College of Agronomy, State Key Laboratory of Wheat and Maize Crop Science, and Center for Crop Genome Engineering, Henan Agricultural University, Zhengzhou, China.
State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
Plant Biotechnol J. 2021 May;19(5):897-909. doi: 10.1111/pbi.13512. Epub 2020 Dec 10.
The LRK10-like receptor kinases (LRK10L-RLKs) are ubiquitously present in higher plants, but knowledge of their expression and function is still limited. Here, we report expression and functional analysis of TtdLRK10L-1, a typical LRK10L-RLK in durum wheat (Triticum turgidum L. ssp. durum). The introns of TtdLRK10L-1 contained multiple kinds of predicted cis-elements. To investigate the potential effect of these cis-elements on TtdLRK10L-1 expression and function, two types of transgenic wheat lines were prepared, which expressed a GFP-tagged TtdLRK10L-1 protein (TtdLRK10L-1:GFP) from the cDNA or genomic DNA (gDNA) sequence of TtdLRK10L-1 under the native promoter. TtdLRK10L-1:GFP expression was up-regulated by the powdery mildew pathogen Blumeria graminis f. sp. tritici (Bgt) in both types of transgenic plants, with the scale of the elevation being much stronger in the gDNA lines. Both types of transgenic plants exhibited enhanced resistance to Bgt infection relative to wild type control. Notably, the Bgt defence activated in the gDNA lines was significantly stronger than that in the cDNA lines. Further analysis revealed that a putative MYB transcription factor binding site (MYB-BS, CAGTTA) located in TtdLRK10L-1 intron I was critical for the efficient expression and function of TtdLRK10L-1 in Bgt defence. This MYB-BS could also increase the activity of a superpromoter widely used in ectopic gene expression studies in plants. Together, our results deepen the understanding of the expression and functional characteristics of LRK10L-RLKs. TtdLRK10L-1 is likely useful for further dissecting the molecular processes underlying wheat defence against Bgt and for developing Bgt resistant wheat crops.
LRK10 样受体激酶(LRK10L-RLKs)广泛存在于高等植物中,但对其表达和功能的了解仍很有限。在这里,我们报告了杜伦小麦(Triticum turgidum L. ssp. durum)中典型的 LRK10L-RLK TtdLRK10L-1 的表达和功能分析。TtdLRK10L-1 的内含子包含多种预测的顺式元件。为了研究这些顺式元件对 TtdLRK10L-1 表达和功能的潜在影响,制备了两种类型的转基因小麦品系,它们分别从 TtdLRK10L-1 的 cDNA 或基因组 DNA(gDNA)序列在其天然启动子的控制下表达 GFP 标记的 TtdLRK10L-1 蛋白(TtdLRK10L-1:GFP)。TtdLRK10L-1:GFP 的表达在两种转基因植物中均被白粉菌病原体 Blumeria graminis f. sp. tritici(Bgt)上调,gDNA 品系中的上调幅度要大得多。与野生型对照相比,两种类型的转基因植物均表现出对 Bgt 感染的增强抗性。值得注意的是,gDNA 品系中激活的 Bgt 防御反应明显强于 cDNA 品系。进一步分析表明,位于 TtdLRK10L-1 内含子 I 中的一个假定的 MYB 转录因子结合位点(MYB-BS,CAGTTA)对于 TtdLRK10L-1 在 Bgt 防御中的有效表达和功能至关重要。该 MYB-BS 还可以增加在植物中外源基因表达研究中广泛使用的超级启动子的活性。总之,我们的研究结果加深了对 LRK10L-RLK 表达和功能特征的理解。TtdLRK10L-1 可能有助于进一步剖析小麦对白粉菌防御的分子过程,并用于开发对白粉菌具有抗性的小麦作物。