RFLP mapping of the sugary enhancer1 gene in maize

RFLP marker data from an F₂₃ population derived from a cross between a sugary1 (su1) and a sugary enhancer1 (su1, sel) inbred were used to construct a genetic linkage map of maize. This map includes 93 segregating marker loci distributed throughout the maize genome, providing a saturated linkage map that is suitable for linkage analysis with quantitative trait loci (QTL). This population, which has been immortalized in the form of sibbed F₂₃ families, was derived from each of the 214 F₂ plants and along with probe data are available to the scientific community. QTL analysis for kernel sucrose (the primary form of sugar) concentration at 20 days after pollination (DAP) uncovered the segregation of seven major QTL influencing sucrose concentration; a locus linked to umc36a described the greatest proportion of the variation (24.7%). Since maltose concentration has previously been reported to be associated with the se1 phenotype, an analysis of probe associations with maltose concentration at 40 DAP was also conducted. The highly significant association of umc36a with maltose and sucrose concentrations provided evidence that this probe is linked to se1. Phenotypic evaluation for the se1 genotype in each F₂₃ family enabled us to map the gene 12.1 cM distal to umc36a. In contrast to previous work where se1 was reported to be located on chromosome four, our data strongly suggest that the sugary enhancer1 locus maps on the the distal portion of the long arm of chromosome 2 in the maize genome.     

Binding-protein expression is subject to temporal,developmental and stress-induced regulation in terminally differentiated soybean organs

Binding protein (BiP) is a widely distributed and highly conserved endoplasmic-reticulum luminal protein that has been implicated in cotranslational folding of nascent polypeptides, and in the recognition and disposal of misfolded polypeptides. Analysis of cDNA sequences and genomic blots indicates that soybeans (Glycine max L. Merr.) possess a small gene family encoding BiP. The deduced sequence of BiP is very similar to that of other plant BiPs. We have examined the expression of BiP in several different terminally differentiated soybean organs including leaves, pods and seed cotyledons. Expression of BiP mRNA increases during leaf expansion while levels of BiP protein decrease. Leaf BiP mRNA is subject to temporal control, exhibiting a large difference in expression in a few hours between dusk and night. The expression of BiP mRNA varies in direct correlation with accumulation of seed storage proteins. The hybridization suggests that maturing-seed BiP is likely to be a different isoform from vegetative BiPs. Levels of BiP protein in maturing seeds vary with BiP mRNA. High levels of BiP mRNA are detected after 3 d of seedling growth. Little change in either BiP mRNA or protein levels was detected in maturing soybean pods, although BiP-protein levels decrease in fully mature pods. Persistent wounding of leaves by whiteflies induces massive overexpression of BiP mRNA while only slightly increasing BiP-protein levels. In contrast single-event puncture wounding only slightly induces additional BiP expression above the temporal variations. These observations indicate that BiP is not constitutively expressed in terminally differentiated plant organs. Expression of BiP is highest during the developmental stages of leaves, pods and seeds when their constituent cells are producing seed or vegetative storage proteins, and appears to be subject to complex regulation, including developmental, temporal and wounding.The mention of vendor or product does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over vendors of similar products not mentioned.Abbreviations BiP binding protein The sequences reported in this paper have been submitted to Gen-Bank and are identified with the accession numbers BiP-A (U08384), BiP-B (U08383), BiP-C (U08382) and -1,3 glucanase (U08405).     

Structure and evolution of teleost mitochondrial control regions

We amplified and sequenced the mitochondrial control region from 23 species representing six families of teleost fish. The length of this segment is highly variable among even closely related species due to the presence of tandemly repeated sequences and large insertions. The position of the repetitive sequences suggests that they arise during replication both near the origin of replication and at the site of termination of the D-loop strand. Many of the conserved sequence blocks (CSBs) observed in mammals are also found among fish. In particular, the mammalian CSB-D is present in all of the fish species studied. Study of potential secondary structures of RNAs from the conserved regions provides little insight into the functional constraints on these regions. The variable structure of these control regions suggests that particular care should be taken to identify the most appropriate segment for studies of intraspecific variation. Correspondence to: T.D. Kocher     

Alternaria infectoria , a Promising Biological Control Agent for the Fig Wax Scale, Ceroplastes rusci (Homoptera: Coccidae), in Egypt

Alternaria infectoria E . G . Simmons was isolated from naturally infected eggs of the fig wax scale insect , Ceroplastes rusci L . (Homoptera: Coccidae) , that were showing different degrees of shriveling . This fungus proved to be pathogenic to eggs , nymphs and adults of this pest . The fungus had significant effects on both the mortality and hatchability of the eggs ( P = 0 . 0001) . Three days after exposure to the fungus , nearly one - third (31 . 5%) of the fungus - treated eggs became discolored and 17% showed moderate shrinking but no mortality . Fifteen days post - application , 91% of the fungus - treated eggs were diseased (74 . 8% of them were dead) due to A. infectoria, while no disease developed on the untreated eggs . Hatching decreased by 39 . 6% over a 15 - day period in the fungus - treated eggs compared to the control . Additionally , crawlers hatching from fungus - treated eggs became infected . The fungus seemed to induce crawlers to enter the settling stage . Seventy - two per cent of the fungus - treated crawlers versus 9% of the untreated controls entered the settling stage 6 days after exposure to the fungal inoculum . The highest level of nymphal mortality attributed to A. infectoria occurred when 30% (wet w:v blended mycelium in water containing 0 . 5% w:v Metamucil) inoculum was applied , and a high relative humidity was maintained for the following 48 h . This is the first record of A. infectoria on the fig wax scale and for the genus Alternaria as an entomopatho genic fungus .     

Biology and Host Range of the Green-seed Weevil, Sibinia fastigiata , for Biological Control of Mimosa pigra

Aspects of the biology and host range of Sibinia fastigiata Clark (Coleoptera: Curculionidae) were studied to assess its safety for release in Australia as a biological control agent of the weed Mimosa pigra L . (Mimosaceae) . Larvae feed on the seeds and adults on open flowers of their host . Adults oviposit on to immature seeds 3 mm long or less and hence seeds of this length and maturity were used in the host range tests and for rearing . Females are shown to avoid previously attacked seeds enhancing their effectiveness as seed destroyers . Survival of adults was higher when provided with open flowers . The host range was determined using laboratory control - choice oviposition tests on excised plant material and , in the field in the native range , no - choice oviposition tests on living plants , surveys of adults on plants , and breeding of insects from pods of plants of various legume species . The control - choice oviposition tests employed a new design in which the control plant alone was offered to the insects followed by a choice of test plants species . Other than M. pigra, only one plant species was acceptable for oviposition , the closely related M. asperata. Larval development also occurs on M. asperata and this host is occasionally used in the field . This insect was approved for release in March 1997 .     

Field Evaluation of a Commercial Formulation of the Spodoptera exigua (Lepidoptera: Noctuidae) Nuclear Polyhedrosis Virus for Control of Beet Armyworm on Vegetable Crops in Thailand

Small - scale field trials were conducted in the Kanchanaburi and Ratchaburi provinces of Thailand between December 1994 and March 1996 to gather efficacy data to support a registration dossier for a commercial formulation of a nuclear polyhedrois virus of beet armyworm , Spodoptera exigua, under the tradename Spod - X . A liquid concentrate formula tion of Spod - X was compared with two commercial standards , Bacillus thuringiensis subsp . aizawai (CenTuri 3 . 5 WDG) and the insect growth regulator tebufenozide (Mimic 20F , a benzoylhydrazine ecdysone agonist) , for control of beet armyworm on garden pea , Chinese kale , shallot and table grape . Spod - X applied at 4 - day intervals at 3 . 1 - 12 . 5 1011 occlusion bodies (OBs) ha 1 in a spray volume of 625 - 1000 l ha 1 provided excellent control of beet armyworm on garden pea and grape (75 - 100 % reductions) after the second application . On Chinese kale and shallot , rates of Spod - X in the rate 5 - 60 1011 OBs ha 1 sprayed every 4 days in a spray volume of 1000 l ha 1 provided significant protection from infestation of beet armyworm over the growing season; however , efficacy was poor to moderate . Tank - mixing Spod - X with the pyrethroid , deltamethrin (Decis3EC) , did not improveefficacy on shallot;however , combinations of virus and B. thuringiensis or tebufenozide achieved commercially acceptable levels of crop protection on Chinese kale compared with virus alone . Lower levels of control and crop pro tection observed with all materials on Chinese kale and shallot reflect , in part , the tendency of S. exigua larvae to feed within sheltered sites and directly damage marketable plant parts . In general , all rates of Spod - X performed as well as , or better than , the B. thuringiensis standard (56 . 3 - 105 g ha 1 a.i.) on all crops tested . In general , reductions of beet armyworm larval populations and plant damage provided by all labeled rates of the chemical standard , tebufenozide (125 - 250 g ha 1 a . i .), were marginally greater than all rates of Spod - X tested . On crops such as pea and grape , where beet armyworm predominately feed on exposed leaf surfaces , our data unequivocally support the use of Spod - X for cost - effective , season - long pest management . Strategies to improve the efficacy of virus on shallot and kale are discussed .     

Comparison of Type 2 Inositol 1,4,5-Trisphosphate Receptor Distribution and Subcellular Ca2+ Release Sites that Support Ca2+ Waves in Cultured Astrocytes

Abstract: We have examined the mechanisms that underlie Ca²⁺ wave propagation in cultured cortical astrocytes. Norepinephrine evoked Ca²⁺ waves in astrocytes that began at discrete initiation loci and propagated throughout the cell by regenerative amplification at a number of cellular sites, as shown by very high Ca²⁺ release rates at these regions. We have hypothesized previously that domains displaying elevated Ca²⁺ release kinetics in astrocytes may correspond to sites of high inositol 1,4,5-trisphosphate receptor (InsP₃R) density. To examine this possibility, we compared the distribution pattern of endoplasmic reticulum (ER) and InsP₃Rs with Ca²⁺ release kinetics in subcellular regions during propagation of norepinephrine-evoked waves. 3,3'-Dihexyloxacarbocyanine iodide staining revealed that the ER in astrocytes exists as a meshwork of membranes extending throughout the cells, including fine processes. A specific antibody directed against type 2 InsP₃Rs (InsP₃R2) detected a 260-kDa band in western blotting of astrocyte membranes. Immunocytochemistry using this antibody stained the entire ER system in a punctate, variegated manner. When Ca²⁺ responses and InsP₃R2 immunofluorescence were compared in the same cell, domains of elevated Ca²⁺ response kinetics (high amplitude and rapid rate of rise) showed significant positive correlation with high local intensity of InsP₃R2 staining. It appears, therefore, that specializations in the ER responsible for discrete local Ca²⁺ release sites that support regenerative wave propagation include increased levels of InsP₃R2 expression.     

Temporal Analysis of Changes in Neuronal c-fos mRNA Levels Induced by Depletion of Endoplasmic Reticulum Calcium Stores: Effect of Clamping Cytoplasmic Calcium Activity at Resting Levels

Abstract: Activation of immediate early gene expression is a key event in stress-induced neuronal cell injury. To study whether changes in cytoplasmic calcium activity are necessary to activate neuronal immediate early gene expression, endoplasmic reticulum (ER) calcium stores of primary neurons were depleted by exposing cells to thapsigargin (Tg), an irreversible inhibitor of ER Ca²⁺-ATPase. Tg-induced rise in [Ca²⁺]_i and the effect of loading neurons with the cell-permeable calcium chelator BAPTA-AM on this increase in [Ca²⁺]_i were measured in fura-2-loaded cells by fluorescence microscopy. Changes in c- fos mRNA levels were evaluated by quantitative PCR. Tg treatment of neurons produced a pronounced rise in c- fos mRNA levels (∼10-fold more than DMSO) which peaked at 1 h after exposure. The Tg-induced rise in c- fos mRNA content was unchanged (hippocampal neurons) or even increased further (cortical neurons) by preloading cells with BAPTA before incubation with Tg. It is concluded that in neuronal cells an increase in cytoplasmic calcium activity is not a prerequisite for a rise in mRNA levels of c- fos . Thus, stress-induced changes in mRNA levels of immediate early genes of neurons may also result from disturbances in ER calcium homeostasis and not necessarily by an overload of cells with calcium ions. The results of the present series of experiments cast further doubt on the widely accepted hypothesis that the stress-induced cytoplasmic overload of neurons with calcium ions is the primary event triggering cell injury.     

Ischemia-Induced Inhibition of Calcium Uptake into Rat Brain Microsomes Mediated by Mg2+/Ca2+ ATPase

Abstract: It is well established that ischemia is associated with prolonged increases in neuronal intracellular free calcium levels. Recent data suggest that regulation of calcium uptake and release from the endoplasmic reticulum is important in maintaining calcium homeostasis. The endoplasmic reticulum Mg²⁺/Ca²⁺ ATPase is the major mechanism for sequestering calcium in this organelle. Inhibition of this enzyme may play a causal role in the loss of calcium homeostasis. In order to investigate the effect of ischemia on calcium sequestration into the endoplasmic reticulum, microsomes were isolated from control and ischemic whole brain homogenates by differential centrifugation. Calcium uptake was measured by radioactive calcium (⁴⁵Ca²⁺) accumulation in the microsomes mediated by Mg²⁺/Ca²⁺ ATPase. Ischemia caused a statistically significant inhibition of presteady-state and steady-state calcium uptake. Duration of ischemia was directly proportional to the degree of inhibition. Decreased calcium uptake was shown not to be the result of increased calcium release from ischemic compared with control microsomes nor the result of selective isolation of ischemic microsomes from the homogenate with a decreased capacity for calcium uptake. The data demonstrate that ischemia inhibits the ability of brain microsomes to sequester calcium and suggest that loss of calcium homeostasis is due, in part, to ischemia-induced inhibition of endoplasmic reticulum Mg²⁺/Ca²⁺ ATPase.     

Impact of the Biological Control Agent Gyranusoidea tebygi Noyes (Hymenoptera: Encyrtidae) on the Mango Mealybug, Rastrococcus invadens Williams (Homoptera: Pseudococcidae), in Benin

The distribution of Rastrococcus invadens among different host plants and the impact of the mealybug on mango growth were investigated on 2067 trees in three surveys across all the ecological zones of Benin. The first survey started in 1989, less than 1 year after the first release of the exotic parasitoid Gyranusoidea tebygi. Within 3 years, G. tebygi had colonized the entire area of infestation, and was found on practically all infested mango trees as well as other infested host plants. By 1991, the incidence of R. invadens on the secondary host plants had declined significantly. The percentage of infested mango trees declined from 31.0% in 1989 to 17.5% in 1991, highest populations being found in the coastal savanna. During the same period, the mean percentage of infested mango trees having indigenous predators declined from 42.3 to 20.9%. Average mealybug densities declined steadily from 9.7 females/48 leaves in 1989, with 3.2% of all mango trees having densities above 100 mealybugs, to 6.4 females/48 leaves in 1991, with 1.3% of all trees having densities above 100 mealybugs. In multiple regression analyses, based on 23 meteorological, agronomic and plant variables, the duration of the parasitoid's presence proved to be a major factor. It influenced mealybug population densities and sooty mould incidence, which, in turn, affected the production of new leaves. In all analyses, the impact of rainfall, for example, on the sooty mould or the mealybug was less important than the effect of G. tebygi. The present study demonstrates for the first time on a large scale the impact of G. tebygi on R. invadens and, indirectly, on its main host plant, mango.