Effector Pt31812 from Puccinia triticina acts as avirulence factor for Lr42-mediated resistance in wheat

As an obligate biotrophic fungus, the leaf rust pathogen Puccinia triticina (Pt) secretes a repertoire of effector proteins into host cells for modulating plant immunity and promoting fungal pathogenesis. Here, we identify the Pt31812 effector and characterize its function in pathogenesis and immune...

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Main Authors: Jianyuan Li, Jiali Li, Jie Wei, Lin Li, Yue Qi, Yue Zhang, Wenxiang Yang, Qian-Hua Shen
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1570072/full
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author Jianyuan Li
Jianyuan Li
Jiali Li
Jie Wei
Jie Wei
Lin Li
Yue Qi
Yue Qi
Yue Zhang
Yue Zhang
Wenxiang Yang
Qian-Hua Shen
author_facet Jianyuan Li
Jianyuan Li
Jiali Li
Jie Wei
Jie Wei
Lin Li
Yue Qi
Yue Qi
Yue Zhang
Yue Zhang
Wenxiang Yang
Qian-Hua Shen
author_sort Jianyuan Li
collection DOAJ
description As an obligate biotrophic fungus, the leaf rust pathogen Puccinia triticina (Pt) secretes a repertoire of effector proteins into host cells for modulating plant immunity and promoting fungal pathogenesis. Here, we identify the Pt31812 effector and characterize its function in pathogenesis and immune-related activity in plants. In the study, Pt31812 was cloned by PCR, and the expression pattern and structure were analyzed by qRT-PCR and online softwares. Subcellular localization of Pt31812 was analyzed using transient expression on Nicotiana Benthamiana. Further functional analysis was conducted using transient expression and host-induced gene silencing (HIGS). The results showed that Pt31812 encodes candidate effector with a predicted signaling peptide (SP) at the N-terminus, and its expression was highly up-regulated during Pt infection of wheat. Subcellular localization analysis revealed that Pt31812 is localized in cytoplasm and nucleus when expressed in N. Benthamiana. Co-expression of Pt31812 and mammalian BAX protein revealed that Pt31812 inhibited BAX-induced cell death in N. Benthamiana, and the fragment of 22–88 aa from the N-terminus of the effector was important for the inhibiting activity. Interestingly, expression of Pt31812 in a panel of wheat differential lines with different Lr resistance genes showed that Pt31812 specifically triggered cell death in a Lr42-harboring wheat line. Furthermore, transient gene silencing of Pt31812 through BSMV-HIGS approach rendered loss of Lr42-mediated resistance against rust race Pt.-THSN and altered the infection type from resistant to susceptible. Our data reveal that Pt31812, as a candidate effector with immune inhibiting activity, acts as an avirulence determinant factor during Pt infection of Lr42-harboring wheat line. These findings highlight immune-related activity of specific Pt effectors and lay the foundation for further investigation into mechanisms of leaf rust fungal pathogenesis and recognition.
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spelling doaj-art-7cd73de29d11454ebc79a796a326b6db2025-08-20T03:51:25ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-07-011610.3389/fmicb.2025.15700721570072Effector Pt31812 from Puccinia triticina acts as avirulence factor for Lr42-mediated resistance in wheatJianyuan Li0Jianyuan Li1Jiali Li2Jie Wei3Jie Wei4Lin Li5Yue Qi6Yue Qi7Yue Zhang8Yue Zhang9Wenxiang Yang10Qian-Hua Shen11Department of Plant Pathology, College of Plant Protection, Agricultural University of Hebei, Biological Control Center for Plant Diseases and Plant Pests of Hebei, Baoding, ChinaCollege of Biological Sciences and Engineering, Xingtai University, Xingtai, ChinaKey Lab of Seed Innovation, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, ChinaDepartment of Plant Pathology, College of Plant Protection, Agricultural University of Hebei, Biological Control Center for Plant Diseases and Plant Pests of Hebei, Baoding, ChinaDepartment of Agriculture and Animal Husbandry Engineering, Cangzhou Technical College, Cangzhou, ChinaCollege of Biological Sciences and Engineering, Xingtai University, Xingtai, ChinaDepartment of Plant Pathology, College of Plant Protection, Agricultural University of Hebei, Biological Control Center for Plant Diseases and Plant Pests of Hebei, Baoding, ChinaKey Laboratory of Plant Resources and China National Botanical Garden, Institute of Botany, Chinese Academy of Sciences, Beijing, ChinaDepartment of Plant Pathology, College of Plant Protection, Agricultural University of Hebei, Biological Control Center for Plant Diseases and Plant Pests of Hebei, Baoding, ChinaDryland Farming Institute, Hebei Academy of Agricultural and Forestry Science, Hengshui, ChinaDepartment of Plant Pathology, College of Plant Protection, Agricultural University of Hebei, Biological Control Center for Plant Diseases and Plant Pests of Hebei, Baoding, ChinaKey Lab of Seed Innovation, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, ChinaAs an obligate biotrophic fungus, the leaf rust pathogen Puccinia triticina (Pt) secretes a repertoire of effector proteins into host cells for modulating plant immunity and promoting fungal pathogenesis. Here, we identify the Pt31812 effector and characterize its function in pathogenesis and immune-related activity in plants. In the study, Pt31812 was cloned by PCR, and the expression pattern and structure were analyzed by qRT-PCR and online softwares. Subcellular localization of Pt31812 was analyzed using transient expression on Nicotiana Benthamiana. Further functional analysis was conducted using transient expression and host-induced gene silencing (HIGS). The results showed that Pt31812 encodes candidate effector with a predicted signaling peptide (SP) at the N-terminus, and its expression was highly up-regulated during Pt infection of wheat. Subcellular localization analysis revealed that Pt31812 is localized in cytoplasm and nucleus when expressed in N. Benthamiana. Co-expression of Pt31812 and mammalian BAX protein revealed that Pt31812 inhibited BAX-induced cell death in N. Benthamiana, and the fragment of 22–88 aa from the N-terminus of the effector was important for the inhibiting activity. Interestingly, expression of Pt31812 in a panel of wheat differential lines with different Lr resistance genes showed that Pt31812 specifically triggered cell death in a Lr42-harboring wheat line. Furthermore, transient gene silencing of Pt31812 through BSMV-HIGS approach rendered loss of Lr42-mediated resistance against rust race Pt.-THSN and altered the infection type from resistant to susceptible. Our data reveal that Pt31812, as a candidate effector with immune inhibiting activity, acts as an avirulence determinant factor during Pt infection of Lr42-harboring wheat line. These findings highlight immune-related activity of specific Pt effectors and lay the foundation for further investigation into mechanisms of leaf rust fungal pathogenesis and recognition.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1570072/fullwheat leaf rusteffectorPt31812avirulence genepathogenesisLr42
spellingShingle Jianyuan Li
Jianyuan Li
Jiali Li
Jie Wei
Jie Wei
Lin Li
Yue Qi
Yue Qi
Yue Zhang
Yue Zhang
Wenxiang Yang
Qian-Hua Shen
Effector Pt31812 from Puccinia triticina acts as avirulence factor for Lr42-mediated resistance in wheat
Frontiers in Microbiology
wheat leaf rust
effector
Pt31812
avirulence gene
pathogenesis
Lr42
title Effector Pt31812 from Puccinia triticina acts as avirulence factor for Lr42-mediated resistance in wheat
title_full Effector Pt31812 from Puccinia triticina acts as avirulence factor for Lr42-mediated resistance in wheat
title_fullStr Effector Pt31812 from Puccinia triticina acts as avirulence factor for Lr42-mediated resistance in wheat
title_full_unstemmed Effector Pt31812 from Puccinia triticina acts as avirulence factor for Lr42-mediated resistance in wheat
title_short Effector Pt31812 from Puccinia triticina acts as avirulence factor for Lr42-mediated resistance in wheat
title_sort effector pt31812 from puccinia triticina acts as avirulence factor for lr42 mediated resistance in wheat
topic wheat leaf rust
effector
Pt31812
avirulence gene
pathogenesis
Lr42
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1570072/full
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