巴东木莲种子贮藏和萌发过程中同工酶分析

以贮藏和萌发过程中的巴东木莲种子为材料,采用非变性聚丙烯凝胶电泳技术分析其种子中淀粉酶(AMY)、酯酶(EST)、超氧化物歧化酶(SOD)、过氧化物酶(POD)同工酶酶谱,并测定其酸性磷酸酶(ACP)和POD的活性,以探讨巴东木莲种子休眠和萌发过程中的生理生化变化特征.结果表明:巴东木莲种子在贮藏和萌发过程中,EST和SOD同工酶在萌发过程中表达增强,并不断有新酶的合成;AMY同工酶在萌发初期表达强度高且酶带数较多,到后期表达水平较低,其可能启动并控制种子萌发快慢;POD同工酶在萌发后期酶的活性增强,且酶的种类也增加,与EST和AMY同工酶的变化相适应.巴东木莲种子ACP和POD活性在储藏条件下以干藏种子最低,在萌发过程中总体上随发育进程呈升高的趋势,与同工酶电泳的结果吻合.因此,EST、AMY、SOD和POD同工酶酶谱变化及表达强弱可作为巴东木莲种子萌发各阶段转变的重要标志.     

'砀山酥'梨果实发育过程中几种酶的谱带和活性变化

幼果期间,‘砀山酥’梨果实中的过氧化物酶（POD）和多酚氧化酶（PPO）的总活性高,同工酶谱带数多,而后,随着果实的生长发育而下降并趋于稳定,花后90d左右出现一条特异性POD同工酶条带;超氧化物歧化酶（SOD）的同工酶谱带和活性在整个发育过程中变化不明显。     

聚丙烯酰胺凝胶电泳配合荧光增白剂显色检测木霉几丁质酶同工酶谱

为提高木霉几丁质酶检测方法的准确性和灵敏度,建立一种快速检测几丁质酶同工酶的方法。采用活性凝胶电泳、变性凝胶电泳、原位显色凝胶电泳结合荧光增白剂（Calcofluor white M2R）显色从绿色木霉LTR-2发酵产物中检测几丁质酶同工酶。活性凝胶电泳在粗酶液浓缩5倍时显示两条活性谱带,变性凝胶电泳在浓缩10倍时显示一条活性谱带,原位显色凝胶电泳在浓缩20倍时显示两条不清晰的活性谱带,SDS-PAGE显示这两条活性谱带的分子量分别为65kDa和42kDa。结果表明活性聚丙烯酰胺凝胶电泳和Calcofluor white M2R显色相结合的方法在几丁质酶上样量为0.47U时具有较好的分辨能力,是检测木霉几丁质酶同工酶的有效的方法。     

磷胁迫诱导大豆叶片酸性磷酸酶同工酶的表达

通过田间试验对两种磷处理的274个大豆基因型叶片酸性磷酸酶活性进行筛选,并将其中8个进行营养液栽培试验以研究磷胁迫对其叶片酸性磷酸酶同工酶表达的影响.结果表明,大豆叶片酸性磷酸酶活性存在着明显的基因型差异,不施磷处理提高了大部分(约60%)供试基因型叶片酸性磷酸酶的活性.营养液栽培试验表明,低磷处理普遍提高了所有8个供试大豆基因型叶片酸性磷酸酶的活性.等电聚焦电泳结果表明,供试大豆基因型的老叶和新叶中均有6条酸性磷酸酶的同工酶带.低磷处理显著增加了叶片酸性磷酸酶酶带的活性,但是没有诱导新的酸性磷酸酶酶带产生.研究发现叶片酸性磷酸酶活性可作为反映大豆磷胁迫的酶学指标;磷胁迫诱导大豆叶片酸性磷酸酶活性的增加是由于已有同工酶活性的提高而不是由于特异性酶带的产生.     

磷胁迫诱导大豆叶片酸性磷酸酶同工酶的表达

通过田间试验对两种磷处理的274个大豆基因型叶片酸性磷酸酶活性进行筛选,行将其中8个进行营养液栽培试验以研究磷胁迫对其叶片酸性磷酸酶同工酶表达的影响。结果表明,大豆叶片酸性磷酸酶活性存在着明显的基因型差异,不施磷处理提高了大部分(约60％)供试基因型叶片酸性磷酸酶的活性。营养液栽培试验表明,低磷处理普遍提高了所有8个供试大豆基因型叶片酸性磷酸酶的活性。等电聚焦电泳结果表明,供试大豆基因型的老叶和新叶中均有6条酸性磷酸酶的同工酶带。低磷处理显著增加了叶片酸性磷酸酶酶带的活性,但是没有诱导新的酸性磷酸酶酶带产生。研究发现叶片酸性磷酸酶活性可作为反映大豆磷肋迫的酶学指标;磷胁迫诱导大豆叶片酸性磷酸酶活性的增加是由于已有同工酶活性的提高而不是由于特异性酶带的产生。     

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.     

镉、铅对蟾蜍精巢毒作用的酶学研究

为研究镉、铅对精巢的生化毒作用机理 ,应用氯化镉、硝酸铅溶液对成年雄性蟾蜍进行腹腔染毒 (按镉计 0 .1、0 .2、0 .4、0 .8mg/ kg体重 ;按铅计 1、2、4、8mg/ kg体重 ) ,连续染毒 7d后活体解剖 ,测定分析精巢中各种酶的活性。结果显示 ,在镉染毒下 ,精巢乳酸脱氢酶 (L DH)和酸性磷酸酶 (ACP)的活性随镉染毒剂量的增加而降低 ,而碱性磷酸酶 (AL P)未发现明显变化 ;在铅染毒下 ,乳酸脱氢酶 (L DH)和碱性磷酸酶 (AL P)的活性随铅染毒剂量的增加而降低 ,而酸性磷酸酶 (ACP)未发现明显变化 ;乳酸脱氢酶 (L DH)同工酶在镉、铅染毒下则主要表现为酶带 L DH1 、L DH2 的抑制或缺失和 L DH5活性的增强。因此 ,镉、铅对蟾蜍的雄性生殖毒性机理可能与酶的活性存在着一定的关系 ,L DH可以考虑作为反映镉、铅中毒对精巢功能影响程度的一种有价值的生化指标     

天麻3种变型过氧化物酶的同工酶研究

采用聚丙烯酰胺凝胶电泳（PAGE）对天麻3种变型的过氧化物酶（POD）的同工酶进行了研究。结果表明,天麻3种变型的过氧化物酶的同工酶活性不同,酶谱条带数在5～7条之间,且具有特征谱带。其中乌天麻和红天麻的酶谱条带数相同,两者的酶谱条带数均多于绿天麻;绿天麻、乌天麻和红天麻的酶谱条数依次分别为5条、7条和7条;其中基本酶带有5条,Rf值分别为0．06、0．24、0．83,0．89、0．98;乌天麻与红天麻的相似度指数为0．86,说明这两个种亲缘关系近;乌天麻与绿天麻的相似度指数为0．71,反映它们之间的亲缘关系相对较远。     

萌发前后燕麦种子内酸性磷酸酶的细胞化学研究

组织化学定位显示,在燕麦盾片和糊粉层细胞内,酸性磷酸酶是普遍地存在着的。种子萌发0,1,2天时,酶活性基本局限在细胞质内;细胞核和壁内则无酶活性。但当种子萌发至3—4天时,酶活性在胞质内增加,显示细胞内开始有新的酶产生。这些新产生出来的酶,还被分泌出细胞,进入胞壁。用组织等电点聚焦,基本上反映了与盾片和糊粉层的组织定位一致的酶变化情形。但从这两种组织所得到的同工酶带谱则并不一样。因为从糊粉层可以得到10条清晰的同工酶带(其中6条在糊粉层不清晰)。其次,胚乳的同工酶带谱,与盾片和糊粉层的同工酶带谱也是不完全一样的。     

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。     

亚麻耐盐性愈伤组织的生理生化特性

以亚麻(双亚5号)耐盐愈伤组织为材,以同种亚麻普通愈伤组织为对照,分别经含0~250 mmol·L^-1 NaC1的培养基培养20 d后,比较含水量、MDA含量、POD酶活、SOD酶活、EST同工酶谱、POD同工酶谱及SOD同工酶谱。结果表明：亚麻耐盐组愈伤组织与对照组愈伤组织在多方面存在明显差异,前者含水量均高于后者;丙二醛含量也高于后者,但变化幅度不大;POD酶活均高于对照组;酯酶同工酶谱不同于对照组,酯酶酶活量均高于对照组;POD同工酶谱比对照组多一条带,且酶活均高于对照组;SOD同工酶谱与对照组一样多,但酶活均高于对照组,且平稳。这为进一步筛选亚麻耐盐突变细胞系奠定了良好的基础。     

木耳栽培菌株酯酶同工酶的酶谱多样性研究

采用聚丙烯酰胺凝胶电泳技术对木耳（Ａｕｒｉｃｕｌａｒｉａａｕｒｉｃｕｌａ（Ｈｏｏｋ）Ｕｎｄｅｒｗ）１０个栽培菌株酯酶同工酶的酶谱多样性进行了研究。１０个供试菌株的幼龄菌丝（７ｄ）中仅检测到２５条酶带,各个菌株分别具有２～３条酶带,１０个菌株仅有２种酶谱类型;老龄菌丝（７２ｄ）中共检测到４４条酶带,各个菌株分别具有３～６条酶带,１０菌株共有８种酶谱类型。研究表明,木耳双核体菌丝中某些酯酶同工酶基因位点在一定的发育时期才开始表达,老龄菌丝酯酶同工酶酶谱在木耳菌株鉴别和遗传育种研究中具有更大的应用价值。     

Study on the Esterase Isozyme from the Related Citrus Plants

In this experiment, the polyacrylamide gel electrophoresis is adopted to test the esterase isozyme of seeds and seedling of the fourteen species of related Citrus plants. The relationship with each other is analysed according to the resutls of zymogram and scanning chart. The data show that there are differences of various degrees in the number of the isozyme bands, the enzyme activity, the migration of enzymes and their chromatogram scanning. The isozyme of the same species has different forms of expression in the different period of development. The esterase isozyme zymogram of the Citrus seed is generally stable. It can be used as the biochemical and genetical index to detemine the genetic ralationship of Citrus.     

柑桔近缘植物酯酶同工酶的研究

本文用聚丙烯酰胺凝胶电泳测定了柑桔近缘植物14个种群的种子及幼苗的酯酶同工酶,根据酶谱及扫描图的异同,分析了彼此的亲缘关系,试验结果表明,柑桔近缘植物种属间的酯酶同工酶的酶带数目,酶活性,迁移率及酶谱扫描均有不同程度的差异,同一品种不同发育时期的同工酶也具有不同表现形式,特别是柑桔种子的酯酶同工酶谱一般较稳定,可以作为柑桔亲缘关系的生化遗传指标。     

Activity of gut alkaline phosphatase,proteases and esterase in relation to diapause of pharate first instar larvae of the gypsy moth,Lymantria dispar

Two distinctly different patterns of gut enzyme activity were noted in relation to diapause in pharate first instar larvae of the gypsy moth, Lymantria dispar. Trypsin, chymotrypsin, elastase, aminopeptidase and esterase activities were low at the initiation of diapause and through the period of chilling needed to terminate diapause. At the completion of a 150 day chilling period, activity of each of these enzymes quickly increased when the pharate larvae were transferred to 25°C. By contrast, activity of alkaline phosphatase (ALP) increased rapidly at the onset of diapause, remained elevated throughout diapause, increased again during postdiapause, and then dropped at the time of hatching. In addition, zymogram patterns of ALP activity differed qualitatively in relation to diapause: several bands were detectable during the pre- and postdiapause periods, but only one band, a band of high mobility, was visible during diapause. The ALP isozyme present in diapausing pharate larvae had a pH optimum of 10.6. Diapause in the gypsy moth can be averted by application of an imidazole derivative, KK-42, and pharate larvae treated with KK-42 showed elevated protease and esterase activity, low ALP activity, and expressed ALP isozymes with low mobility. Thus the overall patterns of gut enzyme activity and the ALP zymogram in KK-42 treated individuals were similar to those observed in untreated individuals at the termination of diapause. Our results suggest a unique pattern of enzyme activity in the gut that is regulated by the diapause program. Arch. Insect Biochem. Physiol. 37:197–205, 1998. © 1998 Wiley-Liss, Inc.     

He-Ne激光对啤酒大麦麦芽同工酶基因表达的影响

采用不同时间He-Ne激光（5 m W/mm2）对＂临啤二号＂大麦种子进行辐照,利用聚丙烯酰胺凝胶电泳（PAGE）技术分析激光辐照对大麦麦芽中超氧化物歧化酶（SOD）同工酶、过氧化物（POD）同工酶、过氧化氢（CAT）同工酶、β-淀粉酶同工酶、三磷酸腺苷（ATPase）同工酶酶谱的影响。结果表明：种子经He-Ne激光辐照后诱导出1条新POD同工酶谱带,且总POD活性在60 s和90 s处有所增加;30 s激光辐照诱导出1条新的CAT同工酶谱带,其它时间处理酶的组分没有变化,但酶活性增加;He-Ne激光辐照使SOD、β-淀粉酶及ATPase同工酶的组分在不同时间下有所减少,但一定时间的激光辐照提高了其同工酶的活性。综合实验结果可知,60 s～90 s He-Ne激光可以促进大麦种子萌发过程中以上同工酶基因的表达,有利于缩短制麦周期,提高麦芽糖化力及抗氧化力。     

Enzymic Mechanism of Starch Breakdown in Germinating Rice Seeds: III. alpha-Amylase Isozymes

The formation of amylase isozymes in germinating rice (Oryza sativa) seeds was studied by isoelectric focusing on polyacrylamide gel disc electrophoresis. Time sequence comparisons of the amylase zymogram were made between extracts from gibberellic acid-treated embryoless and embryo-attached half-endosperm of rice seeds. In both cases, 4 major and 9 to 10 minor isozyme bands were detectable at the maximal stage of the enzyme induction. However, in the embryo-attached half-seeds, bands started to diminish after the 5th day of incubation, in agreement with the results of time sequence analyses of enzyme activities. Nearly identical patterns of amylase isozyme bands on a polyacrylamide gel disc electrophoresis in combination with isoelectric focusing indicate the intrinsic role of gibberellic acid in the starch breakdown in germinating rice seeds. We tentatively assign the newly synthesized enzymes to be α-amylases based on experimental results concerning the lability of the preparation on a prolonged treatment at pH 3.3 and the stability on heat treatment for 15 minutes at 70 C.     

Further studies on the genetic control of chicken plasma alkaline phosphatase isozyme.

The effects of neuraminidase treatment on the electrophoretic pattern of alkaline phosphatase (AP) isozymes and AP activity were investigated in chicken plasma. AP comprised three isozymes. The zymogram of an individual chicken plasma had two bands, either the faster (F) or the slower (S) moving band by isozyme types and the B band irrespective of isozyme types. Mobility of the S band and AP activity in chicken plasma were not affected by neuraminidase treatment. The treatment has a reduced migration rate of the F band equal to that of the S band and the B band of both types closer to the origin. The genetic control of these bands is discussed.     

Further studies on the genetic control of chicken plasma alkaline phosphatase isozyme

The effects of neuraminidase treatment on the electrophoretic pattern of alkaline phosphatase (AP) isozymes and AP activity were investigated in chicken plasma. AP comprised three isozymes. The zymogram of an individual chicken plasma had two bands, either the faster (F) or the slower (S) moving band by isozyme types and the B band irrespective of isozyme types. Mobility of the S band and AP activity in chicken plasma were not affected by neuraminidase treatment. The treatment has a reduced migration rate of the F band equal to that of the S band and the B band of both types closer to the origin. The genetic control of these bands is discussed.     

Immunochemical characterization of human red cell acid phosphatase isozymes

An immunological study was performed on human red cell acid phosphatase (ACP1) isozymes encoded by different alleles, each of which is expressed as an electrophoretically fast (f) isozyme and a slow (s) isozyme. These isozymes reacted as two immunochemically different groups. Allele-specific reactions were not detected between either the f isozymes or the s isozymes. Quantitation of ACP1 isozymes in red cells by crossed immunoelectrophoresis revealed a phenotype-dependent variation in the concentration of isozyme protein. A simple gene dosage effect was indicated and the ordering of the ACP1 alleles (ACP1*A < ACP1*B < ACP1*C < ACP1*E) was identical to that found for enzyme activity levels. Also, an allele effect on the proportion between s and f isozymes (s/f) was observed; the ordering here was ACP1* B < ACP1*A < ACP1*, which is the same as that reported for the susceptibility to modulation with purines. These variations in isozyme protein levels appear to account for the phenotypic differences in the intensity of the isozyme bands, when activity-stained after electrophoresis, and in the red cell enzyme activity levels. Investigation of two carriers of a Null allele showed no evidence of an aberrant protein product, and half-normal concentrations of enzyme protein were observed in the red cells of these individuals.