• Title/Summary/Keyword: cotton (Gossypium hirsutum L.)

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Bioloistic-mediated Transformation of Cotton (Gossypium hirsutum L.): Embryogenic Calli as Explant

  • Haq Ikram-ul;Asad Shaheen;Zafar Yusuf
    • Journal of Plant Biotechnology
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    • v.7 no.4
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    • pp.211-218
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    • 2005
  • Genetic transformation was carried out by using biolistic gun method. The hypocotyl derived embryogenic calli (explants) of cotton (Gossypium hirsutum L.) cv. Cocker-312 were transformed with a recombinant pGreen II plasmid, in which both, bar (selection marker) and GUS (${\beta}$-glucuronidase) reporter genes were incorporated. Explants were arranged on osmoticum-containing medium (0.5M mannitol) 4 hours prior to and 16 hours after bombardment that was resulted into an increase about >80% for GUS stable expression. 3 days after bombardment, GUS assay was performed, which exhibited, $18.36{\pm}1.00$ calli showed blue spots. The transformed embryogenic calli were cultured on selection medium (@ 6 mg/L basta) for 3 months. The putative transgenic plants were developed via selective somatic embryogenesis (@1.50 mg/L basta); maximum $27.58{\pm}1.25$ somatic embryos were obtained while $17.47{\pm}1.00$ embryos developed into plantlets (@ 0.75mg/L basta). In five independent experiments, up to 7.24% transformation efficiency was recorded. The presence of the transgenes was analyzed by using PCR and southern hybridization analysis. The transgenic plants were developed with in 6-7 months, but mostly transformants were abnormal in morphology.

Cloning and expression of glutathione S-transferase (GST) cDNA from Gossypium hirsutum L.

  • Kang, Won-Hee;Kim, Myong-Jo;Lim, Jung-Dae;Yun, Song-Joong;Chung, Ill-Min;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.4
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    • pp.294-297
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    • 2002
  • A gene coding for the GST of cotton (Gh-5) was cloned into Escherichia coli and experssed. The enzyme remained within the cytoplasm of E. coli. An 696 bp open reading frame was in the 988 base pair fragment of the recombinant plasmid pET-30b(+). The deduced protein sequence consists of 232 amino acids and has a molecular mass of 30235.58 Da. The cloned enzyme conjugated reduced glutathione and 1-chloro-2,4-dinitrobenzene (CDNB). Plant GST cDNA was expressed in microbe and produced polypeptide had function as an enzyme.

High Frequency of Callus Induction, its Proliferation and Somatic Embryogenesis in Cotton (Gossypium hirsutum L.)

  • Haq, Ikram-ul;Zafar, Yusuf
    • Journal of Plant Biotechnology
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    • v.6 no.1
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    • pp.55-61
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    • 2004
  • Callus induction and somatic embryogenesis are fundamental to cotton tissue culture biotechnology. An efficient protocol for callus induction, somatic embryogenesis and their maturation have been developed to regenerate plantlets from cotton (Gossypium hirsutum L.) variety coker 312. Embryogenic callus was initiated from hypo-cotyl region that was used as an explant at seedling stage when it was about 7-8 days old. Callus induction was achieved through culturing hypocotyls (5-7mm) on $MS_{1a} medium supplemented with 2,4-D (0.1 mg/L) and KT (0.5 mg/L) for six weeks. A friable, colorless, bulky and well proliferating callus becomes greenish with the addition of NAA (2.0 mg/L), ZT (0.1 mg/L) and removal of 2,4-D (M $S_{1b}$) cultured for two weeks then again transferred to $MS_{1a}. 2,4-dichlorophenoxyacetic acid (2,4-D) promoted the proliferation of embryogenic callus, but had a negative effect on the differentiation and germination of somatic embryos. ZT (0.1mg/L) and activated charcoal (2g/L), both hormones play an important role in differentiation and germination of somatic embryos in hypocotyls derived embryogenic callus but in case of cotton, such a capability have been observed on MS medium with 1.92 g/L $KNO_3$, but it is considered to attain somewhat more improvement. High embryogenesis frequency was achieved through nutrient deficient stress treatment. The frequency of globular embryogenesis (two-three folds) was achieved when well proliferating callus was (from $MS_{1a}$ media) cultured on MS (1/5 strength) medium for four weeks. Here the development of anthocyanins is the best indicator for somatic embryogenesis. However, when embryoid callus was cultured on MS (full strength) medium, the globular embryos were developed into normal plantlets immediately. In this procedure 27.49% cotyledenary embryos were developed. Of that 70% cotyledenary embryos were developed not only into normal plantlets but rooted simultaneously, when cultured on MS (with 0.05 mgg/L giberrelic acid) medium. So complete plants could be regenerated through somatic embryogenesis from hypocotyl explants within 6 months.s.

Enzymatic Hydrolysis of Cotton Fibers in Supercritical $CO_2$

  • Gayrat Muratov;Kim, Chul
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.2
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    • pp.85-88
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    • 2002
  • A study was carried out on the application of supercritical fluid to the hydrolysis of boll fibers of cotton (cultivar Tashkent-6 of Gossypium hirsutum L.) by cellulase enzymes from Trichoderma viride, Trichoderma reesei and Aspergillus niger. Conditions of the enzymatic process were optimized. The stabilities of cellulase enzymes were sustained at the pressure of up to 160 attn for 48 hours at 5$0^{\circ}C$ in supercritical carbon dioxide.

A Novel MAP Kinase Gene in Cotton (Gossypium hirsutum L.), GhMAPK, is Involved in Response to Diverse Environmental Stresses

  • Wang, Meimei;Zhang, Ying;Wang, Jian;Wu, Xiaoliang;Guo, Xingqi
    • BMB Reports
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    • v.40 no.3
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    • pp.325-332
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    • 2007
  • The mitogen-activated protein kinase (MAPK) cascade is one of the major and evolutionally conserved signaling pathways and plays pivotal role in the regulation of stress and developmental signals in plants. Here, a novel gene, termed Gossypium hirsutum MAPK (GhMAPK), was isolated from cotton. The full-length cDNA of GhMAPK encodes for a 372 amino acid protein that contains all 11 of the MAPK conserved subdomains and the phosphorylationactivation motif, TEY. Amino acid sequence alignment revealed that GhMAPK shared high identity with group-C MAPK in plants and showed 83~89% similarities with MAPKs from Arabidopsis, apricot, pea, petunia, and tobacco. Southern blot analysis indicated that the GhMAPK belonged to a multygene family in cotton. Two introns were found within the region of genomic sequence. Northern blot analysis revealed that the transcripts of GhMAPK accumulated markedly when the cotton seedlings were subjected to various abiotic stimuli such as wounding, cold (4$^{\circ}C$), or salinity stress; Furthermore, GhMAPK was upregulated by the exogenous signaling molecules, such as salicylic acid (SA) and hydrogen peroxide ($H_2O_2C$), as well as pathogen attacks. These results indicate that the GhMAPK, which has a high degree of identity with group-C plant MAPKs, may also play an important role in response to environmental stresses.

Molecular Characterization of a Transient Expression Gene Encoding for 1-Aminocyclopropane-1-carboxylate Synthase in Cotton (Gossypium hirsutum L.)

  • Wang, Xia;Zhang, Ying;Zhang, Jiedao;Cheng, Cheng;Guo, Xingqi
    • BMB Reports
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    • v.40 no.5
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    • pp.791-800
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    • 2007
  • Ethylene performs an important function in plant growth and development. 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS), the key enzyme involved in ethylene biosynthesis, has been the focus of most ethylene studies. Here, a cotton ACS gene referred to as Gossypium hirsutum ACS1 (GhACS1), was isolated. The full-length cDNA of GhACS1 encodes for a 476-amino acid protein which harbors seven conserved regions, 11 invariant amino acid residues, and the PLP binding active site, all of which characterize ACC synthases. Alignment analysis showed that GhACS1 shared a high degree of identity with other known ACC synthases from different species. Two introns were detected in the genomic DNA sequence, and the results of Southern blot analysis suggested that there might be a multi-gene family encoding for ACC synthase in cotton. From the phylogenetic tree constructed with 24 different kinds of ACC synthases, we determined that GhACS1 falls into group II, and was closely associated with the wound-inducible ACS of citrus. The analysis of the 5' flanking region of GhACS1 revealed a group of putative cis-acting elements. The results of expression analysis showed that GhACS1 displayed its transient expression nature after wounding, abscisic acid (ABA), and $CuCl_2$ treatments. These results indicate that GhACS1, which was transiently expressed in response to certain stimuli, may be involved in the production of ethylene for the transmission of stress signals.

Use of Remotely-Sensed Data in Cotton Growth Model

  • Ko, Jong-Han;Maas, Stephan J.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.4
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    • pp.393-402
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    • 2007
  • Remote sensing data can be integrated into crop models, making simulation improved. A crop model that uses remote sensing data was evaluated for its capability, which was performed through comparing three different methods of canopy measurement for cotton(Gossypium hirsutum L.). The measurement methods used were leaf area index(LAI), hand-held remotely sensed perpendicular vegetation index(PVI), and satellite remotely sensed PVI. Simulated values of cotton growth and lint yield showed reasonable agreement with the corresponding measurements when canopy measurements of LAI and hand-held remotely sensed PVI were used for model calibration. Meanwhile, simulated lint yields involving the satellite remotely sensed PVI were in rough agreement with the measured lint yields. We believe this matter could be improved by using remote sensing data obtained from finer resolution sensors. The model not only has simple input requirements but also is easy to use. It promises to expand its applicability to other regions for crop production, and to be applicable to regional crop growth monitoring and yield mapping projects.