Genetic Transformation Of PtrRAP2.11 Gene In Populus Trichocarpa

Nitrogen(N)is one of the important mineral elements necessary for plant growth,which affects the growth and development of plants.Nowadays,artificial fertilization is mainly used to improve soil nitrogen deficiency,but it will also produce environmental pollution.In order to reduce environmental pollution and improve plant growth under low nitrogen conditions through genetic engineering technology,it is of great significance for agriculture and forestry production.In this study,the PtrRAP2.11 gene was used as the research object to study its expression characteristics and to carry out genetic transformation of tobacco and 84K poplar.This study will provide reference for genetic engineering breeding in agriculture and forestry.We cloned the PtrRAP2.11 gene from the whole genome of Populus poplar leaves and performed a preliminary analysis using bioinformatics methods.The results showed that the coding length of the gene was 812bpm encoding a protein composed of 270 amino acid residues.the relative molecular mass is 30310.48;the isoelectric point PI=5.60 is acid protein;the coefficient of instability is 61.88;the hydrophobicity is 63.26;the total average hydrophilicity is-0.696.Phylogenetic analysis showed that the amino acid sequence of the gene was similar to RAP2.11 of Populus euphratica and Jatropha curcas,and they had ERF/AP2 transcription factor domains.In order to investigate the response of the PirRAP2.11 gene in the low nitrogen environment(Nitrate nitrogen,Ammonium nitrogen),we construct the plant expression vector of the PtrRAP2.11 gene of poplar poplar and carry out the genetic transformation of tobacco and 84K poplar.The following results were obtained:(1)Transgenic tobacco and wild-type seeds were germinated in a low-nitrogen environment,and the grown seedlings were subjected to Real-time PCR detection,The results showed that the transgenic tobacco had more obvious growth advantage than the wild type tobacco under the low nitrogen environment.It also proved that the PtrRAP2.11 gene responded to the low nitrogen environment and improved the resistance of transgenic tobacco.(2)The genetic transformation of 84K poplar was carried out,and the chlorophyll content and related physiological and biochemical analysis of the transgenic poplar in a low nitrogen environment were performed.The results showed that the gene responded to the low nitrogen environment in the transgenic 84K poplar under low nitrogen conditions.In particular,the nitrogen content and chlorophyll content of transgenic overexpressing lines under low nitrogen treatment were higher than those of wild type.(3)The nitrogen content of leaves treated with low nitrogen in transgenic 84K poplar and wild type 84K poplar was determined.The results showed that nitrate nitrogen,ammonium nitrogen and total nitrogen content were higher in the transgenic lines than in the wild type.It was initially shown that the PtrRAP2.11 gene can increase the nitrogen utilization level of 84K in the low nitrogen environment.Different nitrogen levels had a significant effect on the morphology structure,and physiology of the transgenic lines.Under low nitrogen stress,the transgenic plants showed more obvious growth advantage than the wild type.It shows that the transgenic plants have strong regulation and adaptability to these adverse environments.