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过表达Gbnpr1基因增强大豆对疫霉根腐病的抗性

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分子植物育种 2024年

作者：刘斯祺刘东波邢国杰牛陆张原宇杨向东杨静吉林师范大学生命科学学院吉林省农业科学院吉林省农业生物技术研究所吉林省农业生物技术重点实验室

大豆疫霉根腐病(phytophthora root rot， PRR)是影响大豆生产的重要土传病害之一，一般可造成大豆产量损失10%～30%，严重年份甚至绝产。抗病品种的培育是防治大豆疫霉根腐病最为经济有效的措施。本研究将来源于棉花的广谱抗病基因GbN... 详细信息

大豆疫霉根腐病(phytophthora root rot， PRR)是影响大豆生产的重要土传病害之一，一般可造成大豆产量损失10%～30%，严重年份甚至绝产。抗病品种的培育是防治大豆疫霉根腐病最为经济有效的措施。本研究将来源于棉花的广谱抗病基因Gbnpr1导入大豆栽培品种‘天隆1号’。Southern杂交检测表明，Gbnpr1基因以低拷贝形式(1～3个)整合至大豆基因组；RT-PCR和western杂交检测表明，外源基因能够正确转录和翻译。连续三代(T2-T4)接种疫霉菌(Phytophthora sojae) 1号生理小种后，非转基因植株出现叶片褪绿、植株萎蔫和死亡等典型PRR症状，植物存活率为45.12%～60.24%，而多数转基因植株表现为轻微的叶片褪绿，存活率达66.26%～96.26%，表明Gbnpr1基因过表达显著增强了大豆对疫霉根腐病的抗性，且抗性能够稳定遗传。进一步分析表明，Gbnpr1基因过表达显著提高了水杨酸、茉莉酸等信号途径中下游防卫基因的表达水平，表明Gbnpr1作为植物体内抗病信号通路的节点，通过同时调控多条防卫路径关键基因的表达进而提高植物对疫霉菌的抗性。本研究为拓宽大豆PRR遗传基础，培育广谱抗PRR大豆品种提供科学依据。

关键词：大豆疫霉根腐病 npr1 抗性

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Journal of Genetics and Genomics 2010年第6期37卷 359-369页

作者： Changgen Xie Xiaona Zhou Xingwang Deng Yan Guo College of Life Sciences Peking University Beijing 100871 China National Institute of Biological Sciences Beijing 102206 China

npr1 （Nonexpressor of Pathogenesis-Related gene 1） is a major co-activator of plant defense. Phosphorylations of npr1 play important roles in fine-tuning its activity, however a kinase corresponding to such modification remains uncharacterized. Here, we report that npr1 interacts with PKS5 （SOS2-1ike Protein Kinase 5）. The AKR （AnKyrin Repeats） motif of npr1 is required for this interaction. PKS5 phosphorylates npr1 at the C-terminal region. Expression of PKS5 is induced quickly by Pseudomonas syringae pv. tomato DC3000. Expression level of two npr1 target genes, WRKY38 and WRKY62, is reduced and/or delayed in pks5 mutants. Moreover, the expression of WRKY38 and WRKY62 displays a similar pattern in nprl-lpks5-1 double mutant comparing to that in nprl-1. Our results suggest that PKS5 functions at the upstream of npr1 and might mediate expression of WRKY38 and WRKY62 possibly by interacting with and phosphorylating npr 1.

关键词： phosphorylation kianse AKR co-immunopreeipitation npr1

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npr1 as a transgenic crop protection strategy in horticultural species

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Horticulture Research 2018年第1期5卷 751-761页

作者： Katchen Julliany P.Silva Nasser Mahna Zhonglin Mou Kevin M.Folta Horticultural Sciences Department University of FloridaGainesvilleFL32611USA Department of Horticultural Sciences University of TabrizTabrizIran Department of Microbiology and Cell Science University of FloridaGainesvilleFL32611USA Graduate Program in Plant Molecular and Cellular Biology University of FloridaGainesvilleFL32611USA

The npr1(NONEXPRESSOR OF PATHOGENESIS RELATED GENES1)gene has a central role in the long-lasting,broad-spectrum defense response known as systemic acquired resistance(SAR).When overexpressed in a transgenic context in Arabidopsis thaliana,this gene enhances resistance to a number of biotic and abiotic stresses.Its position as a key regulator of defense across diverse plant species makes npr1 a strong candidate gene for genetic engineering disease and stress tolerance into other crops.High-value horticultural crops face many new challenges from pests and pathogens,and their emergence exceeds the pace of traditional breeding,making the application of npr1-based strategies potentially useful in fruit and vegetable crops.However,plants overexpressing npr1 occasionally present detrimental morphological traits that make its application less attractive.The practical utility of npr-based approaches will be a balance of resistance gains versus other losses.In this review,we summarize the progress on the understanding of npr1-centered applications in horticultural and other crop plants.We also discuss the effect of the ectopic expression of the A.thaliana npr1 gene and its orthologs in crop plants and outline the future challenges of using npr1 in agricultural applications.

关键词： npr1 breeding attractive

The npr1-WRKY46-WRKY6 signaling cascade mediates probenazole/salicylic acid-elicited leaf senescence in Arabidopsis thaliana

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Journal of Integrative Plant Biology 2021年第5期63卷 924-936页

作者： Dingyu Zhang Zheng Zhu Jiong Gao Xin Zhou Shuai Zhu Xiaoyan Wang Xiaolei Wang Guodong Ren Benke Kuai State Key Laboratory of Genetic Engineering and Fudan Center for Genetic Diversity and Designing Agriculture Institute of Plant BiologySchool of Life SciencesFudan UniversityShanghai 200438China Institute of Neurology Huashan HospitalFudan UniversityShanghai 200040China Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering Institute of Biodiversity ScienceFudan UniversityShanghai 200438China

Endogenous salicylic acid(SA) regulates leaf senescence, but the underlying mechanism remains largely unexplored. The exogenous application of SA to living plants is not efficient for inducing leaf senescence. By taking advantage of probenazole(PBZ)-inducedbiosynthesisof endogenous SA, we previously established a chemical inducible leaf senescence system that depends on SA biosynthesis and its core signaling receptor npr1 in Arabidopsis thaliana. Here,using this system, we identified WRKY46 and WRKY6 as key components of the transcriptional machinery downstream of npr1 signaling. Upon PBZ treatment, the wrky46 mutant exhibited significantly delayed leaf senescence. We demonstrate that npr1 is essential for PBZ/SA-induced WRKY46 activation, whereas WRKY46 in turn enhances npr1 expression. WRKY46 interacts with npr1 in the nucleus, binding to the W-box of the WRKY6 promoter to induce its expression in response to SA signaling. Dysfunction of WRKY6 abolished PBZ-induced leaf senescence, while overexpression of WRKY6 was sufficient to accelerate leaf senescence even under normal growth conditions, suggesting that WRKY6 may serve as an integration node of multiple leaf senescence signaling pathways. Taken together,these findings reveal that the npr1-WRKY46-WRKY6 signaling cascade plays a critical role in PBZ/SA-mediated leaf senescence in Arabidopsis.

关键词： leaf senescence npr1 probenazole salicylic acid WRKY46 WRKY6

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A Highway for War and Peace： The Secretory Pathway in Plant-Microbe Interactions

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Molecular Plant 2011年第4期4卷 581-587页

作者： Dong Wang Xinnian Dong Department of Biology Stanford University Stanford CA 94305 USA Department of Biology PO Box 90338 Duke University Durham NC 27707 USA

Secretion of proteins and other molecules is the primary means by which a cell interacts with its surroundings. The overall organization of the secretory system is remarkably conserved among eukaryotes, and many of the components have been investigated in detail in animal models. Plant cells, because of their sessile lifestyle, are uniquely reliant on the secretory pathway to respond to changes in their environments, either abiotic, such as the absence of nutrients, or biotic, such as the presence of predators or pathogens. In particular, most plant pathogens are extracellular, which demands a robust and efficient host secretory system directed at the site of attack. Here, we present a summary of recent advances in our understanding of the molecular details of the secretory pathway during plant-microbe interactions. Secretion is required not only for the delivery of antimicrobial molecules, but also for the biogenesis of cell surface sensors to detect microbes. The deposition of extracellular material is important in the defense against classical bacterial pathogens as well as in the so-called ＂non-host＇ resistance. Finally, boosting the protein secretion capacity is vital for avoiding infection as well as for achieving symbiosis, even though in the latter case, the microbes are engulfed in intracellular compartments. The emerging evidence indicates that secretion provides an essential interface between plant hosts and their associated microbial partners.

关键词： protein secretion defense salicylic acid npr1 symbiosis Arabidopsis legume endoplasmic reticulum SNARE.

Strong Suppression of Systemic Acquired Resistance in Arabidopsis by NRR is Dependent on its Ability to Interact with nprland its Putative Repression Domain

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Molecular Plant 2008年第3期1卷 552-559页

作者： Mawsheng Chern Patrick E. Canlas Pamela C. Ronald Department of Plant Pathology University of California Davis CA 95616 USA

Systemic Acquired Resistance （SAR） in plants confers lasting broad-spectrum resistance to pathogens and requires the phytohormone salicylic acid （SA）. Arabidopsis npr1/NIM1 is a key regulator of the SAR response. Studies attempting to reveal the function of npr1 and how it mediates SA signaling have led to isolation of two classes of proteins that interact with nprI＂ the first class includes rice NRR, Arabidopsis NIMIN1, NIMIN2, and NIMIN3, and tobacco NIMIN2-1ike proteins; the second class belongs to TGA transcription factors. We have previously shown that overexpression of N RR in rice suppresses both basal and Xa21-mediated resistance. In order to test whether NRR affects SA-induced, nprl-mediated SAR, we have transformed Arabidopsis with the rice NRR gene and tested its effects on the defense response. Expression of NRR in Arabidopsis results in suppression of PR gene induction by SAR inducer and resistance to pathogens. These phenotypes are even more severe than those of the nprl-1 mutant. The ability of NRR to suppress PR gene induction and disease resistance is correlated with its ability to bind to npr1 because two point mutations in NRR, which reduce npr1 binding, fail to suppress npr1. In contrast, wild-type and a mutant NRR, which still binds to npr1 strongly, retain the ability to suppress the SAR response. Replacing the C-terminal 79 amino acids of NRR with the VP16 activation domain turns the fusion protein into a transcriptional co-activator. These results indicate that NRR binds to npr1 in vivo in a protein complex to inhibit transcriptional activation of PR genes and that NRR contains a transcription repression domain for active repression.

关键词： NRR npr1 SA SAR disease resistance.

Salicylic Acid and its Function in Plant Immunity

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Journal of Integrative Plant Biology 2011年第6期53卷 412-428页

作者： Chuanfu An Zhonglin Mou Department of Microbiology and Cell Science University of Florida Gainesville FL 32611 USA

The small phenolic compound salicylic acid （SA） plays an important regulatory role in multiple physiological processes including plant im- mune response. Significant progress has been made during the past two decades in understanding the SA-mediated defense signaling network. Characterization of a number of genes functioning in SA biosynthesis, conjugation, accumulation, signaling, and crosstalk with other hormones such as jasmonic acid, ethylene, abscisic acid, auxin, gibberellic acid, cytokinin, brassinosteroid, and peptide hormones has sketched the finely tuned immune response network. Full understanding of the mech- anism of plant immunity will need to take advantage of fast developing genomics tools and bioinformatics techniques. However, elucidating genetic components involved in these pathways by conventional ge- netics, biochemistry, and molecular biology approaches will continue to be a major task of the community. High-throughput method for SA quantification holds the potential for isolating additional mutants related to SA-mediated defense signaling.

关键词： salicylic acid （SA） systemic acquired resistance npr1 crosstalk SA biosensor plant defense.

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Lamin-like Proteins Negatively Regulate Plant Immunity through NAC WITH TRANSMEMBRANE MOTIF1-LIKE9 and NONEXPRESSOR OF PR 3ENES1 in Arabidopsis thaliana

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Molecular Plant 2017年第10期10卷 1334-1348页

作者： Tongtong Guo Xuegao Mao Hui Zhang Yu Zhang Mengdi Fu Zhenfei Sun Peng Kuai Yonggen Lou Yuda Fang National Key Laboratory of Plant Molecular Genetics CAS Center for Excellence in Molecular Plant Sciences Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200032 China University of the Chinese Academy of Sciences Beijing 100049 China State Key Laboratory of Rice Biology Institute of Insect Sciences Zhejiang University Hangzhou 310058 China

Nuclear lamins are involved in multiple biological processes in metazoan cells. The proteins of the CROWDED NUCLEI （CRWN） family are considered lamin-like candidates in Arabidopsis, although the functions of these proteins are largely unknown. In this article we show that crwnl crwn2 double mutant displays an enhanced resistance against virulent bacterial pathogens, and both virulent bacteria and salicylic acid （SA） induce transcription of CRWN1 gene as well as proteasome-mediated degradation of CRWN1 protein. We also show that CRWN1 interacts with NAC WITH TRANSMEMBRANE MOTIF1 -LIKE9 （NTL9）, a NAC transcription factor involved in plant immunity. The interaction between CRWN1 and NTL9 enhances the binding of NTL9 to the promoter of the PATHOGENESIS-RELATED1 （PR1） gene, and inhibits PR1 expression. Further genetic experiments indicated that the defense-related phenotypes of crwnl crwn2 double mutant are dependent on NONEXPRESSOR OF PR GENES1 （npr1）, a transcriptional cofactor of PRI. These findings revealed a regulatory network composed of lamin-like protein CRWN1, NTL9, and npr1 for the regulation of PR1 expression.

关键词： lamin CRWN NTL9 npr1 plant immunity

Pandemonium Breaks Out:Disruption of Salicylic Acid-Mediated Defense by Plant Pathogens

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Molecular Plant 2018年第12期11卷 1427-1439页

作者： Guang Qi Jian Chen Ming Chang Huan Chen Katherine Hall John Korin Fengquan Liu Daowen Wang Zheng Qing Fu State Key Laboratory of Wheat and Maize Crop Science and College of Agronomy Henan Agricultural University Zhengzhou 450002 China lnstitute of Plant Protection Jiangsu Academy of Agricultural Sciences Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology Nanjing 210014 China Department of Biological Sciences University of South Carolina Columbia SC 29208USA These authors contributed equally to this article

Salicylic acid (SA) or 2-hydroxybenoic acid is a phenolic plant hormone that plays an essential role in plant defense against biotrophic and semi-biotrophic pathogens. In Arabidopsis, SA is synthesized from chorismate in the chloroplast through the ICS1 (isochorismate synthase I) pathway during pathogen infectioru The transcription co-activator npr1 (Non-Expresser of Pathogenesis-Related Gene 1), as the master regulator of SA signaling, interacts with transcription factors to induce the expression of anti-microbial PR (Pathogenesis-Related) genes. To establish successful in fections, plant bacterial, oomycete, fun gal, and viral pathogens have evolved at least three major strategies to disrupt SA-mediated defense? The first strategy is to reduce SA accumulation directly by converting SA into its inactive derivatives. The second strategy is to interrupt SA biosynthesis by targeting the ICS1 pathway. In the third major strategy, plant pathogens deploy different mechanisms to interfere with SA downstream signaling. The wide array of strategies deployed by plant pathogens highlights the crucial role of disruption of SA-mediated plant defense in plant pathogenesis. A deeper understanding of this topic will greatly expand our knowledge of how plant pathogens cause diseases and consequently pave the way for the development of more effective ways to corn trol these diseases.

关键词： salicylic acid npr1 PR proteins effectors toxin VIGS