[Haibo Wang, Fushuang Dong, Mengyu Lv, Yanmin Zhang, Zhiheng Ren, Fan Yang, Xinchao Liang, Wenbo Zuo, Xueping Shi, Huanhuan Zhang, Yiping Gao, He Zhao, Xian Xu, Guozhong Sun, Jianfang Chai, Yongwei Liu, Jinyong Zhu, Qiufen Han, Qiang Zhang, Huijie Ma, Zhanwu Wang, Junfeng Guan] CN Shijiazhuang The present invention is a method of shoot apical meristem transformation for monocot plant via sufficient and micro wounding (SMW). The technical process includes: expose the apical meristem by removing the coleoptile away when the shoot grows to 0.2-2 cm after 1-2 days of seed germination; make sufficient and micro wounding transformation to the apical meristem by stabbing and brushing for 2-3 times using the SMW brush having 100-5000 bristles which is 4-20 μm in diameter for each one and 0.5-3 mm in exposed length, and dipped with the Agrobacterium tumefaciens containing binary vector harboring exogenous genes; develop the treated meristems directly to normal plants after co-cultivation; promote the plants to develop big spikes and set more seeds; harvest the seeds of T0 plants separately; detect and identify the transformation results in T1 generation which is bred from each individual T0 plant. The advantages of the invention are independent of tissue culture, unlimited in genotype, unnecessary to carry resistant marker, simple and large scale to perform, and applicable to all monocot plants which can set seeds. The transformation efficiencies for wheat, rice and maize using this method are 49%, 66.3%, and 100%, respectively.

[Haibo Wang, Mengyu Lv, Fushuang Dong, Yanmin Zhang, Zhiheng Ren, Fan Yang, Guozhong Sun, Xian Xu, Xueping Shi, Huanhuan Zhang, He Zhao, Jianfang Chai, Yiping Gao, Yongwei Liu, Jinyong Zhu, Qiufen Han, Qiang Zhang, Huijie Ma, Zhanwu Wang, Junfeng Guan] CN Shijiazhuang The present invention is a method of shoot apical meristem transformation for dicot plant via sufficient and micro wounding (SMW). The technical process includes: germinate the seeds in Petri dish or in the nutrient matrix which can be transplanted with the seedling; expose the shoot apical meristem by removing one cotyledon away; make sufficient and micro wounding transformation treatment in vivo to the apical meristem by stabbing and brushing for 2-3 times using the SMW brush having 100-5000 bristles which is 4-20 μm in diameter for each one and 0.5-3 mm in exposed length, and dipped with the Agrobacterium tumefaciens containing binary vector harboring exogenous genes; after co-cultivation, develop the treated objects directly to normal plants in the nutrient matrix and then transplant the matrix and seedling together into pot or field; promote the plant to develop more branches, pods, bolls, fruits, and seeds; harvest the seeds of each branch of individual T0 plants separately; detect and identify the transformation results in T1 generation from every T0 plant. The advantages of the invention are independent of tissue culture, unlimited in genotype, no need to graft, unnecessary to carry resistant marker, high survival rate in transplantation and seedlings recovered rapidly, easy to manipulate and large scale to perform, and applicable to all dicot plants which can set seeds. The transformation efficiencies for cotton and soybean using this method are 50% and 76.5%, respectively.

[WANG, Haibo, LV, Mengyu, DONG, Fushuang, ZHANG, Yanmin, REN, Zhiheng, YANG, Fan, SUN, Guozhong, XU, Xian, SHI, Xueping, ZHANG, Huanhuan, ZHAO, He, CHAI, Jianfang, GAO, Yiping, LIU, Yongwei, ZHU, Jinyong, HAN, Qiufen, ZHANG, Qiang, MA, Huijie, WANG, Zhanwu, GUAN, Junfeng] LV, Mengyu NO.598, Hepingxilu Shijiazhuang, Hebei 050051;NO.598, Hepingxilu Shijiazhuang, Hebei 050051;;;;;;;;;;;;;;;;;;; The present invention provides a dicotyledon transgenic method for invading growing points of seed sprouts or seedling stems minimally and fully, comprising: removing a cotyledon to expose growing points, and pricking and brushing the growing points by using a minimally-invasive brush of which the diameter of a single brush hair is 4 to 20 micrometers, the length of the brush hair is 0.5 to 3 millimeters and the number of the brush hairs is 100 to 5000 and which is dipped with agrobacterium mediated conversion liquid; and performing identification on T1 generation. The present invention is applicable to all seed bearing dicotyledons. A conversion effect that the conversion rate of cotton is 50%, and the conversion rate of soybeans is 76.5% during the conversion of the cotton and the soybeans is achieved.

[WANG, Haibo, DONG, Fushuang, LV, Mengyu, ZHANG, Yanmin, REN, Zhiheng, YANG, Fan, LIANG, Xinchao, ZUO, Wenbo, SHI, Xueping, ZHANG, Huanhuan, GAO, Yiping, ZHAO, He, XU, Xian, SUN, Guozhong, CHAI, Jianfang, LIU, Yongwei, ZHU, Jinyong, HAN, Qiufen, ZHANG, Qiang, MA, Huijie, WANG, Zhanwu, GUAN, Junfeng] No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051;No.598, Hepingxilu Shijiazhuang, Hebei 050051 The present invention provides a monocotyledon transgenic method for invading growing points of seed buds minimally and fully, comprising: germinating seeds, and when buds extend to be 0.2 to 2 centimeters long, removing coleoptile to expose growing points; pricking and brushing the growing points by using a minimally-invasive brush of which the diameter of a single brush hair is 4 to 20 micrometers, the exposed length is 0.5 to 3 millimeters and the number of the brush hairs is 100 to 5000 and which is dipped with agrobacterium mediated conversion liquid; and performing identification on T1 generation. This present invention is applicable to all seed bearing monocotyledons. A conversion effect that the genetic conversion rate of wheat is 49%, the genetic conversion rate of paddy is 66.3% and the genetic conversion rate of corn is 100% is achieved.