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| 1 | Transcription factor WRKY22 regulates canker susceptibility in sweet orange (Citrus sinensis Osbeck) by enhancing cell enlargement and CsLOB1 expression显示文摘Pathological hypertrophy(cell enlargement)plays an important role in the development of citrus canker,but its regulators are largely unknown.Although WRKY22 is known to be involved in pathogen-triggered immunity and positively regulates resistance to bacterial pathogens in Arabidopsis,rice and pepper,the CRISPR/Cas9-mediated partial knockout of CsWRKY22 improves resistance to Xanthomonas citri subsp.citri(Xcc)in Wanjincheng orange(Citrus sinensis Osbeck).Here,we demonstrate that CsWRKY22 is a nucleus-localized transcriptional activator.CsWRKY22-overexpressing plants exhibited dwarf phenotypes that had wrinkled and thickened leaves and were more sensitive to Xcc,whereas CsWRKY22-silenced plants showed no visible phenotype changes and were more resistant to Xcc.Microscopic observations revealed that the overexpression of CsWRKY22 increased cell size in the spongy mesophyll.Transcriptome analysis showed that cell growth-related pathways,such as the auxin and brassinosteroid hormonal signaling and cell wall organization and biogenesis pathways,were significantly upregulated upon CsWRKY22 overexpression.Interestingly,CsWRKY22 activated the expression of CsLOB1,which is a key gene regulating susceptibility to citrus canker.We further confirmed that CsWRKY22 bound directly to the W-boxes just upstream of the transcription start site of CsLOB1 in vivo and in vitro.We conclude that CsWRKY22 enhances susceptibility to citrus canker by promoting host hypertrophy and CsLOB1 expression.Thus,our study provides new insights into the mechanism regulating pathological hypertrophy and the function of WRKY22 in citrus. | Qin Long Meixia Du Junhong Long Yu Xie Jingyun Zhang Lanzhen Xu Yongrui He Qiang Li Shanchun Chen Xiuping Zou | 2021 | Horticulture Research2021，8，1： | 5 |
| 2 | CitGVD:a comprehensive database of citrus genomic variations显示文摘Citrus is one of the most important commercial fruit crops worldwide.With the vast genomic data currently available for citrus fruit,genetic relationships,and molecular markers can be assessed for the development of molecular breeding and genomic selection strategies.In this study,to permit the ease of access to these data,a web-based database,the citrus genomic variation database(CitGVD,http://gffzz1758061741e3440chbv599x0foq0w6n69.ffgz.tsg.suse.edu.cn/home)was developed as the first citrusspecific comprehensive database dedicated to genome-wide variations including single nucleotide polymorphisms(SNPs)and insertions/deletions(INDELs).The current version(V1.0.0)of CitGVD is an open-access resource centered on 1,493,258,964 high-quality genomic variations and 84 phenotypes of 346 organisms curated from in-house projects and public resources.CitGVD integrates closely related information on genomic variation annotations,related gene annotations,and details regarding the organisms,incorporating a variety of built-in tools for data accession and analysis.As an example,CitGWAS can be used for genome-wide association studies(GWASs)with SNPs and phenotypic data,while CitEVOL can be used for genetic structure analysis.These features make CitGVD a comprehensive web portal and bioinformatics platform for citrus-related studies.It also provides a model for analyzing genome-wide variations for a wide range of crop varieties. | Qiang Li Jingjing Qi Xiujuan Qin Wanfu Dou Tiangang Lei Anhua Hu Ruirui Jia Guojin Jiang Xiuping Zou Qin Long Lanzhen Xu Aihong Peng Lixiao Yao Shanchun Chen Yongrui He | 2020 | Horticulture Research2020，7，1： | 4 |
| 3 | Cloning and Expression Analysis of Citrus Genes Cs GH3.1 and Cs GH3.6Responding to Xanthomonas axonopodis pv. citri Infection显示文摘To study the functions of the early auxin-responsive genes Cs GH3.1 and Cs GH3.6 in citrus resistance against canker disease, we cloned Cs GH3.1and Cs GH3.6 in ‘Newhall' Navel Orange(Citrus sinensis Osbeck). They are 1 797 bp and 1 887 bp and encode 598 and 629 amino acids, respectively. In vitro mature leaves from susceptible ‘Newhall' and resistant Calamondin(C. madurensis) were inoculated by a Xanthomonas axonopodis pv. citri(Xac) bacterial suspension, and expression of Cs GH3.1 and Cs GH3.6 in the two varieties were analyzed using quantitative real-time PCR(q RT-PCR). ‘Newhall' leaves were treated with different hormones for 3 days, inoculated by Xac bacterial suspension, and then the symptoms in these leaves were investigated. We used q RT-PCR to analyze the effect of different hormones on Cs GH3.1 and Cs GH3.6 expression in ‘Newhall'leaves. The expression levels of both Cs GH3.1 and Cs GH3.6 were significantly induced by Xac in ‘Newhall' leaves, compared with levels in Calamondin leaves. 1-naphthy acetic acid(NAA) increased the hypertrophy of infection sites in ‘Newhall' leaves, while naphthyl-phthalamic acid(NPA)had no visible effect on lesion development. NAA hormone greatly improved expression of Cs GH3.1 in ‘Newhall', but not Cs GH3.6. These results indicate that the auxin primary-response gene Cs GH3.1 plays an important role in citrus susceptibility to Xac. | CHEN Min HE Yongrui XU Lanzhen PENG Aihong LEI Tiangang YAO Lixiao LI Qiang ZHOU Pengfei BAI Xiaojing DUAN Minjie JIANG Xueyou JIA Ruirui ZOU Xiuping CHEN Shanchun | 2016 | Horticultural Plant Journal2016，2，4： | 4 |
| 4 | CsWAKL08,a pathogen-induced wall-associated receptor-like kinase in sweet orange,confers resistance to citrus bacterial canker via ROS control and JA signaling显示文摘Citrus bacterial canker(CBC)is a disease resulting from Xanthomonas citri subsp.citri(Xcc)infection and poses a grave threat to citrus production worldwide.Wall-associated receptor-like kinases(WAKLs)are proteins with a central role in resisting a range of fungal and bacterial diseases.The roles of WAKLs in the context of CBC resistance,however,remain unclear.Here,we explored the role of CsWAKL08,which confers resistance to CBC,and we additionally analyzed the molecular mechanisms of CsWAKL08-mediated CBC resistance.Based on systematic annotation and induced expression analysis of the CsWAKL family in Citrus sinensis,CsWAKL08 was identified as a candidate that can be upregulated by Xcc infection in the CBC-resistant variety.CsWAKL08 can also be induced by the phytohormones salicylic acid(SA)and methyl jasmonic acid(MeJA)and spans the plasma membrane.Overexpression of CsWAKL08 resulted in strong CBC resistance in transgenic sweet oranges,whereas silencing of CsWAKL08 resulted in susceptibility to CBC.The peroxidase(POD)and superoxide dismutase(SOD)activities were significantly enhanced in the CsWAKL08-overexpressing plants compared to the control plants,thereby mediating reactive oxygen species(ROS)homeostasis in the transgenic plants.Moreover,the JA levels and the expression of JA biosynthesis and JA responsive genes were substantially elevated in the CsWAKL08 overexpression plants relative to the controls upon Xcc infection.Based on these findings,we conclude that the wall-associated receptor-like kinase CsWAKL08 positively regulates CBC resistance through a mechanism involving ROS control and JA signaling.These results further highlight the importance of this kinase family in plant pathogen resistance. | Qiang Li Anhua Hu Jingjing Qi Wanfu Dou Xiujuan Qin Xiuping Zou Lanzhen Xu Shanchun Chen Yongrui He | 2020 | Horticulture Research2020，7，1： | 3 |
| 5 | An improved procedure for Agrobacterium-mediated transformation of trifoliate orange (Poncirus trifoliata L. Raf.) via indirect organogenesis显示文摘 | Xiuping Zou Demou Li Xiaoying Luo Keming Luo Yan Pei | 2008 | In Vitro Cellular & Developmental Biology - Plant2008，，3： | 1 |
| 6 | Effi- cient auto-excision of a selectable marker gene from transgenic citrus by combining the Cre/loxP system and ipt selection 显示文摘 | Zou Xiuping Peng Aihong Xu Lanzhen | 2013 | Plant Cell Rep2013，32，10： | 1 |
| 7 | Comparative analysis of Wanjincheng orange leaf and root responses to ‘Candidatus liberibacter asiaticus’ infection using leaf-disc grafting显示文摘Citrus Huanglongbing,associated with Candidatus Liberibacter asiaticus(Las),is the serious disease of citrus worldwide.Here,we compared differences between leaf and root responses in Wanjincheng Orange(Citrus sinensis Osbeck)to Las infection using leaf-disc grafting.Trees had no obvious symptoms in the first two months after grafting(MAG),but yellowing leaves,thickened midribs and decayed roots began to appear at 6 MAG.The Las growth rate in roots was greater than inmidribs from 2 to 6 MAG;however,by 10 MAG,it was significantly lower in roots than in midribs.Amicroscopic analysis revealed that starch accumulation,callose deposition,cellwall thickness and the number of cell layers increased in the phloem of infected trees.After Las infection,the starch content in the leaves was significantly higher than that in the roots,while the callose deposition in the midribs was 3–19 times that in the roots.A gene expression analysis showed that transcripts of callose-and starch metabolism-related genes were obviously affected by Las infection,and the pathogenesis-related genes PR1,PR2 and PR5 were significantly upregulated and downregulated in midrib and root,respectively.Our results indicated that the PR-mediated resistance response was repressed in root but activated in midrib,which may result in the more rapid Las growth in roots than in midribs during the early infection stage. | Zhu Xie Ke Zhao Junhong Long Lin Zhen Xiuping Zou Shanchun Chen | 2021 | Horticultural Plant Journal2021，7，5： | 1 |
| 8 | Free-standing SnNb_(2)O_(6)@CSN film as flexible anode for high performance sodium-ion batteries显示文摘Free-standing electrodes are promising candidates for flexible rechargeable batteries, toward the application of flexible energy storage devices, due to their merits of additive-free, lightweight, and high energy density. Herein, we report a free-standing SnNb_(2)O_(6)@CSN flexible film with SnNb_(2)O_(6) encapsulated in 3D carbon skeleton nanofibers by electrospinning and carbonization processes as flexible anode for sodium-ion batteries(SIBs). The 3D carbon skeleton nanofibers serve as ion/electron transport pathway to improve the electrochemical reaction kinetics and meanwhile alleviate the volume changes of SnNb_(2)O_(6) during charge-discharge processes. The as-constructed half-cell(SnNb_(2)O_(6)@CSN‖Na) exhibits excellent cycling stability of 99.2 m Ah/g at 0.5 A/g after 950 cycles(coulombic efficiency of ~100%) and a high rate performance of 108.6 mAh/g at 10 A/g. In addition, the pouch cell can light up the LEDs at different bending angles(0°, 90°, 180°). This research shows a promising anode material for flexible energy storage electronics. | Xuemin Yin Tao Liu Xiuping Yin Xiaochen Feng Yiming Liu Qinhao Shi Xingli Zou Yufeng Zhao | 2023 | Chinese Chemical Letters2023，34，7： | 0 |
| 9 | An endolysin gene from Candidatus Liberibacter asiaticus confers dual resistance to huanglongbing and citrus canker显示文摘The most damaging citrus diseases are Huanglongbing(HLB)and citrus canker,which are caused by Candidatus Liberibacter asiaticus(CaLas)and Xanthomonas citri pv.citri(Xcc),respectively.Endolysins from bacteriophages are a possible option for disease resistance in plant breeding.Here,we report improvement of citrus resistance to HLB and citrus canker using the LasLYS1 and LasLYS2 endolysins from CaLas.LasLYS2 demonstrated bactericidal efficacy against several Rhizobiaceae bacteria and Xcc,according to inhibition zone analyses.The two genes,driven by a strong promoter from Cauliflower mosaic virus,35S,were integrated into Carrizo citrange via Agrobacterium-mediated transformation.More than 2 years of greenhouse testing indicated that LasLYS2 provided substantial and long-lasting resistance to HLB,allowing transgenic plants to retain low CaLas titers and no obvious symptoms while also clearing CaLas from infected plants in the long term.LasLYS2 transgenic plants with improved HLB resistance also showed resistance to Xcc,indicating that LasLYS2 had dual resistance to HLB and citrus canker.A microbiome study of transgenic plants revealed that the endolysins repressed Xanthomonadaceae and Rhizobiaceae populations in roots while increasing Burkholderiaceae and Rhodanobacteraceae populations,which might boost the citrus defense response,according to transcriptome analysis.We also found that Lyz domain 2 is the key bactericidal motif of LasLYS1 and LasLYS2.Four endolysins with potential resistance to HLB and citrus canker were found based on the structures of LasLYS1 and LasLYS2.Overall,the work shed light on the mechanisms of resistance of CaLas-derived endolysins,providing insights for designing endolysins to develop broad-spectrum disease resistance in citrus. | Lanzhen Xu Kaiqing Mo Danlu Ran Juanjuan Ma Lehuan Zhang Yijia Sun Qin Long Guojin Jiang Xiaochun Zhao Xiuping Zou | 2023 | Horticulture Research2023，10，9： | 0 |
| 10 | Production of marker-free transgenic plants from mature tissues of navel orange using a Cre/loxP site-recombination system显示文摘Genetic transformation with mature material as the explants could shorten the transgenic period and avoid seed dependence compared with genetic transformation using the epicotyl seedling stem segments as the receptor. Here, we constructed an Agrobacterium tumefaciensmediated transformation for generation of marker-free transgenic plants from navel orange(Citrus sinensis Osbeck) mature stems using a CreloxP recombination system. To efficiently recover the regenerated buds from mature tissues, five recovery methods were compared: in vitro micrografting of 0.1-0.5(1-2 weeks), > 0.5 cm(3-4 weeks) and > 1 cm long lignified bud and in vitro micrografting of explants with a bud and rooting regenerated bud. The data showed that in vitro micrografting of > 1 cm long regenerated bud with expanded leaves after one month of continuous culture for lignification was the optimal solution for plant recovery from mature tissues. Transgenic plants without selectable marker genes were created from navel orange(Citrus sinensis Osbeck) tissue using a transformation vector PLI-35SPR1aCB containing a Cre/loxP system recombination together with genes encoding the selectable marker isopentenyl transferase(IPT) and an anti-bacterial peptide(PR1aCB).Using IPT positive selection, the transformation efficiency determined by PCR was 0.9%, and in total, 20 transgenic plants were obtained.Southern blotting confirmed further their transgenicity. PCR and sequencing analysis demonstrated that both the Cre and IPT genes had been successfully removed from the transgenic plants(deletion efficiency 100%). Over all, using Cre/loxP system recombination together with the IPT positive selection, marker-free transgenic plants can be recovered efficiently from mature tissues of navel orange(Citrus sinensis Osbeck), which provides a potential method for production of transgenic plants from citrus mature tissue. | Yongrui He Lanzhen Xu Aihong Peng Tiangang Lei Qiang Li Lixiao Yao Guojin Jiang Shanchun Chen Zhengguo Li Xiuping Zou | 2023 | Horticultural Plant Journal2023，9，3： | 0 |
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