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Mortality following cotton defoliation: San Joaquin Valley, California, 1970-1990.

A proportional mortality study comparing the cotton-growing areas of the San Joaquin Valley with the rest of the State of California was performed by the Office of Environmental Health Hazard Assessment as a continuation of earlier studies related to mercaptan-releasing pesticides. This mortality study found a pattern of increased proportion of "respiratory causes" mortality (ICD codes 460-519), statistically significant at less than the .05 probability level, for 15 of 21 years between 1970 and 1990, for the time period during and immediately following cotton defoliation. Defoliants which have the potential to produce acute symptoms include DEF and Folex, both of which release odorous butyl mercaptan gas as a degradation product. This paper tests the hypothesis that exposure to cotton defoliant breakdown products may be associated with a disproportionate increase in mortality. Prediction of the mortality proportions by pounds of DEF and Folex used was not statistically significant in the unadjusted models or in models adjusted for air pollution variables. One air pollution adjustment factor, total suspended particulates, was a statistically significant independent mortality proportion predictor. This finding suggests that total suspended particulates, not defoliants, may be related to mortality differentials during defoliation season. Possible confounding by demographic variation of the counties was not controlled in the analysis.

Air Pollutants↗

Development of a novel subunit vaccine that protects cotton rats against both human respiratory syncytial virus and human parainfluenza virus type 3.

A cotton rat model of experimental human respiratory syncytial virus (RSV) and human parainfluenza virus type 3 (PIV-3) infection was used to examine the efficacy of FRHNP, a novel chimeric glycoprotein which contains the extracellular regions of the fusion glycoprotein of RSV and the attachment glycoprotein of PIV-3, as a single subunit vaccine against these two viruses. This work was prompted by previous cotton rat studies that demonstrated that the major protective antigens of the two viruses were these glycoproteins. FRHNP was expressed in insect cells using a recombinant baculovirus. Vaccination with FRHNP resulted in induction of both RSV and PIV-3 neutralizing antibody and doses of 200 ng completely protected rats from either RSV or PIV-3 challenge. These results demonstrate that in the cotton rat animal model a single chimeric glycoprotein can be an effective vaccine against both RSV and PIV-3.

Amino Acid Sequence↗

Pathology and immunogenicity in the cotton rat (Sigmodon hispidus) model after infection with a bovine adenovirus type 3 recombinant virus expressing the firefly luciferase gene.

The histopathology of adenovirus pneumonia in cotton rats (Sigmodon hispidus) due to bovine adenovirus type 3-luciferase recombinant virus (BAd3-Luc), which has a 0.7 kb deletion from the early region 3 (E3) replaced with the firefly luciferase gene, was compared with that produced by the parental wild-type (wt) bovine adenovirus type 3 (BAd3). After intranasal inoculation of cotton rats with 3 x 10(7) p.f.u. of BAd3-Luc, the infectious virus titres in the lungs at various times post-infection were similar to those of animals infected with the parental virus. Quantitative analysis of histopathological changes and immunohistochemical staining showed that the character and severity of the lesions were indistinguishable in the two infections. Luciferase activity was detected in the lungs of BAd3-Luc-inoculated animals until 4 days post-infection (p.i.). Antibodies to both BAd3 and luciferase were detected in sera collected from BAd3-Luc-infected animals until at least 6 weeks p.i. These results show that Bad3-Luc produces pulmonary lesions in cotton rats similar to those of wt BAd3 and suggest that BAd3-based vectors may be suitable for the development of live recombinant virus vaccines.

Adenoviridae Infections↗

Mucosal immunization with recombinant adenoviruses: induction of immunity and protection of cotton rats against respiratory bovine herpesvirus type 1 infection.

To facilitate the evaluation of vaccines against bovine herpesvirus type 1 (BHV-1), a cotton rat model of intranasal (i.n.) BHV-1 infection was established. Cotton rat lung cells were similar to bovine cells in their ability to support BHV-1 replication in vitro. Furthermore, i.n. inoculation of cotton rats with BHV-1 resulted in pulmonary lesions comparable to BHV-1 infection in cattle. Using this model, the potential of i.n. and gastrointestinal (g.i.) immunization was examined with recombinant human adenoviruses expressing glycoprotein D (gD) of BHV-1 to induce protective immunity against BHV-1. The replication-competent virus (gD-dE3) was more efficient than the replication-defective virus (gD-dE1E3) in inducing gD-specific antibody in the serum and in the respiratory tract. Furthermore, i.n. immunization with gD-dE3 stimulated antigen-specific antibody-secreting cells in the lung 12 weeks following immunization. Protection against BHV-1 challenge correlated with gD-specific antibody levels such that i.n. immunization with gD-dE3 conferred complete protection, while g.i. immunization conferred only partial protection of the lungs of most animals against BHV-1 challenge. In comparison, immunization with gD-dE1E3 by either route resulted in only a partial reduction of BHV-1 titre in the respiratory tract. The results obtained demonstrate that mucosal immunization with replication-competent recombinant adenovirus expressing gD of BHV-1 can induce immunity and protection against BHV-1 challenge.

Animals↗

Biochemical and molecular inhibition of plastidial carbonic anhydrase reduces the incorporation of acetate into lipids in cotton embryos and tobacco cell suspensions and leaves.

Two cDNAs encoding functional carbonic anhydrase (CA) enzymes were recently isolated from a non-photosynthetic, cotyledon library of cotton (Gossypium hirsutum) seedlings with putative plastid-targeting sequences (GenBank accession nos. AF132854 and AF132855). Relative CA transcript abundance and enzyme activity increased 9 and 15 times, respectively, in cotton embryos during the maximum period of reserve oil accumulation. Specific sulfonamide inhibitors of CA activity significantly reduced the rate of [(14)C]acetate incorporation into total lipids in cotton embryos in vivo, and in embryo plastids in vitro, suggesting a role for CA in plastid lipid biosynthesis. CA inhibitors did not affect acetyl-coenzyme A carboxylase activity or total storage protein synthesis. Similar results were obtained for two other plant systems: cell suspensions (and isolated plastids therefrom) of tobacco (Nicotiana tabacum), and chloroplasts isolated from leaves of transgenic CA antisense-suppressed tobacco plants (5% of wild-type CA activity). In addition, tobacco cell suspensions treated with the CA inhibitor ethoxyzolamide showed a substantial loss of CO(2) compared with controls. The rate of [(14)C]acetate incorporation into lipid in cell suspensions was reduced by limiting external [CO(2)] (scrubbed air), and this rate was further reduced in the presence of ethoxyzolamide. Together, these results indicate that a reduction of CA activity (biochemical or molecular inhibition) impacts the rate of plant lipid biosynthesis from acetate, perhaps by impairing the ability of CA to efficiently "trap" inorganic carbon inside plastids for utilization by acetyl-coenzyme A carboxylase and the fatty acid synthesis machinery.

Acetates↗

Spatial Distribution of Photosynthesis during Drought in Field-Grown and Acclimated and Nonacclimated Growth Chamber-Grown Cotton.

Inhomogeneous photosynthetic activity has been reported to occur in drought-stressed leaves. In addition, it has been suggested that these water stress-induced nonuniformities in photosynthesis are caused by "patchy" stomatal closure and that the phenomenon may have created the illusion of a nonstomatal component to the inhibition of photosynthesis. Because these earlier studies were performed with nonacclimated growth chamber-grown plants, we sought to determine whether such "patches" existed in drought-treated, field-grown plants or in chamber-grown plants that had been acclimated to low leaf water potentials (psi(leaf)). Cotton (Gossypium hirsutum L.) was grown in the field and subjected to drought by withholding irrigation and rain from 24 d after planting. The distribution of photosynthesis, which may reflect the stomatal aperture distribution in a heterobaric species such as cotton, was assayed by autoradiography after briefly exposing attached leaves of field-grown plants to (14)CO(2). A homogeneous distribution of radioactive photosynthate was evident even at the lowest psi(leaf) of -1.34 MPa. "Patchiness" could, however, be induced by uprooting the plant and allowing the shoot to air dry for 6 to 8 min. In parallel studies, growth chamber-grown plants were acclimated to drought by withholding irrigation for three 5-d drought cycles interspersed with irrigation. This drought acclimation lowered the psi(leaf) value at which control rates of photosynthesis could be sustained by approximately 0.7 MPa and was accompanied by a similar decline in the psi(leaf) at which patchiness first appeared. Photosynthetic inhomogeneities in chamber-grown plants that were visible during moderate water stress and ambient levels of CO(2) could be largely removed with elevated CO(2) levels (3000 muL L(-1)), suggesting that they were stomatal in nature. However, advanced dehydration (less than approximately 2.0 MPa) resulted in "patches" that could not be so removed and were probably caused by nonstomatal factors. The demonstration that patches do not exist in drought-treated, field-grown cotton and that the presence of patches in chamber-grown plants can be altered by treatments that cause an acclimation of photosynthesis leads us to conclude that spatial heterogeneities in photosynthesis probably do not occur frequently under natural drought conditions.

Journal Article↗

Involvement of Abscisic Acid in Ethylene-Induced Cotyledon Abscission in Cotton Seedlings.

Cotton (Gossypium hirsutum L. cv LG102) seedlings raised from seeds exposed to 100 [mu]M norflurazon (NFZ) during imbibition contained reduced levels of free abscisic acid (ABA) and were visibly achlorophyllous. Exposure of untreated cotton seedlings to ethylene concentrations >1 [mu]L/L for 24 h resulted in cotyledon abscission. In contrast, exposure of NFZ-treated seedlings to concentrations of ethylene [less than or equal to]50 [mu]L/L elicited no cotyledon abscission. Application of ABA, an ABA analog, or jasmonic acid to NFZ-treated seedlings restored ethylene-induced abscission. Isolated cotyledonary node explants prepared from NFZ-treated seedlings exhibited an altered dose-response pattern of ethylene-induced petiole abscission. Endogenous levels of free IAA were unaltered in NFZ-treated seedlings. Ethylene treatment (50 [mu]L/L, 24 h) had no effect on free indoleacetic acid (IAA) levels in either control or NFZ-treated seedlings. Levels of conjugated (ester plus amide) IAA were substantially increased in NFZ-treated seedlings regardless of ethylene treatment. These results indicate that endogenous ABA plays an essential, but physiologically undefined, role in ethylene-induced cotyledon abscission in cotton.

Journal Article↗

An ATP-binding cassette transporter GhWBC1 from elongating cotton fibers.

We have isolated a cDNA (GhWBC1) from cotton (Gossypium hirsutum) that encodes an ATP-binding cassette transporter of the WBC (white/brown complex) subfamily. Members of this subfamily are half-sized transporters and are reported to mediate lipid and drug excretion in human (Homo sapiens). GhWBC1 is highly expressed in developing fiber cells, but transcripts were also detectable in other tissues except roots. The transcript level peaked in rapidly expanding fibers from 5 to 9 DPA and then decreased. The GhWBC1 expression was weak in fiber cells of an li (ligon-lintless) mutant, which is defective in fiber cell elongation. These data indicate that GhWBC1 gene expression correlates with cotton fiber elongation. Transient expression of enhanced green fluorescence protein-GhWBC1 fusion protein in onion (Allium cepa) epidermal cells revealed plasma membrane localization. The GhWBC1 cDNA driven by a constitutive 35S promoter was introduced into Arabidopsis. About 13% of the transformants produced short siliques (SSs), whereas others had normal siliques (long siliques [LSs]). In siliques of SS lines, most embryos were severely shriveled, and only several seeds per silique could be found at maturity. The transgene expression level was higher in SS lines than in LS lines. Expression of AtWBC11, the closest homolog of GhWBC1 in Arabidopsis, was not altered in either SS or LS transgenic plants examined. These data suggest that GhWBC1 interferes with substance translocation that is required for Arabidopsis seed and silique development. Characterization of Arabidopsis WBC members, particularly AtWBC11, will help to dissect the role of GhWBC1 in cotton fiber development and elongation.

ATP-Binding Cassette Transporters↗

Volatile Semiochemicals Released from Undamaged Cotton Leaves (A Systemic Response of Living Plants to Caterpillar Damage).

Cotton plants (Gossypium hirsutum L.), attacked by herbivorous insects release volatile semiochemicals (chemical signals) that attract natural enemies of the herbivores to the damaged plants. We found chemical evidence that volatiles are released not only at the damaged site but from the entire cotton plant. The release of volatiles was detected from upper, undamaged leaves after 2 to 3 d of continuous larval damage on lower leaves of the same plant. Compounds released systemically were (Z)-3-hexenyl acetate, (E)-[beta]-ocimene, linalool, (E)-4,8-dimethyl-1,3,7-nonatriene, (E)-[beta]-farnesene, (E,E)-[alpha]-farnesene, and (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene. All systemically released compounds are known to be induced by caterpillar damage and are not released in significant amounts by undamaged plants. Other compounds, specifically indole, isomeric hexenyl butyrates, and 2-methylbutyrates, known to be released by cotton in response to caterpillar damage, were not released systemically. However, when upper, undamaged leaves of a caterpillar-damaged plant were damaged with a razor blade, they released isomeric hexenyl butyrates, 2-methylbutyrates, and large amounts of constitutive compounds in addition to the previously detected induced compounds. Control plants, damaged with a razor blade in the same way, did not release isomeric hexenyl butyrates or 2-methylbutyrates and released significantly smaller amounts of constitutive compounds. Indole was not released systemically, even after artificial damage.

Journal Article↗

The Differential Expression of Sucrose Synthase in Relation to Diverse Patterns of Carbon Partitioning in Developing Cotton Seed.

Developing cotton (Gossypium hirsutum L.) seed exhibits complex patterns of carbon allocation in which incoming sucrose (Suc) is partitioned to three major sinks: the fibers, seed coat, and cotyledons, which synthesize cellulose, starch, and storage proteins or oils, respectively. In this study we investigated the role of Suc synthase (SuSy) in the mobilization of Suc into such sinks. Assessments of SuSy gene expression at various levels led to the surprising conclusion that, in contrast to that found for other plants, SuSy does not appear to play a role in starch synthesis in the cotton seed. However, our demonstration of functional symplastic connections between the phloem-unloading area and the fiber cells, as well as the SuSy expression pattern in fibers, indicates a major role of SuSy in partitioning carbon to fiber cellulose synthesis. SuSy expression is also high in transfer cells of the seed coat facing the cotyledons. Such high levels of SuSy could contribute to the synthesis of the thickened cell walls and to the energy generation for Suc efflux to the seed apoplast. The expression of SuSy in cotyledons also suggests a role in protein and lipid synthesis. In summary, the developing cotton seed provides an excellent example of the diverse roles played by SuSy in carbon metabolism.

Journal Article↗

Role of IAA-Oxidase in Abscission Control in Cotton.

The potential role of indoleactic acid (IAA)-oxidase as an in vivo abscission regulating system in the cotton (Gossypium hirsutum L.) cotyledonary explant was investigated. Phenols (usually monophenols), which are cofactors of cotton IAA-oxidase in vitro, accelerated abscission. Phenols (usually orthodihydroxyphenols), which inhibit cotton IAA-oxidase in vitro, inhibited abscission. Inhibition or stimulation of abscission was accomplished by phenols both with and without IAA. Results were similar when treatments were applied as lanolin pastes to the cut petiole ends or as solutions in which explants were submerged. An abscission accelerating phenol stimulated the decarboxylation of IAA-1-(14)C by explants and an abscission inhibiting phenol inhibited the decarboxylation of IAA-1-(14)C.The mechanism of abscission regulation by the phenolic compounds was concluded to involve auxin destruction via IAA-oxidase.In addition to the direct relationship of this study to abscission, the results support the more general hypothesis that IAA-oxidase acts in vivo to regulate auxin levels.

Journal Article↗

Oxidative and phosphorylative activities of mitochondria isolated from cotton hypocotyls.

Mitochondria isolated from 2 strains of cotton plant hypocotyls (Gossypium hirsutum L. var. Rex smooth leaf and Rex glandless) were examined for their oxidative phosphorylation activities. Bovine serum albumin at a relatively high concentration was essential in the extraction medium for the isolation of oxidatively active mitochondria from both strains of cotton. Phosphorylation was obtained only with Rex glandless cotton mitochondria. This activity was low in comparison to the mitochondria isolated from soybeans (Glycine max L. var. Lee). The endogenous gossypol content was found to be much higher in the Rex smooth leaf tissue than in the Rex glandless tissue. In turn, comparable gossypol differences were found associated with their respective mitochondrial fractions. Exogenous gossypol uncoupled succinate oxidation with active mitochondria isolated from soybeans. Gossypol as a possible uncoupler is discussed and compared to carbonyl cyanide, m-chlorophenyl hydrazone and 2,4-dinitrophenol.

Journal Article↗

Effects of salt secretion on psychrometric determinations of water potential of cotton leaves.

Thermocouple psychrometers gave lower estimates of water potential of cotton leaves than did a pressure chamber. This difference was considerable for turgid leaves, but progressively decreased for leaves with lower water potentials and fell to zero at water potentials below about -10 bars. The conductivity of washings from cotton leaves removed from the psychrometric equilibration chambers was related to the magnitude of this discrepancy in water potential, indicating that the discrepancy is due to salts on the leaf surface which make the psychrometric estimates too low. This error, which may be as great as 400 to 500%, cannot be eliminated by washing the leaves because salts may be secreted during the equilibration period. Therefore, a thermocouple psychrometer is not suitable for measuring the water potential of cotton leaves when it is above about -10 bars.

Journal Article↗

Malate dehydrogenase isoenzymes from cotton leaves: molecular weights.

Malate dehydrogenase isolated from leaves of the cotton plant (Gossypium hirsutum L.) appears in the form of several isoenzymes. Four of the isoenzymes found in cotton leaf extracts appear to be charge isomers with a molecular weight of approximately 60,000. A fifth malate dehydrogenase isoenzyme found in leaf extracts has a molecular weight of approximately 500,000. Under appropriate conditions it is possible to form this high molecular weight isoenzyme from at least one of the smaller isoenzymes. In addition, malate dehydrogenase isoenzymes of approximately 700,000 and 130,000 molecular weight have been observed under some conditions, although these isoenzymes do not appear in the crude cotton leaf preparations. The relationship of this heterogeneity with respect to size and to the discrepancies in the number and size of malate dehydrogenase isoenzymes reported from plant tissues may be significant.

Journal Article↗

The development of isocitric lyase activity in germinating cotton seed.

In cotyledons of germinating cotton (Gossypium hirsutum L. var. Stoneville 213) seedlings, in the dark, isocitric lyase (EC 4.1.3.1) activity peaks after 2 days and thereafter slowly declines to a negligible value after 8 days. The maximum activity of this enzyme in cotyledons of 2-day-old seedlings was 16.2 mumoles of glyoxylate formed/15 min.10 cotyledon pairs. Actinomycin D at a concentration of 10 mug/ml, if added to the imbibing solution, completely prevents the development of isocitric lyase activity in these germinating seed. In cotyledons of germinating cotton seedlings, in the light, isocitric lyase activity peaks after 2 to 3 days and sharply declines to a negligible value after 4 days. The maximum activity of this enzyme in cotyledons of 2- to 3-day-old seedlings was 13.2 mumoles of glyoxylate formed/15 min.10 cotyledon pairs. Actinomycin D at a concentration of 10 mug/ml, if added to the imbibing solution, severely inhibits the development of enzyme activity.In germinating seed, in the light, the synthesis of chlorophyll and glyceraldehyde-3-P dehydrogenase is also limited by the addition of low concentrations of actinomycin D. The new synthesis of fructose-1, 6-diP aldolase, which is detectable after 1 to 2 days of germination, is inhibited by 10 mug/ml of actinomycin D. We, therefore, conclude that the synthetic events leading to the development of chlorophyll, some glyoxysomal and chloroplast enzymes in germinating cotton seedlings depend on newly transcribed mRNA.

Journal Article↗

Changes in biochemical composition of the cell wall of the cotton fiber during development.

The composition of the cell wall of the cotton fiber (Gossypium hirsutum L. Acala SJ-1) has been studied from the early stages of elongation (5 days postanthesis) through the period of secondary wall formation, using cell walls derived both from fibers developing on the plant and from fibers obtained from excised, cultured ovules. The cell wall of the elongating cotton fiber was shown to be a dynamic structure. Expressed as a weight per cent of the total cell wall, cellulose, neutral sugars (rhamnose, fucose, arabinose, mannose, galactose, and noncellulosic glucose), uronic acids, and total protein undergo marked changes in content during the elongation period. As a way of analyzing absolute changes in the walls with time, data have also been expressed as grams component per millimeter of fiber length. Expressed in this way for plant-grown fibers, the data show that the thickness of the cell wall is relatively constant until about 12 days postanthesis; after this time it markedly increases until secondary wall cellulose deposition is completed. Between 12 and 16 days postanthesis increases in all components contribute to total wall increase per millimeter fiber length. The deposition of secondary wall cellulose begins at about 16 days postanthesis (at least 5 days prior to the cessation of elongation) and continues until about 32 days postanthesis. At the time of the onset of secondary wall cellulose deposition, a sharp decline in protein and uronic acid content occurs. The content of some of the individual neutral sugars changes during development, the most prominent change being a large increase in noncellulosic glucose which occurs just prior to the onset of secondary wall cellulose deposition. Methylation analyses indicate that this glucose, at least in part, is 3-linked. In contrast to the neutral sugars, no significant changes in cell wall amino acid composition are observed during fiber development.Compositional analyses of cell walls derived from culture-grown fibers indicate that these walls are remarkably similar to those derived from fibers grown on the plant, both in terms of composition and in terms of relative changes in composition during development.A comparison of our results on total cell wall composition and linkages of sugars as determined by a preliminary methylation analysis of unfractionated fiber walls indicates that the primary cell wall of cotton fibers is similar to that of primary cell walls of other dicotyledons and of gymnosperms as reported in the literature.

Journal Article↗

Effects of Irradiance during Growth on Adaptive Photosynthetic Characteristics of Velvetleaf and Cotton.

We grew velvetleaf (Abutilon theophrasti Medic.) and cotton (Gossypium hirsutum L. var. Stoneville 213) at three irradiances and determined the photosynthetic responses of single leaves to a range of six irradiances from 90 to 2000 mueinsteins m(-2)sec(-1). In air containing 21% O(2), velvetleaf and cotton grown at 750 mueinsteins m(-2)sec(-1) had maximum photosynthetic rates of 18.4 and 21.9 mg of CO(2) dm(-2)hr(-1), respectively. Maximum rates for leaves grown at 320 and 90 mueinsteins m(-2)sec(-1) were 15.3 and 10.3 mg of CO(2) dm(-2)hr(-1) in velvetleaf and 12 and 6.7 mg of CO(2) dm(-2)hr(-1) in cotton, respectively. In 1 O(2), maximum photosynthetic rates were 1.5 to 2.3 times the rates in air containing 21% O(2), and plants grown at medium and high irradiance did not differ in rate. In both species, stomatal conductance was not significantly affected by growth irradiance. The differences in maximum photosynthetic rates were associated with differences in mesophyll conductance. Mesophyll conductance increased with growth irradiance and correlated positively with mesophyll thickness or volume per unit leaf area, chlorophyll content per unit area, and photosynthetic unit density per unit area. Thus, quantitative changes in the photosynthetic apparatus help account for photosynthetic adaptation to irradiance in both species. Net assimilation rates calculated for whole plants by mathematical growth analysis were closely correlated with single-leaf photosynthetic rates.

Journal Article↗

Lipid and Surface Wax Synthesis in Water-stressed Cotton Leaves.

The incorporation of [2-(14)C]malonate and [1-(14)C]acetate into internal lipid and surface wax by cotton leaves (Gossypium hirsutum L. ;Deltapine') having water potentials of -8 to -15 bars (controls) and -19 to -32 bars (water-stressed) was compared. Lipid from stressed leaves contained a mean of 57% more radioactivity than corresponding controls for five experiments. Acetyl coenzyme A carboxylase was not limiting to fatty acid synthesis in water-stressed cotton leaves at the water potential levels tested, whereas fatty acid synthetase was stimulated. In four of six experiments, wax from stressed leaves contained a mean of 38% less radioactivity than nonstressed leaves when incubated 24 hours after rehydration. Evidence is presented to show that after a suitable period of rehydration, previously stressed cotton leaves produce more wax than leaves prior to stressing.

Journal Article↗