To Cultivate Transgenic Lea Gene Upland Rice

In recent years, the global shortage of water resources has produced a big influence, which will restrict the development of agriculture, industry, and human life, so developing dryland farming is the key to solve the water shortage problem. Upland rice, a kind of rice, accounts for a very significant position in food crops, and upland rice has a lot of advantages compared with rice because of less water requirement and strong drought resistance. Upland rice is one of the best food crops of the implementation of dry land farming. Upland rice solves the problem of soil drought when rice planting, which will reduce the cost of agricultural production greatly. Thus it can replace rice to be developed for large scale cultivation.This project chooses “kunhan 1” as the target plant to be modified, transfers the bar gene and lea gene into the upland rice cells by Agrobacterium-mediated method and Particle bombardment method. The experiment gives light on the impact of various factors towards the two different transformation methods. T2 generation transgenic plants are identified for plant physiology, efficient upland rice transformation system and iden--tification system are established. The homozygote of transgenic plants which can resistant salt and drought have been gotten finally.The experimental results show that the most suitable medium for callus induction is NB+2,4-D(without proline), sugar source is maltose, and the results also show that when the concentration of the Agrobacterium is 0.2 of the OD600, the conversion effect is the best. Adding the concentration 40 mg/L herbicide in screening medium can screen out the herbicide-resistant callus. Selecting the milk white resistant callus carry on 5 days pre-differentiation culture, which can improve the differentiation efficiency. Using the optimized transformation system, the conversion rate can be 37.15% by molecular detection of T0 generation transgenic plants. Using 0.8 mg/L herbicide solution spray T2 generation transgenic plants, if all plants in the same line show herbicide resistance, the experiment will detecte the resistant plants by PCR to select homozygote. Identification the homozygous plants’ salt and drought tolerance, under the moderate drought environ--ment, transgenic plants grow better than wild type, when the salt concentration is 0.75%, transgenic plants grow better than wild type obviously.