Extrachromosomal DNA‐mediated glyphosate resistance in Italian ryegrass
Extrachromosomal DNA‐mediated glyphosate resistance in Italian ryegrass
复制标题
意大利黑麦草染色体外 DNA 介导的草甘膦抗性
DOI：
10.1002/ps.7626
复制
发表时间：
2023
影响因子：
4.1
通讯作者：
Gill, Bikram S.
中科院分区：
文献类型：
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作者：
Koo, Dal‐Hoe;Ju, Yoonha;Putta, Karthik;Sathishraj, Rajendran;Burgos, Nilda;Jugulam, Mithila;Friebe, Bernd;Gill, Bikram S.
背景来自美国阿肯色州的一个多花黑麦草群体由于5-烯醇式莽草酸-3-磷酸合酶（EPSPS）基因扩增而产生了草甘膦抗性。本研究以该群体中约70个EPSPS拷贝的多花黑麦草为材料，对EPSPS基因扩增拷贝进行了物理定位，以确定多花黑麦草EPSPS基因扩增赋予草甘膦抗性的可能机制。具有一个EPSPS拷贝的多花黑麦草（GS）植株显示单一的非特异性EPSPS杂交信号，该信号与5S rDNA位点沿着共定位，在染色体外侧几乎没有额外的信号。脉冲场凝胶电泳（PFGE），然后DNA印迹usingEPSPS基因作为探针确定了一个abundentEPSPS杂交周围的400 kb区域的GR DNA样品，但不是在GS DNA samples.ConclusionWe报告extrachromosomal DNA介导的草甘膦抗性在多花黑麦草。利用FISH技术对多花黑麦草EPSPS基因的扩增拷贝进行了物理定位，结果表明EPSPS基因的扩增拷贝可能存在于染色体外DNA元件中。进一步的PFGE和DNA印迹分析表明，长穗苋含有EPSPs的染色体外DNA大小约为400 kb，与eccDNA复制子的大小相似。© 2023化学工业学会。
BackgroundAn Italian ryegrass population from Arkansas, USA developed glyphosate resistance due to5‐enolpyruvylshikimate‐3‐phosphate synthase(EPSPS) gene amplification. The plants in this population with approximately 70EPSPScopies were used in the present study for the physical mapping of amplified copies ofEPSPSgene to determine the possible mechanism ofEPSPSgene amplification conferring glyphosate resistance in Italian ryegrass.ResultFluorescencein situhybridization (FISH) analysis of glyphosate resistant (GR) Italian ryegrass plants with approximately 70EPSPScopies displayedEPSPShybridization signals randomly on most of the metaphase chromosomes. Glyphosate susceptible (GS) Italian ryegrass plants with oneEPSPScopy displayed single prominentEPSPShybridization signal, which was co‐localized with 5S rDNA locus along with few additional signals on the outside of chromosomes. Pulsed‐field gel electrophoresis (PFGE) followed by DNA blot usingEPSPSgene as a probe identified a prominentEPSPShybridization around the 400 kb region in GR DNA samples, but not in GS DNA samples.ConclusionWe report the extrachromosomal DNA‐mediated glyphosate resistance in Italian ryegrass. Physical mapping of amplified copies ofEPSPSgene in Italian ryegrass by FISH gives us a clue that the amplified copies ofEPSPSgene may be present in the extrachromosomal DNA elements. Further analysis by PFGE followed by DNA blotting revealed that the extrachromosomal DNA containingEPSPSis approximately 400 kb similar in size with that of eccDNA replicon inAmaranthus palmeri. © 2023 Society of Chemical Industry.