Transfoemation Of Two Spring Wheat Cultivar With KN2and PYL5Genes

Wheat is one of the major food crops in the world. Most of the wheat growing areas inthe world located in the arid and semi-arid areas, wheat growing areas distributed throughoutall provinces and cities in china. The Huang-Huai winter wheat region, a major wheatproducing areas in china, often faces drought due to lack of rainfall. Hypothermia has alsobecome a limiting factor in the production of wheat, the cultivation of wheat with goodresistance is essential for wheat production. The traditional breeding techniques combinedwith modern agricultural make a great contribution to the development of world agriculture.However, the lack of gene pool and the barriers in multiplication limiting the development ofagriculture, The technology of genetically modified wheat which can break through thebarriers between species become the focus of attentions.In this study, two spring wheat cultivars Mianyang19and Ningchun16were used asmaterials. we transform the KN2or PYL5with Bar gene into wheat immature callus bybiolistic particle. The purpose of this study is to obtain transgenic wheat with KN2or PYL5gene, which lay a foundation for studying the function of these two genes. The main researchcontent and results were as follows:(1) The transformation of two spring wheat with KN2and Bar.The target gene KN2and selection marker gene Bar were used to co-transformationMianyang19(800Callus) and Ningchun16(800Callus) by biolistic particle respectively.Four regenerated plants for Mianyang19and4for Ningchun16were obtained. Analysis ofallele-specific PCR of these T0plants indicated that3regenerated wheat plants were Bar-and KN2-positive plants with in Mianyang19,2regenerated wheat plants were Bar-andKN2-positive plants with in Ningchun16. The seeds of the T0transgenic positive target plantswere planted in greenhouse to test transgenic T1plants, but no positive plants weredetected.(2) The transformation of two spring wheat with PYL5and BarThe target gene PYL5and selection marker gene Bar were used to co-transformationMianyang19(800Callus) and Ningchun16(800Callus) by biolistic particle respectively.Five regenerated plants for Mianyang19and14for Ningchun16were obtained. Analysis of allele-specific PCR of these T0plants indicated that1regenerated wheat plants were Bar-andPYL5-positive plants with in Mianyang19,4regenerated wheat plants were Bar-and PYL5-positive plants with in Ningchun16. The seeds of the T0transgenic positive target plants wereplanted in greenhouse to test transgenic T1plants,3plants were positive plants.