对含水量、地上和地下部干鲜重的生长指标均显著降低。(2)盐碱胁迫下,叶面喷施Spd可显著促进番茄幼苗的生长,抑制叶片内ALA、PBG、UroⅢ的积累,并提高ProtoⅨ、Mg-protoⅨ、Pchl、Chl a、Chl b、Chl含量和相应Pn值。研究表明,盐碱胁迫显著抑制番茄幼苗的生长,叶面喷施Spd可通过缓解UroⅢ到ProtoⅨ的转化受阻程度,促进盐碱胁迫下番茄叶片的叶绿素合成,提高叶绿素含量和净光合速率,减轻盐碱胁迫对幼苗生长的伤害。     

油菜黄化突变体蛋白质组分析：两种蛋白质提取方法比较

以芥菜型油菜黄化突变体L638-y及其野生型L638-g五叶期叶片为材料,用IEF/SDS-PAGE双向凝胶电泳技术对两种不同蛋白质提取方法(TCA/丙酮沉淀法和改进的PEG分级沉淀法)进行了比较,同时在IPG胶条pH范围及SDS-PAGE胶浓度选择上进行了探索与优化．结果表明,以pH 4～7 17 cm的线性IPG胶条进行IEF,11% SDS-PAGE进行第二向电泳,每350 μl体系上样量为180 μg,蛋白质可以得到较好的分离,2-DE图谱质量最佳．用改进的PEG分级沉淀法提取的突变体L638-y叶片总蛋白的2-DE图谱可清晰识别的蛋白质点数目为(1235 ± 6)个,比TCA/丙酮沉淀法多识别出330个蛋白质点;用该方法提取蛋白质时,在突变体L638-y与其野生型L638-g叶片总蛋白2-DE图谱上可识别出差异蛋白质点数目为190个,比用TCA/丙酮沉淀法提取蛋白质时多鉴别出100个差异蛋白质点．由此表明,研究芥菜型油菜黄化突变体L638-y叶片蛋白质组变化,采用改进的PEG分级沉淀法提取蛋白质更为简单有效．     

SO_2胁迫对金色叶植物叶片叶绿素合成代谢的影响

以金叶女贞(Ligustrum×vicaryi)和金森女贞(Ligustrum japonicum‘Howardii’)2种金色叶女贞为试材,采用人工静态熏气的方法,研究SO2胁迫对金色叶植物叶片叶绿素含量及其合成代谢的影响,探讨SO2胁迫影响金色叶植物叶绿素合成的可能位点。结果表明:20和40 mg·m-3的SO2熏气提高了叶绿素、尿卟啉原Ⅲ(UrogenⅢ)、原卟啉Ⅸ(ProtoⅨ)、镁原卟啉Ⅸ(Mg-protoⅨ)的含量,使δ-氨基酮戊酸脱水酶(ALAD)和尿卟啉原Ⅲ合酶(UROS)活性升高,δ-氨基酮戊酸(ALA)和胆色素原(PBG)含量降低;80 mg·m-3的SO2熏气使叶绿素含量降低,引起ALA和PBG含量的积累,降低了UrogenⅢ、ProtoⅨ和Mg-protoⅨ的含量,抑制了ALAD和UROS的活性。研究发现,金叶女贞叶绿素合成关键前体物质含量和相关酶活性随熏气浓度的变化趋势较金森女贞明显,前者对SO2较为敏感,后者抗SO2的能力强于前者,SO2胁迫影响金色叶植物叶绿素合成的可能位点位于UrogenⅢ的合成过程。     

海水胁迫对菠菜（Spinacia olerancea L.）叶绿体活性氧和叶绿素代谢的影响

以海水敏感品种圆叶菠菜和耐海水品种‘荷兰3号'为试材,采用水培方法,研究了海水胁迫对菠菜(Spinacia olerancea L.)叶绿体活性氧(ROS)和叶绿素(Chl)代谢的影响.结果表明,海水胁迫下,2个菠菜品种叶绿体内超氧阴离子(O-·2)产生速率、过氧化氢(H2O2)和丙二醛(MDA)含量显著升高,并且圆叶菠菜的提高幅度大于‘荷兰3号';圆叶菠菜叶绿体内的超氧化物歧化酶(SOD)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)活性和抗坏血酸(AsA)、还原型谷胱甘肽(GSH)含量均低于‘荷兰3号',而过氧化物酶(POD)活性高于‘荷兰3号';光氧化剂甲基紫精(MV)进一步使2个菠菜品种叶绿体内O-·2产生速率加快、H2O2含量提高、膜质过氧化加重,活性氧清除剂AsA明显降低了菠菜叶绿体内ROS水平,缓解了由MV造成的严重的膜质过氧化伤害.海水胁迫下,2个菠菜品种叶片叶绿素b(Chlb)、叶绿素a(Chla)及其合成前体物质原叶绿素酸(Pchl)、镁原卟啉Ⅸ(Mg-protoIX)、原卟啉Ⅸ(ProtoⅨ)和尿卟啉原Ⅲ(UroⅢ)含量明显降低,而胆色素原(PBG)和δ-氨基酮戊酸(ALA)积累,Chl合成受到阻碍,并且圆叶菠菜的受阻程度大于‘荷兰3号';MV进一步加剧了这种受阻程度,而AsA部分缓解了由海水胁迫和MV造成的阻碍作用.海水胁迫明显提高了圆叶菠菜叶片叶绿素酶(Chlase)活性而对‘荷兰3号'没有影响,MV处理对圆叶菠菜Chlase活性的影响程度大于‘荷兰3号',但AsA对2个品种叶片Chlase活性没有明显影响.上述结果说明,海水胁迫下,菠菜叶绿体内ROS与Chl代谢密切相关,不仅通过叶绿体膜的氧化伤害使Chl降解,而且使Chl合成的PBG向UroⅢ转化步骤受阻.耐海水品种‘荷兰3号'叶绿体清除ROS主要通过SOD和AsA-GSH循环系统,清除能力较强,减轻了ROS对叶绿体膜的氧化损伤和Chl合成的受阻程度,并且海水胁迫对其Chlase活性的影响较小;而海水敏感品种圆叶菠菜叶绿体清除ROS主要依赖于SOD和POD,对ROS的清除能力有限,从而导致了ROS大量积累,叶绿体膜的氧化损伤和Chl合成的受阻程度严重,并且海水胁迫显著提高了Chlase活性,加剧了Chl降解.     

外源亚精胺对盐胁迫下菠菜叶绿素合成前体含量的影响

采用营养液水培的方法,以耐盐性较弱的菠菜(Spinacia oleracea L.)品种‘全能菠菜’为试材,研究外源亚精胺(Spd)对盐胁迫下菠菜生长及叶绿素前体含量的影响。结果显示:(1)菠菜植株在250mmol·L-1 NaCl胁迫下生长受到显著抑制,单株总鲜重、总干重、地上部鲜重、地上部干重、地下部鲜重和地下部干重分别较对照降低57.23%、53.08%、63.14%、55.05%、42.22%和42.86%,叶面喷施1.0mmol·L-1Spd使其分别比盐胁迫处理提高62.83%、71.19%、60.57%、71.74%、70.31%和69.23%;(2)250 mmol·L-1盐胁迫使菠菜叶片叶绿素a(Chl a)、叶绿素b(Chl b)及总叶绿素Chl(a+b)含量分别较对照显著降低42.31%、54.55%和44.53%,叶面喷施1.0mmol·L-1 Spd使其分别较单纯盐胁迫处理显著提高46.24%、51.85%和47.11%;(3)盐胁迫使菠菜叶片Chl a、Chl b、叶绿素酸(Pchl)、镁原卟啉Ⅸ(Mg-protoⅨ)、原卟啉Ⅸ(ProtoⅨ)和尿卟啉Ⅲ(UroⅢ)含量均显著降低,而使胆色素原(PBG)和δ-氨基酮戊酸(ALA)含量升高,而叶片Chl a、Chl b、Pchl、Mg-protoⅨ、ProtoⅨ和UroⅢ含量在施用外源Spd后均显著提高,而其PBG和ALA含量有所降低。可见,盐胁迫条件下,菠菜叶片叶绿素合成受阻,受阻位点在PBG向UroⅢ的转化过程,外源Spd能够缓解盐胁迫下菠菜叶片叶绿素合成受阻的程度,促进叶绿素合成,从而提高叶绿素含量。     

叶面喷施亚精胺对高温胁迫下番茄叶绿素合成代谢的影响

以设施番茄栽培品种‘金棚朝冠’幼苗为试验材料,研究叶面喷施0.25 mmol·L~(-1)亚精胺(Spd)对高温胁迫下(38℃/28℃,昼/夜)番茄幼苗生长及叶绿素合成过程中前体物质含量、关键酶活性和叶绿素含量的影响,探讨Spd缓解番茄高温胁迫伤害的生理机制。结果表明:(1)高温胁迫显著抑制了番茄幼苗的生长,叶面喷施Spd可有效缓解高温胁迫对番茄幼苗地上部生长的抑制作用,但对于地下部的生长无显著缓解作用。(2)高温胁迫下番茄叶片叶绿素合成前体物质δ-氨基酮戊酸(ALA)和胆色素原(PBG)的含量显著提高,而尿卟啉原Ⅲ(UroⅢ)、原卟啉Ⅸ(ProtoⅨ)、镁-原卟啉Ⅸ(Mg-protoⅨ)和原叶绿素酸(Pchl)的含量显著降低,证明叶绿素前体由PBG向UroⅢ的转化受阻,导致叶绿素a(Chla)、总叶绿素(Chlt)含量和Chla/Chlb显著降低。(3)叶面喷施Spd能增强高温胁迫下胆色素原脱氨酶(PBGD)活性,有效缓解了高温胁迫对PBG到UroⅢ转化的阻碍作用,促进PBG之后叶绿素合成前体物质的合成,Chla含量和Chla/Chlb显著增加。研究发现,高温胁迫显著抑制番茄幼苗的生长,叶面喷施Spd可通过显著增强PBGD活性来缓解由PBG向UroⅢ的受阻程度,促进高温胁迫下番茄叶片的叶绿素前体合成,提高叶绿素含量和番茄植株的生长量,减轻高温胁迫对番茄幼苗生长的伤害。     

Role for dopamine in malonate-induced damage in vivo in striatum and in vitro in mesencephalic cultures

Defects in mitochondrial energy metabolism have been implicated in the pathology of several neurodegenerative disorders. In addition, the reactive metabolites generated from the metabolism and oxidation of the neurotransmitter dopamine (DA) are thought to contribute to the damage to neurons of the basal ganglia. We have previously demonstrated that infusions of the metabolic inhibitor malonate into the striata of mice or rats produce degeneration of DA nerve terminals. In the present studies, we demonstrate that an intrastriatal infusion of malonate induces a substantial increase in DA efflux in awake, behaving mice as measured by in vivo microdialysis. Furthermore, pretreatment of mice with tetrabenazine (TBZ) or the TBZ analogue Ro 4-1284 (Ro-4), compounds that reversibly inhibit the vesicular storage of DA, attenuates the malonate-induced DA efflux as well as the damage to DA nerve terminals. Consistent with these findings, the damage to both DA and GABA neurons in mesencephalic cultures by malonate exposure was attenuated by pretreatment with TBZ or Ro-4. Treatment with these compounds did not affect the formation of free radicals or the inhibition of oxidative phosphorylation resulting from malonate exposure alone. Our data suggest that DA plays an important role in the neurotoxicity produced by malonate. These findings provide direct evidence that inhibition of succinate dehydrogenase causes an increase in extracellular DA levels and indicate that bioenergetic defects may contribute to the pathogenesis of chronic neurodegenerative diseases through a mechanism involving DA.     

Scurvy in capybaras bred in captivity in Argentine

In order to determine if the absence of vitamin C in the diet of capybaras (Hydrochoerus hydrochaeris) causes scurvy, a group of seven young individuals were fed food pellets without ascorbic acid, while another group of eight individuals received the same food with 1 g of ascorbic acid per animal per day. Animals in the first group developed signs of scurvy-like gingivitis, breaking of the incisors and death of one animal. Clinical signs appeared between 25 and 104 days from the beginning of the trial in all individuals. Growth rates of individuals deprived of vitamin C was considerably less than those observed in the control group. Deficiency of ascorbic acid had a severe effect on reproduction of another population of captive capybaras. We found that the decrease in ascorbic acid content in the diet affected pregnancy, especially during the first stages. The results obtained suggest that it is necessary to supply a suitable quantity of vitamin C in the diet of this species in captivity.     

Changes in lactate dehydrogenase activity in chicken tissues in hyperthyreosis in ontogenesis

The lactate dehydrogenase activity in reactions of lactate oxidation and synthesis was studied in subfractions of the chicken brain, heart and liver at the embryonal, early postembryonal and adult stages of development after thyroxine administration. It has been shown that during embryogenesis thyroxine predominantly enhanced the rate of lactate oxidation in the mitochondrial tissues. A marked increase in the lactate synthesis was found in cytoplasm of the adult chicken tissues. Specificity of enzyme activity alterations was detected in the chicken brain during ontogenesis after thyroxine administration.     

SSTR5 ablation in islet results in alterations in glucose homeostasis in mice

Somatostatin (SST) peptide is a potent inhibitor of insulin secretion and its effect is mediated via somatostatin receptor 5 (SSTR5) in the endocrine pancreas. To investigate the consequences of gene ablation of SSTR5 in the mouse pancreas, we have generated a mouse model in which the SSTR5 gene was specifically knocked down in the pancreatic beta cells (betaSSTR5Kd) using the Cre-lox system. Immunohistochemistry analysis showed that SSTR5 gene expression was absent in beta cells at three months of age. At the time of gene ablation, betaSSTR5Kd mice demonstrated glucose intolerance with lack of insulin response and significantly reduced serum insulin levels. Insulin tolerance test demonstrated a significant increase of insulin clearance in vivo at the same age. In vitro studies demonstrated an absence of response to SST-28 stimulation in the betaSSTR5Kd mouse islet, which was associated with a significantly reduced SST expression level in betaSSTR5Kd mice pancreata. In addition, betaSSTR5Kd mice had significantly reduced serum glucose levels and increased serum insulin levels at 12 months of age. Glucose tolerance test at an older age also indicated a persistently higher insulin level in betaSSTR5Kd mice. Further studies of betaSSTR5Kd mice had revealed elevated serum C-peptide levels at both 3 and 12 months of age, suggesting that these mice are capable of producing and releasing insulin to the periphery. These results support the hypothesis that SSTR5 plays a pivotal role in the regulation of insulin secretion in the mouse pancreas.