玉米花粉形成过程不同阶段特异蛋白变化规律及酯酶同工酶研究

利用薄层胶电泳技术对玉米花粉形成过程不同阶段的酯酶同工酶及可溶性蛋白种类和含量变化进行了研究。结果表明在玉米花粉形成过程中的不同阶段酯酶同工酶的种类和活性均有不同。四分体时期出现两种小分子量酯酶同工酶到小孢子阶段即很快消失,早期小孢子阶段酯酶同工活性及种类达到高峰,以后则逐步递减。玉米花粉可溶性蛋白电泳显示玉米花粉形成过程中不同发育时期蛋白类和含量有很大变化。四分体时期存在有特异性蛋白（ＴＰ）,它     

菊花不同生长阶段不同器官POD和EST同工酶比较

采用过氧化物酶(POD)、酯酶(EST)2个酶系统的12个同工酶位点,分析了4个菊花品种营养生长和生殖生长阶段不同器官(嫩叶、老叶、嫩茎、木质化茎)的同工酶变化.结果表明:(1)4个品种共有16种POD酶带,15种EST酶带;(2)菊花的POD和EST具有组织特异性和阶段特异性,其中以嫩叶的酶带最多,其次为老叶,再次为嫩茎,而木质化茎的酶带最少;(3)与生殖生长阶段相比,营养生长阶段的POD酶带更清晰,更整齐,分离更好,但生殖生长阶段的EST同工酶比营养生长阶段的更清晰;(4)营养生长阶段的嫩叶最适合用于菊花POD同工酶分析,而EST同工酶研究宜取生殖生长时期的嫩叶.     

罗汉果雌雄株同工酶性别鉴定研究

采用电泳技术结合同工酶染色,分析了罗汉果雌雄株叶片的过氧化物酶同工酶、酯酶同工酶、超氧化物歧化酶同工酶、多酚氧化酶同工酶和过氧化氢酶同工酶。结果表明:罗汉果雌雄叶片在同工酶谱上,存在着与性别性状相关的酶带;雌雄间的差异酶带在每一种同工酶中均有一条以上,可作为罗汉果雌雄株间的性别鉴定。此外,还比较了高产、低产、不结果雌株之间同工酶的酶带和活性差别。     

Isozyme Profile During Somatic Embryogenesis and In Vitro Flowering of Bambusa vulgaris

Peroxidase and esterase isozymes were investigated during plant regeneration via somatic embryogenesis in Bambusa vulgaris, The transition of non-embryogenic calli to embryogenic calli, somatic embryo development, germination and subsequent flowering of somatic embryo derived shoots were associated with selective expression or repression of isoforms of peroxidase and esterase. Non-embryogenic callus showed six peroxidase and four esterase bands. During somatic embryogenesis and germination of somatic embryos, some bands were suppressed and new isoforms of peroxidase and esterase appeared. During flowering, in addition to four peroxidase bands, a new unique esterase band ‘a’ appeared. Each developmental stage was thus associated with a definite isozyme profile.     

苜蓿组织培养中球形胚发生时特异蛋白质和同工酶分析

试验在苜蓿组织培养中,对球形胚形成过程中特异蛋白质表达的模式、过氧化物酶及酯酶同工酶酶谱变化进行研究,结果表明：苜蓿组织培养中从胚性愈伤组织到球形胚发育的进程中,顺序消失和出现了11种中小分子量多肽;过氧化物酶同工酶酶谱发生了显著的变化;酯酶同工酶酶谱变化不大,但其总活力对于维持体细胞胚胎发生是必须的。     

Selection of higher regenerative callus and change in isozyme pattern in rice (Oryza sativa L.)

Summary Calli were initiated from seedling roots in rice (Oryza sativa L. var. Tadukan) and subcultured at 45-day intervals on MS medium supplemented with 2 mg/l 2,4-D. Sectors of callus which differentiated shoot meristems (green spots) under the same 2,4-D concentration were selected from the calli subcultured 90 days after initiation. The selection was continued for about 2 years. Responses to 2,4-D between original and selected lines differed considerably, although differentiation was not generally seen in rice callus in the presence of 2 mg/l 2,4-D. After 180 days, calli of the selected line differentiated into numerous shoot-bud primordia and grew out new callus tissues under 2 mg/l 2,4-D concentration; the frequency of the differentiation exceeded 90%. On the other hand, no calli of non-selected line differentiated into shootbuds under 2 mg/l 2,4-D, and the frequency of the shootbud was only about 50% under lower 2,4-D concentration (0.1 mg/l). The pattern and activity of peroxidase isozyme varied markedly between calli of the selected and non-selected lines. First, two strong peroxidase bands which show fast mobility and one intermediate peroxidase band with slow mobility were detected only in the calli of selected line. Secondly, changes in band pattern of proteins separated by SDS-PAGE were observed. In the calli of selected line, there was a loss of the polypeptide bands with molecular weight of 24 and 42 K in the selected calli, but they were clearly present in the unselected line. The appearance of new peroxidase isozyme bands and loss of polypeptide bands, change in response to auxin and increased ability for shoot bud differentiation are closely correlated to each other.     

N+离子注入板栗生物学效应研究

本文用不同能量和剂量的N+离子注入板栗的冬芽,对其嫁接成活率、叶片面积和新梢最大长度等生物学性状进行调查,采用聚丙烯酰胺凝胶电泳技术进行酯酶和过氧化物同工酶电泳分析.结果表明离子注入促进生长,提高了板栗的经济性状,引起酯酶和过氧化物酶在数量、种类和活性上的变化.通过综合比较,发现经30keV能量、20×1015 ion/cm2剂量处理的"蜜蜂球"和25keV能量、20×1015 ion/cm2剂量处理的"粘底板"各有一变异株,并将该剂量作为板栗品种改良的最佳注入剂量.     

Isozyme studies in Indian mustard (Brassica juncea L.)

Summary A comparative study of peroxidase and esterase isozymes was carried out at five developmental stages of siliqua in order to characterize twelve genotypes of Indian mustard. The studies showed nearly the same number of isozyme bands at every stage for peroxidase and a varying number of isozyme bands for esterase. The appearance and disappearance of bands, along with their intensity scores, indicated the role of different isozymes at different stages of siliqua development. It has been ascertained that these patterns, especially the intensity scores, can be successfully used to characterize different Indian mustard genotypes.     

玉米酯酶同工酶和过氧化物酶同工酶的分子量研究

本试验利用聚丙烯酰胺凝胶梯度电泳分步染色法直接对玉米苗期酯酶同工酶和过氧化物酶同工酶各酶带的分子量进行了比较测定。酯酶同工酶 E_1、E_2、E_3~F、E_3~S、a、b、c 各酶带的分子量分别为<20000,35200、33000、38500、29900、28500、34000道尔顿过氧化物酶同工酶 PX_4~F和 PX_4~S酶带的分子量分别为131000和149000道尔顿。根据酶带在均匀胶和梯度胶中的位置变化对各酶带的生化性质作了初步分析,发现 E_3~F和 E_3~S、PX_4~F 和 PX_4~S 在迁移率上的差异主要是分子量的差异。本文为同工酶的分子量测定提供了一个简便的方法。     

羊肚菌氨基酸含量变化及酯酶同工酶研究

本文对羊肚菌氨基酸含量及酯酶同工酶进行了研究,结果表明不同生长时期菌丝体的氨基酸含量变化较大,在4一10天内逐渐升高,15天达到高峰,尔后又有所降低。与此同时酯酶同工酶活性亦呈现相应的变化规律,由此可推测羊肚菌菌丝体生长过程中酯酶同工酶活性跟氨基酸代谢之间存在一定相关性。同工酶分析还发现不同子实体菌株及同一子实体不同单孢菌株间的酯酶同工酶谱基本相同,子实体与菌丝体均具有稳定的9条酶带。     

Isoenzyme Polymorphism in Flowering Plants I. Diffusion of Enzymes Out of Intact Pollen Grains

Esterases, leucine aminopeptidases, catalases, amylases and acid phosphatases diffuse out of intact and ungerminated pollen grains of Oenothera organensis, whether suspended in 1 % sodium chloride or in pollen medium. A total of 15 esterase isozymes are recorded; 5 of them appear within 5 minutes, 8 within 30 minutes, 9 within 2 hours, and 13 within 19 hours. Pollen grains suspended for 19 hours gave much stronger isozyme bands than macerated pollen grains. However, one esterase hand was consistently missing from the 19 hour suspensions, although present in all others. It is suggested as a working hypothesis that the early growth of pollen tubes and possibly even germination of pollen grains may be influenced by the metabolic products of pistillate tissues caused by the diffused pollen enzymes, and that inactivation of these enzymes by stigmatic or stylar components could lead to incompatibility reactions.     

Esterase Isozyme analyses of the Species and Their Varieties of the Genus Gossypium

The isozyme make up of esterases of the seeds from fifteen species and twenty-three cultivar of Gossypium was analyzed by isoelectrofocusing. The experimental results are summarized as follows: 1. Differeces were observed in the number of esterase isozyme bands among the species of different genome groups. The cultivated species, G. hirsutum (AD)1 gave rise to 46 isozyme bands, the most among the species of the genus Gossypium. G. barba- dense (AD)2, G. arboreum (A2) and G. herbaceum (A1) gave rise to 42, 40 and 38 bands, respectively. In wild species, G. australe (C3) had 20 esterase bands, the least in all species of Gossypium. The bands given rise from other wild species ranged from 26 to 40. 2. Each species of every genome groups had its marker bands. The results were in agreement with the traditional classification and provided some biochemical evidence for modern classification of Gossypium. 3. It was clear that all cotton species of different genome groups contain 5 main isozyme bands, viz. PI=3.85, 4.61, 5.48, 5.73 and 5.91 in the zymograms. In other words, these zymograms are common characters of Gossypium. 4. The esterase of 23 cultivers in four cultivated species studied showed that no variation in isozyme patterns existed within one species, except the disease-resistant variety Hea-7124 which differs from other 4 cultivars of G. barbadense.     

Studies on the Molecular Weights of Esterase and Peroxidase Isozymes of Maize

The molecular weights of esterase and peroxidase isozymes of maize seedlings were directly determined by improved polyacrylamide gradient gel electrophoresis. The different isozyme bands developed in polyacrylamide slab gel electrophoresis (uniform gel) were identified in polyacrylamide gradient gel electrophoresis by means of isozyme variants. The molecular weights of esterase isozymes E1, E2, E3F, E3S, a, b, c, named according to isozyme patterns in uniform gel, are <20000, 35200, 33000, 38500, 29900, 28500, 34000 doltons respectively. The molecular weights of peroxidase isozymes PX4F and PX4S are 131000 and 149000 doltons respectively. According to the band location in uniform gel and in gradient gel, some biochemical properties of the isozyme bands and relationships between the isozyme bands were analyzed. The possible errors in the determination of smaller molecular weight isozymes are discussed.     

The Resumption of Cambial Activity and the Change of Peroxidase Isozymes in Broussonetia papyrifera

Five Broussonetia papyrifera (L.) Vent. trees were selected in a natural stand located on the campus of Peking University, Beijing, China. The trees were ca. 5-6 years old, 3-4 m tall,and had diameters of about 3 cm measured 1.2 m above ground level. They were samplied at monthly intervals between January 28 and March 25, then at ten-day intervals between March 25 and May 20,1991. On each occasion, one 3-year-old shoot was cut from the tree. Two blocks (about 1 cm ×1 cm) contained peridern,phloem,cambium and wood with more than one annual ring were cut from every shoot,fixed in FAA,and then were prepared for anatomical studies. And on each occasion,7 layers of tissues (from periderm to mature xylem)were scraped off from the shoots and 100 mg of separate tissues were randomly extracted in 0.1 ml of 20% sucrose. The extracts were used for isoelectric-focusing in polyacrylamide gel slabs (85 mm × 60 mm × 1 mm). Benziding and odianisidine was used as substrate. After electrophoresis the gel slabs were placed in the substrate buffer until the isozyme bands were visible. Owing to the ring-porous structure of the wood of Broussonetia papyrifera, the cambial activity was comparable with that in the most ring-porous dicots. The cambium activity started about ten days before bud sprouting. On April 4,the dormant cambial zone consisted of ca. 4 cell layers. The trees did not sprout until April 16,but ca. 2 cell layers of immature xylem and phloem were formed concomitantly. Ten days later, 8-9 cell layers of xylem and ca. 5 cell layers of phloem were formed. The formation of immature phloem cells continued to increase slowly between April 4 and May 20, whereas that of immature xylem cells increased rapidly between April 4 and April 26,and then decreased between April 26 and May 20. It was suggested that differentiation of immature xylem into mature xylem lasted ca. 10 days,whereas that of immature phloem into mature one lasted ca. 20 days. There were totally 6 peroxidase isozyme bands in dormant cambial region and functional phloem. Variation of zymogram in cambial region occurred before cambial activity activated which is followed by more or less minor changes of bands in all other tissues. These indicated that several significant changes were related to the level of endogenous IAA and differentiation of vascular tissues.     

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.     

以靛酚乙酸酯为底物的酯酶同工酶显色方法的研究

建立一种以靛酚乙酸酯为底物的酯酶同工酶的显色新方法。酯酶样品的聚丙烯酰胺凝胶电泳（PAGE）凝胶用磷酸缓冲液漂洗约10min后,浸入含有0.002%靛酚乙酸酯的溶液显色5～10min,可显出清晰的蓝色酯酶带。先将酯酶凝胶板浸于有机磷农药溶液中,然后再用靛酚乙酸酯显色液显色,比较同工酶谱,从同工酶带由深蓝色变为浅蓝色的颜色变化,可以看出对有机磷农药敏感的同工酶所受到的抑制程度。     

Electrophoretic variation in esterases and peroxidases of native greek diploidAegilops species(Ae. caudata andAe. comosa,Poaceae)

The esterase and peroxidase patterns in five varieties ofAegilops caudata (genome type C) andAe. comosa (genome type M) were studied in order to elucidate the phylogenetic relationships within and between the two groups. The electrostarch gel electrophoresis technique was applied to extracts of shoot and root of 4-day-old seedlings, and the electropherograms were evaluated by gel densitometer traces. Inspite of considerable isozyme polymorphism, closer relationships in the banding patterns were found between different varieties of a single species than between varieties of the two different species. Esterase and peroxidase patterns of the twoAe. caudata varieties (caudata andpolyathera) are very similar and prove their close phylogenetic relationship. The isozyme affinities withinAe. comosa varieties are illustrated by the seriessubventricosa—biaristata—thessalica. The latter endemic variety has quite a number of characteristic bands and is relatively isolated. Altogether, the electrophoretic data agree well with morphological and cytological similarities (Zhukovsky 1928,Eig 1929,Karataglis 1973, 1975b).     

金针菇品系间酯酶同工酶标记筛选研究

采用聚丙烯酰胺凝胶垂直板状电泳,研究了不同生长发育期,不同组织对金针菇酯酶同工酶电泳表型的影响,筛选出不受生长发育期及常规培养条件等影响的酯酶标记区带。标记区带分所有品系的出现的基本带和部分品系出现的识别带。酯酶同工酶标记区带电泳表型显示出多态性。     

EFFECTS OF X IRRADIATION AND INDOLEACETIC ACID ON SPECIFIC PEROXIDASE ISOZYMES IN PITH TISSUE OF A NICOTIANA AMPHIPLOID

Two electrophoretically anodal peroxidase bands, designated A3 and A4, which normally develop in zymograms of incubated pith tissue of a Nicotiana amphiploid, were inhibited by the inclusion of 0.10 mg indoleacetic acid (IAA) in a liter of culture medium. The visibility of the bands was not inhibited, however, if the pith was irradiated or if unirradiated pith blocks were incubated in either irradiated IAA-containing media or in media supplemented with irradiated IAA. Irradiation inactivates IAA in media, and according to this isozyme test, damages the capability in irradiated tissue of using exogenously supplied IAA. Radiationinduced damage to IAA metabolism caused radiation-induced peroxidase stimulation, shown in the more rapid appearance of isozyme bands A3 and A4.     

荞麦属种质资源发芽种子酯酶同工酶研究

用聚丙烯酰胺凝胶电泳对荞麦属8个种(含大粒组7个和小粒组1个)33份材料发芽种子的酯酶同工酶进行了研究。结果表明:酯酶同工酶不同酶带合计22条,不同物种的酶带数4到8条。其中,甜荞有8条带,苦荞为7条。酶带及聚类分析表明,大粒组荞麦种的谱带与细野荞等小粒组荞麦种间差异极大,甜荞和苦荞酶带分别与大野荞和毛野荞相似,并分别与F.megaspartanium和F.pilus聚类最近,支持F.megaspartanium和F.pilus可能分别是甜荞和苦荞祖先种的假说。