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Interplant movement of Heliothis virescens (Lepidoptera: Noctuidae) larvae in pure and mixed plantings of cotton with and without expression of the Cry1Ac delta-endotoxin protein of Bacillus thuringiensis Berliner.

Laboratory and field studies were conducted during 1993 and 1994 to quantify interplant movement of Heliothis virescens (F.) larvae in pure and mixed plantings of cotton, Gossypium hirsutum L., with ('Event 531') and without ('Coker 312') the expression of Cry1Ac delta-endotoxin protein of Bacillus thuringiensis Berliner. Field studies were conducted with neonate, 4-, and 7-d-old larvae placed on 3-plant experimental units and observed at 24, 48, 72, and 96 h after inoculation of larvae. Combining larval movement across observations of neonates, 4-, and 7-d-old larvae, an estimated 52% of the larvae on pure plantings of Coker 312 had moved at least 1 plant by the cumulative time required to reach the age of 10 d. More larvae placed on Event 531 cotton moved to an adjacent plant (13% of the neonates had moved at least 1 plant within 24 h) than those placed on Coker 312 (0% of the neonates had moved at least 1 plant within 24 h). When larvae were placed on Event 531 plants, an estimated 82% of the larvae had moved to an adjacent plant by cumulative age of 10 d. Collectively, these data indicate that movement of larvae from plant to plant increases with larval age and occurs more rapidly for larvae placed on Event 531 cotton than on Coker 312. Previous studies have suggested that resistance to B. thuringiensis could develop more rapidly in insects exposed to seed mixtures of plants with and without endotoxin if larvae move between plants and if an external refuge exists. These data provide evidence of larval movement between plants in seed mixtures.

Animals↗

Spatial processes in the evolution of resistance in Helicoverpa zea (Lepidoptera: Noctuidae) to Bt transgenic corn and cotton in a mixed agroecosystem: a biology-rich stochastic simulation model.

A simulation model is developed to examine the role of spatial processes in the evolution of resistance in Helicoverpa zea populations to Bt corn and Bt cotton. The model is developed from the stochastic spatially explicit Heliothis virescens model described by Peck et al. (1999), to accommodate a spatial mix of two host crops (corn and cotton), and to reflect the agronomic practices, as well as the spatial and temporal population dynamics of H. zea, in eastern North Carolina. The model suggests that selection for resistance is more intense in Bt cotton fields than in Bt corn fields. It further suggests that local gene frequencies are highly dependent on local deployment levels of Bt crops despite the high mobility of the adult insects. Region-wide average gene frequencies depend on the region-wide level of Bt deployment, so incomplete technology adoption slows the rate of resistance evolution. However, on a local scale, H. zea populations in clusters of fields in which Bt use is high undergo far more rapid evolution than populations in neighboring clusters of fields in which Bt use is low. The model suggests that farm-level refuge requirements are important for managing the risk of resistance. The model can be used as an aid in designing plans for monitoring for resistance by suggesting the appropriate distribution of monitoring locations, which should focus on areas of highest Bt crop deployment. The findings need to be placed in the context of the input parameters, many of which are uncertain or highly variable in nature, and therefore, a thorough sensitivity analysis is warranted.

Agriculture↗

Field and laboratory evaluations of transgenic cottons expressing one or two Bacillus thuringiensis var. kurstaki Berliner proteins for management of noctuid (Lepidoptera) pests.

Field studies were conducted from 1999 to 2001 to evaluate the efficacy of the transgenic cotton, Gossypium hirsutum (L.), genotype, Bollgard II (Monsanto 15985), which expresses two Bacillus thuringiensis Berliner (Bt) proteins (Cry1Ac + Cry2Ab) that are active against lepidopterous pests. Bollgard II was compared with Bollgard (DP50B), which expresses only one Bt protein (Cry1Ac), and, in all tests, the conventional variety, DP50, was used as a non-Bt control. Larval populations of the bollworm, Helicoverpa zea (Boddie), and the soybean looper, Pseudoplusia includens (Walker), were significantly lower in Bollgard II than in Bollgard and conventional cotton, and the proportion of fruit damaged by H. zea was also lower. Fall armyworm, Spodoptera frugiperda (J. E. Smith), populations were lower in Bollgard II than in Bollgard, although not significantly. Field tests were supplemented with laboratory bioassays in 2001 to compare mortality of S. frugiperda, and beet armyworms, Spodoptera exigua (Hübner), feeding on these genotypes. Mortality of both species was significantly greater on Bollgard II plant material than on either Bollgard or conventional cotton. This study demonstrated that the dual-toxin Bollgard II genotype is highly effective against lepidopterous pests that are not adequately controlled by the current single-toxin Bollgard varieties. If toxin expression in future Bollgard II varieties remains consistent with that of Monsanto 15985, supplemental insecticides will be reduced, and may be eliminated for lepidopterous pests in South Carolina.

Animals↗

Injury to preflowering and flowering cotton by brown stink bug and southern green stink bug.

The impact of brown stink bug, Euschistus servus (Say), and southern green stink bug, Nezara viridula (L.), injury was evaluated on preflowering and flowering cotton, Gossypium hirsutum L., plants in no-choice tests. Vegetative stage cotton seedlings and reproductive structures, including flower buds (square) and bolls, were infested with adults and/or nymphs of both species. There were no significant differences in height, height to node ratio, square retention, and flower initiation for cotton seedlings or plants with a match-head square between southern green stink bug adult- or brown stink bug adult-infested and noninfested treatments. Abscission for individual large squares (precandle) and multiple squares (medium and small square on the same sympodial branch) was not significantly different among infested and noninfested treatments for the following species and developmental stages: brown stink bug adults, southern green stink bug adults, and third and fourth to fifth instar southern green stink bug nymphs. In boll infestation studies, the relationship between boll maturity, expressed as heat units beyond anthesis, and boll growth, abscission, hard locked carpels, seedcotton yield, and seed germination was measured. Brown stink bug induced abscission in bolls that had accumulated > 0-350 heat units beyond anthesis. Boll growth and seedcotton yield was significantly lower for bolls infested with brown stink bug through 266.5 and 550 heat units beyond anthesis, respectively. The proportion of hard locked carpels per boll was significantly greater for the infested treatment in a cohort of bolls that accumulated from 51 to 400 heat units beyond anthesis. Seed germination in bolls infested with brown stink bug was significantly lower in bolls aged 101-600 heat units beyond anthesis.

Animals↗

The mitochondrial genome of allotetraploid cotton (Gossypium L.).

Knowledge of patterns of organellar inheritance are crucial for analyses of organellar genetics and population or phylogenetic analyses based on organellar genes. While it has previously been shown that chloroplast DNA is maternally inherited in cotton (Gossypium L.), no, studies have assessed the mitochondrial genome complement of the allotetraploid species of cotton or addressed inheritance of mitochondrial DNA. In this article we report Southern hybridization analyses that indicate that the AD-genome allotetraploid species have an A-genome-like mitochondrial genome and provide evidence that mitochondrial genomes are also maternally inherited in cotton.

Blotting, Southern↗

Evaluating the impacts of refuge width on source-sink dynamics between transgenic and non-transgenic cotton.

Resistance management for Bt-transgenic crops relies in part on the production of sufficient numbers of susceptible insects in non-toxic refuges. Simulation models suggested that source-sink dynamics could interact with the structure of refuges to impact the production of insects in these areas. We tested the hypothesis that altering isolation between refuges and transgenic cotton by manipulating the width of refuges embedded within cotton fields would alter the density of Heliothis virescens and Helicoverpa zea eggs oviposited in refuges. Three categories of refuge widths were tested over two years: they included narrow (16-24 m wide), medium (32-48 m wide) and wide (80-96 m wide) refuges. Isolation between the two habitats increased as refuge width increased. In 1996, eggs of H. virescens from H. zea were not distinguished, but a significant increase in the density of eggs and a significant decrease in relative yield (refuge yield compared to the yield from immediately surrounding Bt-cotton) was found as refuge width increased. In 1997, eggs from H. virescens were analyzed separately from H. zea using an ELISA test. The density of H. virescens eggs increased with increasing refuge width, and there was a significant decline in density of H. virescens eggs with increasing distance from the refuge. In contrast, there was no impact of refuge width on the density of H. zea eggs, nor was the slope of a regression of egg density and distance from the refuge significantly different from zero. We suggest that these differences reflect differences in the biology of the two insects.

Animals↗

Rapid and inexpensive recovery method of DNA fragments from agarose and polyacrylamide gels by a cotton-wool column tube.

We developed a rapid, convenient, simple, and inexpensive method for isolating pure DNA from agarose and polyacrylamide gels using cotton wool tubes. DNA fragments ranging in size from 193-23,130 bp can be easily recovered within 2 hours by centrifugation through cotton wool from gel slices. The recovery rate of this method is 35% to 50%, when estimated for isolation of lambda DNA-HindIII fragments. We have also recovered 700-bp polymerase chain reaction (PCR) products using cotton wool tubes from electrophoresis on both a 0.8% agarose gel and a 6% polyacrylamide gel, in which satisfactory yields of more than 50% were obtained. The DNA thus recovered in this way is biologically active and can be used as a substrate for further experimental procedures without additional purification steps.

Bacteriophage lambda↗

Byssinosis in the cotton waste industry.

A cross-sectional survey was carried out on 772 workers in 27 mills in the North of England involved in the processing of cotton waste. The overall prevalence of byssinosis as defined by Schilling's criteria was 9.8 per cent with 5.4 per cent having grades 2 and 3. The prevalence in workers who had only ever had cotton waste exposure (5 per cent) was significantly less than for those who had mixed waste and raw cotton exposure. There was no clear relationship between prevalence of byssinosis and years of exposure or dust levels.

Adolescent↗

Hydrolysis of cotton cellulose by Exo- and endo-type cellulases from Irpex lacteus: differential scanning calorimetric study.

The mode of hydrolysis of cotton cellulose by two highly purified exo- and endo-type cellulases from Irpex lacteus was investigated by differential scanning calorimetry, to measure changes in the size of the amorphous region in cotton fibers with the enzymatic reaction. The cellulases induced entirely different changes in the size of the amorphous region, particularly at earlier stages of reaction. Exo-type cellulase gradually reduced the amorphous region with release of cellobiose from the initial stage of hydrolysis, but began to increase the amorphous region at more advanced stages of hydrolysis. By contrast, endo-type cellulase caused no liberation of reducing sugar at the initial stage of hydrolysis but caused a sharp increase in the amorphous region, and it thereafter caused a rapid decrease of the amorphous region, accompanied with the production of various kinds of cellooligosaccharides. The rate of size reduction of the amorphous region caused by endo-type cellulase was much higher than that by exo-type cellulase. Convergence of the decrease in the size of amorphous region during hydrolysis by endo-type cellulase is followed by the increase in this region being influenced by further hydrolysis of remained crystalline region. Substantial changes in the morphology of cotton occurred with the two cellulases after the hydrolysis stages at which the size of the amorphous region was minimum.

Calorimetry, Differential Scanning↗

Differential evolutionary dynamics of duplicated paralogous Adh loci in allotetraploid cotton (Gossypium).

Levels and patterns of nucleotide diversity vary widely among lineages. Because allopolyploid species contain duplicated (homoeologous) genes, studies of nucleotide diversity at homoeologous loci may facilitate insight into the evolutionary dynamics of duplicated loci. In this study, we describe patterns of sequence diversity from an alcohol dehydrogenase homoeologous locus pair (AdhC) in allotetraploid cotton (Gossypium, Malvaceae). These data are compared with equivalent information from another homoeologous alcohol dehydrogenase gene pair (AdhA, Small, Ryburn, and Wendel 1999. Mol. Biol. Evol. 16:491-501) which has an overall slower evolutionary rate than AdhC. As expected from the predicted correlation between nucleotide diversity and evolutionary rate, nucleotide diversity was higher for AdhC than for AdhA. In addition, nucleotide diversity is higher in the D-subgenome of allotetraploid cotton for AdhC, confirming earlier observations for AdhA. These observations indicate that for these two pairs of Adh loci, the null hypothesis of equivalent evolutionary dynamics for duplicated genes in allotetraploid cotton is rejected.

Alcohol Dehydrogenase↗

Identification and expression of cotton (Gossypium hirsutum L.) plastidial carbonic anhydrase.

Four carbonic anhydrase (CA) cDNA clones were isolated from a 48 h dark-grown cotton (Gossypium hirsutum L.) seedling cDNA library. Nucleotide sequence analysis revealed two different CA isoforms designated GhCA1 and GhCA2. The encoded polypeptides possess N-terminal serine/threonine-rich regions indicative of plastid transit peptides, and approximately 80% sequence identity to other plant plastidial beta-CAs. The GhCA1 cDNA encodes a nearly complete preprotein of 323 amino acids with a molecular mass of 34.9 kDa and a predicted mature protein of 224 amino acids with a molecular mass of 24.3 kDa. Eleven nucleotide differences within ORFs of GhCA1 and GhCA2 result in 5 conservative amino acid substitutions. The 3' GhCA2 untranslated region contains five additional substitutions and one single nucleotide addition. GhCA1 clones, nearly full-length or with 70% of the transit peptide deleted, were expressed as LacZ alpha fusion proteins in E. coli. Lysates of these strains contained 9-fold higher levels of CA activity as compared to untransformed controls and this activity was inhibited by CA-specific inhibitors. Sulfanilamide, acetazolamide, ethoxyzolamide, each at 10 mM, inhibited recombinant CA activity approximately 50%, 65%, and 75%, respectively. In plant tissue homogenates these inhibitors reduced CA activity by 50%, 70%, and 95%, respectively. Although CA activity was bighest in extracts of mature cotton leaves, probing total RNA with GhCA1 revealed CA transcript levels to be highest in the cotyledons of dark-grown cotton seedlings. Collectively, our data indicate the presence of a plastid-localized CA in cotyledons of germinated seeds, suggesting a role for CA in postgerminative growth.

Amino Acid Sequence↗

Hmg-coA reductase gene family in cotton (Gossypium hirsutum L.): unique structural features and differential expression of hmg2 potentially associated with synthesis of specific isoprenoids in developing embryos.

As a first step towards understanding the biosynthesis of isoprenoids that accumulate in specialized pigment glands of cotton at the molecular level, two full-length genes (hmg1 and hmg2) were characterized encoding hmg-coA reductase (HMGR; EC 1.1.1.34), the enzyme that catalyzes the formation of a key isoprenoid precursor. Cotton hmgr genes exhibited features typical of other plant genes, however, hmg2 encodes the largest of all plant HMGR enzymes described to date. HMG2 contains several novel features that may represent functional specialization of this particular HMGR isoform. Such features include a unique 42 amino acid sequence located in the region separating the N-terminal domain and C-terminal catalytic domain, as well as an N-terminal hydrophobic domain that is not found in HMG1 or other HMGR enzymes. DNA blot analysis revealed that hmg1 and hmg2 belong to small subfamilies that probably include homeologous loci in allotetraploid cotton (Gossypium hirsutum L.). Ribonuclease protection assays revealed that hmg1 and hmg2 are differentially expressed in a developmentally- and spatially-modulated manner during morphogenesis of specialized terpenoid-containing pigment glands in embryos. Induced expression of hmg2 coincided with a possible commitment to sesquiterpenoid biosynthesis in developing embryos, although other developmental processes also requiring HMGR cannot be excluded.

Base Sequence↗

Overexpression of the Arabidopsis 14-3-3 protein GF14 lambda in cotton leads to a "stay-green" phenotype and improves stress tolerance under moderate drought conditions.

The Arabidopsis gene GF14 lambda that encodes a 14-3-3 protein was introduced into cotton plants to explore the physiological roles that GF14 lambda might play in plants. The expression level of GF14 lambda under the control of the cauliflower mosaic virus 35S promoter varied in transgenic cotton plants, and lines that expressed GF14 lambda demonstrated a "stay-green" phenotype and improved water-stress tolerance. These lines wilted less and maintained higher photosynthesis than segregated non-transgenic control plants under water-deficit conditions. Stomatal conductance appears to be the major factor for the observed higher photosynthetic rates under water-deficit conditions. The stomatal aperture of transgenic plants might be regulated by GF14 lambda through some transporters such as H(+)-ATPase whose activities are controlled by their interaction with 14-3-3 proteins. However, since 14-3-3 proteins interact with numerous proteins in plant cells, many metabolic processes could be affected by the GF14 lambda overexpression. Whatever the mechanisms, the traits observed in the GF14 lambda-expressing cotton plants are beneficial to crops under certain water-deficit conditions.

14-3-3 Proteins↗

Differential induction of 3-hydroxy-3-methylglutaryl CoA reductase in two cotton species following inoculation with Verticillium.

Gossypium barbadense cottons are typically more resistant to wilt pathogens than are Gossypium hirsutum cultivars. Both species make terpenoid phytoalexins in response to infection, implicating isoprenoid biosynthesis as a factor in resistance. Conserved regions in plant 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR), the first enzyme in the terpene biosynthesis pathway, were used to design polymerase chain reaction primers for cloning a fragment of a cotton HMGR gene. The clone was used as a probe on Northern blots to show that induction of HMGR mRNA following introduction of Verticillium dahliae spores into the vascular system is much more rapid in Seabrook Sea Island, a restant G. barbadense cotton, than it is in Rowden, a susceptible G. hirsutum. The amount of HMGR mRNA returned to near control levels in 4 days in the former variety but continued to accumulate in the latter. Specific enzyme activity of HMGR also increased more rapidly in stele extracts of Seabrook Sea Island than in Rowden.

Amino Acid Sequence↗

Semenogelin II gene is replaced by a truncated line 1 repeat in the cotton-top tamarin.

The human seminal vesicles secrete two proteins, semenogelin I and semenogelin II, at very high concentrations. It has previously been shown that the cotton-top tamarin (Sanguinus oedipus), a New World monkey, is lacking the semenogelin II gene. We have now determined the nucleotide sequence of DNA located 5--13 kilobases (kb) downstream of the tamarin semenogelin I gene---a region that in man is occupied by the semenogelin II gene. Two regions with homology to the human semenogelin II gene were identified in the tamarin DNA. The first region, of 3.5 kb, is homologous to DNA upstream of the human gene, and the second region, of 0.6 kb, is mainly derived from the second intron. Between these regions, equivalent to 594 base pairs (bp) upstream of the transcription initiation site to 12 bp downstream of the stop codon in the human semenogelin II gene, the cotton-top tamarin DNA carries a truncated LINE1 repeat. In another set of experiments, the tamarin DNA hybridizing to the mouse semenoclotin gene was investigated. It was concluded that hybridization is with the second intron of the semenoclotin gene, but very likely, the material does not represent a cotton-top tamarin semenoclotin gene. Thus, a mammalian ancestor probably carried a single gene that in the rodent lineage developed into the semenoclotin gene and in the primate lineage into a progenitor of the semenogelin genes.

Base Sequence↗

Synchronous eruption of cotton-induced granulomas 45 years after exposure: report of an unusual case.

A 58-year-old woman developed multiple, recurrent abscesses and subcutaneous nodules over both buttocks during a 12-year period. Microbiological studies of surgical biopsy specimens were inconclusive for a specific infectious cause, but histochemical preparations on several separate occasions revealed an exuberant foreign body reaction to raw cotton fibers. Interestingly, the patient gave a detailed history of sliding down piles of cotton hulls during her childhood, some 45 years earlier. With a probable cause-and-effect relationship established, we postulate a dramatically delayed foreign body granulomatous reaction to cotton fibers.

Buttocks↗

Cotton-wool spots.

A series of 24 consecutive patients presenting with a fundus picture characterized by a predominance of cotton-wool spots, or a single cotton-wool spot, is reported. Excluded were patients with known diabetes mellitus. Etiologic conditions found included previously undiagnosed diabetes mellitus in five patients, systemic hypertension in five patients, cardiac valvular disease in two patients, radiation retinopathy in two patients, and severe carotid artery obstruction in two patients. Dermatomyositis, systemic lupus erythematosus, polyarteritis nodosa, leukemia, AIDS, Purtscher's retinopathy, metastatic carcinoma, intravenous drug abuse, partial central retinal artery obstruction, and giant cell arteritis were each found in one patient. In only one patient did a systemic workup fail to reveal an underlying cause. The presence of even one cotton-wool spot in an otherwise normal fundus necessitates an investigation to ascertain systemic etiologic factors.

Adult↗

Nerve fiber bundle visual field defect resulting from a giant peripapillary cotton-wool spot.

Cotton-wool spots are the clinical manifestation of focal infarcts of the retinal nerve fiber layer. They rarely cause significant visual field loss. A large idiopathic cotton-wool spot in a 34-year-old healthy woman caused a nerve fiber bundle visual field defect and an afferent pupillary defect that remained after the cotton-wool spot had disappeared and the retina and optic nerve appeared normal.

Adult↗