PHYLOGENETIC IMPLICATIONS OF DIFFERENCES IN NUMBER OF PLASTID PHOSPHOGLUCOSE ISOMERASE ISOZYMES IN NORTH AMERICAN COREOPSIS (ASTERACEAE: HELIANTHEAE: COREOPSIDINAE)

Enzyme electrophoresis was employed to ascertain the number of loci encoding plastid phosphoglucose isomerase (PGI) in species representing all sections of North American Coreopsis. Several species from each of the closely related genera Bidens, Coreocarpus, Cosmos, and Thelesperma were also examined. Species in nine of the 11 sections of North American Coreopsis have two isozymes for plastid PGI, and nearly all species examined in the four other genera also have two (one species has three) isozymes. Since most diploid vascular plants have one plastid PGI isozyme, a gene duplication probably occurred in an ancestor that is common to Coreopsis and the other four genera. That is, two isozymes represent the ancestral number for Coreopsis. The two sections (Electra and Anathysana) apparently lacking the duplication are closely related woody plants restricted largely to Mexico. One gene encoding plastid PGI ostensibly was silenced in a common ancestor of these two sections. This is concordant with other data suggesting a close relationship between the two sections, i.e., they appear to represent a monophyletic group. The electrophoretic data also indicate that 1) the enigmatic monotypic section Silphidium is more closely related to eastern North American sections and not derived from section Electra; and 2) section Anathysana is not ancestral to the three California sections Leptosyne, Pugiopappus, and Tuckermannia; rather, it represents a terminal element closely related to and possibly derived from section Electra.     

Degradation of extractive-free lignocelluloses by Coriolus versicolor and Poria placenta

Summary The wood-decay fungi Coriolus versicolor, a white-rot fungus, and Poria placenta, a brown-rot fungus, were grown on an extractive-free lignocellulose prepared from quackgrass (Agropyron repens). Their abilities to decompose this lignocellulose were compared to their abilities to decompose softwood (Picea pungens) and hardwood (Acer rubrum) lignocelluloses. The two fungi were grown on malt-extract dampened lignocelluloses at 28°C for up to 12 weeks. Replicate cultures were periodically harvested and lignocellulose decomposition was followed by monitoring substrate weight loss, lignin loss, and carbohydrate loss. Coriolus versicolor decomposed the lignin and carbohydrate components of the grass lignocellulose as efficiently as the softwood and hardwood lignocelluloses. Poria placenta, however, was not an efficient degrader of either lignin or carbohydrate in the grass lignocellulose. Poria placenta readily decomposed carbohydrate components of the softwood lignocellulose but not the hardwood lignocellulose.Paper number 81520 of the Idaho Agricultural Experiment Station     

Adenylate cyclase of human articular chondrocytes. Responsiveness to prostaglandins and other hormones.

Adenylate cyclase [ATP pyrophosphate lyase (cyclizing), EC 4.6.1.1] was shown to be present in cultured human articular chondrocytes. Optimal conditions of incubation time, protein and substrate concentrations and pH were determined in whole cell lysates. Maximal activity occurred at pH 8.5 with no decrease in activity up to pH 10.0. Adenylate cyclase activity of particulate membrane preparations was enhanced by the addition of crude cytosol preparations. The prostaglandins E1, E2, F1 alpha, F2 alpha, D2, B1, B2, A1 and A2, as well as adrenaline and isoprenaline, stimulated adenylate cyclase derived from either adult or foetal chondrocytes. No significant stimulation was observed in the presence of human calcitonin or glucagon. Bovine parathyroid hormone always significantly stimulated the adenylate cyclase derived from foetal chondrocytes, but not from adult chondrocytes. Preincubation of the chondrocytes in culture with indomethacin and with or without supernatant medium from cultured mononuclear cells increased the responsiveness of the adenylate cyclase to prostaglandin E1.     

Sources, Fluxes, and Sinks of Nitrogen during Early Reproductive Growth of Maize (Zea mays L.)

A study was designed to (a) identify sources and sinks of N in the maize (Zea mays L.) shoot, by estimating net N fluxes for each of seven parts of the shoot, (b) determine effects of N entering the plant upon fluxes of N absorbed before reproductive growth, and (c) determine the effects of the opaque-2 gene on N fluxes in the maize shoot during early reproductive growth. Plants of a maize hybrid (Pioneer 3369A) and its opaque-2 counterpart (Pioneer L3369) were grown in a greenhouse using nutrient solution/sand culture, with NO₃⁻ as the N source during the vegetative growth phase. Beginning at the time of pollination, the same nutrient regime was continued, except that some plants received no N, and others received 3.75 millimolar ¹⁵N as NO₃⁻-N.

Stalk and leaves were found to be primary N sources for the grain, while shank, husk, and cob acted first as N sinks, then as N sources during reproductive growth. Net fluxes of N for each plant part were estimated by calculating the first derivatives of regression equations used to fit data for N contents of each plant part as functions of time. All parts of the shoot were sinks for exogenous N (absorbed after pollination). Thirty-six days after pollination, the grain contained 60% endogenous N (absorbed before pollination) when 3.75 millimolar NO₃⁻-N was supplied after pollination. Rates of total N influx to the grain were identical whether or not N was supplied in the nutrient solution during reproductive growth. At 36 days after pollination, less N had accumulated in the grain of the opaque-2 genotype, but otherwise there were no differences in N contents or dry weights of the shoots due to the opaque-2 gene. Absence of N from the rooting medium significantly affected N fluxes throughout the shoot during reproductive growth, but there were no detectable effects of the opaque-2 gene on N fluxes in parts of the plant other than the grain.

     

Comparison between mutagenesis in normal and transformed syrian hamster fibroblasts: Difference in the temporal order of HPRT gene replication

A highly tumorigenic subdiploid cell line, BP6T, derived in our laboratory from Syrian hamster embryo (SHE) cells, is amenable to studies of somatic mutation in vitro. Cellular and biochemical characterization of clonally derived BP6T cells resistant to 6-thioguanine (TG^r) or ouabain (Oua^r) demonstrated these mutants to be similar qualitatively to mutants of SHE cells characterized previously (Barrett et al., 1978). BP6T TG^r mutants resistant to 6-thioguanine are cross-resistant to 8-azaguanine, lack HPRT activity, exhibit a low frequency of reversion and arise spontaneously at a rate of 5 × 10⁻⁷ mutants per cell per generation. BP6T Oua^r mutants were shown to be highly resistant to ouabain-mediated inhibition of ⁸⁶Rb influx, indicating an alteration in the Na⁺/K⁺ ATPase. These studies on the BP6T cell line provide the experimental basis for a comparative study of the mutagenic responses of normal, diploid SHE cells versus those of related, but transformed aneuploid cells. Highly synchronized cultures of these 2 cells were mutagenized by pulse treatment with BrdU during different periods of S phase, followed immediately by near-UV irradiation. The induced mutation frequencies so obtained provided information about the temporal order of replication of genes encoding HPRT and Na⁺/K⁺ ATPase in both SHE and BP6T cells. The temporal pattern of replication of Na⁺/K⁺ ATPase gene loci is similar in both cell types, but the temporal order of replication of the HPRT gene is significantly different between SHE and BP6T cells (mid-late S phase, versus early S phase, resp.). This observed difference emphasizes the caution required in the study of mutagenesis and DNA replication using transformed, aneuploid cells under the assumption that the underlying mechanisms are the same for normal, diploid cells.     

Nucleotide sequence of the trpB gene in Escherichia coli and Salmonella typhimurium

We have obtained the entire nucleotide sequence of the penultimate gene of the tryptophan operon, trpB, in Escherichia coli and Salmonella typhimurium. The amino acid sequence deduced for the E. coli gene product is in agreement with earlier, fragmentary protein sequence results. The trpB nucleotide sequences for the two bacterial species are perfectly colinear and show 85% identity. Most of the nucleotide differences found are without consequence for the amino acid sequence, which shows greater than 96% identity. The degree of conservation of both the nucleotide and amino acid sequences is significantly greater than for trpA, the adjacent gene encoding the other subunit of the same enzyme. When synonymous third codon position nucleotide differences are examined, they seem to be distributed at random throughout trpB and trpA, except for one completely conserved 66 basepair long region within trpB.     

The function of tail fibers in triggering baseplate expansion of bacteriophage T4

Normal particles of bacteriophage T4 have six long tail fibers attached to a hexagonal baseplate. T4 particles having various complements of tail fibers were prepared by in vitro addition of fibers to fiberless particles, and the infectivity of the particles was determined. Particles having fewer than six fibers (partially fibered) were found to have a decreased probability of infection. Partially fibered particles having T4 fibers were completed by addition of T6 fibers, and the infectivity was determined on a host that lacked the T6 tail fiber receptor. Attachment of the additional fibers increased the infectivity even though the T6 fibers could not bind to the host cell. The infectivity of particles having mixtures of T4 and T6 fibers was determined on cells having only one type of receptor. The results indicated that particles bound by only three fibers have a low probability of infection. The effect of thermolabile baseplate mutations was also examined. Studies of partially fibered particles and particles with mixtures of fibers indicated that particles with altered baseplates have a less stringent requirement for binding of the tail fibers for infection.     

Ordering tryptophan synthase genes of Pseudomonas aeruginosa by cloning in Escherichia coli.

The Pseudomonas aeruginosa tryptophan synthase genes, trpA and trpB, which are induced by their substrate indoleglycerol phosphate, were cloned along with their controlling region into the BamHI site of pBR322 to produce the 10.7-megadalton plasmid pZAZ5. SalI partial digestion and ligation yielded a smaller plasmid, pZAZ167, with the chromosomal insert reduced in size from 8.1 to 3.4 megadaltons. Both pZAZ5 and pZAZ167 display Pseudomonas-like regulation of the trpA and trpB genes. Deletion of an EcoRI fragment or a BglII fragment from pZAZ167 yielded plasmids pZAZ168 and pZAZ169; the former expresses trpB but not trpA, and the latter has lost both activities. A deleted form of pZAZ5 designated pZAZ101 was obtained by excising a BglII-BamHI segment and religating the trip gene segment in the opposite orientation. This plasmid expresses trpA and trpB constitutively. The physical maps of these plasmids establish the gene order: promoter-trpB-trpA.     

Flavonoid chemistry of arceuthobium (viscaceae)

Flavonoid compounds from 36 of the 38 known taxa of the genusArceuthobium (dwarf mistletoes) were examined. The flavonoid chemistry of the genus is rather uniform, all taxa producing 3-O-glycosides of the flavonols quercetin and myricetin. No infraspecific chemical variation was encountered, and in those instances where subspecific taxa are recognized, their chemistry was uniform. At the subgeneric level, members of subgenusArceuthobium synthesize primarily glucosides, whereas galactosides are more common in subgenusVaginata. In two of the four Old World species of subgenusArceuthobium (A. juniperi- procerae andA. oxycedri) only myricetin 3-O-glucoside was detected. There are no absolute flavonoid differences between subgenera, sections, or series. On the other hand, flavonoids are useful in several instances at the species level. In several cases, chemical data lend support to the recognition of species which in the past have been considered doubtfully distinct on the basis of morphology.