Physiological and molecular mechanisms of glyphosate tolerance in an in vitro selected cotton mutant |
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| Affiliation: | 1. Department of Agronomy, College of Agricultural and Biotechnology, Zhejiang University, Hangzhou 310029, PR China;2. Department of Biotechnology and Genetic Engineering, Kohat University of Science and Technology (KUST), 26000 NWFP, Pakistan;1. Key Laboratory of Food Quality and Safety of Jiangsu/Province-State Key Laboratory Breeding Base, Institute of Food Quality Safety and Detection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;2. Huaihua Vocational and Technical College, Huaihua 418007, China;1. College of Horticulture and Plant Protection, Yangzhou University, Yangzhou 225009, China;2. Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, China;3. Joint International Research Laboratory of Agriculture & Agri-Product Safety (Yangzhou University), Jiangsu, Yangzhou 225009, China;4. State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Institute for Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 320021, China;1. Department of General Biology, Londrina State University, Londrina, PR, Brazil;2. Embrapa Soja, Rodovia Carlos João Strass, Acesso Orlando Amaral, Warta, PO. Box 231, 86001-970, Londrina, PR, Brazil;3. Embrapa Instrumentação, Rua XV de Novembro, 1452, 13560-970, São Carlos, São Paulo, Brazil;4. São Carlos Institute of Chemistry, University of São Paulo, Avenida Trabalhador São Carlense, 400, 13566-590, São Carlos, Brazil;5. Japan International Research Center for Agricultural Sciences (JIRCAS), Tsukuba, Ibaraki, 305-8686, Japan;6. Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo, 113-8657, Japan;1. Department of Chemistry, University of Louisiana, Lafayette, LA 70504, USA;2. Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19128, USA;3. School of Chemistry, University of Bristol, Bristol BS8 1TS, UK;1. College of Pharmacy, Keimyung University, 1095 Dalgubeoldaero, Dalseo-gu, Daegu 704-701, Republic of Korea;2. National Forensic Service, 10, Ipchun-ro, Wonju 220-170, Gangwon-do, Republic of Korea;3. School of Pharmacy, Sungkyunkwan University, 2066 Seobu-ro, Jangan-gu, Suwon 440-746, Gyeonggi-do, Republic of Korea |
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| Abstract: | We have selected an upland cotton (Gossypium hirsutum L.) cell line (R1098) that is highly tolerant to glyphosate. This cell line was developed by in vitro selection with gradually increasing glyphosate concentrations, and its mechanisms conferring glyphosate tolerance were studied. Based on a whole-plant dose–response bioassay, R1098 plants were tolerant to glyphosate at a concentration of 1500 g ae ha−1 glyphosate (1.5× the recommended field rate) whereas the control plants (Coker 312) were unable to survive at 150 g ae ha−1 glyphosate. Coker 312 accumulated 13.1 times more shikimate in leaves at 5 days after glyphosate treatment (1500 g ae ha−1) than that of R1098. Two distinct cDNAs for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), EPSPS-1 and EPSPS-2, were isolated from R1098. Both cDNAs were 97.7% identical within the common protein-coding region and the predicted sequences of the mature proteins were greater than 83% identical with EPSPS proteins from other known higher plants. In comparison to the glyphosate-susceptible cotton Coker 312, sequence analysis of the EPSPS-1 gene indicated that R1098 has an alanine insertion at nucleotide position 1216 resulting in frameshift. It leads to two copy functional EPSPS genes in R1098. There was no difference between R1098 and Coker 312 in EPSPS mRNA levels before glyphosate treatment. However, its treatment caused a 2–4 times increase in the basal EPSPS mRNA level in R1098. |
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