Qi Yue, Li Jianyuan, Mapuranga Johannes, Zhang Na, Chang Jiaying, Shen Qianhua, Zhang Yue, Wei Jie, Cui Liping, Liu Daqun, Yang Wenxiang
Department of Plant Pathology, Agricultural University of Hebei/Technological Innovation Center for Biological Control of Plant Diseases and Insect Pests of Hebei Province/National Engineering Research Center for Agriculture in Northern Mountainous Areas, Baoding, China.
Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing, China.
Front Plant Sci. 2023 Jan 16;13:1098549. doi: 10.3389/fpls.2022.1098549. eCollection 2022.
Wheat leaf rust, caused by Eriks. (), is a global wheat disease threatening wheat production. Dissecting how effector proteins interact with wheat has great significance in understanding the pathogenicity mechanisms of . In the study, the cDNA of 13-5-72 interacting with susceptible cultivar Thatcher was used as template to amplify Pt13024 gene. The expression pattern and structure of were analyzed by qRT-PCR and online softwares. The secretion function of Pt13024 signal peptide was verified by the yeast system. Subcellular localization of Pt13024 was analyzed using transient expression on . The verification that Pt13024 inhibited programmed cell death (PCD) was conducted on and wheat. The deletion mutation of Pt13024 was used to identify the virulence function motif. The transient transformation of wheat mediated by the type III secretion system (TTSS) was used to analyze the activity of regulating the host defense response of Pt13024. gene silencing was performed by host-induced gene silencing (HIGS). The results showed that Pt13024 was identified as an effector and localized in the cytoplasm and nucleus on the . It can inhibit PCD induced by the Bcl-2-associated X protein (BAX) from mice and infestans 1 (INF1) from on , and it can also inhibit PCD induced by DC3000 on wheat. The amino acids 22 to 41 at N-terminal of the Pt13024 are essential for the inhibition of programmed cell death (PCD) induced by BAX. The accumulation of reactive oxygen species and deposition of callose in near-isogenic line TcLr30, which is in Thatcher background with , induced by Pt13024 was higher than that in 41 wheat leaf rust-resistant near-isogenic lines (monogenic lines) with different resistance genes and Thatcher. Silencing of Pt13024 reduced the leaf rust resistance of Lr30 during the interaction between Pt and TcLr30. We can conclude that Pt13024 is avirulent to TcLr30 when interacts with TcLr30. These findings lay the foundation for further investigations into the role of Pt effector proteins in pathogenesis and their regulatory mechanisms.
由埃里克氏菌（Eriks. ()）引起的小麦叶锈病是一种威胁小麦生产的全球性小麦病害。剖析效应蛋白与小麦的相互作用方式对于理解（此处原文中“.”指代不明，无法准确翻译）的致病机制具有重要意义。在该研究中，以与感病品种撒切尔（Thatcher）相互作用的13 - 5 - 72的cDNA为模板扩增Pt13024基因。通过qRT - PCR和在线软件分析（此处原文中“.”指代不明，无法准确翻译）的表达模式和结构。利用酵母系统验证Pt13024信号肽的分泌功能。使用（此处原文中“.”指代不明，无法准确翻译）上的瞬时表达分析Pt13024的亚细胞定位。在（此处原文中“.”指代不明，无法准确翻译）和小麦上进行Pt13024抑制程序性细胞死亡（PCD）的验证。利用Pt13024的缺失突变来鉴定毒力功能基序。使用III型分泌系统（TTSS）介导的小麦瞬时转化来分析Pt13024调节宿主防御反应的活性。通过宿主诱导基因沉默（HIGS）进行（此处原文中“.”指代不明，无法准确翻译）基因沉默。结果表明，Pt13024被鉴定为一种效应蛋白，在（此处原文中“.”指代不明，无法准确翻译）上定位于细胞质和细胞核。它可以抑制小鼠的Bcl - 2相关X蛋白（BAX）和（此处原文中“.”指代不明，无法准确翻译）的致病疫霉1（INF1）诱导的PCD，并且也可以抑制小麦上DC3000诱导的PCD。Pt13024 N端的第22至41位氨基酸对于抑制BAX诱导的程序性细胞死亡（PCD）至关重要。在与（此处原文中“.”指代不明，无法准确翻译）的撒切尔背景近等基因系TcLr30中，由Pt13024诱导的活性氧积累和胼胝质沉积高于41个具有不同抗性基因的小麦抗叶锈近等基因系（单基因系）和撒切尔。在Pt与TcLr30相互作用期间，Pt13024的沉默降低了Lr30的叶锈抗性。我们可以得出结论，当（此处原文中“.”指代不明，无法准确翻译）与TcLr30相互作用时，Pt13024对TcLr30无毒。这些发现为进一步研究Pt效应蛋白在致病过程中的作用及其调控机制奠定了基础。
