Transcription factor BnaA9.WRKY47 contributes to the adaptation of Brassica napus to low boron stress by up-regulating the boric acid channel gene BnaA3.NIP5;1
Transcription factor BnaA9.WRKY47 contributes to the adaptation of Brassica napus to low boron stress by up-regulating the boric acid channel gene BnaA3.NIP5;1
复制标题
转录因子 BnaA9.WRKY47 通过上调硼酸通道基因 BnaA3.NIP5 促进甘蓝型油菜适应低硼胁迫；1
DOI：
10.1111/pbi.13288
复制
发表时间：
2019-11-19
影响因子：
13.8
通讯作者：
Xu, Fangsen
中科院分区：
文献类型：
--
作者：
Feng, Yingna;Cui, Rui;Xu, Fangsen
缺硼是甘蓝型油菜（B. napus）生长抑制和产量下降的主要原因之一。然而，甘蓝型油菜低B适应的分子机制在很大程度上是未知的。本研究发现51个BnaWRKY转录因子对甘蓝型油菜B缺乏有反应，其中BnaAn。WRKY26 BnaA9。WRKY47 BnaA1。WKRY53和bnan。在酵母单杂交实验中，WRKY57与B转运相关基因BnaNIP5启动子中含有W盒的保守序列具有较强的结合活性；1s和BnaBOR1s。与靶蛋白融合的绿色荧光蛋白证实了BnaA9.WRKY47的核定位。CRISPR/ cas9介导的BnaA9敲除系。与野生型植物相比，甘蓝型油菜WRKY47对低B和低B含量的敏感性更高。相反，BnaA9的过表达。WRKY47增强了植物对低B的适应性，其组织中B含量高于野生型植物。与表型反应和B积累一致，BnaA3.NIP5的转录活性；B效率候选基因1在敲除系中表达量减少，而在低B条件下过表达系中表达量显著增加。电泳迁移率转移实验、烟草瞬时表达实验和原位杂交实验表明，BnaA9。WRKY47直接激活BnaA3.NIP5；通过绑定到特定的顺式元素。综上所述，我们的研究结果支持BnaWRKYs作为低B和BnaA9反应的新参与者。WRKY47通过上调BnaA3.NIP5参与甘蓝型油菜对B缺乏的适应；1表达以促进B的有效摄取。
Boron (B) deficiency is one of the major causes of growth inhibition and yield reduction in Brassica napus (B. napus). However, the molecular mechanisms of low B adaptation in B. napus are largely unknown. Here, fifty-one BnaWRKY transcription factors were identified as responsive to B deficiency in B. napus, in which BnaAn.WRKY26, BnaA9.WRKY47, BnaA1.WKRY53 and BnaCn.WRKY57 were tested in yeast one-hybrid assays and showed strong binding activity with conserved sequences containing a W box in the promoters of the B transport-related genes BnaNIP5;1s and BnaBOR1s. Green fluorescent protein fused to the target protein demonstrated the nuclear localization of BnaA9.WRKY47. CRISPR/Cas9-mediated knockout lines of BnaA9.WRKY47 in B. napus had increased sensitivity to low B and lower contents of B than wild-type plants. In contrast, overexpression of BnaA9.WRKY47 enhanced the adaptation to low B with higher B contents in tissues than in wild-type plants. Consistent with the phenotypic response and B accumulation in these transgenic lines, the transcription activity of BnaA3.NIP5;1, a B efficiency candidate gene, was decreased in the knockout lines but was significantly increased in the overexpressing lines under low B conditions. Electrophoretic mobility shift assays, transient expression experiments in tobacco and in situ hybridizations showed that BnaA9.WRKY47 directly activated BnaA3.NIP5;1 expression through binding to the specific cis-element. Taken together, our findings support BnaWRKYs as new participants in response to low B, and BnaA9.WRKY47 contributes to the adaptation of B. napus to B deficiency through up-regulating BnaA3.NIP5;1 expression to facilitate efficient B uptake.