金鱼草愈伤组织过氧化物酶活性及其同工酶变化

在不同激素比例的MS培养基上 ,金鱼草 (Antirrhinummajus)茎愈伤组织主要有三种分化状态 :a愈伤组织不分化 ;b以器官发生途径分化出不定芽和不定根 ;c通过胚胎发生途径形成胚状体。过氧化物酶活性与愈伤组织分化程度呈正相关。未分化愈伤组织与分化芽、分胚状体及分化芽和根的愈伤组织酶活性呈线性递增关系 ,相对酶活力为 1:3:5 :6。在其同工酶谱中 ,三种状态的愈伤组织都具有酶带I和II,表明这两种同工酶与细胞的增殖生长有联系。分化的愈伤组织比未分化愈伤组织多出现酶带Ⅲ、Ⅳ和Ⅴ ,说明这三条酶带是影响分化的特异蛋白质。在分化芽的愈伤组织中 ,还存在酶带Ⅳ ,说明它是催化器官发生途径的特有同工酶。     

金鱼草愈伤组织过氧化的酶活性及其同工酶变化

在不同激素比例的MS培养基上,金鱼草（Antirrhinum majus）茎愈伤组织主要有三种分化状态：a愈伤组织不分化;b以器官发生途径分化出不定芽和不定根;c通过胚胎发生途径形成胚状体。过氧化物酶活性与愈伤组织分化程度呈正相关。未分化愈合组织与分化芽,分胚状体及分化芽和根的愈伤组织酶活性呈线性递增关系,相对酶活力为1：3：5：6。在其同工酶谱中,三种状态的愈伤组织都具有酶带和Ⅰ和Ⅱ,表明这两种同工酶与细胞的增殖生长有联系。分化的愈伤组织经未分化愈伤组织多出现酶带Ⅲ,Ⅳ和Ⅴ,说明这三条酶带是影响分化的特异蛋白质。在分化芽的愈伤组织中,还存在酶带Ⅳ,说明它是催化器官发生途径的特有同工酶。     

荞麦愈伤组织分化与过氧化物酶活性及其同工酶关系的研究

在附加不同外源激素的培养基中,龙山荞的子叶愈伤组织表现出不同的分化状态:(1)不分化;(2)分化根;(3)分化芽;(4)分化全苗。四种愈伤组织的过氧化物酶活性的相对比值为1:1.47:5.95:7.78,并且过氧化物酶同工酶谱也存在明显差异。酶带4'为分化愈伤组织所特有;酶带2'为未分化和分化愈伤组织所共有;在分化根和未分化愈伤组织中缺少酶带1';未分化愈伤组织比分化愈伤组织多出酶带3'。     

荞麦愈伤组织分化与过氧化物酶活性及其同工酶关系的研究

在附加不同外源激素的培养基中,龙山荞的子叶愈伤组织表现出不同的分化状态：（１）不分化;（２）分化根;（３）分化芽;（４）分化全苗。四种愈伤组织的过氧化物酶活性的相对幽会为１：１．４７：５．９５：７．７８,并且过氧化物酶同工酶谱也存在明显差异。酶带４’为分化愈伤组织所特有;酶带２’为未分化和分化愈伤组织所共有;在分化根和未分化愈伤组织中缺少酶带１’;未分化愈伤组织比分化愈伤组织多出酶带３’。     

吸胀对水稻种子愈伤组织诱导及其性状的影响

水稻种子经浸种吸胀处理后,浸种介质的pH下降,种子细胞透性增加;种子胚芽生长加快,种子愈伤组织诱导和生长均受抑制。种子可溶性蛋白谱和愈伤组织过氧化物酶同工酶谱亦发生变化,对所诱导的种子愈伤组织则有促进分化的后效应。此种后效应因品种及所采用的诱导培养基成分不同而异。     

芦苇胚性愈伤组织的形成及植株的再生

以芦苇种子为外植体,其愈伤组织的诱导率最高。叶鞘和叶片不发生脱分化。培养基中最合适的蔗糖浓度为4%。维生素 B 类、肌醇对愈伤组织的生长起促进作用。而酵母提取物对愈伤组织的诱导和生长具有明显的抑制作用。这种抑制效应,将随酵母提取物浓度的提高而增大。愈伤组织的继代培养,随培养基中2,4-D 浓度的提高,其平均鲜重明显降低。脱分化培养基中2,4-D 浓度对胚性愈伤组织的诱导形成具有一定的相关性。胚性愈伤组织经30代继代培养依然具有90%的分化频率,只是每块愈伤组织的分化苗数减少。反之,非胚性愈伤组织则完全丧失形态发生的能力。对两类愈伤组织进行扫描电镜的观察,发现其表面结构有很大差异。其过氧化物酶、酯酶同工酶谱以及可溶性蛋白的含量均有明显的差别。     

红掌遗传转化受体系统的建立Ⅰ离体高效培养体系的建立

以红掌叶片和叶柄为材料,研究了影响红掌愈伤组织的诱导和分化的因素.结果表明,基因型对愈伤组织诱导有显著的影响,Pink Champion愈伤组织诱导率最高,为90%.基本培养基对愈伤组织诱导影响显著.在改良MS培养基上,愈伤组织诱导率为85%,出愈伤多,质量高.消毒时间和接种方式对愈伤组织诱导率亦有显著影响.初展开叶片用氯化汞消毒8-10min,背面向下接入培养基,愈伤组织诱导率高.外源激素对愈伤组织的诱导和芽分化影响显著.单独使用BA不能诱导红掌叶片产生愈伤组织,高浓度BA、低浓度2,4-D时,愈伤组织诱导率高.在MS+BA0.5 mg·L-1+NAA 0.5 mg·L-1+CM 5%培养基上,芽分化率为97.5%,平均芽分化数为4.5个/块,芽粗壮.将分化出的芽转入1/2 MS+NAA 0.2 mg·L-1+AC 1 g·L-1培养基上诱导生根,生根率达100%.通过上述研究建立了红掌离体高效培养系统.     

资源植物罗布麻的愈伤组织诱导和植株再生研究

以罗布麻（Apocynum venetum L.）无菌苗的叶、茎和根作为外植体,在不同激素与浓度组合的MS培养基上进行愈伤组织诱导和植株再生研究。结果表明,在多个激素浓度配比的培养基中,罗布麻叶和茎的愈伤组织诱导率均可达到100%;但进一步将愈伤组织转接到含有不同激素组合的培养基进行不定芽分化时,叶和茎来源的愈伤组织的不定芽分化率有很大不同,茎来源的愈伤组织虽可在多个培养基中有不定芽分化,但最高诱导率仅达20%;而叶片来源的愈伤组织虽仅有4个激素组合培养基中有不定芽的分化,但最高分化率可达到35%。因此,构建罗布麻再生体系的最佳外植体应选择叶片。愈伤组织诱导与不定芽分化的最佳培养基均为：MS＋NAA0.1 mg/L＋6-BA 1.0 mg/L;不定芽在1/2MS＋NAA 0.2 mg/L生根培养基的生根效果最佳,不仅根群质量好,且生根率也高达100%;此再生苗的移栽成活率也最高,在适宜条件下可达75%。     

尾叶桉的不同类型愈伤组织芽分化与某些相关生理生化指标的关系

切取8d苗龄的尾叶桉幼苗下胚轴,培养7d后愈伤组织的诱导率达100%。8周后,愈伤组织分化成褐色、白色和浅绿色3种类型。其中只有浅绿色愈伤组织能分化出不定芽,分化率达57%。浅绿色愈伤组织的过氧化物酶（POD）、超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和多酚氧化酶（PPO）的活性最高。除SOD外,浅绿色愈伤组织的POD、CAT和PPO活性与其他两类愈伤组织相比具有显著性差异（P〈0.05）。H2O2和丙二醛含量则是褐色愈伤组织的最高。     

尾巨桉不同类型愈伤组织抗性相关酶活性差异与不定芽分化关系研究

本实验以尾巨桉无性系32-29无菌苗的茎段为外植体,应用不同植物生物调节剂处理,诱导愈伤组织形成,分别测量4种类型愈伤组织的蛋白质含量及过氧化物酶(POD)、多酚氧化酶(PPO)、超氧化物歧化酶(SOD)等保护酶的指标活性,比较不同愈伤组织的酶活性强弱与不定芽分化的关系。结果表明:不同处理间愈伤组织诱导率差异达到极显著水平,且不同愈伤组织蛋白质含量及3种酶活力差异明显,POD、SOD活力高的愈伤组织不定芽诱导率也高,而PPO含量高的愈伤组织诱芽率低。     

4D和4R对小麦三种同工酶合成的影响

利用聚丙烯酰胺凝胶电泳对小麦品种天选15号、天选15号—4D缺体,黑麦品种德国白粒及利用“缺体回交法”培育的小麦(4D)一黑麦(4R)异代换系幼苗的过氧化物酶同工酶、细胞色素氧化酶同工酶及酯酶同工酶进行了研究,结果表明4D染色体对小麦的一种过氧化物酶同工酶和一种细胞色素氧化酶同工酶量的合成具有控制作用,在小麦4D缺体的遗传背景下,黑麦4R染色体能够补偿由于4D缺失引起的这两种同工酶合成降低的效应。4D对小麦幼苗期酯酶同工酶的合成没有明显的作用,4R在小麦4D缺体遗传背景下对酯酶同工酶的合成也没有明显的影响     

不同波长的激光对水稻胚芽中可溶性蛋白质及同工酶的作用

用波长0.3371、0.6328μm和1.06μm的激光器,用不同剂量(5、15、50min和3、7、9个脉冲),照射水稻干胚,使其萌动生长72hr后,再用聚丙烯酰胺凝胶电泳法,分析胚芽中可溶性蛋白质,酯酶同工酶和过氧化物酶同工酶。结果证明,波长0.3371～1.06μm的激光辐照水稻种胚,都能使同一生物大分子的活性和分子结构产生变异。由此认为,不能用辐照离体制备液所得结果去预言在活体上的反应。     

Isozyme patterns in callus cultures and in plants regenerated from calli ofCereus peruvianus (Cactaceae)

Electrophoretic patterns for isocitrate dehydrogenase (IDH; EC 1.1.1.42), acid phosphatase (ACP; EC 3.1.3.2), peroxidase (PER; EC 1.11.1.7), and esterase (EST; EC 3.1.1.1) isozymes were determined inCereus peruvianus tissues and used as markers of genetic uniformity of calli and of the plants regenerated from callus cultures. One IDH, six ACP, six PER, and six EST isozymes were induced in cultured callus tissues in medium containing three 2,4-dichlorophenoxyacetic acid and kinetin combinations. Four ACP, two PER, and three EST isozymes were still present in all regenerated plantsin vitro and therefore can be used as markers of theC. peruvianus plants regenerated from callus tissues. The differential patterns of ACP and IDH isozymes and the similar zymograms for PER and EST isozymes presented by callus tissues were used in a comparison of callus tissues cultured for 2 years. The comparative analysis of zymograms within each enzyme system indicated a mean heterogeneity coefficient of 0.33 forC. peruvianus calli cultured for 2 years. Because of the isozyme variations, which developed in culture medium and were transferred to the regenerated plants, the IDH, ACP, PER, and EST enzyme systems can be considered to be good markers for investigating possible genetic variations in plant populations ofC. peruvianus obtainedin vitro from callus culture.This research was supported by the CNPq     

家蝇幼虫同工酶研究

本通过聚丙烯酰凝胶电泳法对家蝇幼虫体内的过氧化物酶、乳酸脱氢酶、苹果酸脱氢酶、酯酶和超氧化物歧化酶,以及对超氧化物歧化酶的特性进行了研究。实验结果为家蝇幼虫资源开发利用提供了理论依据。     

瘿螨总科科间亲缘关系的初步研究

以聚丙烯酰胺凝胶电泳,测定了瘿螨总科中分属3个科的5种瘿螨的酯酶同工酶,根据电泳谱带和所测5种瘿螨的形态特征,计算出欧氏距离和相似系数,绘制谱系图,表明它们的亲缘关系顺序为:桧三毛瘿螨,金钱松博氏瘿螨,雪柳顶冠瘿螨,女贞刺瘿螨和金银木大嘴瘿螨。其中以雪柳顶冠瘿螨和女贞刺瘿螨的亲缘关系最近,桧三毛瘿螨和金钱松博氏瘿螨亦属较近缘的两个种,金银木大嘴瘿螨则独立于外。这种亲缘关系,构成了纳氏瘿螨科→瘿螨科→大嘴瘿螨科的科间进化关系。     

Alcohol dehydrogenase (EC 1.1.1.1) isozymes as markers at 2,4-dichlorophenoxyacetic acid × kinetin combinations in callus cultures ofCereus peruvianus (Cactaceae)

Alcohol dehydrogenase (ADH; EC 1.1.1.1) isozymes were investigated in tissue ofCereus peruvianus cultured in different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin. Five ADH isozymes were detected in starch gel and showed different patterns in seeds, seedlings, calli cultured at 32 and 22°C, and plants regenerated from calli cultured in three 2,4-D and kinetin combinations. Four phenotypes formed by different combinations of ADH-2, ADH-3, ADH-4, and ADH-5 were detected in calli cultured at 32°C and in plants regenerated from calli. ADH-1 isozyme was detected only in calli subcultured for 1 or 2 weeks at 22°C and was indicated as a marker of stress conditions that affect the growth ofC. peruvianus callus tissues in culture. ADH phenotypes with either a higher or a lower number of isozymes were detected in different proportions in the callus tissues cultured in media containing different 2,4-D and kinetin ratios. ADH isozyme patterns were found to be sensitive markers at the highest kinetin concentration or at high kinetin/2,4-D ratios. The results indicate a high correlation between the ADH isozyme patterns and the capacity for regeneration. Thus, ADH isozymes are indicated as good biochemical markers and as a powerful tool for monitoring studies ofC. peruvianus callus cultures.This research was supported by the CNPq.     

Malate dehydrogenase (MDH; EC 1.1.1.37) isozymes in tissues and callus cultures ofCereus peruvianus (cactaceae)

Malate dehydrogenase (MDH; EC 1.1.1.37) isozymes were investigated in seeds and in seedlings and calli cultures ofC. peruvianus to determine if the changes in MDH isozyme banding patterns could be used as biochemical markers to identify the origin of regenerated plants from callus tissues. Four cytoplasmic MDH isozymes (sMDH), five mitochondrial MDH isozymes (mMDH), and one glyoxysomal MDH isozyme (gMDH) were detected and showed tissue- and stage-specific expression. A relationship of mMDH and gMDH isozyme patterns with callus tissues subcultured in three hormonal combinations and with the plants regenerated from these callus tissues was demonstrated. Furthermore, temperature and mechanical stress were found to be closely related to mMDH-1 activity in callus culture. Therefore, the different patterns of MDH isozymes in the various tissues ofC. peruvianus can be used as biochemical markers for the study of gene expression during development and as powerful tools in monitoring studies on callus cultures. This research was supported by the CNPq.     

团头鲂的胚胎及成体组织中八种同工酶系统的研究

用垂直的淀粉凝胶电泳方法分析了胚胎发育阶段(0—105小时)和成体6种组织中的乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)、谷氨酸脱氢酶(GDH)、葡萄糖—6—磷酸脱氢酶(G6PD)、醇脱氢酶(ADH)、异柠檬酸脱氢酶(IDH)、酯酶(EST)和碱性磷酸酶(AKP)等8种同工酶系统的酶带。共约有23个基因座位在其胚胎发育期和成体组织中表达。所分析的大多数同工酶在团头鲂个体发生过程中表达的情况大致可分为3种类型：①胚胎发育期间持续存在的同工酶类,它们在成体组织中无特异性分布;②到胚胎发育后期才开始表达的同工酶类,它们往往和特定的组织或器官的形态发生或机能分化紧密相关,并在成体组织中呈特异性分布;③在成体组织中都没有被发现而只在胚胎发育期所特有的同工酶。团头鲂的MDH同工酶类之间以及它们和G6PD同工酶活性变化之间存在着相关性,二者可能存在共同的调控机制。与许多其他鱼类不同,团头鲂的GDH同工酶在整个胚胎发育过程中都有活性,但在其成体组织中无特异性分布。基因调控系统的时空精确性是保证团头鲂胚胎发育正常代谢活动的必要条件。     

激光对香石竹愈伤组织蛋白质和同工酶谱的影响

用氦氖激光处理香石竹愈伤组织,可使材料中的34kD和28kD蛋白含量高于对照,并产生分子量约为22kD的新的蛋白带。而用氩离子激光处理后,则出现一条分子量约为45kD的新的蛋白带。此外,激光处理亦明显影响愈伤组织的过氧化物酶同工酶谱及其活性,其中氦氖激光处理后产生了Rf为0.17和0.26的新酶带,而用氩离子激光处理则只产生Rf为0.17的新酶带。激光对香石竹愈伤组织的酯酶同工酶无影响。     

Isozyme modifications during morphogenesis of callus from barley and wheat

Callus cultures fron non-organogenic, young and one-year old, and morphogenic calli were used to assess the value of isozymes analysis for the prediction of morphogenic capacity by studying esterase, peroxidase and acid phosphatase. Basic isozyme patterns of each enzyme for the callus were retained in all the callus stages and in the callus which has differentiated into shoots. With the development of shoot and/or root some conspicuous isozymes appeared for esterase and acid phosphatase and some disappeared for peroxidase. As the isozyme changes became apparent only after shoot or root initiation these enzymes could not be used as markers to distinguish between morphogenic and non-morphogenic calli.