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。     

Monomorphism of isozyme loci in natural and cultivated amaranth (<Emphasis Type="Italic">Amaranthus</Emphasis> L.) populations

Electrophoretic spectra of alcohol dehydrogenase (ADH), glutamate dehydrogenase (GDH), malate dehydrogenase (MDH), isocitrate dehydrogenase (IDH), and malic enzyme (ME) in different amaranth populations has been studied using a starch gel electrophoresis. 93 populations and 4 cultivars of amaranth have been analyzed. Some populations have been proved to be polymorphic that provided a possibility of a genetic control of the above-mentioned enzymes. The isozyme variability of the studied amaranth populations is low; all studied loci are found to be monomorphic for 73 populations and 4 cultivars. Some populations demonstrate a polymorphism in separate loci (Adh, Mdh 2, Gdh, Idh 1, Idh 2, and Mod 2). The obtained results evidence the presence of a genetic monomorphism in amaranth concerning the loci studied.     

Comparison of Populations of Pratylenchus brachyurus Based on Isozyme Phenotypes

Enzymes from females of five Pratylenchus brachyurus populations and one P. scribneri population were analyzed by isoelectric focusing electrophoresis. Of the 18 enzyme systems investigated, only malate dehydrogenase (MDH), phosphoglucomutase (PGM), and phosphoglucose isomerase (PGI) were detected from all five P. brachyurus populations and P. scribneri. Faint bands were detected for isocitrate dehydrogenase and phosphogluconate dehydrogenase from one P. brachyurus population. Three distinct phenotypic groups were found in the MDH and PGM systems for P. brachyurus populations, but only a single electromorph was detected for PGI. Multiple electromorphs for MDH, PGM, and PGI were detected for P. scribneri; there was no similarity among these patterns with those from P. brachyurus. No phenotypic differences in PGI were observed between females and mixed juveniles of one population of P. brachyurus.     

山西省六个负蝗种群的遗传多样性

用等位酶电泳分析方法对短额负蝗(A tractom orpha sinensis)和奇异负蝗(A tractom orpha p ereg rina)各3个自然种群10种酶(AAT,CK,G 3PD,HEX,IDH,LDH,M DH,M E,PG I,PGM)进行检测。结果显示:两种负蝗在某些基因座上共享常见的等位基因,如A a t-1-b、A a t-2-b、G 3p d-a、Ck-1-b和Ldh-b;除个别基因座在部分种群符合H ardy-W e inberg平衡外,两种负蝗的大多数基因座的基因型频率显著偏离H ardy-W e inberg平衡。此外,奇异负蝗M e-c(0.318~0.740)、短额负蝗H ex-1-a(0.800~1.000)及Ldh-b(0.487~0.750)等位基因频率呈现出由北向南递增的趋势,表明M e和H ex、Ldh基因座上的等位基因频率与地理分布存在一定的相关关系。短额负蝗平均每个基因座的等位基因数(A)为1.9~2.3、多态基因座百分率(P)为56.3%~68.8%、平均观察杂合度(Ho)为0.072~0.096;而奇异负蝗的相应值依次为A=1.7~2.2,P=43.8~56.3%,Ho=0.070~0.107。从A、P和Ho3个参数可知,短额负蝗遗传多样性明显高于奇异负蝗。6个负蝗种群的平均观察杂合度均明显低于H ardy-W e inberg平衡预期值,表明6个负蝗种群均出现了杂合体缺乏现象。短额负蝗3种群I值为0.971~0.996,奇异负蝗3种群I值为0.982~0.995,短额负蝗与奇异负蝗I值为0.379~0.451,表明种内遗传相似度明显高于种间,从种间I值可知奇异负蝗和短额负蝗属于近缘种。根据R oger's遗传距离进行的聚类分析表明,两种负蝗可分为两支,且两种负蝗的遗传距离与地理距离均存在明显的相关趋势。两种负蝗的平均FST值都不显著偏离0值(奇异负蝗FST=0.087,p>0.05,短额负蝗FST=0.045,p>0.05),表明该两种负蝗种群间的分化不明显。     

Genetic control of enzyme polymorphisms in the California vole,Microtus californicus

Inheritance of 15 polymorphic isozymes was investigated in captive Microtus californicus. Eleven of the isozymes show patterns consistent with a Mendelian model of inheritance: glycerol-3-phosphate dehydrogenase (GPD), lactate dehydrogenases A and B (LDH-A and LDH-B), malic enzyme 2 (ME-2), isocitrate dehydrogenase 1 (ICD-1), phosphogluconate dehydrogenase (PGD), glutamate-oxaloacetate transaminase 1 (GOT-1) phosphoglucomutase 2 (PGM-2), leucine aminopeptidase (LAP), glucosephosphate isomerase (GPI), and esterase 2 (ES-2, from kidney). four of the isozymes show patterns that cannot be interpreted by a simple genetic model: esterases 1 and 4 (ES-1, ES-4, from hemolysate), esterase 3 (ES-3, from plasma), and protein 1 (PT-1). The following pairs of loci are assorting independently: LAP and PGD, LAP and PGM-2, GOT-1 and PGD, GOT-1 and GPD, LAP and GPD, GPD and PGD, GPI and PGD. Data from one test cross mating indicate that GPD and PGM-2 are loosely linked with recombination about 30%. Additional data are needed to confirm this relationship.This study was conducted while the first author was the recipient of an NIH Traineeship. The Departments of Genetics and Zoology provided financial support for the maintenance of the animal colony. This work was supported in part by an NIH-Biomed grant (3-S05-RR-07006-08S1) to W. Z. Lidicker.     

Isozyme Variation among Biological Species in the Gibberella fujikuroi Species Complex (Fusarium Section Liseola)

Isozyme phenotypes were determined for 101 strains of Gibberella fujikuroi and 2 strains of Gibberella nygamai that represent seven biological species (mating populations) isolated from a variety of plant hosts in dispersed geographic locations. Fourteen enzymes were resolved in one or more of three buffer systems. Two of the enzymes, arylesterase and acid phosphatase, were polymorphic within two or more biological species and are suitable for intraspecific studies of population variation. Six enzymes, alcohol dehydrogenase, aspartate aminotransferase, glucose-6-phosphate dehydrogenase, mannitol dehydrogenase, phosphoglucomutase, and phosphogluconate dehydrogenase, were monomorphic in all of the isolates examined. The remaining six enzymes, fumarase, glucose phosphate isomerase, glutamate dehydrogenase (NADP), isocitrate dehydrogenase (NADP), malate dehydrogenase, and triose-phosphate isomerase, could potentially be used to distinguish the different biological species. Mating populations C and D are the most similar, since the mating population C isolates examined had the same isozyme phenotype as did a subset of the isolates in mating population D. Mating population E is the least similar to the other taxa examined. Unique isozyme phenotypes are present but are composed of banding patterns shared among the biological species. This finding supports the hypothesis that these biological species, with the possible exception of mating populations C and D, are reproductively isolated from one another and that no significant gene flow is occurring between them. Isozyme analysis is a useful method to distinguish these closely related biological species. Examination of isozyme phenotypes is more rapid than the present technique, which is based on sexual crosses; can be applied to strains that are not sexually fertile; and is more sensitive than traditional morphological characters, which cannot distinguish more than three or four morphological groups among the seven biological species. While emphasizing the discreteness of the mating populations as biological entities, our isozyme data also reaffirm the close genetic relationship among these groups.     

Use of Cellulose Acetate Electrophoresis in the Taxonomy of Steinernematids (Rhabditida,Nematoda)

A steinernematid nematode was isolated from soil samples collected near St. John''s, Newfoundland, Canada. On the basis of its morphometry and RFLPs in ribosomal DNA spacer, it was designated as a new strain, NF, of Steinernema feltiae. Cellulose acetate electrophoresis was used to separate isozymes of eight enzymes in infective juveniles of S. feltiae NF as well as four other isolates: S. feltiae Umeå strain, S. feltiae L1C strain, Steinernema carpocapsae All strain, and Steinernema riobravis TX strain. Based on comparisons of the relative electrophoretic mobilities (μ) of the isozymes, one of the eight enzymes (arginine kinase) yielded zymograms that were distinctive for each of the isolates, except for the Umeå and NF strains of S. feltiae, which had identical banding patterns. Four enzymes (fumarate hydratase, phosphoglucoisomerase, phosphoglucomutase, and 6-phosphogluconate dehydrogenase) yielded isozyme banding patterns that were characteristic for all isolates, except for the L1C and NF strains of S. feltiae, which were identical. Two enzymes (aspartate amino transferase and glycerol-3-phosphate dehydrogenase) yielded zymograms that permitted S. carpocapsae All strain to be discriminated from the other four isolates, while the remaining enzyme (mannose-6-phosphate isomerase) was discriminatory for S. riobravis TX strain. Except for one enzyme, the isozyme banding pattern of the NF isolate of S. feltiae was the same as in the L1C strain, isolated 13 years previously from Newfoundland. Cellulose acetate electrophoresis could prove invaluable for taxonomic identification of isolates of steinernematids, provided that a combination of enzymes is used.     

Biochemical genetic relationship of Thailand and Hawaii isolates of Parastrongylus cantonensis (Nematoda: Angiostrongylidae)

The genetic relationship of the Thailand and Hawaii isolates (strains) of the rat lungworm Parastrongylus (=Angiostrongylus) cantonensis was investigated by gene–enzyme systems using vertical polyacrylamide slab gel electrophoresis. Six gene–enzyme systems were successfully determined, with each being represented by two presumptive loci. Glucose phosphate dehydrogenase, glucose phosphate isomerase, lactate dehydrogenase, malate dehydrogenase and malic enzyme were monomorphic at both loci, and the respective bands exhibited similar mobility in both isolates implying absence of genetic variation.     

Isozyme analysis in a genetic collection of amaranths (Amaranthus L)

In various populations of the cultivated and weedy amaranth species, the electrophoretic patterns of alcohol dehydrogenase (ADH), glutamate dehydrogenase (GDH), malate dehydrogenase (MDH), isocitrate dehydrogenase (IDH) and malic enzyme (Me) were studied. In total, 52 populations and two varieties (Cherginskii and Valentina) have been examined. Allozyme variation of this material was low. Irrespective of species affiliation, 26 populations and two varieties were monomorphic for five enzymes; a slight polymorphism of three, two, and one enzymes was revealed in three, nine, and fourteen populations, respectively. A single amaranth locus, Adh, with two alleles, Adh F and Adh S, controls amaranth ADH. Two alleles, common Gdh S and rare Gdh F, control GDH; no heterozygotes at this locus were found. The MDH pattern has two, the fast- and slow-migrating, zones of activity (I and II, respectively). Under the given electrophoresis conditions, the fast zone is diffuse, whereas slow zone is controlled by two nonallelic genes, monomorphic Mdh 1 and polymorphic Mdh 2 that includes three alleles: Mdh 2-F, Mdh 2-N, and Mdh 2-S. Low polymorphism of IDH and Me was also found, though their genetic control remains unknown.     

Genetic variation in cultivars of diploid ryegrass,Lolium perenne andL. multiflorum,at five enzyme systems

Summary Samples of approximately 100 plants from each of 22 populations ofLolium perenne representing 15 cultivars, and from 13 populations ofLolium multiflorum representing six cultivars were scored for iso-zyme variants in five enzyme systems, PGI, GOT, ACP, PGM and 6-PGD. From the individual banding patterns a genetic interpretation of the variation was formulated and population studies of the resulting six polymorphic enzyme loci were performed. No strong indications of partial selfing was found since at four of the six loci,Pgi 2, Got 3, Pgm 1 andPgd 1, the genotypic proportions were in correspondence with the Hardy-Weinberg expectations. This indicated, further, that the genetical interpretations of the banding patterns might be correct. Deviations from Hardy-Weinberg proportions forAcp 1 andGot 2 indicated presumably selection working on the linkage group including these loci. Gametic phase disequilibrium was observed betweenPgi 2 andPgd 1 for populations of one cultivar. These results were discussed in relation to the variation expected within a cultivar.     

Isoenzyme studies in one Brazilian and two Venezuelan strains of Schistosoma mansoni.

1. Enzyme polymorphism, analyzed by starch gel electrophoresis, was found to be zero for acid phosphatase, phosphoglucomutase, phosphoglucose isomerase, glucose 6-phosphate dehydrogenase, lactate dehydrogenase, malate dehydrogenase and malic enzyme, in one Brazilian and two Venezuelan strains of Schistosoma mansoni. 2. All loci studied were monomorphic within strains, but the isoenzymic patterns were, however, different among the strains. 3. Results suggest a drastic loss of the genetic variability usually found in natural populations.     

Characterization of Enterobacter cloacae and E. sakazakii by electrophoretic polymorphism of acid phosphatase, esterases, and glutamate, lactate and malate dehydrogenases

Acid phosphatase, esterases, and glutamate, lactate and malate dehydrogenases of 34 strains of Enterobacter cloacae and 22 strains of Enterobacter sakazakii were analysed by horizontal polyacrylamide agarose gel electrophoresis and by isoelectrofocusing in thin-layer polyacrylamide gel. The two species could be separated on the basis of distinct electrophoretic patterns of all enzymes analysed. Glutamate dehydrogenase and acid phosphatase were detected exclusively in E. cloacae, whereas esterase bands were more intensively stained in E. sakazakii. For each species, two zymotypes could be distinguished, on the basis of electrophoretic mobilities of malate dehydrogenase and banding patterns of esterase for E. cloacae, and by both isoelectric point and electrophoretic mobilities of an esterase and of lactate and malate dehydrogenases for E. sakazakii. The high degree of enzyme polymorphism within the two species permitted precise identification of strains. The variations in electrophoretic patterns might therefore provide useful epidemiological markers.     

Genetic diversity of six populations of Atractomorpha sinensis and Atractomorpha peregrina in Shanxi Province, China

Ten enzymes (AAT,CK,G3PDH,HEX,IDH,LDH,MDH,ME,PGI,PGM)were examined using horizontal starch gel electrophoresis to estimate the levels of genetic variation within and among six natural populations of two grasshopper species Atractomorpha sinensis and A.peregrina from Shanxi,China.The collecting sites were geographically distant from each other from south to north:Quwo district,Linfen city;Xiangyuan county,Changzhi;Jinyuan district,Taiyuan city;Yuanping county,Xinzhou city and Fanshi county of Xinzhou.A.sinensis showed 43 alleles at 16 loci but A.peregrine showed 39 alleles at 15 loci (ldh-1 was deficient).The zymograms showed that some common alleles were shared at several loci in these two species (Aat-1-b,Aat-2-b,G3pdh-a,Ck-1-b and Ldh-b).However,Hex-1-a,Hex-2-a,Hex-3-a,Idh-2-b,Mdh-2-b,Mdh-1-f Pgi-b,Pgm-b had common alleles in A.sinensis and Hex-1-b,Hex-2-b,Hex-3-b,Idh-2-a,Mdh-2-a,Mdh-1-d,Pgi-a,Pgm-c were of high frequency in A.peregrine instead.Most of the observed genotype frequencies were found to significantly deviate from the Hardy-Weinberg expectations in both species.A tendency of clinal distribution of allele frequency was observed at three loci.The frequency of the moderately migrating allele Me-c (0.318-0.740)in A.peregrina,Hex-1-a (0.800-1.000)and Ldh-b (0.487-0.750)in A.sinensis demonstrated increased frequency from north to south.Such tendency suggests that the allele frequency in these three loci may be correlated with the species'geographic distributions.A.sinensis showed higher genetic diversity than A.peregrina as indicated by higher mean number of alleles per locus (A=1.9-2.3 in A.sinensis and 1.7-2.2 in A.peregrina),percentage of polymorphic loci (56.3%-68.8%in A.sinensis and 43.8%-56.3%in A.peregrina),and the observed heterozygosities (Ho=0.072-0.096 in A.sinensis and 0.070-0.107 in A.peregrina).The observed heterozygosities of the six populations were all noticeably lower than the Hardy-Weinberg expectations,mostly due to heterozygote deficiency in the populations of both species.The overall mean Fsr were small (FST=0.045,P＞0.05 in A.sinensis populations and 0.087,P＞0.05 in A.peregrina populations).Nei's genetic identity (I)estimates indicate low intraspecific (＞0.95)but higher interspecific (0.377-0.447)genetic diversity.The cluster analysis based on modified Roger's genetic distance (D)showed that the two species were divided into two branches.Both species are of limited dispersal capacity and a moderate geographical barrier might significantly restrict the gene exchange among populations,resulting in accumulation of local genetic differentiations.The A.sinensis populations used in this study were separated from each other by 155.2 to 271.4 km and the A.peregrina populations were separated from each other by 78.8 to 174.9 km with observable physical barriers.The aUozyme data showed only minimal genetic differentiation at population level,most likely as a result of gene exchange.It is reasoned that natural factors and human agricultural activities might have facilitated migration and dispersal for the two species.     

Isozyme Analysis in a Genetic Collection of Amaranths (<Emphasis Type="Italic">Amaranthus</Emphasis> L.)

In various populations of the cultivated and weedy amaranth species, the electrophoretic patterns of alcohol dehydrogenase (ADH), glutamate dehydrogenase (GDH), malate dehydrogenase (MDH), isocitrate dehydrogenase (IDH) and malic enzyme (Me) were studied. In total, 52 populations and two varieties (Cherginskii and Valentina) have been examined. Allozyme variation of this material was low. Irrespective of species affiliation, 26 populations and two varieties were monomorphic for five enzymes; a slight polymorphism of three, two, and one enzymes was revealed in three, nine, and fourteen populations, respectively. A single amaranth locus, Adh, with two alleles, Adh F and Adh S, controls amaranth ADH. Two alleles, common Gdh S and rare Gdh F, control GDH; no heterozygotes at this locus were found. The MDH pattern has two, the fast- and slow-migrating, zones of activity (I and II, respectively). Under the given electrophoresis conditions, the fast zone is diffuse, whereas slow zone is controlled by two nonallelic genes, monomorphic Mdh 1 and polymorphic Mdh 2 that includes three alleles: Mdh 2-F, Mdh 2-N, and Mdh 2-S. Low polymorphism of IDH and Me was also found, though their genetic control remains unknown.     

Electrophoretic studies on blood proteins of Arvicanthis niloticus

Electrophoretic patterns of blood proteins were compared among groups of Arvicanthis niloticus from widely distant origin: Senegal, Haute-Volta, Egypt. Four blood proteins/enzymes were studied: albumin, transferrin, esterase and 6-phosphogluconate dehydrogenase. The results demonstrate two levels of difference: (a) inter-population differences; and (b) intra-species genetic polymorphism. The latter was not observed for all populations nor for all loci. The differences between populations suggest that some process of specification could be under way among Arvicanthis.     

泡沙参同工酶基因位点的遗传分析

利用聚丙烯酰胺凝胶电泳技术 ,对来自天然群体 (居群 )的泡沙参 (Adenophora potaninii Korsh.)及其人工杂交子代进行了 8种同工酶的电泳检测和谱带遗传分析 ,以确定编码这些酶系统的基因位点和等位基因。选用 4种不同的凝胶缓冲系统 ,对下列不同酶系统进行了酶谱的遗传分析 :天冬氨酸转氨酶 (AAT)、酯酶 (EST)、甲酸脱氢酶 (FDH)、谷氨酸脱氢酶 (GDH)、异柠檬酸脱氢酶 (IDH)、乳酸脱氢酶(LDH)、苹果酸酶 (ME)和超氧化物歧化酶 (SOD)。结果表明 ,这 8种酶系统至少由 1 8个基因位点编码 ,其中 1 2个位点为遗传稳定的等位酶位点 ,是可靠的遗传标记。酶谱的分离式样表明 ,EST为单聚体结构 ,AAT、FDH、IDH、SOD为二聚体结构 ,GDH为六聚体结构。最后对同工酶的器官和发育特异性以及同工酶基因位点的遗传分析进行了讨论     

Esterase patterns in species of the triatome bug Rhodnius

The aim of the present study was to analyse esterase patterns in three triatomine species of Rhodnius genus. Four loci, Est 1, Est 2, Est 3 and Est 4, were found. The corresponding enzymes were characterized as carboxylesterases (E.C. 3.1.1.1) or cholinesterases (E.C. 3.1.1.8) based on inhibitory experiments, using eserine sulphate, malathion, mercury chloride, p-chloromercuribenzoate (pCMB) and iodoacetamide. Low genetic variability was observed: Est 1, Est 2 and Est 3 were monomorphic in Rhodnius domesticus , Rhodnius robustus and Rhodnius neivai , whereas locus Est 4 was polymorphic in the first two species. The UPGMA analysis based on esterase genotypic frequencies indicated greater similarity between R. domesticus and R. robustus when compared with R. neivai . The present study expands our knowledge about genetic variability among triatomines and accords with the hypothesis that R. domesticus is a species derived from R. robustus .     

Enzyme variation among clones of Trypanosoma cruzi

The genetic relationships within and between stocks of Trypanosoma cruzi were studied by the in vitro isolation of clones and sub-clones and by the comparison of their isoenzyme patterns in thin-layer starch-gel electrophoresis. Altogether 13 clones and 36 sub-clones were isolated from stocks CL89 and Y207 of T. cruzi. Employing the enzymes L-alanine aminotransferase (ALAT), L-aspartate aminotransferase (ASAT), glucose phosphate isomerase (GPI), malic enzyme (ME), malate dehydrogenase (MDH), glucose-6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), and phosphoglucomutase (PGM), two zymodemes, B and C, emerged among the clones with distinct banding patterns. These zymodemes were consistently distinguished by ALAT, ASAT, GPI, G6PD, 6PGD, and PGM and the differences in enzyme profiles were simultaneously reflected in all six enzyme systems. That the enzymic characters as genetic determinants are stable was demonstrated after recloning and successive replica-platings, i.e., the distinct enzyme patterns remained consistent and homogeneous, the siblings retained the enzyme profiles of their parental clones, and no segregation of the enzyme patterns was observed. Our data from clone and sub-clone examinations show that the isoenzymes act as labels to characterize T. cruzi stocks. Furthermore, enzyme variation was demonstrated among clones isolated from stock CL89.     

Allozyme polymorphism and interspecific relationships in the Common starling (Sturnus vulgaris) and Spotless starling (S. unicolor) (Aves: Sturnidae)

This paper reports a genetic study of Sturnus vulgaris and S. unicolor (Sturnidae), two similar bird species which have recently become sympatric in north-eastern Spain. Seven enzyme systems (alcohol dehydrogenase, esterase, glutamate dehydrogenase, glucose-6-phosphate dehydrogenase, leucin aminopeptidase, L-lactate dehydrogenase and superoxide dismutase) were analyzed, resolving 22 presumed genetic loci with 28 alleles; 27 of them were found in both species. Gene flow was estimated to be number of migrants per generation (Nm) = 6, and genetic identities among populations were high, ranging from I = 0.96 to I = 0.99. On the whole, the results demonstrate that these Sturnus populations share a gene pool, showing slight genetic differences between the two starling speices.