TY - JOUR
T1 - Virus-Induced Gene Silencing in Cultivated Cotton (Gossypium spp.) Using Tobacco Rattle Virus
AU - Mustafa, Roma
AU - Shafiq, Muhammad
AU - Mansoor, Shahid
AU - Briddon, Rob W.
AU - Scheffler, Brian E.
AU - Scheffler, Jodi
AU - Amin, Imran
N1 - Funding Information:
The authors are grateful to Dr. Ping He (Department of Biochemistry and Biophysics, Institute of Plant Genomics and Biotechnology, Texas A&M University, Texas, USA) for providing the TRV-GrCLA1 construct and Dr. Mehboob-ur-Rehman (NIBGE, Faisalabad) for providing cotton seeds. This material is based upon work supported by the "Pak-US cotton productivity enhancement program" of the International Center for Agricultural Research in the Dry Areas (ICARDA) funded by United States Department of Agriculture (USDA), Agricultural Research Service (ARS), under Agreement No. 58-6402-0-178F. Any opinions, findings, conclusions or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the views of the USDA or ICARDA.
Publisher Copyright:
© 2015, Springer Science+Business Media New York.
PY - 2016/1/1
Y1 - 2016/1/1
N2 - 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.
AB - 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.
KW - Chloroplastos alterados 1
KW - Gene silencing
KW - Gossypium
KW - Tobacco rattle virus vector
KW - Virus-induced gene silencing
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U2 - 10.1007/s12033-015-9904-z
DO - 10.1007/s12033-015-9904-z
M3 - Article
C2 - 26603442
AN - SCOPUS:84954402675
SN - 1073-6085
VL - 58
SP - 65
EP - 72
JO - Molecular Biotechnology
JF - Molecular Biotechnology
IS - 1
ER -