Pentose phosphate metabolism during dormancy breakage in Corylus avellana L.

Commercially obtained fruits of Corylus avellana exhibit the characteristic loss of dormancy of this seed following chilling under moist conditions. The activities of cytosolic and organellar enzymes of pentose phosphate pathway in cotyledonary tissue were assayed throughout stratification and over a similar period in damp vermiculite at 20° C. Glucose-6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconic acid dehydrogenase (6PGDH) were both found in cytosolic extracts in all treatments; only 6PGDH was present in the organellar fraction.The enzyme activities monitored in seeds at 20° C remained relatively constant over the course of the investigation except in the case of cytosolic 6PGDH where it is suggested an inhibitor of the enzyme accumulated. This inhibitor was removed by the partial purification procedure. Increases in the activities of the enzymes occurred during stratification, the major increase coinciding exactly with dormancy breakage but prior to the initiation of germination. The marked increase in G6PDH and 6PGDH concurrent with the change in germination potential of the chilled seed may have considerable biochemical significance in breaking down the dormant state.Abbreviations G6P glucose-6-phosphate - G6PDH glucose-6 phosphate dehydrogenase - NADP nicotinamide adenine dinucleotide phosphate - 6 PGDH 6-phosphogluconic acid dehydrogenase - PPP pentose phosphate pathway     

VARIATION IN ALCOHOL DEHYDROGENASE ACTIVITY AND FLOOD TOLERANCE IN WHITE CLOVER,TRIFOLIUM REPENS

Flooding results in induction of anaerobic metabolism in many higher plants. As an important component of anaerobic energy production, alcohol dehydrogenase (ADH) activity increases markedly in response to flooding in white clover, Trifolium repens. Significant inter-individual variation in flood-induced ADH activity exists in natural populations of T. repens. The genetic basis of this variation was analyzed by offspring-midparent regression of data from 75 greenhouse reared families; the estimated heritability of flood-induced ADH activity was 0.55 (±0.13). Genetic variation in flood-induced ADH activity has pronounced effects on physiological response and flood tolerance in this species. ADH activity is positively correlated with the rate of ethanol production, indicating that observed in vitro activity differences are manifested in in vivo physiological function. T. repens plants with higher ADH activities during flooding have greater flood tolerance (measured as growth rate when flooded/unflooded growth rate). Variation in ADH activity during flooding accounts for more than 79% of the variance in flood tolerance. On the basis of a limited field survey of populations occupying three sites differing in exposure to flooding conditions, individuals from site C, the most frequently flooded site, expressed significantly higher average ADH activity when flooded than individuals from site A, a site with no history of flooding. Since ADH activity levels are not correlated with electrophoretic mobility variation in T. repens, this work supports previous suggestions that regulatory variation in enzyme activity may play a central role in biochemical adaptations to environmental stress.     

Changes in the activities of carbon metabolizing enzymes with pod development in lentil (<Emphasis Type="Italic">Lens culinaris</Emphasis> L.)

Activities of some key enzymes of carbon metabolism sucrose synthase, acid and alkaline invertase, phosphoenol pyruvate carboxylase, malic enzyme and isocitrate dehydrogenase were investigated in relation to the carbohydrate status in lentil pods. Sucrose remained the dominant soluble sugar in the pod wall and seed, with hexoses (glucose and fructose) present at significantly lower levels. Sucrose synthase is the predominant sucrolytic enzyme in the developing seeds of lentil (Lens culinaris L.). Acid invertase was associated with pod elongation and showed little activity in seeds. Sucrose breakdown was dominated by alkaline invertase during the development of podwall, while both the sucrose synthase and alkaline invertase were active in the branch of inflorescence. A substantial increase of sucrolytic enzymes was observed at the time of maximum seed filling stage (10–20 DAF) in lentil seed. The pattern of activity of sucrose synthase highly paralleled the phase of rapid seed filling and therefore, can be correlated with seed sink strength. It seems likely that the fruiting structures of lentil utilize phosphoenol pyruvate carboxylase for recapturing respired carbon dioxide. Higher activities of isocitrate dehydrogenase and malic enzyme in the seed at the time of rapid seed filling could be effectively linked to the deposition of protein reserves.     

长期水淹对‘中山杉118’幼苗呼吸代谢的影响

中山杉(Taxodium ‘Zhongshansha’)具有极强的耐淹性, 但其耐淹机理仍没有明确。该研究以‘中山杉118’ (Taxodium ‘Zhongshansha 118’)幼苗为对象, 在经过93天不同水淹处理(对照、水浸、浅淹、深淹)后测定中山杉叶片和根系的无氧呼吸酶活性、淀粉及可溶性糖含量、生物量以及根系活力, 从能量消耗的角度初步探索了中山杉的耐淹性。结果表明: 长期水淹使中山杉叶片与根系中3种无氧呼吸酶(乙醇脱氢酶、丙酮酸脱羧酶、乳酸脱氢酶)活性显著增加, 且叶片与根系的乙醇脱氢酶活性均高于乳酸脱氢酶活性, 中山杉的根系和叶片是通过加强以酒精发酵为主的无氧呼吸适应长期缺氧环境; 不同水淹处理的叶片中3种无氧呼吸酶活性均高于根系, 叶片对缺氧环境更加敏感; 中山杉叶片和根系淀粉、可溶性糖含量均随水淹深度的增加显著增加, 根系淀粉含量显著高于叶片, 可溶性糖含量低于叶片; 中山杉根系淀粉含量高是其能够长期忍受水淹的重要原因, 且中山杉适应长期水淹的策略为忍耐型; 经受长期水淹后中山杉根茎结合部长出气生根及茎基部膨大, 同时根系外壁的木质化能将根系与外部水淹环境隔离, 具有很强的耐淹性, 可作为湿地生态修复、消落带生物治理的优良植物材料。     

PEG模拟干旱胁迫对栓皮栎种子萌发及生长生理的影响

为了阐明栓皮栎种子萌发期对水分胁迫的耐受性,以当年采集的栓皮栎种子为材料,采用培养皿滤纸萌发的方法研究了不同浓度（0%、5%、10%、20%、30%）PEG-6000溶液对其种子萌发、生长、保护酶活性和有机渗透调节物质的影响,并对种子萌发率、胚根生长速率等与PEG浓度进行了回归分析。结果表明:（1）随着干旱胁迫程度的加剧,栓皮栎种子的发芽率、发芽指数、胚根长度、胚根生长速率和活力指数均呈先升高后下降趋势,说明低浓度（5%、10%）PEG胁迫可以促进栓皮栎种子萌发及生长,而高浓度（20%、30%）则具有抑制作用。（2）建立水分胁迫条件下种子萌发率、胚根生长速率等与PEG浓度的回归方程发现,栓皮栎种子在PEG胁迫下的萌发临界值和极限值分别为32.01%和36.99%,胚根生长速率的临界值和极限值分别为30.1%和33.35%。（3）随着PEG浓度的升高,栓皮栎种子中3种保护酶（SOD、POD、CAT）活性和3种有机渗透调节物质（可溶性蛋白、脯氨酸、可溶性糖）含量呈现不同的变化,但均有不同程度升高趋势。由此可见,栓皮栎种子可通过增加保护酶活性和体内渗透调节物质来适应水分胁迫环境,使自身萌发和生长受抑制程度降到最低。     

Pod photosynthesis and seed dark CO2 fixation support oil synthesis in developingBrassica seeds

Rate of photosynthesis and activities of photosynthetic carbon reduction cycle enzymes were determined in pods (siliqua), whereas rate of dark CO₂ fixation, oil content and activities of enzymes involved in dark CO₂ metabolism were measured in seeds ofBrassica campestris L. cv. Toria at different stages of pod/seed development. The period between 14 and 35 days after anthesis corresponded to active phase of seed development during which period, seed dry weight and oil content increased sharply. Rate of pod photosynthesis and activities of photosynthetic carbon reduction cycle enzymes were maximum in younger pods but sufficiently high levels were retained up to 40 days after anthesis. The rate of dark¹⁴CO₂ fixation in seeds increased up to 21 days after anthesis and declined thereafter but maintaining sufficiently high rates till 35 days after anthesis. Similarly various enzymes viz., phosphoenolpyruvate carboxylase, NAD⁺-malate dehydrogenase and NADP⁺-malic enzyme, involved in dark CO₂ metabolism retained sufficient activities during the above period. These enzyme activities were more than adequate to maintain the desired supply of malate which mainly arises from dark CO₂ fixation in seeds and further translocated to leucoplasts for onward synthesis of fatty acids. Enzyme localization experiments revealed phosphoenolpyruvate carboxylase and enzymes of sucrose metabolism to be present only in cytosol, whereas enzymes of glycolysis were present both in cytosolic and leucoplastic fractions. These results indicated that oil synthesis in developingBrassica seeds is supported by pod photosynthesis and dark CO₂ fixation in seeds as the former serves as the source of sucrose and the latter as a source of malate     

The expression and anaerobic induction of alcohol dehydrogenase in cotton

The alcohol dehydrogenase (ADH) system in cotton is characterized, with an emphasis on the cultivated allotetraploid speciesGossypium hirsutum cv. Siokra. A high level of ADH activity is present in seed of Siokra but quickly declines during germination. When exposed to anaerobic stress the level of ADH activity can be induced several fold in both roots and shoots of seedlings. Unlike maize andArabidopsis, ADH activity can be anaerobically induced in mature green leaves. Three major ADH isozymes were resolved in Siokra, and it is proposed that two genes,Adh1 andAdh2, are coding for these three isozymes. The genes are differentially expressed. ADH1 is predominant in seed and aerobically grown roots, while ADH2 is prominent in roots only after anaerobic stress. Biochemical analysis demonstrated that the ADH enzyme has a native molecular weight of approximately 81 kD and a subunit molecular weight of approximately 42 kD, thus establishing that ADH in cotton is able to form and is active as dimers. Comparisons of ADH activity levels and isozyme patterns between Siokra and other allotetraploid cottons showed that the ADH system is highly conserved among these varieties. In contrast, the diploid species of cotton all had unique isozyme patterns.This work was generously supported by an Australian Cotton Research Council Postgraduate Studentship.     

Can Hypoxia Tolerance Explain Differences in Distribution of Two Co-Occurring North Temperate Sunfishes?

Many small, isolated north temperate waterbodies experience hypoxic conditions and winterkill events. Although such waterbodies are found in the natural ranges of two congeneric sunfishes (pumpkinseed, Lepomis gibbosus and bluegill, L. macrochirus), many contain pumpkinseed but no bluegill; a biogeographic pattern that has remained unexplained. To test whether a greater hypoxia tolerance in pumpkinseeds could explain these differences in distribution, we conducted hypoxia tolerance trials by subjecting each species to declining oxygen concentrations over ca. 16 h in aquaria. We also measured the activities of key metabolic enzymes in the muscle of wild individuals. Pumpkinseeds showed significantly higher tolerance to hypoxic stress than bluegills, as indicated by dissolved oxygen concentration at the time of equilibrium loss. Consistent with this result, white muscle from pumpkinseed had higher levels of lactate dehydrogenase, a marker enzyme for anaerobic capacity. There was no difference between species in the activity level of pyruvate kinase, suggesting that pumpkinseed do not display a general upregulation of glycolysis, but rather an upregulation of anaerobic capacity. Our results support the hypothesis that evolved differences in winter hypoxia tolerance can act as a macrohabitat partitioning mechanism in North American sunfishes.     

Engineering of enhanced glycine betaine synthesis improves drought tolerance in maize

Glycine betaine plays an important role in some plants, including maize, in conditions of abiotic stress, but different maize varieties vary in their capacity to accumulate glycine betaine. An elite maize inbred line DH4866 was transformed with the betA gene from Escherichia coli encoding choline dehydrogenase (EC 1.1.99.1), a key enzyme in the biosynthesis of glycine betaine from choline. The transgenic maize plants accumulated higher levels of glycine betaine and were more tolerant to drought stress than wild-type plants (non-transgenic) at germination and the young seedling stage. Most importantly, the grain yield of transgenic plants was significantly higher than that of wild-type plants after drought treatment. The enhanced glycine betaine accumulation in transgenic maize provides greater protection of the integrity of the cell membrane and greater activity of enzymes compared with wild-type plants in conditions of drought stress.     

Enzyme reactions involved in anaerobic cyclohexanol metabolism by a denitrifying Pseudomonas species

The enzymes involved in the anaerobic degration of cyclohexanol were searched for in a denitrifying Pseudomonas species which metabolizes this alicyclic compound to CO₂ anaerobically. All postulated enzyme activities were demonstrated in vitro with sufficient specific activities. Cyclohexanol dehydrogenase catalyzes the oxidation of the substrate to cyclohexanone. Cyclohexanone dehydrogenase oxidizes cyclohexanone to 2-cyclohexenone. 2-Cyclohexenone hydratase and 3-hydroxycyclohexanone dehydrogenase convert 2-cyclohexenone via 3-hydroxycyclohexanone into 1,3-cyclohexanedione. Finally, the dione is cleaved by 1,3-cyclohexanedione hydrolase into 5-oxocaproic acid. Some kinetic and regulatory properties of these enzymes were studied.     

Enzyme activities during imbibition and germination of seeds of tamarack (Larix laricina)

Activities of six enzymes from extracts of separated embryos and gametophytes of tamarack [ Larix laricina (Du Roi) K. Koch] seeds were assayed at various stages of imbibition and germination. On a per seed part basis, activities of 6-phosphogluconate dehydrogenase (6-PGD, EC 1.1.1.44), glucose-6-phosphate dehydrogenase (G-6-PD, EC 1.1.1.49), malate dehydrogenase (NAD⁺–MDH, EC 1.1.1.37), isocitrate dehydrogenase (NADP⁺–IDH, EC 1.1.1.42), soluble peroxidase (PER, EC 1.11.1.7), and acid phosphatase (ACP, EC 3.1.3.2) from both the embryo and gametophyte tissues generally increased slowly, following cold stratification for 30 days and imbibition under germinating conditions for 5 days, but then increased at a faster rate with emergence of the radicle and subsequent growth of the seedling. The rate of increase of enzyme activity was highest for PER. Soluble protein levels also increased with imbibition and germination, with about 3 times greater levels present in the gametophyte than in the embryo. Heat inactivation experiments showed that, except for G-6-PD, activities were stable up to 40°C. Inactivation occurred at lower temperatures for G-6-PD, while higher temperatures were required for PER. Incubation of extracts for 7 days at 4°C indicated that loss of enzyme activity was greatest for G-6-PD (3.9% remaining) and least for PER and ACP (94 and 95% remaining, respectively).     

干旱胁迫对鹿角杜鹃种子萌发和幼苗生理特性的影响

为探明鹿角杜鹃种子萌发和幼苗生长期的耐旱性,以鹿角杜鹃干种子和90d苗龄幼苗为材料,采用聚乙二醇(PEG-6000)模拟不同程度的干旱胁迫,研究干旱胁迫对其种子萌发、早期幼苗生长及幼苗的细胞膜透性、MDA含量、有机渗透调节物质和抗氧化酶活性的影响,并对种子萌发率、早期幼苗生长量与PEG胁迫浓度间进行了回归分析。结果表明:(1)5%~25%PEG胁迫范围内,随着干旱胁迫程度的增加,鹿角杜鹃种子的发芽启动时间推迟,发芽持续时间延长,发芽率、发芽势、发芽指数、活力指数和幼苗生长量显著降低;重度干旱胁迫(25%PEG)下,鹿角杜鹃种子完全未萌发。(2)发芽率、发芽势、发芽指数、活力指数以及幼苗生长量的变化均与干旱胁迫程度呈极显著负相关关系,回归分析求得鹿角杜鹃种子萌发的半致死PEG干旱胁迫浓度为15.68%、半致矮PEG干旱胁迫浓度为15.37%。(3)随着PEG胁迫浓度的增加,鹿角杜鹃幼苗叶片SOD活性呈先升后降的趋势,但各胁迫处理仍显著高于CK(0%PEG);细胞膜透性、MDA、脯氨酸、可溶性糖含量、POD和CAT活性则在中度(15%~20%PEG)和重度胁迫下显著升高,与干旱胁迫程度呈极显著正相关关系。研究表明,干旱胁迫显著抑制了鹿角杜鹃种子萌发和早期幼苗生长,使其细胞膜受到损伤,同时鹿角杜鹃可通过体内渗透调节物质和抗氧化酶活性的增加来适应干旱环境,使得自身受抑制、损伤程度降到最低。     

The effects of recessive lethal Notch mutations of Drosophila melanogaster on flavoprotein enzyme activities whose inhibitions cause Notch-like phenocopies

The biochemical action of the Notch locus whose mutants cause morphological aberrations in flies, viz., notches of wings and bristle multiplication, has been analyzed (1) by the addition to the food medium of enzyme inhibitors causing phenocopies of Notch and (2) by comparison of enzyme activity patterns of Notch mutants with different degrees of phenotypic expression. Notch phenocopies were induced by inhibitors of enzyme activities in two biochemical pathways: (1) the de novo pyrimidine synthesis by 5-methylorotate (inhibitor of dihydroorotate dehydrogenase) and (2) the choline shunt by amobarbital (inhibits choline dehydrogenase) and methoxyacetate (inhibits sarcosine dehydrogenase). The inhibition of de novo pyrimidine synthesis prevents the production of deoxyuridine-5-phosphate, the substrate for the synthesis of thymidine-5-phosphate via thymidylate synthase, whereas the inhibition of the choline shunt prevents the production of HCHO groups and glycine, both of which are involved in the synthesis of 5,10-methylenetetrahydrofolate, which is a cofactor of thymidylate synthase. It was already known that the inhibition of the latter enzyme in vivo induces Notch phenocopies. Notch mutants with a strong morphological expression show low enzyme activities for dihydroorotate dehydrogenase and choline dehydrogenase. Both are flavoprotein enzymes linked to the respiratory chain. The correspondence between the low enzyme activities in Notch mutants with a strong morphological expression and the phenocopying effect of antimetabolites on these enzymes in the two biochemical pathways involved strongly suggests that the morphological effects of Notch on flies are a consequence of lowered activities of choline dehydrogenase and dihydroorotate dehydrogenase.     

Metabolic adaptation to flooding stress in upland and lowland rice seedlings

Ability of metabolic adaptation in upland and lowland rice (Oryza sativa L.) seedlings to flooding stress was compared. Flooding stress increased alcohol dehydrogenase (ADH) activity and ethanol concentration in shoots and roots of the upland and lowland rice seedlings. The difference in ADH activity and ethanol concentration in shoots between the upland and lowland rice was not apparent. However, both ADH activity and ethanol concentration in roots of the lowland rice were 2-fold greater than those in roots of the upland rice, suggesting that flooding-induction of ethanolic fermentation in lowland rice roots may be significantly greater than that in the upland rice roots. Since flooding often causes the anaerobic conditions in rooting zone than aerial part of plants and ethanolic fermentation is essential to survive in the anaerobic conditions, the ability of metabolic adaptation in lowland rice seedlings to flooding stress may be greater than that in upland rice seedlings.     

Variation for Root Aerenchyma Formation in Flooded and Non-Flooded Maize and Teosinte Seedlings

Morphological and anatomical factors such as aerenchyma formation in roots and the development of adventitious roots are considered to be amongst the most important developmental characteristics affecting flooding tolerance. In this study we investigated the lengths of adventitious roots and their capacity to form aerenchyma in three- and four-week-old seedlings of two maize (Zea mays ssp. mays, Linn.) inbred accessions, B64 and Na4, and one teosinte, Z. nicaraguensis Iltis & Benz (Poaceae), with and without a flooding treatment. Three weeks after sowing and following a seven day flooding treatment, both maize and teosinte seedlings formed aerenchyma in the cortex of the adventitious roots of the first three nodes. The degree of aerenchyma formation in the three genotypes increased with a second week of flooding treatment. In drained soil, the two maize accessions failed to form aerenchyma. In Z. nicaraguensis, aerenchyma developed in roots located at the first two nodes three weeks after sowing. In the fourth week, aerenchyma developed in roots of the third node, with a subsequent increase in aerenchyma in the second node roots. In a second experiment, we investigated the capacity of aerenchyma to develop in drained soil. An additional three teosinte species and 15 maize inbred lines, among them a set of flooding-tolerant maize lines, were evaluated. Evaluations indicate that accessions of Z. luxurians (Durieu & Asch. Bird) and two maize inbreds, B55 and Mo20W, form aerenchyma when not flooded. These materials may be useful genetic resources for the development of flooding-tolerant maize accessions.     

Metabolic changes in tomato fruits and seeds after viral, bacterial and fungal infections

The metabolic changes in tomato fruits and seeds separately infected with cucumber mosaic virus, Pseudomonas syringae pv. tomato or Botrytis cinerea were investigated cytochemically. The changes of peroxidase (E.C. 1.11.1.7) and β-glucosidase (E.C. 3.2.1.21) were investigated biochemically as well. Tomato fruits were involved in the study because of their high economic value. Tomato seeds were investigated since they have been used most extensively as a model system for studying the physiology and biochemistry of seed development. The diseases caused by the pathogens under study are of special importance for yield reduction in tomato. The three pathogens provoked local changes in the activities of enzymes under study that affect the infected pericarp tissues and neighboring seeds. It was established non-specific and specific responses. The non-specific responses of invaded tissues were expressed as a local enhancement of peroxidase activity in both pericarp tissues and seeds as well as a decrease in activities of: i. enzymes taking part in aerobic and anaerobic respiration, ii. hydrolases esterase and acid phosphatase involved in the basic metabolism as well as an enhancement of their activities in neighboring tissues. Furthermore, it was observed an enhancement of α-galactosidase activity in infected area was observed. The specific responses depending on the type of the pathogen consisted in an enhancement of glucose-6-phosphate dehydrogenase activity by virus infection and an increase of β-glucosidase activity by fungal invasion.     

Mitochondrial enzymes in aerobically and anaerobically germinated seedlings of Echinochloa and rice

Activities of tricarboxylic acid (TCA) cycle enzymes in seedlings of barnyard grass (Echinochloa phyllopogon (Stapf.) Koss) and rice (Oryza sativa L.) germinated under aerobic and anaerobic conditions were investigated. In E. phyllopogon, development of TCA-cycle enzyme activities during 10 d of anoxia generally paralleled those in air, although at lower rates. After 5 d, E. phyllopogon seedlings germinating under N₂ exhibited 50–80% of the activity of seedlings grown in air, except for 2-oxoglutarate dehydrogenase (EC 1.2.4.2) and fumarate reductase (EC 1.3.1.6) which exhibited only 25–35% of aerobic activity. In anaerobically germinated rice, development of TCA-cycle enzyme activities also paralleled those in air except for aconitase (EC 4.2.1.3), isocitrate dehydrogenase (EC 1.1.1.41), and 2-oxoglutarate dehydrogenase. Those enzymes did not increase in activity under anoxia. Development of maximum enzyme activities generally occurred more rapidly and persisted longer in E. phyllopogon compared to rice. The data indicate that mitochondria of E. phyllopogon function better during anaerobiosis than those of rice and this factor may contribute to the successful biochemical strategy of this weed in rice paddies throughout the world.Abbreviation TCA tricarboxylic acid This work was supported by U.S. Department of Agriculture Competitive Research grant No. 87-CRCR1-2595 and a Herman Frasch Foundation grant in Agricultural Chemistry to R.A.K.     

New isozyme systems for maize (Zea mays L.): Aconitate hydratase,adenylate kinase,NADH dehydrogenase,and shikimate dehydrogenase

Electrophoretic variation and inheritance of four novel enzyme systems were studied in maize (Zea mays L.). A minimum of 10 genetic loci collectively encodes isozymes of aconitate hydratase (ACO; EC 4.2.1.3.), adenylate kinase (ADK; EC 2.7.4.3), NADH dehydrogenase (DIA; EC 1.6.99.—), and shikimate dehydrogenase (SAD; EC 1.1.1.25). At least four loci are responsible for the genetic control of ACO. Genetic data for two of the encoding loci,Aco1 andAco4, demonstrated that at least two maize ACOs are active as monomers. Analysis of organellar preparations suggests that ACO1 and ACO4 are localized in the cytosolic and mitochondrial subcellular fractions, respectively. Maize ADK is encoded by a single nuclear locus,Adk1, governing monomeric enzymes that are located in the chloroplasts. Two cytosolic and two mitochondrial forms of DIA were electrophoretically resolved. Segregation analyses demonstrated that the two cytosolic isozymes are controlled by separate loci,Dia1 andDia2, coding for products that are functional as monomers (DIA1) and dimers (DIA2). The major isozyme of SAD is apparently cytosolic, although an additional faintly staining plastid form may be present. Alleles atSad1 are each associated with two bands that cosegregate in controlled crosses. Linkage analyses and crosses with B-A translocation stocks were effective in determining the map locations of six loci, including the previously described but unmapped locusAcp4. Several of these loci were localized to sparsely mapped regions of the genome.Dia2 andAcp4 were placed on the distal portion of the long arm of chromosome 1, 12.6 map units apart.Dia1 was localized to chromosome 2, 22.2 centimorgans (cM) fromB1. Aco1 was mapped to chromosome 4, 6.2 cM fromsu1. Adk1 was placed on the poorly marked short arm of chromosome 6, 8.1 map units fromrgd1. Less than 1% recombination was observed betweenGlu1 (on chromosome 10) andSad1. In contrast to many other maize isozyme systems, there was little evidence of gene duplication or of parallel linkage relationships for these allozyme loci. This work was supported by grants from Pioneer Hi-Bred International, Inc., of Johnston, Iowa, the National Institute of Health (Research Grant GM11546), and the United States Department of Agriculture (Competitive Research Grant 83-CRCR-1-1273). This is Paper No. 11372 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh.     

Oxidative and reductive acetyl CoA/carbon monoxide dehydrogenase pathway in Desulfobacterium autotrophicum

It has been proposed that in some anaerobic facultatively autotrophic bacteria the acetyl CoA/CO dehydrogenase pathway is operating both in the reductive and in the oxidative direction, depending on the growth conditions. One of these anaerobes, the Gram-negative sulfate-reducing cubacterium Desulfobacterium autotrophicum, was examined for enzymes of the proposed pathway. All the required enzyme activities were present in sufficient amounts both in autotrophically and in heterotrophically grown cells, provided that the cellular tetrahydropterin rather than tetrahydrofolate was used as cosubstrate in some of the enzyme assays. The question arises whether two sets of enzymes are operating in the reductive and oxidative direction, respectively. The key enzyme of this pathway, CO dehydrogenase, which was reasonably oxygen stable, was analysed by native polyacrylamide gel electrophoresis and anaerobic activity staining. Extracts from heterotrophically grown cells exhibited five enzyme activity bands. Extracts from autotrophically grown cells showed the same pattern but an additional activity band appeared.