Low root temperature effects on soybean nitrogen metabolism and photosynthesis

The influences of low root temperature on soybeans (Glycine max [L.] Merr. cv. Wells) were studied by germinating and maintaining plants at root temperatures of 13 and 20 C through maturity. At 42 days from the beginning of imbibition, 13 and 20 C plants were switched to 20 and 13 C, respectively. Plants were harvested after 63 days. Control plants (13 C) did not nodulate, whereas those switched to 20 C did and at harvest had C₂H₂ reduction rates of 0.2 micromoles per minute per plant. Rates of C₂H₂ reduction decreased rapidly in plants switched from 20 to 13 C; however, after 2 days, rates recovered to original levels (0.8 micromoles per minute per plant) and then began a slow decline until harvest. Arrhenius plots of C₂H₂ reduction by whole plants indicated a large increase in the energy of activation below the inflection at 15 C. Highest C₂H₂ reduction rates (1.6 micromoles per minute per plant) were at 58 days for the 20 C control. Root respiration rates followed much the same pattern as C₂H₂ reduction in the 20 C control and transferred plants. At harvest, roots from 13 C-treated plants had the highest activities for malate dehydrogenase, glutamate oxaloacetate transaminase, and phosphoenolpyruvate carboxylase. Roots from transferred plants had intermediate activities and those from the 20 C treatment the lowest activities. Newly formed nodules from plants switched from 13 to 20 C had much higher glutamate dehydrogenase than glutamine synthetase activity.     

I. LIFE HISTORY FEATURES OF THREE SEXUAL MORPHS OF ATRIPLEX CANESCENS (CHENOPODIACEAE) CLONES GROWN IN A COMMON GARDEN

Reproductive and growth parameters of Atriplex canescens were examined in a common garden study. Cloned ramets of male, female, and hermaphrodite individuals from two natural populations were planted in irrigated and nonirrigated portions of the garden. Sexual differences in growth and flowering schedules suggest tradeoffs in resource allocation between growth and reproduction. Males flowered with the greatest frequency and intensity while producing the smallest growth parameters. Females had the largest measures of vegetative growth, but flowered with the least frequency and intensity. Hermaphrodites appear to have a biology distinct from males and females, being more like females in growth and more like males in reproduction.     

Regulation of endothelial argininosuccinate synthase expression and NO production by an upstream open reading frame

Argininosuccinate synthase (AS) catalyzes the rate-limiting step in the recycling of citrulline to arginine, which in endothelial cells, is tightly coupled to the production of nitric oxide (NO). In previous work, we established that endothelial AS mRNA can be initiated from multiple start sites, generating co-expressed mRNA variants with different 5'-untranslated regions (5'-UTRs). One of the 5'-UTRs, the shortest form, represents greater than 90% of the total AS mRNA. Two other extended 5'-UTR forms of AS mRNA, resulting from upstream initiations, contain an out-of-frame, upstream open reading frame (uORF). In this study, the function of the extended 5'-UTRs of AS mRNA was investigated. Single base insertions to place the uORF in-frame, and mutations to extend the uORF, demonstrated functionality, both in vitro with AS constructs and in vivo with luciferase constructs. Overexpression of the uORF suppressed endothelial AS protein expression, whereas specific silencing of the uORF AS mRNAs resulted in the coordinate up-regulation of AS protein and NO production. Expression of the full-length of the uORF was necessary to mediate a trans-suppressive effect on endothelial AS expression, demonstrating that the translation product itself affects regulation. In conclusion, the uORF found in the extended, overlapping 5'-UTR AS mRNA species suppresses endothelial AS expression, providing a novel mechanism for regulating endothelial NO production by limiting the availability of arginine.     

Infectious bronchitis virus 3a protein localizes to a novel domain of the smooth endoplasmic reticulum

All coronaviruses possess small open reading frames (ORFs) between structural genes that have been hypothesized to play important roles in pathogenesis. Infectious bronchitis virus (IBV) ORF 3a is one such gene. It is highly conserved among group 3 coronaviruses, suggesting that it has an important function in infection. IBV 3a protein is expressed in infected cells but is not detected in virions. Sequence analysis predicted that IBV 3a was a membrane protein; however, only a fraction behaved like an integral membrane protein. Microscopy and immunoprecipitation studies demonstrated that IBV 3a localized to the cytoplasm in a diffuse pattern as well as in sharp puncta in both infected and transfected cells. These puncta did not overlap cellular organelles or other punctate structures. Confocal microscopy demonstrated that IBV 3a puncta lined up along smooth endoplasmic reticulum (ER) tubules and, in a significant number of instances, were partially surrounded by these tubules. Our results suggest that IBV 3a is partially targeted to a novel domain of the smooth ER.     

Persistence of wild-type virus and lack of temporal structure in the latent reservoir for human immunodeficiency virus type 1 in pediatric patients with extensive antiretroviral exposure

Although highly active antiretroviral therapy (HAART) for human immunodeficiency virus type 1 (HIV-1) infection can reduce levels of HIV-1 RNA in plasma to below the limit of detection, replication-competent forms of the virus persist in all infected individuals. One form of persistence involves a stable reservoir of latent but potentially infectious virus that resides in resting memory CD4(+) T cells. The mechanisms involved in maintaining this latent reservoir are incompletely understood. In the present study, we examined the dynamic characteristics of this reservoir in a cohort of children who developed drug-resistant HIV-1 as a result of extensive exposure to inadequately suppressive one- or two-drug regimens prior to the advent of HAART. We have previously shown that drug-resistant viruses selected by nonsuppressive pre-HAART regimens can enter and persist in this reservoir. We have extended these findings here by demonstrating that archival wild-type HIV-1 persists in this reservoir despite the fact that in these patients drug-resistant mutants have been favored by the selective conditions for many years. Phylogenetic analysis of replication-competent viruses persisting in resting CD4(+) T cells revealed a striking lack of temporal structure in the sense that isolates obtained at later time points did not show greater sequence divergence than isolates from earlier time points. The persistence of drug-sensitive virus and the lack of temporal structure in the latent reservoir provide genetic evidence for the idea that HIV-1 can persist in a latent form free of selective pressure from antiretroviral drugs in long-lived resting memory CD4(+) T cells. Although there may be other mechanisms for viral persistence, this stable pool of latently infected cells is of significant concern because of its potential to serve as a lasting source of replication-competent viruses, including the infecting wild-type form and all drug-resistant variants that have arisen subsequently.     

Regulation of chicken gizzard ecto-ATPase activity by modulators that affect its oligomerization status

The major ectonucleoside triphosphate phosphohydrolase in the chicken gizzard smooth muscle membranes is an ecto-ATPase, an integral membrane glycoprotein belonging to the E-ATPase (or E-NTPDase) family. The gizzard ecto-ATPase is distinguished by its unusual kinetic properties, temperature dependence, and response to a variety of modulators. Compounds that promote oligomerization of the enzyme protein, i.e., concanavalin A, chemical cross-linking agent, and eosin iodoacetamide, increase its activity. Compounds that inhibit some ion-motive ATPases, e.g., sulfhydryl reagents, xanthene derivatives, NBD-halides, and suramin, also inhibit the gizzard ecto-ATPase, but not another E-ATPase, the chicken liver ecto-ATP-diphosphohydrolase, which contains the same conserved regions as the ecto-ATPase. Furthermore, inhibition of the gizzard ecto-ATPase by these compounds as well as detergents is not prevented by preincubation of the membranes with the substrate, ATP, indicating that their interaction with the enzyme occurs at a locus other than the catalytic site. On the other hand, the inhibitory effect of these compounds, except suramin, is abolished or reduced if the membranes are preincubated with concanavalin A. It is concluded that these structurally unrelated modulators exert their effect by interfering with the oligomerization of the ecto-ATPase protein. Our findings suggest that, under physiological conditions, the gizzard smooth muscle ecto-ATPase may exhibit a range of activities determined by membrane events that affect the status of oligomerization of the enzyme.     

Genetic Regulation of Seed Dormancy in Purshia tridentata(Rosaceae)

Seeds of Purshia tridentata, a shrub of semi-arid North America,require chilling to become non-dormant. Using seeds producedfrom controlled crosses in a common garden, we examined effectsof ovule parent, pollen parent, and year of production on germinationpercentage in response to chilling for 2 weeks at 2°C. Differencesamong ovule parents accounted for most of the variance in chillingresponse, and these differences (2 to 83% germination) wereconsistent across years. Differences among pollen parents werealso significant, producing a two- to five-fold difference inmean germination percentage. Differences among years were significantbut small. Ovule parent by year interactions showed that among-yearvariation in ripening environment did not affect ovule parentsequally. There was no significant pollen parent by year interaction,suggesting that the effect of maturation environment was mediatedthrough maternal tissues. In reciprocal crosses, two plantsthat showed contrasting dormancy levels as ovule parents producedseeds with similar dormancy as pollen parents, indicating thatthe genetic difference between them was at the testa level.Two plants that produced seeds with contrasting dormancy aspollen parents showed a similar but stronger pattern of contrastas ovule parents, showing that the genetic difference betweenthem was at both embryo and endosperm or testa level. Testaor endosperm genotype was primarily responsible for chillingresponse of intact seeds, while embryo genotype affected chillingresponse and also exercised primary control over low temperaturegermination rate, whether of excised embryos or of intact seeds.Copyright 2000 Annals of Botany Company Antelope bitterbrush, bitterbrush, common garden study, germination