| Brassica napus L, which is one of the oldest oil crops in the world and has the largest planting area in China, has a long history in China and India. At present, rape has been one of the four major oil crops (soybean, sunflower, rapeseed, peanuts) in the world. Since1980s, as important sources of vegetable oil and feed, the rape has a soaring period of development. Water shortages become severely and arouse serious problem in the world, especially in China. Therefore, breeding new species of drought-resistant rape in China is an extremely urgent task in subsequent investigation.With the rapid development of transgenic technology, it provides the most effective way for biological directed reconstruction and molecular breeding. The transformation including Agrobacterium-mediated method, electric shock, PEG-mediated DNA uptake, microinjection, pollen-tube pathway, gene gun and ion beam mediated method and so on. But Agrobacterium-mediated methods are the preferred choice over others because of its the simple operation and low cost, Agrobacterium tumefaciens is also the most widely used genetic transformation method.In this study, we made the zhongyou821as test material and several main factors affecting transformation efficiency which included Kan antibiotic concentration, bacterium liquid concentration, infection time, cultivation time and administrations of acetyl clove ketone (AS) as well as silver nitrate were investigated. We created an efficient Agrobacterium-mediated genetic transformation system. On the basis of the plant expression vector of PBI121-SHN1/WIN1constructed by Chai Lingyan in2009, with the purposes of prevention of excessive water lost and maintaining water in rape, we inverted the SHN1/WIN1which regulates the thickness of cutinase into Rape cell with Agrobacterium-mediated Transformation so as to enhance the drought, cold and disease resistance abilities of rape. Eventually, we obtained drought resistance transgenic rape zhongyou821. |
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