Isolation of five rice nonendosperm tissue-expressed promoters and evaluation of their activities in transgenic rice.
Isolation of five rice nonendosperm tissue-expressed promoters and evaluation of their activities in transgenic rice.
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五种水稻非胚乳组织表达启动子的分离及其在转基因水稻中的活性评价
DOI：
10.1111/pbi.12858
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发表时间：
2018-06
影响因子：
13.8
通讯作者：
Yang J
中科院分区：
文献类型：
--
作者：
Li H;Li J;Xu R;Qin R;Song F;Li L;Wei P;Yang J
利用在非胚乳组织中表达的启动子来激活特定植物组织或器官中的目的基因，而这些基因在胚乳中的表达非常有限，是作物转基因工程中的一种有吸引力的方法。本文从水稻基因组中克隆了5个可能的非胚乳组织表达启动子，分别命名为PO S启动子1、PO S启动子2、PO S启动子3、PO S启动子4和PO S启动子5。通过对转基因水稻中GUSplus报告基因表达的定性和定量检测，发现PO S Net1-PO S nete5在水稻不同发育阶段的根、叶、茎、鞘和圆锥花序中都有活性，但在胚乳中没有活性。此外，PO S NET2、PO S NET4和PO S NET5在水稻胚中也处于失活状态。在这些启动子中，PO S启动子4和PO S启动子5在所有受试组织中都表现出较高的活性，其在茎、叶、根和鞘中的活性高于或接近水稻Actin1启动子的活性。我们还对PO-S启动子1-PO-S-启动子5在两代单拷贝系中的活性进行了逐步检测，发现这些启动子在世代间稳定表达。用PO S网衣4号和PO S网衣5号分别驱动Bt基因MC ry1Ab，获得转基因水稻。Bt蛋白在供试植物中的表达范围为1769.4~4428.8 ng/g鲜叶，而在胚乳中几乎检测不到Bt蛋白。总体而言，我们的研究确定了五个新的非胚乳组织表达启动子，它们可能适合于水稻基因工程，并可能减少潜在的社会对转基因作物安全性的担忧。
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 PO s NETE 1, PO s NETE 2, PO s NETE 3, PO s NETE 4 and PO s NETE 5. By qualitatively and quantitatively examining GUSplus reporter gene expression in transgenic rice plants, PO s NETE 1‐PO s NETE 5 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, PO s NETE 2, PO s NETE 4 and PO s NETE 5 were also inactive in rice embryos. Among these promoters, PO s NETE 4 and PO s NETE 5 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 PO s NETE 1‐PO s NETE 5 in two generations of single‐copy lines and found that these promoters were stably expressed between generations. Transgenic rice was produced using PO s NETE 4 and PO s NETE 5 to drive a modified Bt gene, mC ry1Ab. 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.
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