光/暗对玉米叶片果糖-6-磷酸激酶-2和果糖-2,6-二磷酸酯酶活力的影响

比较了照光和黑暗条件下玉米叶片果糖—6—磷酸激酶—2(PFK-2)和果糖—2,6—二磷酸酯酶(FBPase-2)的活力变化。当玉米植株从暗中转入光下后,其叶片PFK—2的活力随光照时间的延长而逐渐降低,而FBPase-2活力变化不明显;从光下转入暗后叶片PFK-2活力明显上升,FBPase-2活力仍无明显变化;其PFK-2/FBPase-2比值在光处理时下降,暗处理时上升。同时叶片中果糖—2,6—二磷酸的含量与PFK-2/FBPase-2活力比值的变化趋势一致。连续光照 20 h,PFK-2活力持续下降,表明PFK-2的光钝化现象与玉米植株的昼夜节律变化无关。     

玉米幼苗叶片保护酶活性的昼夜变化

人为控制环境下,对玉米幼苗叶片中电解质外渗率、MDA含量以及保护酶活性的昼夜变化同步进行了测定。结果表明：叶片电解质外渗率的昼夜变化呈现双峰曲线,分别在光照4 h和光照后黑暗4 h达到峰值;叶片中MDA含量表现为单峰的波形曲线,黑暗开始的4 h内最高;随着光照时间的延长,SOD酶、POD酶和CAT酶活性均表现出不同程度的降低,但均在16：00（光照8 h）至20：00（光照12 h）之间达到一昼夜内的最低点。随着光照后黑暗时间的延长,各保护酶活性均增加,但增加的速率不同,至次日8：00所有保护酶活性均接近光照前水平。光照和黑暗条件下,各保护酶活性差异极显著。保护酶活性的昼夜变化与叶片中的可溶性蛋白质含量并无显著的相关关系。     

大鼠视交叉上核与松果体中Clock基因转录的昼夜节律性及不同光反应性

本研究旨在观察和比较视交叉上核（suprachiasmatic nucleus,SCN）与松果体（pineal gland,pG）中Clock基因内源性昼夜转录变化规律以及光照对其的影响。Sprague-Dawley大鼠在持续黑暗（constant darkness,DD）和12h光照：12h黑暗交替（12hourlight：12hour-darkcycle,LD）光制下分别被饲养8周（n=36）和4周n=36）后,在一昼夜内每隔4h采集一组SCN和PG组织（n=6）,提取总RNA,用竞争性定量RT-PCR测定不同昼夜时点（circadian times．CT or zeitgeber times．ZT）各样品中Clock基因的mRNA相对表达量,通过余弦法和ClockLab软件获取节律参数,并经振幅检验是否存在昼夜节律性转录变化。结果如下：（1）SCN中Clock基因mRNA的转录在DD光制下呈现昼低夜高节律性振荡变化（P〈0．05）,PG中Clock基因的转录也显示相似的内源性节律外观,即峰值出现于主观夜晚（SCN为CTl5,PG为CT18）,谷值位于主观白天（SCN为CT3,PG为CT6）（P〉0．05）。（2）LD光制下SCN中Clock基因的转录也具有昼夜节律性振荡（P〈0．05）,但与其DD光制下节律外观相比,呈现反时相节律变化（P〈0．05）,且其表达的振幅及峰值的mRNA水平均增加（P〈0．05）,而PG中Clock基因在LD光制下转录的相应节律参数变化却恰恰相反（P〈0．05）。（3）在LD光制下,光照使PG中Clock基因转录的节律外观反时相于SCN（P〈0．05）,即在SCN和PG的峰值分别出现于光照期ZT10和黑暗期ZT17,谷值分别位于黑暗期ZT22和光照期ZT5。结果表明,Clock基因的昼夜转录在SCN和PG中存在同步的内源性节律本质,而光导引在这两个中枢核团调节Clock基因昼夜节律性转录方面有着不同的作用。     

光／暗对玉米叶片果糖—6—磷酸激酶—2和果糖—2,6—二磷酸酯酶活…

比较了照光和黑暗条件下玉米叶片果糖－６－磷酶激酶－２和果糖－２,６－二磷酸酯酶的活力变化。当玉米植株从暗中转入光下后,其叶片ＰＦＫ－２的活力随光照时间的延长而逐渐降低,而ＦＢＰａｓｅ－２活力变化不明显;从光下转入暗后叶片ＰＦＫ－２活力明显上升,ＦＢＰａｓｅ－２活力仍无明显变化;其ＰＦＫ－２／ＦＢＰａｓｅ－２比值在光处理时下降,暗处理时上升。同时叶片中果糖－２,６－二磷酸的含量与ＰＦＫ－２／ＦＢＰａ     

毛地黄叶离体培养中光质对强心苷含量的影响

用相近辐度但不同光质的光进行光照及黑暗处理,光质对愈伤组织中强心苷的含量有明显的影响,绿光下强心苷含量最高,黑暗处理强心苷含量最低。     

黄化水稻幼苗转绿期AOX1基因家族的表达与功能分析

完全黄化的水稻幼苗叶片在持续光照下总呼吸速率、交替途径的速率以及交替途径在总呼吸中的比值均上升,但以水稻AOX1基因家族3个成员的特异性片段为探针,仅观察到其中AOX1c转录本的增加。交替途径的专一性抑制剂SHAM可以降低水稻幼苗在持续光照过程中的相对光合放氧速率与叶绿素含量。同时,水稻黄化幼苗光照前黑暗处理时间越长,在恢复光照后交替途径能力的增加越显著,表现了转绿进程与交替途径之间的相关性。推测加强交替途径可能是植物缓和能量和物质需求矛盾的一个重要调控机制。     

大鼠下丘脑交叉上核中CREB的昼夜节律

利用凝胶迁移率变化的实验方法,对饲养在光照－黑暗循环的条件和持续黑暗的条件下Wistar雄性大鼠下丘脑交叉上核中CREB含量的昼夜间变化进行了分析,发现CREB在交叉上核中具有内源性的昼夜节律.     

脱落酸和己糖激酶抑制剂对高表达C4-PEPC转基因稻苗耐旱性的影响

为了研究高表达转玉米C₄-磷酸烯醇式丙酮酸羧化酶（phosphoenolpyruvate carboxylase,PEPC）基因水稻（PC）的耐旱性机制,本研究以PC和未转基因野生型原种kitaake为材料,分别在光照和黑暗预处理24 h,其中光照处理中光强为600 μmol·m^-2·s^-1,预处理后稻苗再在15％聚乙二醇-6000模拟干旱胁迫下,同时使用药理学的方法,通过加入脱落酸和己糖激酶的专一性抑制剂100 μmol·L^-1去甲二氢愈创木酸和10 mmol·L^-1葡萄糖胺,观察两种水稻4~5叶期稻苗耐旱表现。结果发现,与WT水稻相比,在PEG-6000处理后,经过光预处理的PC水稻叶片相对含水量下降较少,始终比WT的高,而且丙二醛含量则较少,脯氨酸诱导增加,表现耐旱;而经过暗预处理后PC植株显著降低这个优势,表明光预处理有利于PC耐旱性的表现;黑暗预处理均显著下调了2供试材料植株叶片中可溶性糖的含量,而对可溶性蛋白的含量影响不显著;而光预处理后PC水稻叶片内可溶性糖含量比WT增加,而在黑暗预处理则PC的显著低于WT的,其中葡萄糖胺对光预处理下PC的可溶性糖含量的下调作用最显著;暗预处理逆转或消除了NO,H₂O₂和钙离子含量变化趋势,这些变化与暗预处理减少了两材料叶片蔗糖和葡萄糖含量变化同步;光暗预处理对两材料的PEPC酶活性的差异影响不大,表明外源玉米C₄-PEPC在水稻中是组成型表达。可见PC叶片可部分通过糖组分,参与内源糖介导ABA和HXK信号途径,缓解干旱处理对叶片的伤害,稳定光合能力。     

丛毛垂叶榕种子萌发的光敏感性及其生态意义

以丛毛垂叶榕（Ficns benjamina L．var．nuda）种子为实验材料,研究了温度、光照、植物激素（GA3、6-BA、乙烯）和含氮化合物（硝普钠、亚硝酸盐和硝酸盐）对种子萌发的影响,讨论了光在丛毛垂叶榕种子萌发中的生态意义。在交替光照下（14h光照,10h黑暗,12μmol m^-2s^-1）,种子在15℃、20℃、25℃、30℃、35℃、40℃和30℃,20℃下的最终萌发率分别为87．5％、100％、100％、100％、98％、89％和100％;种子的平均萌发时间分别为34．7d、16．3d、5．6d、4．8d、6．4d、9d和6．3d。黑暗条件下,种子在15℃、20℃、25℃、30℃、35℃、40℃和30℃／20℃下萌发35d,种子的萌发率为零;添加交替光照后,种子迅速萌发。0．5～20μmol m^-2s^-1的光照强度显著地增加种子的最终萌发率,不同的光照强度对种子的最终萌发率没有明显的影响,但影响种子的萌发速率。在24h光暗周期下,种子的最终萌发率随着光期长度的增加而显著提高。在连续光照下处理24h、36h和48h后能够促进种子在随后的连续黑暗中萌发。不同浓度的GA3、6-BA、乙烯、硝普钠、NaNO3和NaNO2处理不能代替光照在黑暗条件下促进种子萌发,添加交替光照后,种子迅速获得萌发能力。丛毛垂叶榕种子的上述萌发行为与其长期适应热带雨林的生境密切相关。     

光/暗对高粱叶片磷酸烯醇式丙酮酸羧化酶基因表达的调节

高粱幼苗黄化叶片经照光转绿后,其PEP-Case活性提高4～15倍,mRNA含量提高了1.03倍,并测定出PEPCase mRNA的分子量为3.4kb。以等量的总RNA及mRNA进行体外翻译,发现转绿后PEPCase专一性翻译活性提高了51％～53％。这表明光照可以在转录水平上调节PEP-Case的基因表达。     

Differential Involvement of the Circadian Clock in the Expression of Genes Required for Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Synthesis,Assembly, and Activation in Arabidopsis thaliana

We have investigated the role of the circadian clock in the regulation of expression of genes required for ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) synthesis, assembly, and activation. Circadian oscillations in RCA (the gene encoding Rubisco activase) and RBCS (the gene encoding Rubisco small subunit) mRNA accumulation, with peak abundance occurring soon after dawn, occur in Arabidopsis thaliana grown in a light-dark (LD) photoperiod. These oscillations persist in plants that have been transferred from LD to either continuous darkness (DD) or continuous light (LL). In contrast, CPN60[alpha] (the gene encoding [alpha]-chaperonin) and CPN60[beta] (the gene encoding [beta]-chaperonin) mRNA abundance oscillates in a diurnal, but not in a circadian, fashion. Although rapid damping of the circadian oscillation in RCA mRNA abundance is observed in Arabidopsis that have been grown in LD and then transferred to DD for 2 d, the circadian oscillations in RCA and RBCS mRNA abundance persist for at least five continuous cycles in LL, demonstrating the robustness of the circadian oscillator.     

光和糖对水稻Rubisco活化酶基因表达的影响

水稻黄化苗在光照2h内其Rubisco。活化酶的mRNA和蛋白量明显增加,然后维持在相对稳定的水平。光对水稻Rubisco活化酶的基因表达的诱导作用主要在转录水平上。Rubisco活化酶主要在绿叶中表达,这与Rubisco基因表达的器官特异性完全一致。用等渗葡萄糖喂养成熟的水稻叶片1h,促使水稻Rubisco大、小亚基和Rubisco活化酶可翻译mRNA含量下降。同样蔗糖对Rubisco小亚基和Rubisco活化酶的表达也有抑制,其作用弱于葡萄糖。     

Effect of light on the development of the circadian rhythm of motor activity in the mouse

In previous experiments, we found that rats raised in constant light (LL) manifested a more robust circadian rhythm of motor activity in LL and showed longer phase shifts after a light pulse in constant darkness (DD) than those raised under constant darkness. In addition, we observed that the effects produced by constant light differed depending on the time of postnatal development in which it was given. These results suggest that both sensitivity to light and the functioning of the circadian pacemaker of the rat could be affected by the environmental conditions experienced during postembryonic development. Thus, the present experiment aimed to study whether postnatal exposure to light could also affect the circadian system of the mouse. Three groups of mice were formed: One group was raised under constant darkness during lactation (DD group), the second under constant light (LL group), and the third under light-dark cycles (LD group). After lactation, the three groups were submitted first to constant light of high intensity, then to LD cycles, and finally to constant darkness. In the DD stage, a light pulse was given. Finally, mice were submitted to constant light of low intensity. We observed that the circadian rhythm of the DD group was more disturbed under constant light than the rhythm of the LL group, and that, when light intensity increased, the period of the rhythm of the DD group lengthened more than that of the LL group. No significant differences among the groups were found in the phase shift induced by the light pulse. Therefore, it appears that DD mice are more sensitive to light than their LL counterparts. However, at present there is no evidence to affirm that the light environment experienced by the mouse during postnatal development affects the circadian pacemaker. (Chronobiology International, 18(4), 683-696, 2001)     

EFFECT OF LIGHT ON THE DEVELOPMENT OF THE CIRCADIAN RHYTHM OF MOTOR ACTIVITY IN THE MOUSE

In previous experiments, we found that rats raised in constant light (LL) manifested a more robust circadian rhythm of motor activity in LL and showed longer phase shifts after a light pulse in constant darkness (DD) than those raised under constant darkness. In addition, we observed that the effects produced by constant light differed depending on the time of postnatal development in which it was given. These results suggest that both sensitivity to light and the functioning of the circadian pacemaker of the rat could be affected by the environmental conditions experienced during postembryonic development. Thus, the present experiment aimed to study whether postnatal exposure to light could also affect the circadian system of the mouse. Three groups of mice were formed: One group was raised under constant darkness during lactation (DD group), the second under constant light (LL group), and the third under light-dark cycles (LD group). After lactation, the three groups were submitted first to constant light of high intensity, then to LD cycles, and finally to constant darkness. In the DD stage, a light pulse was given. Finally, mice were submitted to constant light of low intensity. We observed that the circadian rhythm of the DD group was more disturbed under constant light than the rhythm of the LL group, and that, when light intensity increased, the period of the rhythm of the DD group lengthened more than that of the LL group. No significant differences among the groups were found in the phase shift induced by the light pulse. Therefore, it appears that DD mice are more sensitive to light than their LL counterparts. However, at present there is no evidence to affirm that the light environment experienced by the mouse during postnatal development affects the circadian pacemaker. (Chronobiology International, 18(4), 683–696, 2001)     

Cytoplasm Rescues an Arrhythmic Mutant on the Circadian Rhythm of Mating Reactivity in Paramecium bursaria

Cells of an unusual Paramecium bursaria stock (Sj2) expressed rhythmic mating reactivity in a light/dark cycle (LD) and under continuous illumination (LL). When placed in continuous darkness (DD), did not show rhythmicity but rather demonstrated a continuous high mating reactivity. However, mating reactivity was reduced following exposure to a 6-h light pulse interrupting the DD, and then recovered to its former condition. Genetic analysis showed the arrhythmicity in DD to be a dominant character inherited in a Mendelian ratio. On the other hand, a clone (MCIw) that did not show the rhythmicity in either DD or LL was isolated from the parent stock Sj2w following a 5-h treatment with 2 μg/ml nitrosoguanidine (MNNG). The MCIw cells expressed weak rhythmicity in LD, but were insensitive to a 6-h light pulse in DD. The arrhythmicity in LL was inherited cytoplasmically. In addition to this, rhythmicity in LL could be recovered by injection of cytoplasm from the wild-type cell when the recipient cell was homozygous for the wild-type nuclear gene (+/+). The cytoplasmic components or factors are assumed to control the functional circadian system and genetically determine the rhythmicity of mating reactivity.     

Antisense inhibition of Rubisco activase increases Rubisco content and alters the proportion of Rubisco activase in stroma and thylakoids in chloroplasts of rice leaves

BACKGROUND AND AIMS: Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase (RCA) is a nuclear-encoded chloroplast protein that modifies the conformation of Rubisco, releases inhibitors from active sites, and increases enzymatic activity. It appears to have other functions, e.g. in gibberellin signalling and as a molecular chaperone, which are related to its distribution within the chloroplast. The aim of this research was to resolve uncertainty about the localization of RCA, and to determine whether the distributions of Rubisco and RCA were altered when RCA content was reduced. The monocotyledon, Oryza sativa was used as a model species. METHODS: Gas exchange and Rubisco were measured, and the sub-cellular locations of Rubisco and RCA were determined using immunogold-labelling electron microscopy, in wild-type and antisense rca rice plants. KEY RESULTS: In antisense rca plants, net photosynthetic rate and the initial Rubisco activity decreased much less than RCA content. Immunocytolocalization showed that Rubisco in wild-type and antisense plants was localized in the stroma of chloroplasts. However, the amount of Rubisco in the antisense rca plants was greater than in the wild-type plants. RCA was detected in both the chloroplast stroma and in the thylakoid membranes of wild-type plants. The percentage of RCA labelling in the thylakoid membrane was shown to be substantially decreased, while the fraction in the stroma was increased, by the antisense rca treatment. CONCLUSIONS: From the changes in RCA distribution and alterations in Rubisco activity, RCA in the stroma of the chloroplast probably contributes to the activation of Rubisco, and RCA in thylakoids compensates for the reduction of RCA in the stroma, allowing steady-state photosynthesis to be maintained when RCA is depleted. RCA may also have a second role in protecting membranes against environmental stresses as a chaperone.