Study On Glucose Oxidase Gene Was Introduced Into Indica Restorer Line

The study on the glucose oxidase gene was introduced into O.sativa ssp.Indica shuhui162 from transgenic Oryza sativa L.ssp.japonica.Nipponbare by the way of sexual hybridization.The RT-PCR molecular identification and the color reaction with starch—KI showed the GO gene introduced into rice Shuhui 162,and Utilizing transgenic Shuhui 162F₁ sexual hybridization with Lehui 188 and Jinhui21,respectively. Physiological and biochemical methods were used to detect the reestablishment of catalase(CAT),Peroxidase(POD),superoxide dismutase(SOD) activity and isozyme analysis with transfered glucose oxidase gene Rice Shuhui 162F₁ material leaves and multiple cross F₁(GO gene Shuhui 162F₁ and Lehui 188) material leaves and multiple cross F₁(GO gene Shuhui 162 F₁ and Jin Hui 21 ) material leaves.The Resistance identification to transfered glucose oxidase gene Shuhui 162 F₁ and their multiple cross F₁ after Magnaporthe grisea inducing.The results are as follows:(1) The RT-PCR test result showed that the purpose fragment was existent,the hydrogen peroxide content in plants was increased.It proved the GO gene can be transfered between lndica and japonica.The GO gene was introduced into Shuhui 162 and expressed successful.(2) Whether it was induced by Magnaporthe grisea or not,the POD,CAT,SOD activity of the transfer of glucose oxidase gene Rice Shuhui 162F₁ material leaves and their multiple cross F₁ material leaves were higher than comparison;compared to susceptible plant material,the POD,CAT,SOD activity of not susceptible plant material leaves were higher;isozyme analysis of the transgenic plants with a deeper color line.It showed that the glucose oxidase gene had expressed in the level of protein and exerted its function in shuhui162 rice and GO gene was transferred into the multiple cross F₁ with resistance.(3) Identification of resistance to leaf rice blast:transgenic rice Shuhui 162F₁ resistance was MR,but comparison was HS.The resistance R accounts for 80%in the Nipponbare(GO)/Shuhui162//Lehui188 F₁ plants,the comparison accounts for 79%with HS.The resistance R accounts for 78%in the Nipponbare(GO)/Shuhui162//Jinhui21 F₁ plants,the comparison accounts for 25%with MS,31%with S and 22%with HS.By the way of DPS analysis,besides one group,others groups are distinct difference to the comparison groups.The leaf rice blast index of the Nipponbare(GO)/Shuhui162 //Jinhui21 F₁ was higher than Nipponbare(GO)/Shuhui162//Lehui188 F1.It showed that the transgenic rice resistance is obvious,and the GO gene was transfered into the Lehui188 and Jinhui21,but it is different in both the ratio of success and the resistance due to the different varieties.(4) Identification of resistance to neck rice blast:The GO gene plants were stronger to resistant than non-GO gene plants in future generations plants.The non-GO gene plants got more rice blast than GO gene plants.The GO gene in different genetic background has the different strength of express in future generations plants.(5) This research is the successful application of integrating genetic engineering with conventional breeding.