Effects of temperature on germination and mitochondrial dehydrogenases in two soybean (Glycine max) cultivars

The effects of eight germination temperatures from 10°C to 35°C on germination and dehydrogenase activities of two soybean (Glycine max [L.] Merr.) cultivars were investigated after 48 h of seedling growth. Axis fresh weights of cv. Chippewa increased as germination temperature increased from 10°C to 35°C. In contrast, axis fresh weights for the cv. Wells increased more slowly with increasing temperature and reached a maximum at c. 25°C. In general, in vitro activities of glutamate dehydrogenase (GDH), NADP-isocitrate dehydrogenase (NADP-ICDH), and malate dehydrogenase (MDH) from the axes of cv. Chippewa correlated well with increases in axis fresh weights. GDH and MDH activities from axes of the cv. Wells also reflected increases in axis fresh weights although the correlation was not as evident as for the cv. Chippewa. NADP-ICDH activity from ‘Wells’ axes was highest at 35°C even though germination was poor at this temperature. GDH and MDH activities from cotyledons of both cultivars were not correlated with axis weight increases. No GDH activity was detected in ‘Wells’ cotyledons from seeds germinated at 35°C.     

Isozyme Diversity in Cassava Cultivars (Manihot esculenta Crantz)

Isoenzyme electrophoresis was used as a method to determine genetic diversity in various M. esculenta cultivars collected in the Southwestern (SW) and Northwestern (NW) regions of the State of Parana, in the South region of Brazil, and in cultivars produced at the Agronomic Institute of Campinas (IAC), S~ao Paulo State, Southeastern region of Brazil. The cultivars have been maintained by vegetative propagation for 5 years and are useful in production programs. A total of 28 loci in the acid phosphatase (ACP; EC 3.1.3.2), esterases (EST; EC 3.1.1.1), malate dehydrogenase (MDH; EC 1.1.1.37), and shikimate dehydrogenase (SKDH; EC 1.1.1.15) isozymes was analyzed. The proportion of polymorphic loci for NW, SW, and IAC cultivars was 57.14, 50.0, and 53.6%, respectively. Genetic diversity calculated by Nei's genetic identity (I) showed high I values for the three M. esculenta subpopulations. The high degree of polymorphism expressed by cassava cultivars is highly relevant to stimulate breeding programs with M. esculenta species.     

栽培黄瓜与野黄瓜正反杂交的几种同工酶分析

运用天冬氨酸转氨酶(AAT)、苹果酸脱氢酶(MDH)以及酯酶(EST)3种同工酶对栽培黄瓜"长春密刺"Cucumis sativus cv. Changchunmici (2n=14)与野黄瓜C. hystrix (2n=24)的正反交种间杂种F1 (正交: 野黄瓜×栽培黄瓜"长春密刺", 反交: 栽培黄瓜"长春密刺"×野黄瓜)及其双亲进行鉴定和比较研究。结果表明: 正反交种间杂种F1主要表现为双亲酶带的互补, 同时还形成4个杂合带(Aat-1-94、Aat-2-104、Mdh-3-102和Est-5-102)。上述3种酶均能准确地鉴定种间杂种的真实性。研究还发现正反交杂种F1的AAT和MDH的酶谱分别在酶带数目和强弱上表现出一定的差异, 进一步证实了野黄瓜与栽培黄瓜杂交存在正反交差异。     

Biochemical and Genetic Polymorphisms for Carboxylesterase and Acetylesterase in Grape Clones of Vitis vinifera L. (Vitaceae) Cultivars

Native polyacrylamide gel electrophoresis (PAGE) was employed to show the highest number of esterase loci and to detect alpha- and beta-esterase polymorphisms in leaf buds of Vitis vinifera cultivars. A total of 16 esterase isozymes were detected in leaf buds from 235 plants including Italia, Rubi, Benitaka, and Brasil cultivars. Biochemical characterization of the grape esterases using ester substrates revealed alpha-, beta-, and alpha/beta-esterases with inhibitor tests distinguishing both carboxylesterases (EST-2, EST-3, EST-5, EST-6, EST-7, EST-8, EST-9, EST-10, and EST-16 isozymes) and acetylesterases (EST-4, EST-11, EST-12, EST-13, EST-14, EST-15 isozymes). No allele variation for alpha-, beta-, and alpha/beta-esterases was detected; however, EST-3 alpha-carboxylesterase was absent in 61.7% of vines, and EST-4 alpha/beta-acetylesterase was absent in one vine of Rubi cv. Null EST-3 carboxylesterase phenotype (61.7%) cannot be explained in this article, but the high genetic polymorphism in four V. vinifera clones is a positive aspect for genetic selection and development of new clones with different characteristics.     

Isozymatic Differentiation in Local Population of Glycine soja Sieb. & Zucc.

The biochemical genetic structure and variation among local population of Glycine soja Sieb. & Zucc. were investigated based on isozyme analysis using the techniques of polyacrylamide gel electrophoresis. The isoenzyme zymography of 6 enzymes viz malate dehydrogenase (MDH), peroxidase (PER), adenosine triphosphatase (ATPase), amylase (AMY), esterase (EST) and isocitric dehydrogenase (IDH) of 14 culture seedlings were respectively compared. Isozymatic analysis revealed high genetic variation in the population of G. soja. MDH, PER, ATPase, AMY are polymorphic. ATPase has the highest polymorphic index (PI=O. 1582). EST and IDH are monomorphic for all populations. The average population heterozygosity (He) was 0. 3141, and the average genetic distance (Da) among the 14 samples is 0. 1512. Cluster analysis and canonical analysis showed no correlation existed between the population's biochemical genetic structure and its environment. It was concluded that mutation could be the major cause of the high enzymatic polymorphism in population; and the mechanism that keeps the polymorphism could be random drift sampling strategy for conservation of crop genetic resources was also put forward.     

Comparison of malate dehydrogenase isoenzymes in someAllium species by isoelectric focusing

Malate dehydrogenase isoenzymes were studied in tenAllium species and in six cultivars ofA. cepa by isoelectric focusing in polyacrylamide gel with Ampholine pH 3.5–10.0. Using this method better resolution was obtained than by polyacrylamide gel electrophoresis. The number of MDH isoenzymes obtained by isoelectric focusing is from five to ten in the range of pH 3.65 to 6.75. MDH isoenzymes can be used for characterization on the level of species and cultivars (inA. cepa), but its use on the level of sections and subgenera is questionable.     

Isozyme expression in F1 hybrids between carp and goldfish

Interspecific genetic differences in malate dehydrogenase (MDH), lactate dehydrogenase (LDH), superoxide dismutase (SOD), and esterase (EST) isozymes in carp (Cyprinus carpio) and goldfish (Carassius auratus) were used to examine the allelic expressions in the hybrid between these species. A unique liver SOD and muscle LDH phenotype unambiguously identifies all presumed hybrid individuals. There was no evidence of F₂ or backcross phenotypes in hybrid individuals. Liver MDH and EST phenotypes in hybrids show a preferential expression of goldfish isozymes. Variation in the levels of carp liver MDH isozymes may result from the polymorphism of a regulatory mutation affecting isozyme expression, leading to gene silencing after duplication.This work was supported through NSERC (Canada) grants to James P. Bogart and John F. Leatherland.     

黑鲷不同组织的9种同工酶谱

对黑鲷（Sparus macrocephalus）9种组织的9种同工酶采用垂直聚丙烯酰胺平板电泳技术进行研究。结果表明,乳酸脱氢酶（LDH）、醇脱氢酶（ADH）、苹果酸脱氢酶（MDH）、苹果酸酶（ME）、超氧化物歧化酶（SOD）、过氧化物酶（POD）、酯酶（EST）、半乳糖脱氢酶（GAD）、甲酸脱氢酶（FDH）等酶在表型、分布和活性上组织特异性明显。还对眼和肠组织同工酶表达特性的生理意义进行了讨论。     

Relations between heterosis and enzymatic polymorphism in populations of cultivated sunflowers (Helianthus annuus L.)

Nine polymorphic isoenzymatic systems were studied in 39 cultivated sunflower populations originating from ten countries. Analysis of combining abilities with four tester lines was also performed on these populations for seed yield, seed moisture and seed oil content. The MDH, PGI, PGD and GOT systems appeared to provide the best discrimination of specific combining ability effects with the four testers. The MDH and GOT systems provided a between-population structure that was consistent with the country of origin.Abbreviations MDH Malate dehydrogenase - PGD phosphogluconate dehydrogenase - PGI phosphoglucoisomerase - PGM phosphoglucomutase - ACO aconitase hydratase - ADH2 alcohol dehydrogenase - GOT glutamate oxaloacetate transaminase - LAP leucine amino peptidase - EST esterases     

Effect of tannery sludge amendments on the activity of soil enzymes and phytoremediation potential of two economically important cultivars of geranium (Pelargonium graveolens)

A field experiment was conducted to observe the effect of TS amendments on soil enzymes and phytoremediation potential of two economically important cultivars of geranium. Different doses of TS were applied in soil to examine threshold limit of HMs where geranium cultivars can be grown successfully in contaminated sites. Treatment variation significantly affected pH, EC, OC, N, P, K and HM content in soil after 50 days of incubation. After harvest, both cultivars were examined to assess the impact of various treatments on their fresh herb, dry matter, essential oil yield and HM accumulation. C/G ratio close to 1 was observed at 50 tha^?1 sludge treatment in both cultivars. Urease and β-glucosidase activities in soil were maximum at 50 tha^?1 whereas dehydrogenase and phosphatase activities were maximum at 100 tha^?1 in both cultivars. β-glucosidase, acid and alkaline phosphatase, urease and dehydrogenase activities were relatively high after 85 days over 45 days in both cultivars. Maximum metal uptake was found in roots of cv. Bourbon followed by leaves. Geranium was observed to be a good candidate for phytoremediation as it mitigates metal toxicity by root absorption and cv. Bourbon is better candidate for the same.     

棉铃虫地理种群的等位酶变异

利用聚丙烯酰胺梯度凝胶电泳检测了棉铃虫Helicoverpa armigera的13种等位酶：α-磷酸甘油脱氢酶(α-GPDH)、酸性磷酸酯酶(ACPH)、碱性磷酸酯酶(ALP)、醛氧化酶(AO)、酯酶(EST)、谷氨酸草酰乙酸转氨酶(GOT)、己糖激酶(HEX)、亮氨酸氨肽酶(LAP)、乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)、苹果酸酶(ME)、磷酸葡萄糖变位酶(PGM)和黄嘌呤脱氢酶(XDH),染色采用双染法。对其中9种等位酶的遗传变异进行了分析,包括13个位点,6个位点表现出多态性,7个位点是单态的,其中多态性位点比例为46.15%。AO、GOT、LAP、LDH、ME和XDH计算出棉铃虫的平均杂合度为0.1160,南京、成都、武穴、衡阳和哈密5个种群的平均遗传距离为0.0008～0.0293,平均遗传相似度为0.9707～0.992。棉铃虫种群内存在很高的遗传多态性,而已测定的种群间遗传分化程度较小,种群间没有基因交流的障碍。迁飞阻碍了不同地理种群间的遗传分化。     

Producing commercially attractive,uniform true potato seed progenies: the influence of breeding scheme and parental genotype

The production of attractive, uniform true potato seed (TPS) progenies was investigated. Four breeding schemes were compared: intercrossing tetraploid cultivars (cv x cv); doubled dihaploids x cultivars (ddh x cv); cultivars x diploid unreduced-gamete producers (cv x FDR) and doubled dihaploids x diploid unreduced-gamete producers (ddh x FDR). Fifty three progenies and five clones were grown in a glasshouse in a randomised complete block design with three replicates of 25 plants per progeny and clone. Each plant's tubers were counted and the colour, shape, quality of skin finish, flesh colour, and commencai attractiveness (which includes yield) recorded. The most uniform progenies were also selected by visual comparison with the clones. For mean attractiveness, differences (P < 0.001) between breeding schemes and between progenies within breeding schemes were detected. The cv x cv and cv x FDR progenies were more attractive than clonal controls. There were significant additive and non-additive effects for attractiveness in all breeding schemes except cv x FDR. There were between-progeny differences (P < 0.001) for uniformity for all characters. Progenies uniform for one character could be variable for other traits. Breeding schemes gave different levels of uniformity (P < 0.001) for all characters except shape and flesh colour, but none gave low levels of variation for all traits. Doubled-dihaploid parents increased the variation in progenies. There were uniformity differences (P < 0.001) between progenies within breeding schemes for all characters. Evidence of additive and nonadditive genetic variation for uniformity in all traits was detected. In each breeding scheme, parents with good general combining ability (GCA) for uniformity in several characters were identified. Visually selected uniform progenies had parents with good GCAs for uniformity in a range of traits and high specific combining abilities (SCAs) for several traits. A desynaptic first-division restitution (FDR) clone and a male-sterile doubled-dihaploid clone had the best GCAs for tuber uniformity in TPS progenies. Achieving multitrait uniformity in TPS is problematic but may be aided by the selection of parents with GCAs for uniformity coupled with progeny testing to allow for non-additive effects.     

The palmitoleate: a natural selective denaturant of enzymes

A study has been carried out in order to explain the enzyme-palmitoleate interaction. The highly purified and crystalline enzymes representative of fundamental metabolic pathways were: alcohol dehydrogenase (ADH), lactate dehydrogenase (LDH), malate dehydrogenase (MDH), isocitrate dehydrogenase (ICDH), glucose-6-phosphate dehydrogenase (G6P-DH), alkaline phosphatase. The enzyme-palmitoleate interaction was studied as a phenomenon time-independent (inhibition) and time-dependent (inactivation). Palmitoleate inhibited remarkably LDH, MDH, ICDH and G6P-DH. A kinetic analysis of the inhibitory action of palmitoleate on LDH and MDH was also carried out. Inactivation studies have shown that ADH and alkaline phosphatase are not sensitive to palmitoleate action, unlike the other enzymes. A comparison was made between the action of palmitoleate and that of a synthetic anionic detergent, sodium dodecyl sulfate (SDS).     

乳酸脱氢酶与酯酶同工酶同板染色法

介绍一种在同一块凝胶板上染乳酸脱氢酶(LDH)与酯酶(EST)的染色方法. 该同板染色法利用两种同工酶显色反应互不干扰和颜色不同的特点, 先染LDH, 后染EST, 可以在同一块胶板上得到两种同工酶清晰的酶带, 每一种酶的酶带与单板染色的酶带完全一样. 这种染色法, 能节省同工酶分析所需的试剂、时间和经费, 也便于样品的鉴定与比较, 是一种经济有效的方法. 此方法, 同样适用于苹果酸脱氢酶(MDH)与酯酶等同工酶的同板染色.     

Effect of salt on malondialdehyde and antioxidant enzymes in seedling roots of Jerusalem artichoke (<Emphasis Type="Italic">Helianthus tuberosus</Emphasis> L.)

Two cultivars of Jerusalem artichoke (Helianthus tuberosus L.) differing in genotype, Red skin (cv. R., salt-tolerant but low-yield) and White skin (cv. W., salt-sensitive but high-yield), were used to investigate malondialdehyde (MDA) content and antioxidant enzyme activity changes in their roots under a hydroponic culture system with 250 mM NaCl. The results showed that MDA contents in roots of the two genotypes increased, but MDA content of cv. R. was higher than that of cv. W. Changes in all antioxidant enzymes in roots of both varieties exhibited a similar trend, namely increased initially and then decreased. However, there were still some differences existing between the two cultivars. In other words, activities of the other two antioxidant enzymes except catalase (CAT) and peroxidase (POD) in roots of cv. R. were less than controls at 48 h, while all others except ascorbate peroxidase (APX) in roots of cv. W. were greater than controls. The peak of superoxide dismutase (SOD) activity of cv. W. was observed to appear earlier than that of cv. R. CAT activity of cv. W. was significantly greater than the value of cv. R. and the latter showed a moderate trend. POD activity of cv. R. obtained the maximum at 6 h, whereas the peak of cv. W. displayed at 24 h. APX activity of cv. R. declined more than that of cv. W. These results suggested that there was a lower efficiency of scavenging reactive oxygen species (ROS) in cv. R. roots. Concomitantly, salt stress caused more severe damage to roots of cv. R. Antioxidant enzymes in roots were inadequate to elucidate salt-tolerance mechanisms of the whole plant.     

High throughput analysis of grape genetic diversity as a tool for germplasm collection management

Using 20 SSR markers well scattered across the 19 grape chromosomes, we analyzed 4,370 accessions of the INRA grape repository at Vassal, mostly cultivars of Vitis vinifera subsp. sativa (3,727), but also accessions of V. vinifera subsp. sylvestris (80), interspecific hybrids (364), and rootstocks (199). The analysis revealed 2,836 SSR single profiles: 2,323 sativa cultivars, 72 wild individuals (sylvestris), 306 interspecific hybrids, and 135 rootstocks, corresponding to 2,739 different cultivars in all. A total of 524 alleles were detected, with a mean of 26.20 alleles per locus. For the 2,323 cultivars of V. vinifera, 338 alleles were detected with a mean of 16.9 alleles per locus. The mean genetic diversity (GDI) was 0.797 and the level of heterozygosity was 0.76, with broad variation from 0.20 to 1. Interspecific hybrids and rootstocks were more heterozygous and more diverse (GDI?=?0.839 and 0.865, respectively) than V. vinifera cultivars (GDI?=?0.769), Vitis vinifera subsp. sylvestris being the least divergent with GDI?=?0.708. Principal coordinates analysis distinguished the four groups. Slight clonal polymorphism was detected. The limit between clonal variation and cultivar polymorphism was set at four allelic differences out of 40. SSR markers were useful as a complementary tool to traditional ampelography for cultivar identification. Finally, a set of nine SSR markers was defined that was sufficient to distinguish 99.8% of the analyzed accessions. This set is suitable for routine characterization and will be valuable for germplasm management.     

我国松阳和宽甸地区卫氏并殖吸虫的同工酶分析

作者采用Disc-PAGE电泳对我国浙江松阳和辽宁宽甸夹皮沟二倍体型和三倍体型卫氏并殖吸虫的乳酸脱氨酶(LDH)、苹果酸脱氢酶(MDH)、酯酶(EST)同工酶进行了比较。结果显示两型卫氏并殖吸虫的LDH、MDH和EST同工酶酶带的数目、Rf值、优势同工酶带的位置存在明显的差异。这些差异是否属种的特征尚待进一步研究。     

Sodium dodecyl sulfate, concurrent inhibitor of several dehydrogenases

The effect of sodium dodecyl sulfate on the activity of highly purified or crystalline enzymes has been studied. The enzymes were: lactate dehydrogenase (LDH), malate dehydrogenase (MDH). isocitrate dehydrogenase (ICDH), glucose-6-phosphate dehydrogenase (G6P-DH), lipase, alkaline phosphatase. Sodium dodecyl sulfate, always under the critical micellar concentration, shows a selective inhibitory effect. A kinetic analysis of the inhibitory action on LDH, MDH, ICDH and G6P-DH was also carried out.