几种同工酶与猕猴桃品种对溃疡病抗性关系的研究

应用聚丙烯酰胺凝胶电泳、同工酶分析技术分别研究了猕猴桃植株体内过氧化物酶(POD)、多酚氧化酶(PPO)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、酯酶(EST)同工酶谱带的变化,结果表明:自然感染溃疡病前后,此6种同工酶谱带特征在不同抗感品种中表现出一定的差异.未感染溃疡病菌前,抗(感)品系枝条、叶片POD同工酶均有2条酶带,PPO同工酶有3条酶带,但感病品种酶带颜色深且粗,而抗病品种酶带颜色浅且细,叶片酶带颜色深于枝条;SOD、CAT同工酶谱带均为1条,Rf值分别为0.38、0.28,感性品种较抗耐品种谱带亮度高活性强;自然发病后,抗(感)品系POD、PPO同工酶谱带数都增加,分别为4、3条和5、4条,且抗病品种新酶带出现较感病品种早且酶带粗颜色深活性强,感病品系虽也有新酶带出现,但酶带少活性弱,抗病品系枝条、叶片POD、PPO同工酶新谱带的Rf值分别为0.63、0.67和0.85、0.87;抗感病品种SOD、CAT同工酶都被诱导产生了1条新的同工酶谱带,Rf分别为0.32和0.27,新酶带现色时间迟,且酶带颜色浅活性弱,但抗耐品种较感性品种谱带亮且活性强;EST同工酶于自然发病前后变化不大,与抗病性关系不很明显.     

哈茨木霉对水稻过氧化物酶及多酚氧化酶活性的影响*

哈茨木霉不仅对病原微生物有拮抗作用,而且能够诱导植物产生抗病性。采用酶动力学扫描及垂直板凝胶电泳技术,对水稻喷施木霉发酵液后,测定多酚氧化酶(PPO)、过氧化物酶(POD)活性和同工酶带图谱的动态变化。与对照组相比,哈茨木霉不仅能够提高PPO和POD酶的活力,而且还能诱导产生新的同工酶谱带,但不同浓度发酵液对诱导合成的酶作用不同。稀释50倍的发酵液对POD活力有较大的阳性效应。高浓度的发酵液对PPO活性有先抑制后出现提高的趋势,表明了该发酵液对PPO活性调节的程序性。     

干旱胁迫下内生真菌感染对黑麦草叶内几种同工酶的影响

以内生真菌感染(endophyte-infected,EI)与不感染(endophyte-free,EF)的黑麦草(Lolium perenne L．)种子建立实验种群,分别对其施加长时间不同强度的干旱胁迫,通过比较黑麦草体内过氧化物酶(POD)、超氧化物歧化酶(SOD)、多酚氧化酶(PPO)活性及其同工酶谱的变化以探讨保护酶系统在内生真菌——植物共生体的抗旱性方面所作的贡献。研究结果表明,水分胁迫和内生真菌对黑麦草3种酶的影响不仅表现在总量上而且表现在同工酶的酶谱及各区带的酶活力上。就总酶活力而言,EI和EF植株中POD、SOD和PPO的活性均随着干旱胁迫强度的增加而增加,进一步将EI和EF植株的酶活力进行比较,发现与EF植株相比,EI植株中POD和PPO的活性相对较低,而SOD的活性相对较高。从同工酶的谱带数量和强弱来看,POD同工酶各区带活力均随干旱胁迫强度的增加而增加,EI植株叶片增加的幅度高于EF叶片,而且EI叶片在重度胁迫下出现了1条新带SOD同工酶各区带活力在EI叶片中有随干旱胁迫增加而增加的趋势,而在EF叶片中有些区带酶活力增强,有些区带酶活力减弱,且EI叶片在中度胁迫下出现了1条新带;PPO同工酶随干旱胁迫的增强,EI和EF叶片均表现为有些区带酶活力增强,有些区带酶活力减弱。总之,内生真菌的感染虽然没有显著提高宿主植物黑麦草POD、SOD和PPO的活性,但使宿主黑麦草对干旱胁迫的反应更为迅速,其中既包括POD、SOD等酶活力的迅速升高,也包括新酶带的产生。     

甘蓝幼苗受桃蚜危害后叶片中部分酶活性的变化

对桃蚜（Myzus persicae Sulzer）危害后甘蓝（Brassica oleracea L.）幼苗叶中多酚氧化酶（PPO）、苯丙氨酸解氨酶（PAL）、过氧化物酶（POD）活性及同工酶进行了研究。结果表明,随桃蚜危害时间延长,处理叶片PPO、PAL和POD的活性与对照相比均表现出升高的趋势,方差分析表明与对照之间差异显著。同工酶电泳结果表明,POD同工酶的部分谱带随危害时间而发生变化,但EST同工酶在接虫前后则无明显的变化。     

3个桉树无性系过氧化氢酶活性及同工酶比较研究

本文研究杂交桉树(Eucalyptus urophylla×Eucalyptus camaldulensis)LH21无性系、LH22无性系和尾叶桉(Eucalyptus urophylla)U6无性系的过氧化氢酶(CAT)活性及同工酶的差异。结果表明,3个桉树无性系同一器官的CAT活性存在差异,同一个无性系中不同器官的CAT活性差异很显著。采用聚丙烯酰胺不连续凝胶垂直板电泳比较分析CAT同工酶,发现3个无性系的CAT同工酶存在着一定的差异,其中LH21和LH22叶片有相同的谱带,但根的谱带与叶片有差异;而U6各器官的CAT谱带与LH21和LH22有差异。3个无性系的CAT同工酶都在一定程度上具有器官的特异性。     

5种樱桃属植物的POD、CAT和SOD同工酶分析

采用聚丙烯酰胺垂直平板凝胶电泳技术对5种樱桃属植物的过氧化物酶(POD)同工酶、过氧化氢酶(CAT)同工酶和超氧化物歧化酶(SOD)同工酶的酶谱特征进行分析,结果表明:5种樱桃属植物共电泳出12条POD同工酶酶带、4条CAT同工酶酶带和8条SOD同工酶酶带;其中,POD同工酶酶谱具有5条共同谱带,CAT同工酶4条,SOD同工酶5条。冬季休眠期的樱桃属植物,POD与SOD同工酶谱带的多样性比较丰富,不同植物之间的谱带差异较大;而CAT同工酶谱带差异不明显。     

Cucumis属3种不同倍性种间杂交后代的同工酶分析

以3种不同倍性的Cucum is属种间杂交后代[异源四倍体C.hy tivus(2n=4x=38,HHCC)、异源三倍体(2n=3x=26,HCC)和正反交种间杂种F1(2n=2x=19,HC/CH)]及其双亲为试材,比较研究了过氧化物酶(POD)和酯酶(EST)同工酶在不同器官中的酶谱表达特性.结果表明,花蕾中的POD和EST同工酶谱带都比叶片中的丰富,表现出明显的组织特异性.在相同器官的同一酶系统中,3种不同倍性种间杂交后代的酶谱基本一致,主要表现为互补双亲的酶带,同时出现了双亲所没有的酶带(POD4c和EST3b等),表明远缘杂交扩大了黄瓜的遗传基础.此外,在幼叶和花蕾的POD同工酶中,大部分酶带活性随染色体倍性增加而减弱,表明基因剂量与POD同工酶酶谱的表达呈负相关.     

几种薏苡的过氧化物酶和多酚氧化酶同工酶分析

用聚丙烯酰胺凝胶电泳（PAGE）的方法,对薏苡,野生薏苡和水生薏苡的几个不同种质分别进行了过氧化物酶（POD）和多酚氧化酶（PPO）的同工酶酶谱分析,结果表明：POD和PPO含量丰富,活性强,酶谱复杂,酶带清晰,稳定,种间差异明显。结果可以作为薏苡研究种质亲缘关系的基础。     

大别山区六种黄精属植物的五种同工酶分析

采用聚丙烯酰胺凝胶电泳法对皖西大别山区六种黄精属植物的抗坏血酸氧化酶(AOD)、多酚氧化酶(PPO)、超氧化物歧化酶(SOD)、酯酶(EST)和过氧化物酶(POD)五种同工酶进行了比较分析。结果表明:(1)五种同工酶共显示出66条酶带,其中有3条是黄精属的特征酶带;并且来源于不同种的同一种酶的谱带数、相对迁移率、酶活性均不相同,呈现多样性;(2)五种同工酶谱的模糊聚类分析结果与形态学分类的结果一致,利用酶谱差异可以将六种黄精初步区分;(3)在六种黄精属植物中,湖北黄精、轮叶黄精、多花黄精和金寨黄精是较进化类型,玉竹和长梗黄精是较原始类型。     

GB对低温胁迫黄瓜叶绿体及SOD、POD同工酶的影响

采用电镜观察和聚丙烯酰胺垂直板电泳法,研究了GB对低温胁迫下黄瓜幼苗叶绿体超微结构和SOD、POD同工酶的影响。结果表明,低温胁迫下,GB可使黄瓜幼苗叶绿体膜结构保持完好,而对照的叶绿体基粒破坏严重;与对照相比,GB处理对SOD、POD同工酶谱带数目均无影响,但POD同工酶的P2(Rf0．24)、P3(Rf0．30)、P4(Rf0．36)酶带的活性增加,SOD同工酶的S1(Rf0．14)、S5(Rf0．58)酶带活性较高,保持了黄瓜幼苗体内相对较高的抗氧化酶活性。GB可保护膜的完整性和稳定性,从而提高黄瓜幼苗的抗冷性。     

杂交水稻金优63幼苗期SOD和POD特性研究

对杂交水稻金优63幼苗不同时期的根、茎、叶进行SOD同工酶电泳分析,并测定SOD、POD活性。结果表明,自播种后第7天到第13天,幼苗的SOD同工酶在根、茎、叶中有明显的器官特异性,且SOD活性叶 >茎 >根。相同器官不同时期的SOD同工酶电泳谱带条数及SOD活性都有变化,且SOD活性强弱与SOD同工酶电泳谱带中有无Mn-SOD同工酶带有一定的关系。幼苗的POD活性在根、茎、叶中也有明显的器官特异性,茎中POD活性明显高于根和叶,且POD活性变化与SOD活性变化有一定的关系。     

Tissue browning of in vitro cultures of Scots pine: Role of peroxidase and polyphenol oxidase

Callus cultures from shoot tips of mature Scots pine ( Pinus sylvestris L.) were characterized by rapid browning and an inability to regenerate. The peroxidase (POD) and polyphenol oxidase (PPO) activities and relationship to browning in such cultures were compared with embryogenic and non-embryogenic cultures of Scots pine, started from immature embryos of three different pine clones. The browning in callus cultures derived from pine buds was visible approximately after 2 weeks of culture, and continued thereafter until the callus was dark brown and poorly growing. The non-embryogenic cultures induced from immature embryos showed either light yellow coloring or browning, whereas the embryogenic cultures showed browning. POD activity increased during the first 4 weeks in callus tissue initiated from pine buds, and was significantly higher than in pine buds or cultures derived from immature embryos. The ability of cultures initiated from pine buds to oxidize catechol was notably high compared with cultures initiated from immature embryos, regardless of the time of measurement. Addition of catalase revealed that both POD and PPO were able to use catechol as substrate. An antibody raised against broad bean ( Vicia faba ) chloroplast PPO was used to recognize PPO. One polypeptide with a molecular mass of 50 kDa was detected in all pine samples on SDS-PAGE and non-denaturing PAGE. Another polypeptide with a molecular mass of 70 kDa was shown exclusively in the light-yellow non-embryogenic cultures. The results suggest that especially the high POD activities in callus tissues started from mature trees cause rapid and early browning and possibly subsequent cell death.     

红豆杉组织培养中SINENXANS高产细胞系Ts19的几种同工酶动态比较研究

对从中国红豆杉的茎来源的愈伤组织经筛选而得的ｓｉｎｅｎｘａｎｓ高产细胞系Ｔｓ１９的过氧化物酶（ＰＯＤ）、酯酶（ＥＳＴ）、细胞色素氧化酶（ＣＯＤ）、淀粉酶（ＡＭＬ）、多酚氧化酶（ＰＰＯ）及超氧化物歧化酶（ＳＯＤ）的同工酶,可溶性蛋白的含量及电泳谱带、超氧化物歧化酶、多酚氧化酶和苯丙氨酸解氨酶（ＰＡＬ）的活性作了比较研究。并与培养过程中ｓｉｎｅｎｘａｎｓ含量的动态变化相比较,探索了这几种同工酶的酶谱和两种酶活性与ｓｉｎｅｎｘａｎｓ的生物合成的关系。旨在为建立紫杉醇生物合成的中间代谢模型奠定基础。     

绞股蓝属三种植物的POD、EST和SOD同工酶研究

采用聚丙烯酰胺凝胶电泳法对绞股蓝属的五柱绞股蓝、广西绞股蓝和绞股蓝3种植物的POD、EST和SOD同工酶进行了比较研究。结果表明,3种绞股蓝的3种同工酶谱带不同。3种同工酶在广西绞股蓝上表现最强。3种同工酶在同一物种不同器官中的表达具有一定差异,以叶中的表达最强,茎次之,根最弱。由同工酶推断的3种绞股蓝的亲缘关系为绞股蓝与广西绞股蓝亲缘关系较近,与五柱绞股蓝亲缘关系较远。     

Characterisation of phenol oxidase and peroxidase from maize silk

Silk of some maize genotypes contains a high level of phenolics that undergo enzymatic oxidation to form quinones, which condense among themselves or with proteins to form brown pigments. Two phenolic oxidizing enzymes, peroxidase (POD; EC 1.11.1.7) and polyphenol oxidase (PPO; EC 1.10.3.1), from maize (Zea mays L.) silk were characterised with respect to their preferred substrate, different isoforms and specific effectors. One browning silk sample with high, and two non‐browning samples with low phenolic content were investigated. Although POD oxidizes a wide range of phenolic substrates in vitro, its activity rate was independent of silk phenolic content. PPO activity, detected with o‐diphenolic substrates, was abundant only in browning silk, and low or absent in non‐browning silk. Pollination increased POD but not PPO activity. Isoelectric‐focusing (IEF) and specific staining for POD and PPO showed a high degree of polymorphism that varied with silk origin. The IEF pattern of POD revealed a number of anionic and several cationic isoenzymes, with the most pronounced having neutral pI 7 and a basic isoform with pI 10. Detected isoforms of PPO were anionic, except for one neutral form found only in browning silk, and occupied positions different from those of POD. Different inhibitory effects of NaN₃, EDTA, KCN, and L‐cysteine, as well as different impacts of a variety of cations on the oxidation of chlorogenic acid, mediated by PPO or POD, were detected. The findings are discussed in terms of a possible roles of these enzymes in defence and pollination.     

橄榄果实中PPO和POD活性抑制研究

本文研究温度和几种抑制剂对橄榄果实多酚氧化酶(PPO)和过氧化物酶(POD)活性的影响。结果表明,橄榄果实PPO活性最强的温度为27℃,POD活性最强的温度为45℃;低温对PPO活性的抑制较POD显著;抗坏血酸和亚硫酸钠可有效地抑制PPO活性,而抗坏血酸和氯化镁则对POD活性有明显的抑制作用。     

Effects of putrescine and ethephon on some oxidative stress enzyme activities and proline content in salt stressed spinach leaves

The effects of putrescine and ethephon on peroxidase (POD; EC 1.11.1.7), polyphenol oxidase (PPO; EC 1.14.18.1), catalase (CAT; EC 1.11.1.6) activities and proline content in spinach leaves under saline stress were investigated. In control conditions, putrescine increased PPO and CAT activities and proline content, but decreased POD activity. Ethephon increased these three enzyme activities but did not affect proline content. In saline conditions, putrescine increased POD and CAT activities and proline content, while it decreased PPO activity. Ethephon increased both PPO and CAT activities and proline content, but decreased POD activity. Putrescine and ethephon have opposite effects on the enzyme activities and proline accumulation because they acts as antagonists.