The Relationship Between Senescence and Ca2+-ATPase Activity of Microsomal Membrane and Lipid Peroxidation in Harvested Peach Fruit

Peach fruit easily soften and have a short storage time at normal temperature. In this study, peach fruit (Prunus persica sieb et Zucc cv. Yingqing) were picked and stored at 25 and 4℃ to investigate the senescence in correlation with Ca2+- ATPase activity of microsomal membrane and lipid peroxidation during ripening and senescence. In comparison with that stored at 25℃, the fruit stored at 4℃ exhibited a higher flesh firmness, lower respiration rate, and generated the late bigger peak value of Ca2+-ATPase activity as well as maintained the higher activity of the enzyme. Meanwhile, the lower levels of super oxygen radical (O2-) production and content of malondialdehyde (MDA), a product of membrane lipid peroxidation were observed. Sodium orthovanadate (SO) and erythrosin B (EB), as Ca2+-ATPase inhibitors, could stimulate the respiration rate. The results suggested that the slower senescence rate of peach fruit was closely related to the higher peak value and longer duration of Ca2+-ATPase activity in microsomal membrane, with the slighter membrane lipid peroxidation and lower O2(-) production rate.     

Relationship Between Ca2+-CaM and Ethylene-Induced PG Activity in Tomato Fruit

Polygalacturonase (PG) was studied during ripening and senescence of postharvest tomatofruit at pink stage at low and normal temperature. The results showed that the PG activity increased, thendecreased during ripening and senescence of tomato. Low temperature inhibited but ethylene enhanced PGactivity. Ethylene also enhanced caimodulin content, which was dependent on Ca2+ concentration in cell.When EGTA(Ca2+ chelator), verapamil (Vp) and LaCl3 (Ca2+ channel blockers), trifluoperazine and chloro-promaize (two CaM antagonisms) were used to treat tomato fruit at green mature stage with ethylene, theycould reverse ethylene-induced increase in PG activity, but Vp, chloropromaize (CPZ), trifluoperazine(TFP) could not directly influence PG activity, which indirectly indicated that influx of Ca2+ from the ex-tracellular space including the cell wall via the Ca2+ channel localized in plasma membrane and CaM were re-quired for ethylene-induced PG activity increase and that ethylene signal transduction may be related to Ca2+- CaM messenger system.     

Effects of Ca2+/CaM Messenger System Inhibitors on Ethylene-Induced Increase in Lycopene Content

The changes of lycopene content during ripening and senescence of tomato fruit and the relationship between ethylene glycol-bis (EGTA, Ca2+ chelator), verapamil (Vp, Ca2+ channel blockers), trifluoperazine (TFP), chloropromaize (CPZ) (CaM antagonism) and ethylene-induced increase in lycopene content in tomato fruit were investigated. Lycopene content accumulated obviously during ripening and senescence of tomato fruit after harvest at pink stage. Low temperature inhibited but ethylene enhanced the lycopene content.Meanwhile, ethylene also promoted calmodulin (CaM) content in tomato fruit, which was related to the concentration of ethylene. When EGTA, Vp, TFP and CPZ with ethylene were used to treat tomato fruit, ethylene-induced increase in lycopene content could be reversed, indicating that blocking Ca2+ channel in plasma membrane or chelating extracellular Ca2+ or inhibiting the activity of CaM could decrease the action of ethylene, and suggesting that Ca2+-CaM messenger system may be involved in lycopene increase inducedby ethylene.     

Changes of Reactive Oxygen Species and Related Enzymes in Mitochondria Respiratory Metabolism During the Ripening of Peach Fruit

The fruits of peach cultivar Yuhua 3 were used as materials to investigate the changes of active oxygen and related enzymes in mitochondria respiratory metabolism during ripening of peach fruit, involving their influence on the proceeding of peach fruit senescence. The results showed that the large decrease in firmness occurred between maturity II and IV. The decrease in firmness coincided with an increase in respiratory intensity. Obvious peaks of respiratory intensity lagging to the rapid change of fruit firmness could be shown during peach ripening. Reactive oxygen species （ROS） had a cumulative process and positively correlated with respiratory intensity. During peach ripening, the content of Ca^2＋ increased, the activities of succinic dehydrogenase （SDH）, cytochrome C oxidase （CCO）, H＋-ATPase, and Ca^2＋-ATPase decreased varying in different degree at the later step of ripening. These suggested a close relationship existed between ROS metabolism and mitochondrial respiration, namely, both ROS metabolism and mitochondrial respiration probably played important roles in ripening and senescing of peach fruit.     

NO处理延缓采后枇杷果实木质化劣变及其与能量代谢的关系

【目的】枇杷果实采后生命活动旺盛,衰老速度快,常温下极易变质腐烂。低温贮藏虽然可以有效延长贮藏期,减少腐烂,但会出现果皮难以剥离、果肉木质化并褐变、质地糙硬少汁等品质劣变现象,这是造成冷藏枇杷商品性丧失和损失的主要原因,已成为其市场拓展的限制因素,是当前枇杷果实在冷链集散和流通中急需解决的关键问题。探讨外源NO处理对冷藏枇杷果肉木质化劣变进程的作用机制,并分析木质化劣变与能量代谢的关系,以期为进一步研究采后枇杷果实低温品质劣变进程调控的分子生物学机理和贮运保鲜技术奠定基础。【方法】将‘解放钟’枇杷（Eriobotrya japonica Lindl.）果实在密闭容器中用0（对照组）、15和25 μL•L-1 NO熏蒸2 h后,取出通风20 min,然后将各处理果实置于5℃、相对湿度85%条件下贮藏,测定冷藏期间各处理组果实细胞膜透性、硬度、出汁率、木质素含量、ATP含量、ADP含量、AMP含量、能荷值及琥珀酸脱氢酶（SDH）、细胞色素氧化酶（CCO）、H+-ATPase和Ca2+-ATPase活性的变化,并分析NO处理后木质素含量与能荷值间的相关性。【结果】随着贮藏时间的延长,枇杷果实细胞膜透性和硬度逐渐上升,出汁率逐渐下降,贮藏10 d后木质素含量迅速上升,果实冷害症状明显。与对照组相比, NO处理能延缓细胞膜透性和硬度的上升及出汁率的降低,显著抑制木质素的合成,较好地保持细胞膜的完整性,从而减轻果实冷害的发生。冷藏期间,枇杷果实ATP含量逐渐下降,贮藏前10 d ADP含量迅速下降并最终维持在较低水平,贮藏中后期（15-30 d）SDH、CCO、H+-ATPase和Ca2+-ATPase活性急剧下降,表明线粒体功能受损导致枇杷果实能荷水平迅速下降。与对照组相比,NO处理可以延缓ATP、ADP含量的下降,且显著抑制贮藏中后期SDH、CCO、H+-ATPase和Ca2+-ATPase活性的降低,保持枇杷果实较好的线粒体功能;贮藏30 d后,15 和25 μL•L-1 NO处理的枇杷果实能荷值分别比对照组高11.8%和12.9%。相关性分析表明,15和25 μL•L-1 NO处理的枇杷果实能荷值和木质素含量呈极显著负相关,相关系数（r）分别为 -0.715**、-0.598**。【结论】低温条件下,能量代谢失调与枇杷果实木质化劣变密切相关,NO处理可以通过调节SDH、CCO、H+-ATPase、Ca2+-ATPase线粒体代谢相关酶活性,维持较高的能量水平,从而有效地提高冷藏枇杷果实抗低温的能力,进而延缓果实木质化进程,其中以25 μL•L-1 NO处理效果较好。     

温度逆境交叉适应过程中葡萄幼苗质膜Ca2+-ATPase的细胞化学定位与活性变化

【目的】研究植物交叉适应现象及其生理生化机制。【方法】以京秀葡萄（Vitis vinifera L. cv. Jingxiu）为试材，研究了38℃/10 h的高温锻炼预处理和8℃/2.5 d的低温锻炼预处理分别对随后0℃低温和45℃高温胁迫期间葡萄叶片细胞质膜Ca2+-ATPase的影响。从生化水平测定了定位在质膜、液泡膜上的Ca2+-ATPase的活性，并用氯化铈沉淀的电镜细胞化学方法，定位观察了质膜Ca2+-ATPase的活性。【结果】高温锻炼预处理提高了微粒体膜上的Ca2+-ATPase酶的活性，并且在随后的低温胁迫条件下，保持了酶活性的稳定性；低温锻炼提高了微粒体在高温胁迫下的ATP酶活性。【结论】高温锻炼诱导的抗冷性和低温锻炼所诱导的耐热性的提高与微粒体膜上Ca2+-ATPase活性的变化有关，且高温锻炼和低温锻炼具有相同的调控机制。     

套袋红富士苹果果实发育过程中Ca~(2+)-ATPase活性变化

以红富士苹果(Malus domesticaBorkh.cv.‘Red Fuji’)为试材,研究了发育过程中以及成熟后期套袋果实Ca2+-ATPase的活性变化。结果表明,发育初期,苹果果实的Ca2+-ATPase活性较高,随着果实生长发育,Ca2+-ATPase活性逐渐下降,在发育后期Ca2+-ATPase活性有所提高。与对照相比,套袋红富士苹果果实Ca2+-AT-Pase的活性呈现先降低后升高又降低的变化趋势。发育后期,摘除果袋后苹果果实的Ca2+-ATPase活性与对照相比有显著增加。与对照相比,套袋降低苹果果实的Ca2+浓度。由此表明,套袋可能通过影响钙素的吸收运转和Ca2+-ATPase活性而调控苹果果实的生长发育和品质形成。     

南海鸢乌贼肌原纤维蛋白加热过程中理化性质的变化

为研究南海鸢乌贼Symplectoteuthis oualaniensis肌原纤维蛋白在加热过程中理化性质的变化,以南海鸢乌贼胴体肌肉为原料,对其肌原纤维蛋白在加热过程中浊度、溶解度、Ca~(2+)-ATPase活性和总巯基(-SH)含量进行了测定,并对该蛋白质在整个加热过程中SDS-PAGE电泳图的变化进行了研究。结果表明:在25~35℃时,鸢乌贼肌原纤维蛋白的浊度、溶解度、Ca~(2+)-ATPase活性、总巯基含量的变化均比较缓慢;在35℃后继续升温,浊度、Ca~(2+)-ATPase活性、总巯基含量均发生明显变化;总巯基含量在40~50℃时出现短暂升高现象后又急剧降低,溶解度在50℃时开始明显降低;在25~35℃时,随着温度的逐渐升高,鸢乌贼肌原纤维蛋白的肌球蛋白重链(MHC)略有减少,副肌球蛋白(PM)和肌动球蛋白(Actin)基本上未发生变化,但在35℃后,鸢乌贼肌原纤维蛋白开始产生大分子物质,逐渐发生变性。     

Changes of Reactive Oxygen Species and Related Enzymes in Mitochondrial Respiration During Storage of Harvested Peach Fruits

Peach fruits [Prumus persica (L.) Batsch,cv.Yuhuasanhao] were used as materials to investigate the changes of reactive oxygen species (ROS) and related enzymes in mitochondria respiration during storage and then their influence on senescence of harvested Peach fruits was studied.The results showed that low temperature (50C) strongly inhibited the reduction of firmness and the increase in respiration rate.During storage at ambient temperature (200C),ROS had a cumulative process while malondialdehye (MDA) content continued to increase in associated with enhanced membrane lipid peroxidation.Lipoxygenase (LOX) activity was strongly inhibited under the low temperature condition.The activities of succinic dehydrogenase (SDH),cytochrome C oxidase (CCO),and Caz+-ATPase declined to a certain extent at ambient temperature,while they showed higher activities at low temperature,which may be related to lower membrane lipid peroxidation at low temperature.Higher Ca~(2+) content at ambient temperature may be responsible for impairment of mitochondrial function,thus,leading to fruit senescence.The results showed that under low temperature condition,the low accumulation of ROS and the low level of membrane lipid peroxidation could maintain the function of mitochondria that would help to delay the senescence of peach fruits.These suggested a close relationship existed between ROS metabolism and mitochondrial respiration.It can be inferred that the low temperature helps to delay senescence of peach fruits via suppression of ROS and related enzymes,maintain better homeostasis of Ca~(2+) in mitochondria and thus better mitochondrial functions.     

喷钙对红富士苹果果实Ca2+-ATPase活性及品质的影响

研究了不同钙肥对红富士苹果果实品质和Ca2 -ATPase活性的影响。结果表明,喷钙后单果重增加,果形指数增大,Vc含量提高,可溶性固形物和花青苷含量增大,果实的叶绿素和可滴定酸含量下降。不同钙肥间,喷巨金钙的效果最好,其次为氨基酸钙,再次为翠康钙宝,最后为钙宝2000。钙肥对Ca2 -ATPase活性影响极为显著,其活性明显高于对照,而不同钙肥间的变化趋势与果实品质的变化趋势相同。     

The Relationship Between Senescence and Ca^2＋-ATPase Activity of Microsomal Membrane and Lipid Peroxidation in Harvested Peach Fruit

Peach fruit easily soften and have a short storage time at normal temperature. In this study, peach fruit （Prunus persica sieb et Zucc cv. Yingqing） were picked and stored at 25 and 4℃ to investigate the senescence in correlation with Ca^2＋- ATPase activity of microsomal membrane and lipid peroxidation during ripening and senescence. In comparison with that stored at 25~C, the fruit stored at 4℃ exhibited a higher flesh firmness, lower respiration rate, and generated the late bigger peak value of Ca^2＋-ATPase activity as well as maintained the higher activity of the enzyme. Meanwhile, the lower levels of super oxygen radical （O2^-.） production and content of malondialdehyde （MDA）, a product of membrane lipid peroxidation were observed. Sodium orthovanadate （SO） and erythrosin B （EB）, as Ca^2＋-ATPase inhibitors, could stimulate the respiration rate. The results suggested that the slower senescence rate of peach fruit was closely related to the higher peak value and longer duration of Ca^2＋-ATPase activity in microsomal membrane, with the slighter membrane lipid peroxidation and lower O2^-. production rate.     

毛白杨花粉败育过程中Ca^2＋-ATPase的异常变化

为研究木本植物花粉败育过程中Ca2+ ATPase的影响,本文利用氯化铈沉淀和电镜细胞化学方法,对不同育性毛白杨花药在小孢子发生发育过程中Ca2+ ATPase进行了定位。结果表明:Ca2+ ATPase在可育花药小孢子母细胞时期大量分布于小孢子母细胞质膜、液泡膜及内膜系统,花药内壁细胞内膜系统和绒毡层细胞质膜,随后Ca2+ ATPase在上述细胞中均减少甚至消失,并于小孢子时期和成熟花粉时期分别在小孢子和花药内壁细胞中再次沉积;不育花药小孢子母细胞以及此时期绒毡层细胞质膜上并无明显Ca2+ ATPase,花药表皮、药室内壁以及中层的Ca2+ ATPase都高于同一时期的可育花药,其绒毡层细胞解体不完全。由上述结果推测:毛白杨不育花药内小孢子母细胞和药壁中Ca2+ ATPase分布异常,影响细胞内钙离子的主动转运,可能导致钙离子异常累积,进而通过影响小孢子母细胞和药室内壁细胞的正常代谢、绒毡层细胞的及时降解而导致花粉败育。      

茉莉酸甲酯诱导的采后香蕉果实耐冷性与活性氧信号的关系

 【目的】探讨茉莉酸甲酯（MeJA）对采后香蕉果实耐冷诱导的效果及其对活性氧代谢、Ca2+-ATPase活性的影响。【方法】香蕉果皮用10 µmol•L-1 MeJA处理1 min,在20℃恒温培养1 d（取样作第0天）,然后置于7℃下离体培养10 d,测定果皮冷害指数、细胞膜透性、过氧化氢和超氧阴离子含量、过氧化氢酶（CAT）、抗坏血酸过氧化物酶（APX）、NADPH氧化酶以及Ca2+-ATPase活性的变化。【结果】与对照香蕉果皮相比,10 µmol•L-1 MeJA处理的冷害指数较低、细胞膜透性增加缓慢;MeJA处理诱导了果皮H2O2含量和 产生速率的提高,并维持了前4 d的高水平,而MeJA处理降低了CAT和APX的活性,并在前4 d维持了较低的水平;研究还发现,MeJA处理还诱导了NADPH氧化酶和Ca2+-ATPase活性的升高。【结论】MeJA诱导的活性氧猝发可能作为信号分子参与诱导了香蕉果皮冷害和耐冷诱导,并可能与钙信号相关。     

温度预锻炼和Ca~(2+)对温度胁迫下黄连膜脂过氧化及保护酶活性的影响

为了明确温度预锻炼和Ca2+对温度胁迫下黄连生理指标的影响,以2年生黄连为材料,以25℃为对照,设计2组试验,探索温度胁迫下黄连膜脂过氧化及保护酶活性的变化。结果表明:在低温(-5～15℃)或高温(35℃)胁迫下,黄连丙二醛(MDA)、脯氨酸(Pro)含量增加,Pr(蛋白质)含量降低;超氧化物歧化酶(SOD)和过氧化物酶(POD)活性在低温胁迫下升高,在高温胁迫下降低,而过氧化氢酶(CAT)活性在低温和高温胁迫下都升高;进行12h温度预锻炼或CaCl2溶液浇灌,可以有效降低黄连MDA和Pro增加量,促使黄连Pr含量上升、SOD和POD活性升高。在受试的3个CaCl2浓度中,以100mmol/L CaCl2对膜系统的保护作用最佳。这表明,温度预处理和Ca2+能在一定程度上降低温度胁迫对黄连的伤害,可作为提高黄连抗逆性的一种途径。     

AMP和MH对菊花绿蕾期叶片叶绿体Ca~(2+)-ATPase、Mg~(2+)-ATPase及超微结构的影响

在秋菊姹紫嫣红品种的营养生长期,以氨卞西林钠(AMP,1 000mg/L)和不同浓度马来酰肼(MH 100mg/L、200mg/L和300mg/L)的水溶液进行叶面喷雾处理,初步研究了AMP和MH对菊花绿蕾期叶片光合速率、叶绿体Ca2+-ATPase、Mg2+-ATPase活性和叶绿体超微结构的影响。结果表明,AMP处理使菊花绿蕾期叶片上述生理指标均明显高于对照,叶绿体片层结构数量和密集程度明显增加,嗜锇颗粒减少。MH处理叶绿体Ca2+-ATPase和Mg2+-ATPase活性比对照明显降低,且随处理升高呈先降后升的变化趋势;在100~200mg/L MH浓度范围内,光合速率和叶绿体片层结构数量和密集程度均高于对照,嗜锇颗粒减少,且处理效应随处理浓度升高而有所减弱,在300mg/L处理下,光合速率低于对照且叶绿体片层结构趋于解体,嗜锇颗粒数量明显增多。     

钾与钙·硼混合喷施对龙眼果实品质及耐贮性的影响

在果园土壤低硼条件下,以储良龙眼为试材,作了连续两年研究.结果表明,在龙眼假种皮发育期间,树冠分别喷施0.3%KH2PO4、0.3%KH2P04+1%CaCl2、0.3%KH2PO4+0.2%H3803溶液3次,可显著增大成熟龙眼果型、提高果实组织鲜重,增加果实的可食率、果肉可溶性固形物和糖含量,并促进果实组织对Ca、B的吸收积累,显著提高果实组织K含量;在30±2℃条件下贮藏,三处理均降低了贮藏期间果实失重率、提高好果率,减缓了果肉营养成分含量的下降.可见钾与钙、硼混合喷施对于提高龙眼果实的品质与耐贮性和维持贮藏过程中果实品质有一定作用.     

蜘蛛头胸部钙含量与其活动能力的相关性研究

以拟环纹豹蛛头胸部为材料,利用微波消解法对其进行处理,采用火焰原子吸收光谱(FAAS)法对样品中的总钙离子含量进行检测,分析不同温度下蜘蛛头胸部钙、镁离子含量的变化;同时采用田间调查法研究拟环豹纹蛛在不同温度下空间生态位宽度的变化。试验结果表明:在一定范围内,随着温度的升高,蜘蛛头胸部Ca2+含量升高,导致神经冲动传递速度加快,范围加广,新陈代谢加强,活动力及兴奋性加强,空间生态位幅度变宽。     

青椒果实低温贮藏及冷害生理的研究

本试验研究了辣椒果实采后在10℃、7℃、4℃、1℃四个不同温度下的生理生化变化、贮藏效果、以及低温下冷害发生的规律。结果表明:10℃是辣椒果实冷藏的安全温度,采后初期的低温虽明显地抑制了辣椒果实的后熟,但随贮期的延长,7℃及以下的低温均对果实产生了不同程度的冷害,表现为果实的呼吸强度、MDA含量、相对电导率异常开高;冷害指数的变化与腐烂指数的变化是显著的正相关关系;且4℃处理的辣椒果实冷害发生更早,程度更严重。     

钙处理对苹果梨叶片钙调蛋白含量及过氧化物酶、多酚氧化酶活性的影响

以延边地区苹果梨树为试材,探索苹果梨树体生长期进行钙处理后对叶片钙调蛋白(CaM)含量及POD、PPO活性的影响。结果表明:钙处理对苹果梨叶片的CaM含量及POD、PPO活性有明显影响。其中幼果期、果实迅速膨大期、果实成熟期3个时期都喷施质量分数为0.5%CaCl2+20 mg/L NAA的处理对增加叶片CaM含量的效果最好,果实迅速膨大期喷施质量分数为0.5%CaCl2对降低叶片POD、PPO活性的效果最好。