中文摘要：利用非胚乳组织中表达的启动子在特定的植物组织或器官中激活目标基因是转基因作物工程中的一个有效方法。本文从水稻基因组中克隆了五个假定的非胚乳组织表达启动子，并命名为P-OsNETE1、P- OsNETE2、P-OsNETE3、P-OsNETE4和P-OsNETE5。在转基因水稻中，通过对GuSPLUS报告基因表达的定性和定量检测，发现在不同发育阶段植物的胚根、叶、茎、鞘和穗中都含有P-OsNETE1-P-OsNETE5基因，而在胚乳中则不存在。此外，P- OsNETE2、P-OsNETE4和P-OsNETE5在水稻胚胎中也不活跃。在这些启动子中，P-OsNETE4和P-OsNETE5在所有被测组织中均表现出较高的活性，其在茎、叶、根和鞘中的活性均高于或与水稻AcTiN1启动子的活性相媲美。我们还对两代单拷贝系中POSNETE1-POSNETE5的活性进行了监测，发现这些启动子在世代间稳定表达。利用P-OsNETE4和P-OsNETE5可驱动改良的Bt基因McIL1Ab以制备转基因水稻。本文的研究确定了五个新的非胚乳组织表达启动子，其可能适合于水稻基因工程，并可能减少转基因作物的潜在安全性。
外文摘要：Using promoters expressed in nonendosperm tissues to activate target genes in specific plant tissues or organs with very limited expression in the endosperm is an attractive approach in crop transgenic engineering. In this article, five putative nonendosperm tissue-expressed promoters were cloned from the rice genome and designated P-OSNETE1, P-OsNETE2, P-OsNETE3, P-OsNETE4 and P-OsNETE5. By qualitatively and quantitatively examining GUSplus reporter gene expression in transgenic rice plants, P-OsNETE1-P-OsNETE5 were all found to be active in the roots, leaves, stems, sheaths and panicles but not in the endosperm of plants at different developmental stages. In addition, POsNETE2, POsNETE4 and POsNETE5 were also inactive in rice embryos. Among these promoters, POsNETE4 and POsNETE5 exhibited higher activities in all of the tested tissues, and their activities in stems, leaves, roots and sheaths were higher than or comparable to those of the rice Actin1 promoter. We also progressively monitored the activities of POsNETE1-POsNETE5 in two generations of single-copy lines and found that these promoters were stably expressed between generations. Transgenic rice was produced using POsNETE4 and POsNETE5 to drive a modified Bt gene, mCry1Ab. Bt protein expressed in the tested plants ranged from 1769.4 to 4428.8 ng/g fresh leaves, whereas Bt protein was barely detected in the endosperm. Overall, our study identified five novel nonendosperm tissue-expressed promoters that might be suitable for rice genetic engineering and might reduce potential social concern regarding the safety of GMO crops.
外文关键词：Bacillus thuringiensis, GUSplus, nonendosperm tissue-expressed promoters, transgenic rice
作者：Li, Hao; Li, Juan; Xu, Rongfang; Qin, Ruiying; Song, Fengshun; Li, Li; Wei, Pengcheng; Yang, Jianbo
作者单位：Anhui Acad Agr Sci
期刊名称： PLANT BIOTECHNOLOGY JOURNAL
期刊影响因子：0.0
出版年份：2018
出版刊次：7
点击下载：转基因水稻中五个水稻非胚乳组织表达启动子的分离及其活性评价