in vitro mutagenesis protocol for the production of sugarcane tolerant to the herbicide imazapyr

• Published in: In vitro cellular & developmental biology. Plant Vol. 48; no. 4; pp. 417 - 427
• Main Authors: Koch, Aimée C, Ramgareeb, Sumita, Rutherford, R. Stuart, Snyman, Sandra J, Watt, M. Paula
• Format: Journal Article
• Language: English
• Published: New York Springer-Verlag 01.08.2012; Springer Science + Business Media; Springer Nature B.V
• Subjects: aboveground biomass; Acclimatization; acetolactate synthase; Biomedical and Life Sciences; Biosynthesis; Callus; Cell Biology; Crops; Cultivars; Deoxyribonucleic acid; Developmental Biology; DNA; Enzymes; ethyl methanesulfonate; Genetic mutation; Genotypes; Herbicide resistance; Herbicides; imazapyr; Life Sciences; Mutagenesis; Mutagens; Mutation; Plant Breeding/Biotechnology; Plant Genetics and Genomics; Plant Sciences; Plantlets; Plants; somaclonal variation; SOMATIC CELL GENETICS; Sugar cane; Sugarcane; Chemical mutagenesis; Acetohydroxyacid synthase; spp; Herbicide tolerance; Somatic embryogenesis
• ISSN: 1054-5476; 1475-2689
• DOI: 10.1007/s11627-012-9448-x

A protocol was developed to induce and identify imazapyr tolerance in sugarcane, which involved induction of somaclonal variation via exposure to 8 or 16 mM ethyl methanesulfonate for 4 h, followed by a stepwise increase in imazapyr concentration in the medium from 0.08 to 0.16 μM. The regenerated plantlets were then acclimatized for 3 mo after which they were sprayed with 182 g a.i. ha−1 imazapyr, and the above-ground biomass was determined after 47 d. Following a 1-mo waiting period for the putative tolerant plants to regrow, acetohydroxyacid synthase (AHAS; EC 2.2.1.6) enzyme assays of the plants that survived and showed a normal growth pattern were undertaken. Based on the enzymatic I 50 values, three imazapyr-tolerant genotypes were identified with an AHAS activity of 2.8 to 4.0 times that in sensitive sugarcane plants.