利用Real-time PCR对4个转Bt基因水稻的非预期效应研究  

The Exploration of Unintended Effects in Four Transgene Rice for Using Real-time PCR

作　　者：孙玥 王春雷 欧阳林娟[1] 周大虎 傅军如[1] 朱昌兰[1] 贺浩华[1] 彭小松[1] SUN Yue;WANG Chunlei;OUYANG Linjuan;ZHOU Dahu;FU Junru;ZHU Changlan;HE Haohua;PENG Xiaosong(Collaboration Center for Double-season Rice Modernization Production/Research Center of Super Rice Engineering and Technology,Jiangxi Province/Key Laboratory of Crop Physiology,Ecology and Genetic Breeding,Ministry of Education/College of Agronomy,Jiangxi Agricultural University,Nanchang,Jiangxi 33004)

机构地区：[1]江西农业大学农学院/作物生理生态与遗传育种教育部重点实验室/江西省超级稻工程技术研究中心/双季稻现代化生产协同中心,江西南昌330045

出　　处：《核农学报》2018年第11期2107-2118,共12页Journal of Nuclear Agricultural Sciences

基　　金：国家转基因生物新品种培育重大专项(2016ZX08001001)

摘　　要：转基因水稻中外源基因的转入可能会形成新的代谢产物,造成农艺性状的改变等,这些均不利于农业生产的非预期效应。为探究转基因水稻是否存在非预期效应,本研究以江西农业大学选育的4个转Bt基因恢复系为试验材料,通过反向PCR扩增侧翼序列并比对,用216对SSR引物对供试材料进行全基因组背景分析,再以Real-time PCR检测外源基因插入位点上下游100 kb内所有基因表达差异,并对光合色素含量及9个主要农艺性状进行显著性分析。结果表明,cry1C侧翼序列片段大小为1 276bp,定位在水稻第11号染色体上,cry2A侧翼序列片段大小为948 bp,定位在水稻第12号染色体上;昌恢121T、昌恢891T、昌恢T025T、R205选T的遗传背景回复率分别为95.60%、88.43%、93.52%、94.44%;基因表达分析显示试验组和对照组在cry1C、cry2A插入位置上下游100 kb内所有侧翼基因的表达水平均无显著性差异;进一步调查发现,试验组和对照组的光合色素含量及9个农艺性状无显著性差异。综上,cry1C和cry2A插入水稻基因组没有改变侧翼100 kb内基因表达水平,且4个转基因水稻并未产生非预期效应。本研究结果为探究转基因水稻的非预期效应评价提供了一定的理论参考。Insertion of exogenous gene in transgenic rice may form new metabolites and change agronomic traits,the unintended effects bad for agricultural production have attracted the attentions of researchers greatly. Four trans-Bt gene restorer line bred by Jiangxi Agricultural university bred were used as the experimental materials to explore whether exist unintended effects of transgene rice,with inverse PCR,flankingsequences were amplified and blast,216 SSR primer marker were used to analyze genetic background of the whole genomic,real-time PCR was used to detected the expression of all flanking genes located within 100 kb up-and down-stream of exogenous gene,and photosynthetic pigment content and nine primary agrinomic trait were compared. The results showed that the flanking sequence of cry1 C gene was 1276 bp and located on chromosome 11,flanking sequence of cry1 C gene has 948 bp and located on chromosome 12; Genetic background recovery rate of changhui 121 T、changhui 891 T、changhui T025 T、R205 xuan T were 95. 60% 、88. 43% 、93.52% 、94. 44%,respectively; The gene expression analysis indicated that there were no significant differences in the expression level of flanking genes located within 100 kb up-and down-stream of exogenous genes between test and control group; Further studies showed that photosynthetic pigment content and nine primary agrinomic trait have no significant difference between test and control group. The study indicated that cry1 C and cry2 A insertions in genome in transgenic rice don＇t change the expression level of flanking genes and no unintended effects were found in for transgenic rice. In addition,our study provides a clue for safety assessment of unintended effects in GM plants.

关 键 词：转基因水稻 非预期效应 REAL-TIME PCR 侧翼序列 遗传背景回复率 

分 类 号：S511[农业科学—作物学]