Transgenic Anti Rice Stem Borer Japonica Cultivation Through Agrobacterium Mediated

To improve the insect-resistant of rice in Heilongjiang Province, japonica rice(Oryza sativa L.)varieties “Songjing9”,“Longdao8” and “Longdao9” widely cultivated in the first and secondaccumulated temperture zones were utilized as starting materials for rice transformation viaAgrobacterium mediated co-cultivation method. A variety of insect-resistant plasmids (containingone anti-insect gene cry1Ab or two anti-insect genes cry1Ac/sck)were transformed into the currentrice cultivars to obtain the new japonica rice cultivars with multi-resistance and high quality.11rice cultivars were tested to investigate the regeneration capacity and culture conditions ofcallus induced from mature seeds. The results indicated:1)The time of scutellum enlargementshows no differences between the japonica and indica rice, while the speed of callus inducementfrom indica rice embryos is faster than the japonica ones.2) The callus rate is related to the qualityof seeds. The scutellum-derivedcallus rate of seeds with low germination rate is accordinglylow.3)Light could not improve callus rate remarkly.The suitable temperature of callus inducementis26℃.The frequency of subculture.as great effect on resistant callus production and plantlet regeneration，which are inducedfrom mature embryos, callus rate decreases while the times of subculture increases.4) Callusinduced on N6medium and MS medium are easy to brown，while callusinduced on NB mediumare hardly browning。The cry1Ac/sck genes had integrated into the genome of three japonica ricecultivars“Songjing9”,“Longdao8” and “Longdao9”via Agrobacterium mediated co-cultivationmethod. At present, we obtained9T1trangenic rice lines (about230individuals) of “Songjing9”,and T2transgenic rice lines (Hpt-,strip test+) are about360transgenic plants of5transgenic lines.T0transgenic rice plants of “Longdao8” are605, and strip test positive seedlings are157; T0transgenic rice plants of “Longdao9” are746, and strip test positive seedlings are45.The cry1Ab gene had integrated into the genome of three japonica rice cultivars“Songjing9”,“Longdao8” and “Longdao9” via Agrobacterium mediated co-cultivation method. At present, weobtained200T1transgenic rice plants (Hpt-,strip test+) of “Songjing9”; T2transgenic rice linesare about2070seedlings of7transgenic lines, and the screened seedlings (Hpt-,strip test+) are2320seedlings of7lines. T0transgenic rice plants of “Longdao8” are801, and strip test positiveseedlings are210; T0transgenic rice plants of “Longdao9” are1116, and strip test positiveseedlings are839.After four generation detection,3transgenic rice plants were selected under Hpt-BT+andsignle copy which are all from “Longdao9” modified Bt from Cry1Ab.