| Poplars has been widely cultivated due to its rapid growth and adaptability.In order to effectively solve the problem of poplar pests and diseases,transgenic technology is increasingly applied to the research of poplar pests.With the increase of transgenic plants,the requirements for their detection technology are getting higher and higher,and the biosafety issues of transgenic plants were getting more and more attention.In this dissertation,we used the LAMP technology to establish a set of methods for the simple,rapid and accurate detection of foreign genes in poplars with Bt Cry1 Ac gene 741 poplar.High-throughput sequencing was performed on dry,root and rhizosphere soil DNA samples of transgenic Bt Cry1 Ac gene 107 poplar euphratica,and the effect of exogenous genes on the diversity of microbial communities within poplars was explored by comparison with non-transgenic 107 poplars.The high-throughput sequencing of dry,rhizosphere,and rhizosphere soil DNA samples from three different transgenic poplars of the Bt Cry1 Ac gene 107 poplar plantation was performed to investigate the genetically modified receptors of transgenic poplar 107 and its surrounding planting patterns.Effect of microbial community diversity in non-transgenic 107 poplars.The main findings were as follows:(1)Using the LAMP-PCR optimized system established for transgenic poplars,LAMP-PCR and conventional PCR were performed on DNA at different concentrations,ie,2-fold and 10-fold dilution gradients.The results of electrophoresis showed that LAMP-PCR obtained a clearer band than conventional PCR.In addition,the occurrence of a positive reaction can be directly observed by using the precipitation of magnesium pyrophosphate as a by-product of LAMP-PCR,or the LAMP-PCR product can be stained with a Gold View nucleic acid stain,and judged directly based on the color of the product.Therefore,it was demonstrated that LAMP-PCR has higher sensitivity and convenience.(2)Using the LAMP-PCR optimized system established for transgenic poplars,LAMP-PCR technology was used to detect the Bt Cry1 Ac and Npt II genes in five clones of PB1,PB3,PB11,PB17,and PB29 of transgenic 741 poplar.The results of electrophoresis showed that all of the clones could stably amplify a clear diffuse band.Therefore,it was demonstrated that LAMP-PCR has higher accuracy and reliability.(3)High-throughput sequencing was performed on DNA samples from the Pb1 high resistant strain of transgenic 107 poplar with Bt Cry1 Ac gene and the non-transgenic 107poplar(endophyte,root and rhizosphere),and the sequencing results of microbial community composition were exported.Cluster analysis,Alpha diversity analysis,Beta diversity analysis,LEf Se analysis,etc.showed that there was no significant difference between the two lines of transgenic and non-transgenic.(4)High-throughput sequencing was performed on DNA samples from endophyte,root and rhizosphere of non-transgenic 107 poplar in three different cultivation renewal modes(transplantation,initiation,and grafting)in Bt Cry1 Ac gene 107 poplar test plots.The sequencing results of microbial community composition were analyzed by OTU clustering analysis,Alpha diversity analysis,Beta diversity analysis,and LEf Se analysis.The results showed that there were no significant differences among the three different cultivation renewal modes.(5)High-throughput sequencing of microbial communities in endophyte,root,and rhizosphere of poplar in longitudinal synthesis(3)and(4)yields the composition of microbial communities in the root and rhizosphere DNA samples.The degree of similarity was extremely high,and both were significantly different from the composition of the microbial community in the endophyte DNA sample.Relatively speaking,the diversity of microorganisms in the root and rhizosphere was higher than the diversity of microorganisms in the endophyte. |
