Production of Herbicide-Resistant Crops by Cell-Engineering

Research Project

Project/Area Number	05660057
Research Category	Grant-in-Aid for General Scientific Research (C)
Allocation Type	Single-year Grants
Research Field	植物保護
Research Institution	Kansai University
Principal Investigator	MATSUNAKA Shooichi Kansai University, Dept.of Biotecnology, Professor, 工学部, 教授 (50116258)
Project Period (FY)	1993 – 1995
Project Status	Completed (Fiscal Year 1995)
Budget Amount *help	¥2,100,000 (Direct Cost: ¥2,100,000) Fiscal Year 1995: ¥600,000 (Direct Cost: ¥600,000) Fiscal Year 1994: ¥600,000 (Direct Cost: ¥600,000) Fiscal Year 1993: ¥900,000 (Direct Cost: ¥900,000)
Keywords	Herbicide-Resistant Crops / Glyphosate / Paraquat / Bent Putter / Sunflower / Horseweed / Cell Culture / Cell Fusion / 除草剤抵抗性作物作出 / 細胞工学的選抜法 / ベントパタ- / 除草剤グリホセ-ト / 除草剤パラコート / ベントグラス / 除草剤グリホセート
Research Abstract	Long time and much money are required for the development of a new selective herbicide for a crop plant. If we can create a crop plant having resistance to a herbicide which is low-toxic to animals, safe to the environment, cheap and has a long practical use history, it is evaluated as the same matter as the development of above-described new selective herbicides for such crop plant. In this study, severaltrials were examined, and as the examples having some possibility for the research purpose, the creation of glyphosate-resistant bent putter and paraquat-resistant sunflower. In the selection of glyphsate-resistant bent butter, callus having resistance to 1,000 uM glyphosate were obtained by the gradual increasing of the concentration of co-existing herbicide from 100 until 1,000 uM.This tolerant callus was transplanted to hormone-free medium and cultured in the light. After 20 days, the generated plants were soaked in 1,000 uM glyphosate solution for 8 hours. After more 20 days the plant regenerated from resistant callus were survival while ones generated from susceptible callus were dead. The sikimic acid content of the resistant one was one sixth of original susceptible one. On the other hand, both protoplasts prepared from leaves of sunflower and paraquat-resistant horseweed by enzyme treatment were fused by a electrofusion system (SHIMAZU SOMATIC HYBRIDIZER,SSH-10). The concentration of both protoplasts was 100 thousands cells per ml. The fusates were cultured on MS medium (NAA 0.2 ppm, BA 0.5 ppm), at 25ﾟC under 3,000 lux, for 2 months and moved to paraquat-containing MS medium. Unfused sunflower colonies could not form any callus upper 1 x 10^<-5>M paraquat, under which condition the fused cells could form callus and regenerated plants.