| As the fourth widest cultivated cereal crop in the world, barely has played a significant role in modem agriculture with a wide range of climate adaptability, short growth period, early maturity, and high yield. Barely, as a model species for genetic research, is widely used in studying the genetically modified crops. But the genotypes and explant source is limited, which hindered using barley gene engineering to cultivate new barley varieties. To establish an efficient regeneration system can lay a foundation for barley genetic transformation. Mature embyros was good trial material for genetic transformation with advantages of operating easily and free of time. But most of the callus induced from mature embryo have poor quality and low differentiation rate. It is very important to establish an efficient regeneration system for barely mature embryo by studying different media, genotypes, different ratio of hormone and metal ions, which can also make a preparation for barley genetic transformation.Salinity, one of the main abiotic stress factors, seriously affect the plant production, photosynthesis, protein synthesis and energy metabolism. Under salt stress, the main strategies for eliminating ion toxicity in the cytosol include sodium extrusion and sodium compartmentation, which is mainly completed by plasma membrane Na+/H+ antiporters and vacuolar Na+/H+ antiporters. Na+/H+ antiporters played an important role in ion balance in the cytosol, salt resistance and the overall course of plant growth and development. So cloning, expression and functional analysis of Na+/H+ antiporters gene is very significant for genetic engineering of plant salt tolerance.As an annual plant, Halogeton glomeratus is widely distributed in gansu, ningxia, qinghai, xinjiang and other regions, and most of them grow in the desert and saline-alkali land. It has a strong salt resistance. So it is good wild resources for studying salt-tolerant genes, and also provides a rich genetic resources to cultivate salt-tolerant crops.The main content of this research and the experimental results are as follows:(I) To optimize the tissue culture conditions of the barley mature embryo1. Five barley varieties ganpi 3, ganpi4, ganpi 6, Belana and marine were used to study the influence on barley mature embryo tissue culture of the different varieties in the different induction media. The results showed that NMB was the more suitable induced medium for barley mature embryo regeneration, but showed some effects of genotype. That is to say, it is not identical to produce barely callus in the different media. In comparison, the callus induction and differentiation of ganpi 3 and ganpi 4 were better in five varieties, which were good genotype for researching genetic transformation of barely.2. The results showed that adding 2.0~5.0mg/L 2, 4-D to media can improve the callus induction. And 4.0 mg/L 2, 4-D is the optimum concentration to generate callus.3. In the process of culturing mature embryos of Ganpi 4, the results indicated that the callus induction was improved by adding Ag NO3 and ABA, which the most suitable concentration inducting callus is respectively 7.5mg/L and 5.0mg/L. And 2.5mg/L Ag NO3 can improve the differentiation rate of ganpi 4.(â…¡) To construct the plant expression vector p BI-Hg NHX1This test successfully cloned Hg NHX1 gene from Halogeton glomeratus and constructed plant expression vector p Bin-Hg NHX1. It was transferred into LBA4404 and used to mediate tobacco leaf. We gained the resistant plants. The results of PCR detection can help preliminary judging that Hg NHX1 has mediated into tobacco, which laid a foundation for further function verification test and also made a preparation for the application of the Hg NHX1 gene in barley genetic breeding. |
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