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An endolysin gene from Candidatus Liberibacter asiaticus confers dual resistance to huanglongbing and citrus canker

Horticulture Research 2023年第9期10卷 165-180页

作者： lanzhen xu Kaiqing Mo Danlu Ran Juanjuan Ma Lehuan Zhang Yijia Sun Qin Long Guojin Jiang Xiaochun Zhao Xiuping Zou Integrative Science Center of Germplasm Creation in Western China(CHONGQING)Science City Citrus Research InstituteSouthwest University/National Citrus Engineering Research CenterChongqing 400712China

The most damaging citrus diseases are Huanglongbing(HLB)and citrus canker,which are caused by Candidatus Liberibacter asiaticus(CaLas)and Xanthomonas citri ***(Xcc),*** from bacteriophages are a possible option for disease resistance in plant ***,we report improvement of citrus resistance to HLB and citrus canker using the LasLYS1 and LasLYS2 endolysins from ***2 demonstrated bactericidal efficacy against several Rhizobiaceae bacteria and Xcc,according to inhibition zone *** two genes,driven by a strong promoter from Cauliflower mosaic virus,35S,were integrated into Carrizo citrange via Agrobacterium-mediated *** 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 ***2 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 *** also found that Lyz domain 2 is the key bactericidal motif of LasLYS1 and *** endolysins with potential resistance to HLB and citrus canker were found based on the structures of LasLYS1 and ***,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.

关键词： citrus canker resistance

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Production of marker-free transgenic plants from mature tissues of navel orange using a Cre/loxP site-recombination system

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Horticultural Plant Journal 2023年第3期9卷 473-480页

作者： Yongrui He lanzhen xu Aihong Peng Tiangang Lei Qiang Li Lixiao Yao Guojin Jiang Shanchun Chen Zhengguo Li Xiuping Zou Key Laboratory of Plant Hormones and Development Regulation of Chongqing School of Life SciencesChongqing UniversityChongqing 401331China National Citrus Engineering Research Center(NCERC) Citrus Research InstituteSouthwest University/Chinese Academy of Agricultural SciencesChongqing 400712China

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 *** 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.

关键词： Citrus sinensis Osbeck Mature stem segment Marker-free Transgene Bud micrografting

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Function and molecular mechanism analysis of CaLasSDE460 effector involved in the pathogenesis of“Candidatus Liberibacter asiaticus”in citrus

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Molecular Horticulture 2023年第1期3卷 197-213页

作者： Shuai Wang Meixia Du Liting Dong Rongrong Qu Danlu Ran Juanjuan Ma xuefeng Wang lanzhen xu Weimin Li Yongrui He Xiuping Zou Integrative Science Center of Germplasm Creation in Western China(CHONGQING)Science City Citrus Research InstituteSouthwest University/National Citrus Engineering Research CenterChongqingPeople’s Republic of China Key Laboratory for Northern Urban Agriculture of Ministry of Agriculture and Rural AffairsBeijing University of AgricultureBeijingPeople’s Republic of China

Citrus Huanglongbing(HLB),caused by Candidatus Liberibacter asiaticus(CaLas),is the most serious disease ***460 was previously characterized as a potential virulence factor of ***,the function and mechanism of CaLasSDE460 involved in CaLas against citrus is still ***,we showed that transgenic expression of CaLasSDE460 in Wanjincheng oranges(*** Osbeck)contributed to the early growth of CaLas and the development of *** the temperature increased from 25℃to 32℃,CaLas growth and symptom development in transgenic plants were slower than those in WT ***-seq analysis of transgenic plants showed that CaLasSDE460 affected multiple biological *** 25℃,transcription activities of the“Protein processing in endoplasmic reticulum”and“Cyanoamino acid metabolism”pathways increased while transcription activities of many pathways decreased at 32℃.124 and 53 genes,separately annotated to plant-pathogen interaction and MAPK signaling pathways,showed decreased expression at 32℃,compared with these(38 for plant-pathogen interaction and 17 for MAPK signaling)at 25℃.Several important genes(MAPKKK14,HSP70b,NCED3 and WRKY33),remarkably affected by CaLasSDE460,were ***,our data suggested that CaLasSDE460 participated in the pathogenesis of CaLas through interfering transcription activities of citrus defense response and this interfering was temperature-dependent.

关键词： Citrus HLB CaLasSDE460 Transcriptome Ectopic expression

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CsWAKL08,a pathogen-induced wall-associated receptor-like kinase in sweet orange,confers resistance to citrus bacterial canker via ROS control and JA signaling

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Horticulture Research 2020年第1期7卷 2053-2067页

作者： Qiang Li Anhua Hu Jingjing Qi Wanfu Dou Xiujuan Qin Xiuping Zou lanzhen xu Shanchun Chen Yongrui He Citrus Research Institute Southwest University/Chinese Academy of Agricultural SciencesChongqing 400712China

Citrus bacterial canker(CBC)is a disease resulting from Xanthomonas citri ***(Xcc)infection and poses a grave threat to citrus production ***-associated receptor-like kinases(WAKLs)are proteins with a central role in resisting a range of fungal and bacterial *** roles of WAKLs in the context of CBC resistance,however,remain ***,we explored the role of CsWAKL08,which confers resistance to CBC,and we additionally analyzed the molecular mechanisms of CsWAKL08-mediated CBC *** 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 ***08 can also be induced by the phytohormones salicylic acid(SA)and methyl jasmonic acid(MeJA)and spans the plasma *** of CsWAKL08 resulted in strong CBC resistance in transgenic sweet oranges,whereas silencing of CsWAKL08 resulted in susceptibility to *** 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 ***,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 *** 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 *** results further highlight the importance of this kinase family in plant pathogen resistance.

关键词： resistance orange citrus

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异氟醚吸入麻醉调控星形胶质细胞活化对围术期小鼠神经认知功能的影响

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武警医学 2022年第3期33卷 192-197页

作者：彭景徐兰真联勤保障部队第901医院麻醉科 230031合肥

目的探讨异氟醚吸入麻醉调控星形胶质细胞活化对围术期小鼠神经认知功能的影响。方法C57BL/6小鼠80只随机分为对照组、2%异氟醚组、手术组、2%异氟醚+手术组,每组20只。Morris水迷宫实验评估小鼠术后1、3、7 d认知功能,测定海马组织炎... 详细信息

目的探讨异氟醚吸入麻醉调控星形胶质细胞活化对围术期小鼠神经认知功能的影响。方法C57BL/6小鼠80只随机分为对照组、2%异氟醚组、手术组、2%异氟醚+手术组,每组20只。Morris水迷宫实验评估小鼠术后1、3、7 d认知功能,测定海马组织炎性反应及凋亡相关因子水平,免疫荧光染色观察星形胶质细胞活化情况,TUNEL法检测细胞凋亡。结果术后1、3、7 d,与2%异氟醚组和手术组比较,2%异氟醚+手术组小鼠逃避潜伏期[(77.36±13.48)s vs.(66.71±4.54)s、(64.47±4.43)s]、[(51.63±8.43)s vs.(46.37±5.69)s、(45.72±5.71)s]、[(30.85±4.56)s vs.(21.13±3.24)s、(22.36±3.18)s]显著延长,目标象限停留时间[(13.16±4.25)s vs.(35.36±4.48)s、(36.14±4.42)s]、[(19.48±6.37)s vs.(37.64±6.33)s、(37.15±6.28)s]、[(23.69±5.54)s vs.(39.15±6.34)s、(38.63±5.43)s]显著缩短,海马组织肿瘤坏死因子-α、白细胞介素-6和半胱氨酸天冬氨酸蛋白酶-3表达水平显著升高,海马组织胶质纤维酸性蛋白阳性细胞数[(11.97±1.35)vs.(14.15±1.48)、(15.67±1.51)]、[(34.74±1.52)vs.(31.92±1.44)、(31.96±1.52)]、[(35.69±1.48)vs.(32.14±1.51)、(31.96±1.52)]显著增加,海马神经元细胞凋亡率[(56.71±1.87)%vs.(48.63±1.45)%、(48.34±1.69)%]、[(59.64±1.57)%vs.(49.63±1.51)%、(49.45±1.58)%]、[(63.49±2.62)%vs.(50.68±2.45)%、(50.38±2.44)%]显著增高(P<0.05)。结论异氟醚吸入麻醉可引起围术期小鼠认知功能下降,其机制可能与促进海马星形胶质细胞活化、诱导神经元凋亡、加重神经炎性反应水平有关。

关键词：异氟醚吸入麻醉围术期神经认知紊乱神经认知功能星形胶质细胞

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CitGVD:a comprehensive database of citrus genomic variations

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Horticulture Research 2020年第1期7卷 2529-2536页

作者： 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 Citrus Research Institute Southwest University/Chinese Academy of Agricultural Sciences400712 ChongqingChina Key Laboratory of Plant Hormones and Development Regulation of Chongqing School of Life SciencesChongqing University401331 ChongqingChina

Citrus is one of the most important commercial fruit crops *** 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 *** this study,to permit the ease of access to these data,a web-based database,the citrus genomic variation database(CitGVD,http://***/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 *** 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 *** 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 *** features make CitGVD a comprehensive web portal and bioinformatics platform for citrus-related *** also provides a model for analyzing genome-wide variations for a wide range of crop varieties.

关键词： database analysis comprehensive

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Riverine sulfate sources and behaviors in arid environment,Northwest China:Constraints from sulfur and oxygen isotopes

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Journal of Environmental Sciences 2024年第3期137卷 716-731页

作者： Yifu xu Wenjing Liu Bing xu Zhifang xu Key Laboratory of Cenozoic Geology and Environment Institute of Geology and GeophysicsChinese Academy of SciencesBeijing 100029China Innovation Academy for Earth Science Chinese Academy of SciencesBeijing 100029China University of Chinese Academy of Sciences Beijing 100049China

The fate of riverine sulfate ion (SO_(4)^(2-)) and its environmental effects in arid environment are difficult to evaluate due to its complicated sources and strongly coupled behaviors with water cycle which is significantly modified by *** understand the sulfur cycle in aquatic systems in arid environment,the chemical and sulfur and oxygen isotopic compositions (δ^(34)S_(SO4)and δ^(18)O_(SO4)) of major rivers around the Badain Jaran Desert,northwestern China,were *** rivers had averaged SO_(4)^(2-)content at 1336μmol/L,over 10times higher than the global *** δ^(34)S_(SO4)and δ^(18)O_(SO4)values ranged from-5.3‰to+11.8‰and+1.6‰to+12.8‰,*** end-member analysis and the inverse model showed that riverine sulfate was mainly derived from evaporites dissolution (0-87%),sulfide oxidation (13%-100%) and precipitation (0-33%),indicating heterogeneity in sulfur sources and behaviors along the river drainage with the lithology variations and climate *** isotopic tools combining with hydro-chemistry compositions could be applied to reveal sulfur cycle in arid *** on the calculation,sulfide oxidation plays the primary role in the headwater and upstream in the Qilian-Mountains area,where sulfide is widely *** the proportion of evaporites dissolution contributing to riverine sulfate is much higher in downstream in a drier ***,less precipitation and higher temperature can lead to more intensive evaporation,affecting the process of sulfide oxidation and enhancing the rates of evaporites dissolution and sulfate precipitation in the basin.

关键词： Sulfur and oxygen isotopes Source apportionment Inverse model Arid environment Inland rivers Northwestern China

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Laboratory evaluation of high-friction thin overlays for pavement preservation

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Frontiers of Structural and Civil Engineering 2024年第6期18卷 936-948页

作者： Ouming xu Rentao xu Lintong JIN School of Highway Chang’an UniversityXi’an 710064China School of Materials Science and Engineering Chang’an UniversityXi’an 710061China

Traditional asphalt concrete(AC)and stone matrix asphalt(SMA),which are used as thin asphalt overlays,are common maintenance strategies to enhancing ride quality,skid resistance,and ***,several studies have used a novel asphalt mixture known as a high-friction thin overlay(HFTO)to improve surface ***,it remains uncertain whether the laboratory properties of HFTO differ significantly from those of conventional *** study aims to evaluate the laboratory properties of HFTO mixtures and compare them with those of AC and *** mixtures with nominal maximum size of 9.5 mm were produced in the laboratory,and performance tests were conducted,including wheel tracking test,low temperature flexural creep test,moisture susceptibility test,Cantabro Abrasion Test,Marshall Test,sand patch test,British pendulum test,and indoor tire-rollingdown *** results showed that the HFTO exhibited a lower tire/pavement noise than the AC and ***,HFTO had superior high-temperature stability,larger macro texture,and higher skid resistance in comparison to those of AC,but lower than those of ***,HFTO mixtures may be considered a suitable replacement for traditional AC mixtures in regions where skid resistance and noise reduction are concerns.

关键词： road engineering pavement maintenance high friction thin overlay performance skid resistance tire/pavement noise

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Aldehyde oxidase mediated enantioselective metabolic health risk of dinotefuran

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Science China Chemistry 2024年第6期67卷 2079-2091页

作者： Liwei xu Xinxin xu Hua Kuang Chuanlai xu Xiaoling Wu State Key Laboratory of Food Science and Resources Jiangnan UniversityWuxi214122China International Joint Research Laboratory for Biointerface and Biodetection and School of Food Science and Technology Jiangnan UniversityWuxi214122China Collaborative Innovation center of Food Safety and Quality Control in Jiangsu Province Jiangnan UniversityWuxi214122China

Chiral pollutants often pose significant differential environmental health *** this study,the biotransformation of chiral dinotefuran(DIN)and its enantioselective metabolic toxicity mechanisms have been systemically ***,reversedphase chromatography-high resolution mass spectrometry was developed to quantify the content of DIN R/S chiral enantiomer with pg level sensitivity,revealing a lower elimination rate constant(K_(e))of S-DIN(0.730 h^(-1))than R-DIN(0.746 h^(-1)).Secondly,the interaction mechanism between DIN metabolism and important endogenous bioactive molecules,such as aldehyde oxidase(AOX)and neurotransmitters,was *** DIN nitro-group was converted into a guanidine group by the reducing site of nearby flavin adenine dinucleotide(FAD)in AOX with the preferred higher affinity of ***,the endogenous tryptophan(Trp)aldehyde metabolic intermediate,5-hydroxyindoleacetaldehyde(5-HIAL),provides a persistent electron donor for DIN reduction via the oxidation-catalyzed site in AOX,resulting in remarkable up-regulation of monoamine neurotransmitters such as serotonin and ***,the higher level of neurotransmitters further mediated dysregulation of oxylipin homeostasis via the serotonergic pathway,where S-DIN exhibited more pronounced liver lipid damage and environmental health risk with the accumulated lipid biomarkers,oxidized triglyceride(OxTG)and oxidized sphingomyelin(OxSM).This study elucidates the AOX-mediated enantioselectivity metabolic pathway of DIN,providing a new analytical method for chiral pollutants and paves the way for their health risk assessments.

关键词： neonicotinoid dinotefuran enantiomer aldehyde oxidase metabolomics health risk

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Skeletal interoception in osteoarthritis

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Bone Research 2024年第2期12卷 241-254页

作者： Dinglong Yang Jiawen xu Ke xu Peng xu Department of Joint Surgery Honghui HospitalXi’an Jiaotong UniversityXi’an 710054China Department of Orthopedic Surgery and Orthopedic Research Institute West China HospitalSichuan UniversityChengdu 610041China

The interoception maintains proper physiological conditions and metabolic homeostasis by releasing regulatory signals after perceving changes in the internal state of the *** its various forms,skeletal interoception specifically regulates the metabolic homeostasis of ***(OA)is a complex joint disorder involving cartilage,subchondral bone,and *** subchondral bone undergoes continuous remodeling to adapt to dynamic joint *** findings highlight that skeletal interoception mediated by aberrant mechanical loads contributes to pathological remodeling of the subchondral bone,resulting in subchondral bone sclerosis in *** skeletal interoception is also a potential mechanism for chronic synovial inflammation in *** this review,we offer a general overview of interoception,specifically skeletal interoception,subchondral bone microenviroment and the aberrant subchondral *** also discuss the role of skeletal interoception in abnormal subchondral bone remodeling and synovial inflammation in OA,as well as the potential prospects and challenges in exploring novel OA therapies that target skeletal interoception.

关键词： homeostasis inflammation releasing

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