Cotton α-Globulin Promoter: Isolation and Functional Characterization in Transgenic Cotton,Arabidopsis, and Tobacco

Globulins are the most abundant seed storage proteins in cotton and, therefore, their regulatory sequences could potentially provide a good source of seed-specific promoters. We isolated the putative promoter region of cotton -globulin B gene by gene walking using the primers designed from a cotton staged embryo cDNA clone. PCR amplified fragment of 1108 bp upstream sequences was fused to gusA gene in the binary vector pBI101.3 to create the test construct. This was used to study the expression pattern of the putative promoter region in transgenic cotton, Arabidopsis, and tobacco. Histochemical GUS analysis revealed that the promoter began to express during the torpedo stage of seed development in tobacco and Arabidopsis, and during cotyledon expansion stage in cotton. The activity quickly increased until embryo maturation in all three species. Fluorometric GUS analysis showed that the promoter expression started at 12 and 15 dpa in tobacco and cotton, respectively, and increased through seed maturation. The strength of the promoter expression, as reflected by average GUS activity in the seeds from primary transgenic plants, was vastly different amongst the three species tested. In Arabidopsis, the activity was 16.7% and in tobacco it was less than 1% of the levels detected in cotton seeds. In germinating seedlings of tobacco and Arabidopsis, GUS activity diminished until it was completely absent 10 days post imbibition. In addition, absence of detectable level of GUS expression in stem, leaf, root, pollen, and floral bud of transgenic cotton confirmed that the promoter is highly seed-specific. Analysis of GUS activity at individual seed level in cotton showed a gene dose effect reflecting their homozygous or hemizygous status. Our results show that this promoter is highly tissue-specific and it can be used to control transgene expression in dicot seeds.     

Cotton Leaf Curl Virus： Ionic Status of Leaves and Symptom Development

In the present study, the relationship between the nutritional status of leaves and the development of symptoms of cotton leaf curl virus （CLCuV） in two cotton （Gossypium hirsutum L.） cuItlvars （I.e. CIM-240 and S-12） was Investigated. The incidence of disease attack was found to be 100% In the S-12 cuItlvar and 16% in the CIM-240 cuItivar. Geminivirus particles in infected leaves were confirmed by transmission electron microscope examination of highly specific geminivirus coat protein antlsera-treated cell sap. The CLCuV Impaired the accumulation of different nutrients in both cuItivars. A marked decrease in the accumulation of Ca^2＋ and K^＋ was observed in infected leaves. However, the disease had no effect on leaf concentrations of Na^＋, N, and P. It was observed that the curling of leaf margins in CLCuV-Infected plants was associated with the leaf Ca^2＋ content; leaf curling was severe in plants with a significant reduction In Ca^2＋ content. Moreover, leaf K＆＋ content was found to be associated with resistance/susceptibility to CLCuV infection.     

Morphometric Variation in Geographic Populations of Cotton Bollworm,Helicoverpa armigera (Hübner) Occurring in South Indian Cotton Ecosystems

Morphometric characterization of twelve geographic populations of cotton bollworm, Helicoverpa armigera occurring in south Indian cotton ecosystems was done at larval, pupal and adult stages over three cropping seasons. Traits such as length and weight of larvae, pupa and length and width of the wing, length of fore-, mid- and hind femur, male reproductive organ-length of genital capsule, valves, and ejaculatory duct, female reproductive organ-length of appendix bursae and ductus bursae at adult stage were measured across three years. Populations significantly differed for most of the traits studied. It was evident that populations from northern parts recorded higher phenotypic attributes compared to those from southern parts of south Indian cotton ecosystem. Besides larval, pupal and adult external phenotypic traits, attributes of male reproductive organ viz., length of genital capsule, valves, and ejaculatory duct and female reproductive organ viz., length of appendix bursae and ductus bursae differed significantly among populations. Information on population structure and differences based on morphometry would be useful in better understanding of population dynamics and management of this pest in cotton and other crops in south India.     

Genotype and Planting Density Effects on Rooting Traits and Yield in Cotton （Gossypium hirsutum L,）

Root density distribution of plants is a major Indicator of competition between plants and determines resource capture from the solh This experiment was conducted in 2005 at Anyang, located in the Yellow River region, Henan Province, China. Three cotton （Gossyplum hlrsutum L.） cultivars were chosen： hybrid Btcultlvar CRI46, conventional Btcultlvars CRI44 and CRI45. Six planting densities were designed, ranging from 1.5 to 12.0 plants/m^2. Root parameters such as surface area, diameter and length were analyzed by using the DT-SCAN Image analysis method. The root length density （RLD）, root average diameter and root area Index （RAI）, root surface area per unit land area, were studied. The results showed that RLD and RAI differed between genotypes; hybrid CRI46 had significantly higher （P 〈0.05） RLD and RAI values than conventlonal cultlvars, especially under low planting densities, less than 3.0 plants/m^2. The root area index （RAI） of hybrid CRI46 was 61% higher than of CRI44 and CRI45 at the flowering stage. The RLD and RAI were also significantly different （P = 0.000） between planting densities. The depth distribution of RAI showed that at Increasing planting densities RAI was Increasingly distributed in the soil layers below 50 cm. The RAI of hybrid CRI46 was for all planting densities, obviously higher than other cultivars during the flowering and boll stages. It was concluded that the hybrid had a strong advantage in root maintenance preventing premature senescence of roots. The root diameter of hybrid CRI46 had a genetically higher root diameter at planting densities lower than 6.0 plants/m^2. Good associations were found between yield and RAI In different stages. The optimum planting density ranged from 4.50 plants/m^2 to 6.75 plants/m^2 for conventional cultlvars and around 4.0-5.0 plants/m^2 for hybrids.     

Fusarium verticillioides (Saccardo) Nirenberg Associated with Hardlock of Cotton

The development of new immune potentiators for human vaccines is an important and expanding field of research. In the present study, the ability of the capsular polysaccharide from Neisseria meningitidis serogroup A (CPS-A), a mannose-containing carbohydrate, to enhance the antibody production against a co-administered model vaccine antigen, is examined. A protein-meningococcal serogroup C capsular polysaccharide (CPS-C) conjugate was selected as the model antigen for this study. After subcutaneous immunization of Balb/C mice, the conjugate mixed with CPS-A induced higher anti-CPS-C IgG and IgG_2a antibody levels and higher anti-meningococcal serogroup C bactericidal titers than the conjugate alone or mixed with CPS-C. The immuno-stimulatory properties exhibited by CPS-A and the fact that vaccines based on purified CPS-A has been safely used during decades to fight the serogroup A meningococcal disease, support the proposal to use CPS-A as immune potentiator for human vaccination studies.     

Salinity Induced Changes in α-Amylase Activity During Germination and Early Cotton Seedling Growth

Salinity induced changes in -amylase activity in three cotton cultivars (NIAB-Karishma, NIAB-86 and K-115) was studied during germination and early seedling growth under controlled conditions. The increase in NaCl concentration resulted in the decrease in -amylase activity and break down of starch into reducing and non-reducing sugars in all cultivars, however, it was more pronounced in NIAB-86. K-115 showed highest germination followed by NIAB-Karishma and NIAB-86.     

Comparative Development of Lint and Fuzz Using Different Cotton Fiber-specific Developmental Mutants in Gossypium hirsutum

A series of fiber-specific mutants, or germplasms, have been recently used in the study of fiber development. In the current study, scanning electron microscopy （SEM） was used to investigate developmental differences in lint and fuzz initiation in different genotypes （Gossypium hirsutum） of upland cotton. These fiber mutants included dominant naked seed N1, recessive naked seed n2, Xuzhou-142 lintless-fuzzless （XZ142WX）, Xinxiangxiaojilintless-fuzzless （XinWX）, Xinxiangxiaojilinted-fuzzless （XinFLM）, with TM-1, the cytogenetic and genetic experimental standard stock, as the control. Characteristics of fiber initiation were analyzed from -1 to ＋1 days post anthesis （dpa） and at 4 and 5 dpa for fuzz initiation. Our data suggested that lint initiation centered on day of anthesis （0dpa）, and elongated significantly at 1dpa, while fuzz initiation began at 4dpa, although the shape of fuzz protrusions differed from that of lint fibers. Fiber initiation occurred first on the ovule funicular crest. Compared to TM-1, there was a noted retardation in development and fiber protrusion in N1 and XinFLM. Microscopy data also demonstrated that lintless-fuzzless mutants （XZ142WX and XinWX） developed irregular protrusions during early developmental stages, which were unable to grow into fiber.     

Cotton cultivation and textile production in the Arabian Peninsula during antiquity; the evidence from Mada��in Salih (Saudi Arabia) and Qal��at al-Bahrain (Bahrain)

The discovery of seeds and textiles from Gossypium (cotton) in Achaemenian levels of the mid-6th–late 4th century b.c. at Qal’at al-Bahrain, Bahrain and in early 1st millennium a.d. at Mada’in Salih, Saudi Arabia, reveals the role played by the Arabian Peninsula as a textile production centre during the centuries before and after the beginning of the Christian era. Both these sites were situated on important trade routes, overseas (Qal’at al-Bahrain) and overland (Mada’in Salih), and it is likely that at least part of the cotton production was intended for trade, complementing and perhaps competing with other sources of cotton textiles in the contemporary Middle East. In the arid climate of the Arabian Peninsula, cotton was probably grown in association with irrigated date palm gardens where a wide array of other crops was grown, as is shown by the analysis of charred seeds and wood from occupation levels at both sites. The present article places these particular finds in the larger context of cotton cultivation in the Middle East and India.     

“Spider Monkey Cotton”: Bridging Waiwai and Scientific Ontologies to Characterize Spider Monkey (Ateles paniscus) Filariasis in the Konashen Community Owned Conservation Area,Guyana

International Journal of Primatology - Zoonotic disease risk is greatly influenced by cultural practices and belief systems. Yet, few studies have investigated how different ways of knowing are...     

Transient Expression of β-Glucuronidase in Embryo Axes of Cotton by Agrobacterium and Particle Bombardment Methods

Transient expression of -glucuronidase (GUS) in zygotic embryo axes of two cotton (Gossypium hirsutum L.) cultivars NHH-44 and DCH-32 was induced by Agrobacterium mediated transformation or by particle bombardment. For Agrobacterium transformation, disarmed A. tumefaciens strain GV 2260/p35SGUSINT was used. In cv. NHH-44, the maximum frequency of transient expression (14.28 %) was achieved on spotting Agrobacterium paste on the apical regions of the split embryo axes. The method resulted in a transformed callus line, which showed strong GUS activity. Integration of NPTII gene was confirmed by Southern analysis. Transgene expression by particle bombardment was achieved with p35SGUSINT and pIBGUS plasmids independently. The maximum frequency of GUS expression in 29.16 % explants was observed in cultivar NHH-44 with gold microcarriers (1.1 µm) when bombarded once with rupture disc of 7586 kPa at target cell distance of 6 cm. A transformed callus line was obtained when explants were bombarded with p35SGUSINT and cultured on Murashige and Skoog's medium supplemented with B₅ vitamins, 0.1 mg dm^–3 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea, 0.01 mg dm^–3 -naphthaleneacetic acid, 3 % glucose + 50 mg dm^–3 kanamycin. High GUS activity was observed in callus tissue as well as in somatic embryo like structures achieved in liquid shake cultures.     

Molecular Cloning of α-Amylases from Cotton Boll Weevil, Anthonomus grandis and Structural Relations to Plant Inhibitors: An Approach to Insect Resistance

Anthonomus grandis, the cotton boll weevil, causes severe cotton crop losses in North and South America. Here we demonstrate the presence of starch in the cotton pollen grains and young ovules that are the main A. grandis food source. We further demonstrate the presence of α-amylase activity, an essential enzyme of carbohydrate metabolism for many crop pests, in A. grandis midgut. Two α-amylase cDNAs from A. grandis larvae were isolated using RT-PCR followed by 5′ and 3′ RACE techniques. These encode proteins with predicted molecular masses of 50.8 and 52.7 kDa, respectively, which share 58% amino acid identity. Expression of both genes is induced upon feeding and concentrated in the midgut of adult insects. Several α-amylase inhibitors from plants were assayed against A. grandis α-amylases but, unexpectedly, only the BIII inhibitor from rye kernels proved highly effective, with inhibitors generally active against other insect amylases lacking effect. Structural modeling of Amylag1 and Amylag2 showed that different factors seem to be responsible for the lack of effect of 0.19 and α-AI1 inhibitors on A. grandis α-amylase activity. This work suggests that genetic engineering of cotton to express α-amylase inhibitors may offer a novel route to A. grandis resistance.     

Population Genetic Structure of the Cotton Bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) in India as Inferred from EPIC-PCR DNA Markers

Helicoverpa armigera is an important pest of cotton and other agricultural crops in the Old World. Its wide host range, high mobility and fecundity, and the ability to adapt and develop resistance against all common groups of insecticides used for its management have exacerbated its pest status. An understanding of the population genetic structure in H. armigera under Indian agricultural conditions will help ascertain gene flow patterns across different agricultural zones. This study inferred the population genetic structure of Indian H. armigera using five Exon-Primed Intron-Crossing (EPIC)-PCR markers. Nested alternative EPIC markers detected moderate null allele frequencies (4.3% to 9.4%) in loci used to infer population genetic structure but the apparently genome-wide heterozygote deficit suggests in-breeding or a Wahlund effect rather than a null allele effect. Population genetic analysis of the 26 populations suggested significant genetic differentiation within India but especially in cotton-feeding populations in the 2006–07 cropping season. In contrast, overall pair-wise F _ST estimates from populations feeding on food crops indicated no significant population substructure irrespective of cropping seasons. A Baysian cluster analysis was used to assign the genetic make-up of individuals to likely membership of population clusters. Some evidence was found for four major clusters with individuals in two populations from cotton in one year (from two populations in northern India) showing especially high homogeneity. Taken as a whole, this study found evidence of population substructure at host crop, temporal and spatial levels in Indian H. armigera, without, however, a clear biological rationale for these structures being evident.     

Of mice and (Viking?) men: phylogeography of British and Irish house mice

The west European subspecies of house mouse (Mus musculus domesticus) has gained much of its current widespread distribution through commensalism with humans. This means that the phylogeography of M. m. domesticus should reflect patterns of human movements. We studied restriction fragment length polymorphism (RFLP) and DNA sequence variations in mouse mitochondrial (mt) DNA throughout the British Isles (328 mice from 105 localities, including previously published data). There is a major mtDNA lineage revealed by both RFLP and sequence analyses, which is restricted to the northern and western peripheries of the British Isles, and also occurs in Norway. This distribution of the 'Orkney' lineage fits well with the sphere of influence of the Norwegian Vikings and was probably generated through inadvertent transport by them. To form viable populations, house mice would have required large human settlements such as the Norwegian Vikings founded. The other parts of the British Isles (essentially most of mainland Britain) are characterized by house mice with different mtDNA sequences, some of which are also found in Germany, and which probably reflect both Iron Age movements of people and mice and earlier development of large human settlements. MtDNA studies on house mice have the potential to reveal novel aspects of human history.     

Synthesis and properties of (2-pyridyl)alkylamine- and (2-pyridyl)alkylamine–amide-coordinated copper(II) complexes: Structures and non-covalent interactions

Reaction of the ligand N-methyl-N-((6-pivaloylamido-2-pyridyl)methyl)-N-(2-pyridylethyl)amine (mpppa) with equimolar amounts of [Cu(H₂O)₆][ClO₄]₂ or CuCl₂ · 2H₂O in MeCN afforded mononuclear copper(II) complexes [Cu(mpppa)][ClO₄]₂ (1) and [Cu(mpppa)Cl₂] (2). Crystal structure analysis reveals CuN₃O (two pyridyl, an aliphatic amine, and an amide oxygen) coordination in 1 and CuN₃Cl₂ (two pyridyl, an aliphatic amine, and two chlorides) coordination in 2. Crystal packing diagram of 1 reveals that one of the perchlorate counteranions provides weak coordination to copper(II) centers and in turn the copper(II) centers assume pseudo-six-coordination, generating 1D chain. Notably, one of the copper(II)-coordinated chloride ions in 2 participates in an intramolecular N–H?Cl interaction. Intermolecular C–H?Cl interactions in the solid state generate helical structure. Spectroscopic (IR, UV–Vis, and EPR) and redox properties of the two complexes have been investigated and compared.     

Graves’ Disease and Stiff-Person (Stiff-Man) Syndrome: Case Report and Literature Review

ObjectiveTo report an association between two autoimmune conditions, Graves’ disease and stiff-person (stiff-man) syndrome, and discuss the relevant literature.MethodsWe present a case of a 52-year-old white woman with stiff-person syndrome who also had Graves’ disease, discuss her management, and review the related literature. Pertinent published reports from 1950 through 2004 were researched with use of MEDLINE and PubMed, and cross-references to other articles were reviewed.ResultsA 52-year-old white woman presented with symptoms of hyperthyroidism due to Graves’ disease. Laboratory data were as follows: thyrotropin < 0.005 μIU/mL, thyroxine 11.1 μg/dL, free thyroxine index (FTI) 10.7, and triiodothyronine 170 ng/dL. Thyroid-stimulating immunoglobulins (TSI) and thyrotropin-binding inhibitory immunoglobulins (TBII) were positive at 1,986% and 82.5 U/L, respectively. The hyperthyroidism was treated with propranolol. She had a long-standing history of musculoskeletal complaints and was ultimately diagnosed with stiff-person syndrome. During her thyroid evaluation, she had severe neurologic deterioration that necessitated hospitalization and treatment with clonazepam, baclofen, intravenous immunoglobulin, and subsequently prednisone and azathioprine for appreciable symptomatic relief. The aggressive immunosuppression had a profound effect on her symptoms of hyperthyroidism, results of thyroid function tests, and thyrotropin receptor antibodies (TRABs). Thyrotropin was 0.52 μIU/mL, thyroxine was 6.9 μg/dL, and FTI was 5.7. The TSI decreased from 1,986% to 248%, and her TBII normalized from 82.5 U/L to < 5 U/L. She was clinically and biochemically euthyroid at last follow-up in May 2004.ConclusionThis case illustrates the association between TRAB-positive Graves’ disease and stiff-person syndrome and the improvement of Graves’ disease with immunosuppressive therapy. (Endocr Pract. 2005;11: 259-264)     

Increase of β-1, 3-Glucanase and Chitinase Activities in Cotton Callus Cells Treated by Salicylic Acid and Toxin of Verticillium dahliae

The different resistance of cotton (Gossypium hirsutum L.) cultivars to crude toxin of Verticillium dahliae (VD) was correlated with the activities of chitinase and β-1, 3-glucanase in callus cells. The activities of chitinase and β-1, 3-glucanase in the callus cells treated with the VD-toxin were increased to the higher level at earlier time point in resistant cultivars than these in the susceptible cultivars. Exogenous salicylic acid (SA) induced the accumulation of chitinase and β-1,3-glucanase, which resulted in the resistance of callus cells to the VD. toxin. Western blot using a polyclonal antibody against β-1,3-glucanase identified 28 kD protein that was induced by VD-toxin, SA, or VD-toxin plus SA.