title: Herbicide-resistant cotton (Gossypium hirsutum) plants: an alternative way of manual weed removal.
abstract: BACKGROUND
  Cotton yield has been badly affected by different insects and weed competition. In Past Application of multiple chemicals is required to manage insects and weed control was achieved by different conventional means, such as hand weeding, crop rotation and polyculture, because no synthetic chemicals were available. The control methods shifted towards high input and target-oriented methods after the discovery of synthetic herbicide in the 1930s. To utilise the transgenic approach, cotton plants expressing the codon-optimised CEMB GTGene were produced in the present study.
  
  
  RESULTS
  Local cotton variety CEMB-02 containing Cry1Ac and Cry2A in single cassette was transformed by synthetic codon-optimised 5-enolpyruvylshikimate-3-phosphate synthase gene cloned into pCAMBIA 1301 vector under 35S promoter with Agrobacterium tumifaciens. Putative transgenic plants were screened in MS medium containing 120 µmol/L glyphosate. Integration and expression of the gene were evaluated by PCR from genomic DNA and ELISA from protein. A 1.4-kb PCR product for Glyphosate and 167-bp product for Cry2A were obtained by amplification through gene specific primers. Expression level of Glyphosate and Bt proteins in two transgenic lines were recorded to be 0.362, 0.325 µg/g leaf and 0.390, 0.300 µg/g leaf respectively. FISH analysis of transgenic lines demonstrates the presence of one and two copy no. of Cp4 EPSPS transgene respectively. Efficacy of the transgene Cp4 EPSPS was further evaluated by Glyphosate spray (41 %) assay at 1900 ml/acre and insect bioassay which shows 100 %mortality of insect feeding on transgenic lines as compared to control.
  
  
  CONCLUSIONS
  The present study shows that the transgenic lines produced in this study were resistant not only to insects but also equally good against 1900 ml/acre field spray concentration of glyphosate.
authors.affiliation: CEMB University of the Punjab, 87 West canal Bank Road, Lahore, 53700, Pakistan. ayeshazeeshan1981@yahoo.com.
authors.forename: Ayesha
authors.initials: A
authors.lastname: Latif
authors.affiliation: CEMB University of the Punjab, 87 West canal Bank Road, Lahore, 53700, Pakistan. qayyumabdul77@yahoo.com.
authors.forename: Abdul Qayyum
authors.initials: AQ
authors.lastname: Rao
authors.affiliation: CEMB University of the Punjab, 87 West canal Bank Road, Lahore, 53700, Pakistan. mazmatullahkhan@yahoo.com.
authors.forename: Muhammad Azmat Ullah
authors.initials: MA
authors.lastname: Khan
authors.affiliation: CEMB University of the Punjab, 87 West canal Bank Road, Lahore, 53700, Pakistan. naila.cemb1@gmail.com.
authors.forename: Naila
authors.initials: N
authors.lastname: Shahid
authors.affiliation: CEMB University of the Punjab, 87 West canal Bank Road, Lahore, 53700, Pakistan. kamrancembian@yahoo.com.
authors.forename: Kamran Shehzad
authors.initials: KS
authors.lastname: Bajwa
authors.affiliation: CEMB University of the Punjab, 87 West canal Bank Road, Lahore, 53700, Pakistan. aleem059@yahoo.com.
authors.forename: Muhammad Aleem
authors.initials: MA
authors.lastname: Ashraf
authors.affiliation: CEMB University of the Punjab, 87 West canal Bank Road, Lahore, 53700, Pakistan. botanist.adil@gmail.com.
authors.forename: Malik Adil
authors.initials: MA
authors.lastname: Abbas
authors.affiliation: CEMB University of the Punjab, 87 West canal Bank Road, Lahore, 53700, Pakistan. azam.tks@gmail.com.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Azam
authors.affiliation: CEMB University of the Punjab, 87 West canal Bank Road, Lahore, 53700, Pakistan. ahmadali.shahid@gmail.com.
authors.forename: Ahmad Ali
authors.initials: AA
authors.lastname: Shahid
authors.affiliation: CEMB University of the Punjab, 87 West canal Bank Road, Lahore, 53700, Pakistan. dr.idrees@gmail.com.
authors.forename: Idrees Ahmad
authors.initials: IA
authors.lastname: Nasir
authors.affiliation: CEMB University of the Punjab, 87 West canal Bank Road, Lahore, 53700, Pakistan. tayyabhusnain@yahoo.com.
authors.forename: Tayyab
authors.initials: T
authors.lastname: Husnain
mesh_terms: D056915:DNA Copy Number Variations; D005998:Glycine / Q000031:analogs & derivatives; D003368:Gossypium / Q000235:genetics; D053209:Herbicide Resistance; D030821:Plants, Genetically Modified; D000097797:Glyphosate
chemical_list: D005998:Glycine; D000097797:Glyphosate