Genetic Transformation of Perennial Ryegrass for Chemical Induced Elimination

transformation;perennial ryegrass

Chen, Xi ; Ron Qu, Committee Chair,Chen, Xi ; Ron Qu ; Committee Chair

University:North Carolina State University

关键词: transformation;    perennial ryegrass;   

Others  :  https://repository.lib.ncsu.edu/bitstream/handle/1840.16/2255/etd.pdf?sequence=1&isAllowed=y

美国|英语

来源: null

【 摘 要 】

Perennial ryegrass (Lolium perenne L.) is an important forage and turfgrass species, widely distributed throughout the world, including North and South America, Europe, New Zealand, and Australia. As a turfgrass, it is widely used in winter overseeding of warm-season turfgrasses. Summer persistence is a major problem for overseeding use of the species.To solve this problem through biotechnology approach, we introduced an E.coli argE gene into perennial ryegrass. ArgE gene encodes N-acetylornithinase (NAO) which is involved in the arginine biosynthesis pathway. It has been shown that NAO can deacetylate N-acetyl-phosphinothricin (N-acetyl-PPT), a chemical which is non-toxic to plants, and produce phosphinothricin (PPT), the active ingredient of herbicide Basta and Finale.The objectives for this project are (1) to develop an efficient transformation system for perennial ryegrass, (2) to introduce argE gene into perennial ryegrass and (3) to determine if transformed plants can be killed using N-acetyl-PPT.Three approaches were used to develop target materials for bombardment transformation: suspension lines from liquid culture, selected embryogenic calli and embryogenic calluse lines from solid culture medium.A total of 200 plates of selected calli induced from mature seeds were bombarded and 51 hygromycin B resistant calli were recovered, among which a total of 47 green plants were regenerated from 22 independently transformed resistant calli. Although a few resistant calli were obtained, no transgenic plants were produced from callus lines and suspension cell lines. Presence of the transgenes into plant genomes was demonstrated by PCR as well as Southern hybridization analysis. When N-acetyl-PPT was painted on transgenic leaves, leaf damage ranged from yellow to completely dead and the whole transgenic plants were killed in N-acetyl-PPT spray experiment. It suggested that the argE gene function as expected in transgenic perennial ryegrass plants.