The Study Of Transformation AtCKX3 Into Maize Inbred Line With Linear DNA Mediated By Cell Penetrating Peptide Through Pollen Tube

Drought is an important issue restricting agricultural production and social development. Temporary or permanent drought can cause more damage than any other adversity factors to reduce maize yield. Corn is the main source food of human,meanwhile,it is an important resource of industry and feed. But maize is very sensitive to drought stress. Maize are affected by drought stress in normal growth and development, which will reduce the yield and quality of maize.In our country, many places are facing the impact of drought and water shortage,because the irregular terrain.So,it is more and more important to improve maize varieties and drought-tolerant of maize by the technology of transgenic. Root is an important organ of plants to absorb water and nutrients.and corn root is closely related to its drought tolerance. Its growth has a significant impact on plant stems and leaves and other parts of the ground and the final yield.So we choose the CKX gene So we chose the CKX gene that is closely related to the maize root growth, in order to obtain drought-tolerant maize plant. CKX3 is a key enzyme to metabolize cytokinin, it can adjust the content of cytokinins in plants to regulate the growth and development of tissues and organs. which can promote the growth of plant roots.The CKX3 gene was transformed into 178 by the pollen tube pathway in order to improve the drought resistance of maize, and 122 transgenic lines were obtained by screening with herbicide. Then the T2 generation seeds were planted in drought shed to test the drought resistance, and the the indicators of roots and seedlings were measured.35Sbar gene and CKX3 gene has been proved to be successfully introduced into the maize genome by PCR detection, and the bar and CKX3 were proved to be expressed in the transgenic plants by RT-PCR and Bar-pat strip detection. The plant height, leaf area, grain weight, single panicle weight and other solid case of transgenic plants and 178 inbred line were monitored. the results showed that transgenic plants were significantly better than the 178 inbred plants under drought conditions. Measured the relevant indicators about root length, and number of lateral roots and so on of eight transgenic plants. the results showed that transgenic plants were significantly better than the 178 inbred plants of root indicators and have reached a significant difference or very significant difference. The results showed that the expression of CKX3 in the roots of transgenic plants,can significantly improve drought resistance compared to 178 inbred plants.PGM-EPSP-AtCKX3 expression vector were constructed,and it is proved to be correct by digestion and sequencing. Obtain the desired linear gene by PCR method. Then recovered and purified and mix to penetrating peptide according to mass ratio of 1:1. This genes was transformed into 572and HZ 178 inbred maize plant by the pollen tube pathway. the To maize plants seedlings were sprayed with Glyphosate,and get 12 transgenic plants. The EPSP gene、CKX3 gene has been proved to be integrated into maize genome by PCR、ddPCR detection in theTo transgenic plants.and we know the EPSP gene is a single copy in 572-3, HZ178-1 plants,and have stable inheritance.