Cloning And Functional Analysis Of Cold Response Gene ZmRanBP1 In Maize

The chilling injury causes the corn-related physiological function to be damaged and the metabolism to be inhibited,so that the maize yield is seriously affected.Therefore,it is very important to improve the adaptability of cold stress of maize.In view of the above problems,this study carried out cloning,functional verification and genetic transformation of Zm RanBP1.The results are as follows:1.The ZmRanBP1 coding sequence of 1467bp was successfully cloned from W9816,and contains 488 amino acids.DNAMAN was used to compare the amino acid sequences of ZmRanBP1 of various plants,and found that the similarity between corn and sorghum was 83%.2.Constructed the expression vector pCAMBIA-3301-35s-ZmRanBP1,and obtained three transgenic arabidopsis plants.The wild-type,homozygous mutant and T₃ generation transgenic plants were tested for cold-resistant and salt-resistant phenotype identification.The result showed that over expression of ZmRanBP1 in Arabidopsis has increased the sensitivity to abiotic stress.3.The genetic transformation of maize was carried out using agrobacterium-mediated method and pollen tube passage method.There were a total of 710regenerated plants of infecting immature embryo,22 were positive for PCR,and the positive rate was 3.09%.There were a total of 846 regenerated plants of infecting callus,35 were positive for PCR,and the positive rate was 4.13%.A total of 2346 T₀seeds was obtained by using the pollen tube channel method,and 5 were positive for PCR.The T₀ generation plants were self-pollinated,and there were 254 T₁ seeds.