Protein differences among the Mediterranean species of the genus Spicara

Protein electrophoresis (PAGE) was used to study the three morphologically different species of Spicara (S. flexuosa, S. maena, S. smaris). Of the 28 enzymatic and additional myogenic loci, five monomorphic loci (LDH-1*, G6PD-1*, PGI-1* and two PMMs*) were species-specific markers of S. smaris with respect to S. flexuosa and S. maena. Four of the 28 enzymatic loci were polymorphic (EST-1*, GLDH*, PEPD*, PGI-2*). Discriminating genetic markers were not identified between S. flexuosa and S. maena. Genetic distance (D) as calculated by Nei's index (1978), between S. smaris v. S. maena and S. flexuosa showed a value, respectively of Z) = 0·137 and 0·141. Between S. flexuosa and S. maena the value was Z)=0-006. From the data it can be inferred that S. flexuosa and S. maena are conspecific, despite morphological differences.     

Biochemical genetic differentiation between Pomatoschistus marmoratus and P. tortonesei

Several diagnostic genetic markers were identified in Pomatoschistus marmoratus and P. tortonesei using polyacrylamide gel electrophoresis (PAGE) of allozymes. Twenty-one loci were resolved, including the electrophoretic pattern of muscle proteins. The MDH*, PGM-,2*, EST-1,2*, FUM* and PGI-2* loci exhibited different alleles which were fixed for the two species being analysed. Genetic distance, as calculated by Nei's index, showed a value of 0·413. Environmental hypersalinity, could have influenced the geographical distribution of P. tortonesei .     

Phosphoglucose isomerase isozymes and allozymes of the brown trout, Salmo trutta L

1. In brown trout the Pgi-1 and Pgi-2 loci are predominantly expressed in white skeletal muscle; Pgi-3 being mainly expressed in most other tissues. 2. Total PGI activity determinations revealed that the allele formerly designated Pgi-2(65) is a null allele, Pgi-2(n). 3. Enzyme kinetic studies on the partially purified PGI homodimeric isozymes revealed that from 5 to 25 degrees C both PGI-1 and PGI-2 had significantly higher mean Km(F6P) values compared to PGI-3. 4. Distinct metabolic roles for the "muscle" (PGI-1, PGI-2) and "liver" (PGI-3) isozymes are proposed. 5. Significant Km (F6P) differences were found among the PGI-2 allozymes and among the PGI-3 allozymes.     

Effect of PGI-2 on uterine activity in vivo in non-pregnant ovariectomized goats (Capra hircus)

Jugular administration of 200 micrograms PGI-2 salt significantly reduced spontaneous uterine activity in ovariectomized, oestrogen-primed goats; the effect was acute and persisted for about 3 h. Peripheral plasma concentrations of 6-keto-PGF-1 alpha, the stable metabolite of PGI-2, decreased to 50% of initial values after 30 min; but at the start of uterine recovery were in excess of 2 ng.ml-1. Uterine reactivity to both oxytocin and PGF-2 alpha after PGI-2 administration was unaffected.     

Effect of infusion of PGI-2, 6-keto-PGF-1 alpha and PGF-2 alpha on luteal function in the pregnant rat

Prostacyclin (PGI-2), 6-keto-PGF-1 alpha and PGF-2 alpha were infused continuously for 6 h into the dorsal aorta of rats 8 days pregnant. PGF-2 alpha (10 micrograms/h) significantly reduced plasma progesterone concentrations by 66% and luteal tissue concentrations of pregnenolone and progesterone by 78% and 95% respectively. Plasma concentrations of 20 alpha-dihydroprogesterone remained unchanged whilst luteal tissue concentrations rose 2-fold. Plasma progesterone concentrations were significantly reduced to 50% by PGI-2 (10 micrograms/h) but were unaffected by 6-keto-PGF-1 alpha (10 or 100 micrograms/h). Neither PGI-2 (10 micrograms/h) nor 6-keto PGF-1 alpha (10 or 100 micrograms/h) had any significant effect on plasma concentrations of 20 alpha-dihydroprogesterone or on luteal tissue concentrations of pregnenolone, progesterone or 20 alpha-dihydroprogesterone. Arterial blood pressure was unaffected by PGF-2 alpha and 6-keto-PGF-1 alpha, but was significantly reduced by PGI-2 at infusion rates greater than or equal to 60 micrograms/h.     

Human apolipoprotein A-I induces cyclooxygenase-2 expression and prostaglandin I-2 release in endothelial cells through ATP-binding cassette transporter A1

High-density lipoprotein (HDL) can induce cyclooxygenase-2 (COX-2) expression and prostacyclin I-2 (PGI-2) release in endothelial cells to exert multiple antiatherogenic functions. This effect has been attributed mainly to the role of sphingosine-1-phosphate (S1P) integrated in HDL. However, whether apolipoprotein A-I (apoA-I), the major apolipoprotein of HDL, could induce COX-2 expression and PGI-2 release still remains unclear. In the present study, we selectively delipidated HDL and confirmed that apoA-I could facilitate COX-2 expression and PGI-2 production in human umbilical vein endothelial cells (HUVECs). ApoA-I, but not trypsinized apoA-I, induced COX-2 expression in a time- and dose-dependent manner consistent with a key role for apoA-I in this process. Additionally, cotreatment of apoA-I with S1P further enhanced COX-2 expression and PGI-2 production in HUVECs. These effects triggered by apoA-I were not inhibited by pertussis toxin, consistent with SIP receptor independent pathway for apoA-I effect. Moreover, we demonstrated that the activation of p38 mitogen-activated protein kinase (MAPK), extracellular receptor kinase (ERK) 1/2, and JAK2 pathways by apoA-I was involved in the expression of COX-2 and the release of PGI-2 in HUVECs, and these effects were inhibited by their specific inhibitors, respectively. Small interfering RNA experiments showed that ATP binding-cassette transporter A1 (ABCA1) was required for COX-2 expression and PGI-2 release induced by apoA-I. Thus our results indicate that apoA-I induces COX-2 expression and PGI-2 release through ABCA1 and the activation of intracellular p38 MAPK, ERK1/2, as well as JAK2 pathways, and apoA-I can reinforce these effects with S1P in HUVECs. These novel effects of apoA-I could in part mediate antiatherogenic effects of HDL.     

High-density lipoprotein induces cyclooxygenase-2 expression and prostaglandin I-2 release in endothelial cells through sphingosine kinase-2

High-density lipoprotein (HDL) has a significant cardioprotective effects. HDL induces cyclooxygenase-2 (COX-2) expression and prostacyclin I-2 (PGI-2) release in vascular endothelial cells, which contributes to its anti-atherogenic effects. However, the underlying mechanisms are not fully understood. In the present study, we observed that HDL-stimulated COX-2 expression and PGI-2 production in human umbilical vein endothelial cells (HUVECs) in a time- and dose-dependent manner. These effects triggered by HDL were inhibited by pertussis toxin (PTX), protein kinase C (PKC) inhibitor GF109203X, and ERK inhibitor PD98059, suggesting that Gαi/Gαo-coupled GPCR, PKC, and ERK pathways are involved in HDL-induced COX-2/PGI-2 activation. More importantly, we found that silencing of sphingosine kinase 2 (SphK-2) also blocked HDL-induced COX-2/PGI-2 activation. In addition, HDL-activated SphK-2 phosphorylation accompanied by increased S1P level in the nucleus. Our ChIP data demonstrated that SphK-2 is associated with CREB at the COX-2 promoter region. Collectively, these results indicate that HDL induces COX-2 expression and PGI-2 release in endothelial cells through activation of PKC, ERK1/2, and SphK-2 pathways. These findings implicate a novel mechanism underlying anti-atherothrombotic effects of HDL.     

Phosphoglucose isomerase gene duplication in the bony fishes: An evolutionary history

Electrophoretic patterns of phosphoglucose isomerase (PGI) in bony fishes provide strong evidence for a model of genetic control by two independent structural gene loci, most likely resulting from a gene duplication. This model is confirmed by a comparison of certain kinetic and molecular properties of the PGI homodimers (PGI-1 and PGI-2) isolated from extracts of the teleost Astyanax mexicanus. In addition, in most higher teleosts examined, the PGI enzymes show a regular pattern of tissue distribution, with PGI-2 predominant in muscle, the heterodimer often strongest in the heart, and PGI-1 predominant in liver and other organs. An examination of 53 species of bony fishes belonging to 38 families indicates a widespread occurrence of duplicate PGI loci and an early origin of the gene duplication, perhaps in the Leptolepiformes. The apparent presence of three PGI loci in trout and goldfish exemplifies how new loci can be incorporated into the genome through polyploidization.This research was supported in part by a NSF graduate traineeship to J.C.A., by the Clayton Foundation for Research in Biochemistry (G.B.K.), by NSF Grant GB-15644 and NIH Grant GM-15769 to Robert K. Selander, and by contract AT(38-1)-310 between the University of Georgia and the U.S. Atomic Energy Commission.     

Inhibition of luteal function by oxytocin antagonist in goats (Capra hircus).

Early corpus luteum development in nonpregnant and pregnant goats was characterized by a steady increase in peripheral plasma concentrations of progesterone and a high release of prostacyclin (PGI-2) but low release of prostaglandin F-2 alpha (PGF-2 alpha). Jugular administration of oxytocin antagonist (OXA) (0.2 microgram/kg/day) on the day of oestrus and for 3 days thereafter to cyclic and mated goats, significantly (P less than 0.001) inhibited progesterone and prostaglandin secretion and reduced conception rate. Co-administration of PGI-2 (200 micrograms/day) with OXA resulted in a steady increase of progesterone and establishment of pregnancy, but co-administration of PGF-2 alpha (175 micrograms/day) with OXA had no effect. It is suggested that oxytocin is required for early development of the corpus luteum and such effects may be mediated via PGI-2 production.     

Linkage between an isozyme marker and a restorer gene in radish cytoplasmic male sterility of rapeseed (Brassica napus L.)

Summary Co-segregation studies of isozyme markers and male fertility restoration showed that a restorer gene from radish was introduced into rapeseed along with an isozyme marker (Pgi-2). The radish chromosome segment carrying these genes was introgressed into rapeseed through homoeologous recombination, substituting for some of the rapeseed alleles. By crossing heterozygous restored plants to male-sterile lines and to maintainers, tight linkage was found between the restorer gene and the marker. The recombination fraction was estimated at 0.25 ± 0.02%. Although few restored plants lacked the radish isozyme marker, it was still possible to distinguish male-fertile from male-sterile plants by their PGI-2 patterns. Furthermore, homozygous and heterozygous restored plants could be separated by specific PGI-2 phenotypes. Thus, the Pgi-2 marker is now currently used in restorer breeding programs.     

0-group cod(Gadus morhua) in captivity: Differential survival of certain genotypes

Genotypes atLDH-3 (lactate dehydrogenase) andPGI-1 (phosphoglucoisomerase) in 263 0-group specimens of Atlantic cod(Gadus morhua) caught in Trondheimsfjorden, Norway, in fall 1983 showed significantly different survival rates during 72 days in captivity. The heterozygote was nominally superior at both loci, and there was a significant accumulation of double heterozygotes among the survivors. Apparently,LDH-3 andPGI-1 are not selectively neutral, and allele frequency differences at these two loci between groups of cod in nature should not be interpreted as markers of reproductive isolation. The results are related to current concepts of cod population structure.Contribution No. 231, Biological Station, N-7000 Trondheim, Norway     

Balancing Selection on Electrophoretic Variation of Phosphoglucose Isomerase in Two Species of Field Cricket: Gryllus Veletis and G. Pennsylvanicus

Two species of crickets, Gryllus veletis and G. pennsylvanicus, share six electrophoretic mobility classes for the enzyme phosphoglucose isomerase (PGI), despite evidence from other genetic markers that the two species are not closely related within eastern North American field crickets. Moreover, the frequencies of the two most common PGI electrophoretic classes (PGI-100 and PGI-65) covary in sympatric populations of these species in the eastern United States, suggesting that PGI may be subject to trans-specific balancing selection. To determine the molecular basis of the electrophoretic variation, we characterized the DNA sequence of the Pgi gene from 29 crickets (15 G. veletis and 14 G. pennsylvanicus). Amino acid substitutions that distinguish the electrophoretic classes are not the same in the two species, and there is no evidence that specific replacement substitutions represent trans-specific polymorphism. In particular, the amino acids that diagnose the PGI-65 allele relative to the PGI-100 allele differ both between G. veletis and G. pennsylvanicus and within G. pennsylvanicus. The heterogeneity among electrophoretic classes that covary in sympatric populations coupled with analysis of patterns of nucleotide variation suggest that Pgi is not evolving neutrally. Instead, the data are consistent with balancing selection operating on an emergent property of the PGI protein.     

Ralstonia solanacearum ��PGI-1 Strain KZR-5 Is Affected in Growth, Response to Cold Stress and Invasion of Tomato

The survival and persistence of Ralstonia solanacearum biovar 2 in temperate climates is still poorly understood. To assess whether genomic variants of the organism show adaptation to local conditions, we compared the behaviour of environmental strain KZR-5, which underwent a deletion of the 17.6?kb genomic island PGI-1, with that of environmental strain KZR-1 and potato-derived strains 1609 and 715. PGI-1 harbours two genes of potential ecological relevance, i.e. one encoding a hypothetical protein with a RelA/SpoT domain and one a putative cellobiohydrolase. We thus assessed bacterial fate under conditions of amino acid starvation, during growth, upon incubation at low temperature and invasion of tomato plants. In contrast to the other strains, environmental strain KZR-5 did not grow on media that induce amino acid starvation. In addition, its maximum growth rate at 28°C in rich medium was significantly reduced. On the other hand, long-term survival at 4°C was significantly enhanced as compared to that of strains 1609, 715 and KZR-1. Although strain KZR-5 showed growth rates (at 28°C) in two different media, which were similar to those of strains 1609 and 715, its ability to compete with these strains under these conditions was reduced. In singly inoculated tomato plants, no significant differences in invasiveness were observed among strains KZR-5, KZR-1, 1609 and 715. However, reduced competitiveness of strain KZR-5 was found in experiments on tomato plant colonisation and wilting when using 1:1 or 5:1 mixtures of strains. The potential role of PGI-1 in plant invasion, response to stress and growth in competition at high and moderate temperatures is discussed.     

Allozyme evidence for reproductively isolated sympatric populations of brown trout Salmo trutta L. in Lough Melvin, Ireland

Electrophoretic studies of five polymorphic enzyme loci ( G-3-PDH-2, LDH-I, LDH-5, PGI-2, PGI-3 ) in brown trout from Lough Melvin in northwestern Ireland have demonstrated that the morphotypes known by the vernacular names of 'ferox', 'gillaroo' and 'sonaghen', are not merely ecophenotypes but represent genetically distinct and reproductively isolated populations. The results suggest that the long life and higher growth potential of ferox trout of this lake, and possibly others, has a genetic basis. These separate demes of brown trout are probably the result of multiple invasions in post-glacial times of allopatrically derived stocks. Lough Melvin's isolated position and absence of pike, Esox lucius , and large cyprinids have probably contributed to its pristine condition. As such it is one of the few remaining examples of what may once have been a widespread situation in Britain and Ireland.     

Inheritance of allozymes in Atlantic herring (Clupea harengus harengus)

Progeny from single pair crosses of Atlantic herring were examined to determine the heritability of genetic variation at seven polymorphic allozyme loci. Mendelian inheritance of codominant autosomal alleles was established for IDH-2, LDH-1, LDH-2, ME-2, PGM-1, and PGI-2. This demonstration of Mendelian inheritance is essential for accurate interpretation of allozymic variation among natural populations of this pelagic species.     

Effects of temperature on physiology and reproductive success of a montane leaf beetle: implications for persistence of native populations enduring climate change

Understanding how climate change impacts natural systems requires investigations of the effects of environmental variation on vulnerable species and documentation of how populations respond to change. The willow beetle Chrysomela aeneicollis is ideal for such studies. It lives in California's Sierra Nevada on the southern edge of its worldwide range. Beetles experience elevated air temperatures during summertime egg laying and larval development. Exposure to these temperatures causes physiological stress, which may reduce reproductive success and endanger populations. The glycolytic enzyme phosphoglucose isomerase (PGI) is a marker of temperature adaptation in C. aeneicollis. PGI allele frequency varies across a latitudinal gradient: allele 1 is common in Rock Creek (RC), which is cooler and to the north, and allele 4 is common in Big Pine Creek (BPC), which is warmer and to the south. In populations that are intermediate in geography and climate (e.g., Bishop Creek [BC]), PGI-4 frequency increases from north to south such that alleles 1 and 4 are in relatively equal frequency in southern BC. Over the past decade, Sierra Nevada beetle populations have colonized high elevations and have become extinct at lower elevations where they were once common. In BC, the magnitude of PGI allele frequency fluctuations among life-history stages is related to maximal air temperature, with the frequency of PGI-4 increasing after the hottest part of summer. To identify mechanisms that may cause shifts in PGI allele frequency, we measured metabolic rate and fecundity for beetles collected at BC. Metabolic rate of males and females was measured at 20 degrees and 36 degrees C using flow-through respirometry. To measure laboratory fecundity, mating pairs were acclimated for 4 h each afternoon at a control temperature (20 degrees C) or at mildly elevated temperatures (26 degrees or 32 degrees C) and number of eggs laid was counted daily for 24 d, after which tissue levels of 70-kD heat shock proteins (Hsp70) were determined. Previous studies had demonstrated differences in Hsp70 expression among PGI genotypes at these temperatures. To measure field fecundity, mating pairs from BC were transplanted to similar elevations in BPC, BC, and RC and were monitored in situ for 24 d. Metabolic rate was higher for PGI 4-4 genotypes than for PGI 1-4 or PGI 1-1 individuals at 36 degrees C but not at 20 degrees C. In contrast, laboratory fecundity was greatest for females possessing PGI-1, independent of acclimation temperature. At the end of the laboratory fecundity experiment, Hsp70 expression was positively related to fecundity, suggesting minimal reproductive cost of upregulation of heat shock proteins in response to mild heat stress. In the field, fecundity was highest for PGI 1-1 and PGI 1-4 individuals in RC and PGI 4-4 individuals in BPC and was similar for all genotypes in BC. Thus, fecundity in nature was greatest for the genotypes that were most common in each area. Taken together, data reported here suggest that hot, dry summers in the Sierra Nevada may result in an increase in frequency of the PGI-4 allele and shifts to higher elevations for C. aeneicollis populations.     

人工促成檀香结香的研究

在自然情况下生长的檀香（ＳａｎｔａｌｕｍａｌｂｕｍＬ．）植株,约１０龄左右开始形成具芳香的心材（通称结香）,约３０—４０年方可砍伐利用．作者采用两年生的幼树,施用植物生长抑制剂ＰＧＩ１进行促成结香试验,结果证明采用１％３ｍｌ生长激素处理的植株,其檀香油和檀香醇的含量一般较对照和用水处理的植株高１—２倍．     

Biochemical Polymorphism in Yellow Catfish, Mystus nemurus (C&V), from Thailand

Yellow catfish, Mystus nemurus (Cuv. & Val.), is becoming one of the major freshwater species farmed by aquaculturists in Southeast Asia. It was of interest to examine levels of genetic subpopulation differentiation among samples of this species obtained from parts of its range, as well as to compare the genetics of wild and hatchery-bred fish. Horizontal starch gel electrophoresis and histochemical staining techniques were used to examine genetic variation within and among eight wild and one hatchery populations of M. nemurus from northern, northeastern, central and southern Thailand. Four tissues (heart, liver, kidney, and muscle) from individual specimens were used to analyze variations at 23 protein-coding loci. Fifteen of the 23 loci examined (65.22%), namely, ACP*, AAT-1*, EST-1*, EST-2*, GPI*, IDH-1*, IDH-2*, MDH-1*, MDH-2*, MDH-3*, ME*, PGM*, 6PGD*, SOD*, and HB*,were polymorphic at the 0.95 level. Observed heterozygosities ranged from 0.041 to 0.111, with an average of 0.068 ± 0.028. Genetic distances ranged from 0.005 to 0.164. The greatest genetic distance was found between the Chainat and the Suratthani populations (0.164), a level indicative of subspecific differentiation in M. nemurus from within Thailand.     

ABSORPTION OF HERBICIDES BY ROOTS

Crafts , A. S., and S. Yamaguchi . (U. California, Davis.) Absorption of herbicides by roots. Amer. Jour. Bot. 47(4): 248—255. Illus. 1960.—Many herbicides are used through soil. When 2,4-D*² was applied to culture-solution barley, bean, cotton, and Zebrina plants, there was evidence that the herbicide is held at high concentration by the roots. Very little of the labeled compound moved into the tops of barley, Zebrina and bean; a fair quantity was found in cotton foliage. Barley seedlings allowed to absorb 2,4-D*, ATA*, MH*, urea*, monuron*, dalapon*, simazin*, P³² and IAA* showed interesting differences. All chemicals were highly sorbed by roots, 2,4-D* was moved to tops in least amount, urea was next; the other seven were moved in larger quantities. ATA*, MH*, IAA*, P³², and dalapon* seemed readily phloem mobile; monuron* and simazin* seemed limited to xylem movement. Results are discussed in relation to the mechanism of root uptake.