The Research Of The Genetically Modified Rice Safety Assessment Based On Transcriptome

With the development of transgenic technology in recent years, crop insect resistance, disease resistance and other issues have been effectively controlled. However, when the exogenous genes transcript into Genetically Modified Organisms(GMO) and gain the target traits, other unexpected effect may occur too. So that the safety of GM products has attracted many attention of the society from all walks of life. So far, the study of the unexpected effect of the GMO still stays on the phenotypic observation, rarely involves the study of molecular level. Therefore, it is necessary to study the unexpected effect of the GMO on the molecular level.This task launched a series of work to assessment the safety of exogenous genes Cry1 Ac and EPSPS on the transcriptome level. We choose the Non-GM rice xiushui 11, the GM rice P1417, P1418 lines with the exogenous EPSPS gene based on xiushui 11, the GM rice P1411, P1415 lines with the exogenous Cry1 Ac and EPSPS genes based on xiushui 11 as the experimental materials, investigate their agronomic traits during their growth process and analysis their phenotypic differences. At the same time, using TAIL-PCR technology and real-time fluorescent quantitative PCR to determine the insertion sites and copy numbers of the foreign genes. And extract the RNA of rice samples to transcriptom sequencing and analysis the results.DNA molecular verification, protein test strips and experiment of spraying glyphosate in vitro made sure the P1411, P1415, P1417 and P1418 lines all have the foreign genes inserted. The lines P1417 and P1418 have EPSPS gene, lines P1411 and P1415 have Cry1 Ac and EPSPS genes. By the TAIL-PCR technology we acquired the flanking sequences of the 4 GM rice lines. Then we confirmed the exogenous gene insertion sites of the 4 GM rice lines were chromosome 3/OSJNBb0016P23, chromosome 3/OSJNBa0004G17, chromosome 12/OJ1388_B05/Os12g0616500, and chromosome 11/OSJNBa0058G18 by the NCBI website. And by the real-time PCR we estimated the inserted copy number of exogenous gene was only one.Transcriptome sequencing results showed the number of common different expression genes of line P1411 and P1415 is 14. They don’t have common significant GO and KEGG enrichment pathways. The number of common different expression genes of line P1417 and P1418 is 20 and they don’t have any significant GO and KEGG enrichment pathways either. The common different-expression genes of the 4 GM rice lines are EPSPS, EPlOSAG00000010309 and OS07G0504200. EPlOSAG00000010309 is non-coding RNA and don’t have GO and KEGG enrichment pathways. OS07G0504200 is coding protein and has one KEGG enrichment pathway. EPSPS is exogenous gene and has 43 GO enrichment pathways and 4 KEGG enrichment pathways. The enrichment pathways of the common different-expression genes are all not significant. Neighborhood genes analysis showed there are 25 genes expressed differently and the difference are small. Relative quantification of two exogenous genes and two different-expression genes showed the genes’ expressions are stable.The statistics of the agronomic traits showed the heading date of the insect-glyphosate-resistant rice P1411 and P1415 were later than the Non-GM rice and the glyphosate-tolerant rice, this result may caused by the specific events of the P1411 and P1415 transformation. However, the plant height, hundred-grain weight and tiller number of the common transformation did not show the common differences.Based on the analysis of the transcriptome results and agronomic traits, we find that unexpected effects of the 4 GM rice lines are caused by foreign genes, but the foreign genes do not cause adverse unexpected effects and the same transformation do not show the same differences. And the numbers of the different expressed genes was tiny, only occupied 0.01%. Therefore, based on the transcriptome, the GM rice with Cry1 Ac and EPSPS genes are not significant differenent with Non-GM rice. All content and the results of this experiment can provide important basis for research and safety assessment of the GM varieties.