title: Virus-Induced Gene Silencing in Cultivated Cotton (Gossypium spp.) Using Tobacco Rattle Virus.
abstract: The study described here has optimized the conditions for virus-induced gene silencing (VIGS) in three cultivated cotton species (Gossypium hirsutum, G. arboreum, and G. herbaceum) using a Tobacco rattle virus (TRV) vector. The system was used to silence the homolog of the Arabidopsis thaliana chloroplastos alterados 1 (AtCLA1) gene, involved in chloroplast development, in G. herbaceum, G. arboreum, and six commercial G. hirsutum cultivars. All plants inoculated with the TRV vector to silence CLA1 developed a typical albino phenotype indicative of silencing this gene. Although silencing in G. herbaceum and G. arboreum was complete, silencing efficiency differed for each G. hirsutum cultivar. Reverse transcriptase polymerase chain reaction (PCR) and real-time quantitative PCR showed a reduction in mRNA levels of the CLA1 homolog in all three species, with the highest efficiency (lowest CLA1 mRNA levels) in G. arboreum followed by G. herbaceum and G. hirsutum. The results indicate that TRV is a useful vector for VIGS in Gossypium species. However, selection of host cultivar is important. With the genome sequences of several cotton species recently becoming publicly available, this system has the potential to provide a very powerful tool for the rapid, large-scale reverse-genetic analysis of genes in Gossypium spp.
authors.affiliation: Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan. romamustafa4@gmail.com.
authors.forename: Roma
authors.initials: R
authors.lastname: Mustafa
authors.affiliation: Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan. shafiq.4721@gmail.com.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Shafiq
authors.affiliation: Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan. shahidmansoor7@gmail.com.
authors.forename: Shahid
authors.initials: S
authors.lastname: Mansoor
authors.affiliation: Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan. rob.briddon@gmail.com.
authors.forename: Rob W
authors.initials: RW
authors.lastname: Briddon
authors.affiliation: Genomics and Bioinformatics Research Unit, PO Box 36, Stoneville, MS, 38776, USA. brian.scheffler@ars.usda.gov.
authors.forename: Brian E
authors.initials: BE
authors.lastname: Scheffler
authors.affiliation: Genomics and Bioinformatics Research Unit, PO Box 36, Stoneville, MS, 38776, USA. jodi.scheffler@ars.usda.gov.
authors.forename: Jodi
authors.initials: J
authors.lastname: Scheffler
authors.affiliation: Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan. imranamin1@yahoo.com.
authors.forename: Imran
authors.initials: I
authors.lastname: Amin
mesh_terms: D029681:Arabidopsis Proteins / Q000096:biosynthesis / Q000235:genetics*; D002736:Chloroplasts / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D003368:Gossypium / Q000235:genetics* / Q000821:virology; D010942:Plant Viruses / Q000235:genetics*; D014026:Nicotiana / Q000821:virology
chemical_list: D029681:Arabidopsis Proteins; C099778:CLA1 protein, Arabidopsis
keywords: Chloroplastos alterados 1; Gene silencing; Gossypium; Tobacco rattle virus vector; Virus-induced gene silencing