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杧果细菌性黑斑病菌hpaXcm基因克隆及其编码蛋白的功能研究

杧果细菌性黑斑病菌hpaXcm基因克隆及其编码蛋白的功能研究

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中国植物病理学会第十一届全国会员代表大会暨2018年学...

作者：周晓韵刘悦黄佳敏刘清桓孙建章蔡新峰刘文波缪卫国海南大学植物保护学院/海南大学热带作物种质资源保护与开发利用教育部重点实验室

杧果细菌性黑斑病是是由野油菜黄单胞杧果致病变种Xanthomonas campestris ***(Xcm)引起,2009年更名为Xanthomonas citri ***(Xcm)。本研究利用PCR技术从Xcm基因组中首次扩增到了hpaXcm基因,该基因具有405bp,其编码的HpaXcm蛋白含有134... 详细信息

杧果细菌性黑斑病是是由野油菜黄单胞杧果致病变种Xanthomonas campestris ***(Xcm)引起,2009年更名为Xanthomonas citri ***(Xcm)。本研究利用PCR技术从Xcm基因组中首次扩增到了hpaXcm基因,该基因具有405bp,其编码的HpaXcm蛋白含有134个氨基酸,大小为13.56kD,GC含量为60.49%,富含甘氨酸,缺乏半胱氨酸。利用GST基因操作系统获得hpaXcm的基因产物HpaXcm,经研究发现,HpaXcm与以前报道的harpin类物质具有相同的性质,是一种新的harpin蛋白。HpaXcm蛋白的最佳诱导条件为IPTG浓度0.1mM,28℃诱导5h。为了进一步对HpaXcm的特性进行研究,对诱导表达的GST融合蛋白进行纯化和酶切,并采用Western blot获得与理论大小相符的HpaXcm蛋白,为进一步研究蛋白功能提供了更为有利的基础。以纯化的HpaXcm与经过煮沸处理的HpaXcm为材料处理烟草叶片,结果表明:无论是否经过煮沸处理,HpaXcm都能够激发烟草过敏性反应并产生明显的坏死斑,并且煮沸处理的HpaXcm诱导产生的坏死斑面积更大;纯化的HpaXcm与经过煮沸处理的HpaXcm都能增强烟草对TMV抗病性,并且喷施了经煮沸处理的HpaXcm的叶片诱导的TMV抗性更强。通过qRT-PCR对HR标志基因hinl、hsr203J和防卫基因chia5、PR-1a、PR-1b、NPR1进行检测,结果表明:HpaXcm能够激发hin1、hsr203J、NPR1的表达,并且煮沸后的HpaXcm所激发的表达量更高,但出现表达量最高峰的时间与未经煮沸处理的HpaXcm有差异,结合以上烟草表型实验推测,煮沸后的HpaXcm激发了不同的信号通路。本研究对首次报道的HpaXcm蛋白特性和功能进行探索,为比较黄单胞菌harpin类蛋白的结构和功能关系提供了基础依据,从而有助于新型微生物蛋白农药的研发。

关键词：杧果细菌性黑斑病菌 HpaXcm 过敏性反应诱导抗病性 qRT-PCR

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Development and application of specific SSR primers for Malus sieversii and cultivated apple

Development and application of specific SSR primers for Malu...

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中国植物病理学会第十一届全国会员代表大会暨2018年学...

作者： Yu Shaoshuai Zhao Wenxia Yao Yanxia Huai Wenxia Xiao Wenfa Key Laboratory of Forest Protection of State Forestry Administration Research Institute of Forest EcologyEnvironment and ProtectionChinese Academy of Forestry

TheSSR primers with specific,stable and polymorphic genes were designed and screened with the encoding gene sequences of Xinjiang wild apple(Malus sieversii) or cultivated apple resistance related proteins according to the complete genome sequencing of the completed genome of the cultivated *** newly developedSSR primers were screened and detected by PCR amplification and gene *** specific primers could be used to amplify specific,stable and polymorphicSSR bands in *** and cultivated apple,and 8 specific primers ofSSR could only amplifySSR stripe specifically in cultivated apples.290 samples of *** and cultivated apple were amplified and total 69 alleles were obtained using the 15 pairs of specificSSR *** is a close genetic relationship between *** and cultivated *** genetic diversity of *** is rich with a genetic differentiation coefficient of 0.248 6 and a gene flow of 0.755 *** *** from Xinyuan and Gongliu,is clustered in an evolutionary *** cultivated apple and the red meat apple collected from Xinjiang are clustered in an evolutionary *** number of alleles amplified by differentSSR loci was distinct and the frequencies of different alleles in the same microsatellite loci in different *** or cultivated apple groups were significantly different.15 pairs of specificSSR primers revealed the genetic diversity and phylogenetic relationship of *** and cultivated apples and the possible effects of genetic variation on the biological functions of *** and cultivated apple,exploring the genetic variation associated with resistance to ***.

关键词： Malus sieversii Microsatellite markers Genetic variation Stress resistance

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Latent infection of Valsa mali on crabapple and apple seeds implies multiple inoculum sources for Valsa canker

Latent infection of Valsa mali on crabapple and apple seeds ...

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中国植物病理学会第十一届全国会员代表大会暨2018年学...

作者：孟祥龙 Qi Xinghua Guo Yongbin Wang Yanan Hu Tongle 曹克强王树桐 College of Plant Protection Hebei Agricultural University

A real-time quantitative PCR assay with a pair of species-specific primers was developed to fast and accurately detect and quantify Valsa mali in crabapple seeds,crabapple seedlings,apple twigs and apple seeds,which is the pathogen of apple valsa *** seeds collected from different regions with 12.87-49.01%were positive for *** detection,and exopleura and endopleura were two major infection sites in crabapple *** fact of infection of crabapple seeds was also confirmed by high-throughput sequencing *** the growth of crabapple seedlings,the concentration of *** in crabapple seedlings generally increased,until eight or more leaf blades *** one-year-old twigs from apple scion nursery can be infected by ***,and only those apple seeds from infected apple trees with obvious valsa canker symptom carried *** conclusion,this study report that crabapple seeds and apple seeds carried *** as latent inoculum *** infected not only apple tissues but also seedlings of crabapple,which is the stock of apple *** study indicated that the inoculum sources for Valsa canker are *** of the novel developed realtime PCR assay may potentially improve the disease management of apple Valsa canker.

关键词： Apple valsa canker(AVC) Valsa mali Real-time quantitative PCR Seeds detection Sources of infection

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Genetic variability and molecular evolution of the untranslated regions of Potato virus Y

Genetic variability and molecular evolution of the untransla...

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中国植物病理学会第十一届全国会员代表大会暨2018年学...

作者： Oswald Nkurikiyimfura Shen Linlin Linda Iradukunda Xie Jiahui Wang Yanping Zhan Jiasui Fujian Key Laboratory of Plant Virology Institute of Plant VirologyFujian Agricultural and Forestry University

Isolates of Potato virus Y(PVY) differ widely in their biological *** properties may depend on the structure of viral RNA populations comprising the different *** a first approach to study the molecular basis of the biological variability,we have compared the sequences of multiple cDNA clones of the two terminal regions of the RNA from different PVY *** this study,the Genetic variability and molecular evolution of PVY in their untranslated region were studied using 270 sequences generated from isolates collected from potato in China and 148 PVY sequences selected from "GeneBank" which classification,country and continent origins have been *** genetic differentiation was found in the populations from the different countriesgrouped by continent,with higher nucleotide diversity in Europe thanAmerica,Asia and the isolates collected in *** demonstrated that although there is high variability at the 5'-UTR among members of the 4 major groups of PVY isolates,this region is mainly conserved among individual members within each group andat the 3'-UTR were more diverse and variable over the host species and the regions where they were *** analyses showed that spatial genetic structure in the untranslated region of PVY populations was likely caused by demographic dynamics of the pathogen and natural selection generated by habitat *** from all groups tended to group together,indicating that nucleotides diversification was maintained,in part,by host-driven adaptation.

关键词： Potato virus Y,Variation Evolution,3'-UTR,5'-UTR

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香蕉枯萎病菌在不同酸度和有机质含量土壤中的变化分析

香蕉枯萎病菌在不同酸度和有机质含量土壤中的变化分析

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中国植物病理学会第十一届全国会员代表大会暨2018年学...

作者：刘继琨饶雪琴李华平华南农业大学

香蕉枯萎病是由尖孢镰刀菌古巴专化型(Fusarium oxysporum ***,Foc)侵染引起的香蕉上的一种毁灭性病害,目前对病菌在土壤中的流行学研究十分有限。该研究通过配制不同酸度(pH值=4.5,pH值=5.5,pH值=6.5,pH值=7.5和pH值=8.5)在灭菌和非灭... 详细信息

香蕉枯萎病是由尖孢镰刀菌古巴专化型(Fusarium oxysporum ***,Foc)侵染引起的香蕉上的一种毁灭性病害,目前对病菌在土壤中的流行学研究十分有限。该研究通过配制不同酸度(pH值=4.5,pH值=5.5,pH值=6.5,pH值=7.5和pH值=8.5)在灭菌和非灭菌条件下和不同有机质含量(16g/kg,32g/kg,64g/kg,96g/kg腐殖酸钾)的蕉园土壤,通过人工接种Foc 4号生理小种(Foc4),定期观察Foc4在不同土壤中的生长和繁殖情况。结果表明,在不同酸度处理中,Foc4的含量,均是随着pH的增大而呈减小趋势,其中在pH值=7.5以上的处理中明显低于pH值=7.5以下的3个酸度的处理;在不同酸度条件下,Foc4的含量在灭菌处理中均显著高于非灭菌处理。在不同有机质含量的土壤中,Foc4的含量随腐殖酸钾含量的增大呈减小趋势,其中在32g/kg以上的3个腐殖酸钾处理中明显低于16g/kg处理。利用平板计数方法分析和比较不同处理类型土壤中各类微生物的多样性和群落结构差异,结果表明,可培养的细菌和放线菌的数量在pH处理组中大致随着pH的增大而增大,在腐殖酸钾处理组中随着腐殖酸钾含量的升高而增多。利用激光共聚焦显微镜对不同处理土壤中Foc4进行观察,结果发现Foc4在灭菌处理土壤的中可观察到的Foc4孢子要明显多于非灭菌处理与有机质处理的土壤,在pH值=4.5处理中的视野内孢子含量最高;在非灭菌pH处理与有机质组合处理中,在pH值=7.5下,腐殖酸钾含量在64g/kg时,Foc4的孢子含量最低;在病菌接种后30d时可观察到厚垣孢子,并随着时间增加,其分生孢子数逐渐减少,而厚垣孢子逐渐增多。进一步的Foc4在不同处理土壤中的变化规律、以及土壤中各类微生物的多样性和群落结构差异研究工作正在进行中。

关键词：香蕉香蕉枯萎病菌 pH值土壤有机质含量土壤微生物群落

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Transcription factor SsSte12 is involved in Sclerotia development and appressoria production in Sclerotinia sclerotiorum

Transcription factor SsSte12 is involved in Sclerotia develo...

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中国植物病理学会第十一届全国会员代表大会暨2018年学...

作者： Lu Dongxu Liu Fengwen Sun Rongze Wang Kangning Zhang Bowen Zhang Yanhua Pan Hongyu College of Plant Sciences Jilin University

Sclerotinia sclerotiorum is a challenging agricultural pathogen for management,causing large global economic losses *** sclerotia and infection cushions are critical for its long-term survival and successful penetration on a wide spectrum of *** mitogen-activated protein kinase(MAPK) cascades serve as central signaling complexes that are involved in various aspects of sclerotia development and *** this study,the putative downstream transcription factor of MAPK pathway,SsSte12,was analyzed in *** SsSte12 in *** resulted in phenotypes of delayed vegetative growth,reduced size of sclerotia,and fewer appressoria ***,the SsSte12 RNAi mutants showed attenuated pathogenicity on the host plants due to the defect compound *** two-hybrid(Y2H) and bimolecular fluorescence complementation(BiFC) assays demonstrated that the SsSte12 interacts with ***,the SsMcm1 expression is independent of the regulation of SsSte12 as revealed by qRT-PCR analysis in SsSte12 RNAi *** with high accumulation of SsSte12 transcripts in the early development of *** results demonstrated that SsSte12 function was essential in the vegetative mycelial growth,sclerotia development,appressoria formation and penetration-dependent ***,the SsSte12-SsMcm1 interaction might play a critical role in the regulation of the genes encoding these traits in ***.

关键词： SsSte12 Sclerotia Appressoria Pathogenicity SsMCM1

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VmE02 from Valsa mali is a new pathogen-associated molecular pattern

VmE02 from Valsa mali is a new pathogen-associated molecular...

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中国植物病理学会第十一届全国会员代表大会暨2018年学...

作者： Nie Jiajun Yin Zhiyuan Li Zhengpeng Wu Yuxing Huang Lili State Key Laboratory of Crop Stress Biology for Arid Areas College of Plant ProtectionNorthwest A&F University

The necrotrophic fungus Valsa mali causes severe necrosis on apple trees,resulting in substantial yield losses in eastern *** the aim to identify cell death-inducing factors of ***,we carried out transient expression of candidate effector proteins in Nicotiana benthamiana,and VmE02,a protein with unknown function was *** analysis revealed that VmE02 is widely spread in fdamentous *** agroinfiltration,we found there exists homologs of VmE02 in both fungi and oomycetes,which can also elicit cell death in *** of VmE02 in *** activated plant immune responses,including ROS burst,callose deposition,and the expression of both salicylic acid and jasmonate signaling ***,VmE02 induced the expression of PTI marker *** virus-induced gene silencing(VIGS) in ***,we found that cell deathinducing activity of VmE02 requires both Brassinosteroid insensitive 1-Associated Kinase 1(NbBAK1),suppressor of BIR1-1(NbSOBIR1),HSP90,and *** out of VmE02 in *** showed no apparent influence on pathogenicity,however,it considerably reduced pathogen ***,VmE02 manipulated resistance to Sclerotinia sclerotiorum and Phytophthora capsici when expressed in *** conclude that VmE02 is a novel PAMP in fungi and oomycetes,and suggest it is also an effector with diverse function.

关键词： Valsamali Cell death Plant immunity Nicotiana benthamiana Filamentous pathogens Pathogen-associated molecular pattern

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Evaluating the role of chloroplasts targeting non-classically secreted effectors in the development of rice blast disease

Evaluating the role of chloroplasts targeting non-classicall...

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中国植物病理学会第十一届全国会员代表大会暨2018年学...

作者： Ammarah Shabbir Lin Lili Chen Xiaomin Justice Norvienyeku Wang Zonghua State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops College of Plant ProtectionFujian Agriculture and Forestry University Fujian Province Key Laboratory of Pathogenic Fungi and Mycotoxins College of Life SciencesFujian Agriculture and Forestry University Minjiang University

Most of the plant pathogens modulate host immunity by secreting effector proteins consist of a signal peptide that directs them into different host compartments like the nucleus,chloroplasts,and mitochondria,*** ER/Golgi pathway depending upon the signal ***,some effector proteins lack an N-terminal signal peptide yet they are secreted,termed as leaderless secretory *** literature elucidates different non-classically secreted proteins in *** studies seek to identify their homologs in the rice blast fungus,Pyricularia oryzae,which has become a serious threat to rice production *** study aimed at identifying non-classically secreted effectors that specifically target host chloroplasts and evaluate their contributions to the pathogenicity and virulence of the rice blast *** chloroplasts besides serving as a site for photosynthesis also functions as an essential regulator of plant immune response during biotic and abiotic stress conditions,hence,we firmly believed that the identification of unconventionally secreted proteins in *** that specifically target host chloroplast during host-pathogen would tremendously enhance our knowledge on how the rice blast deploys these non-classically proteins in manipulating host immunity by bleaching the integrity of chloroplasts in invaded cells.

关键词： ER/Golgi pathway Chloroplast Non-classical effectors P.oryzae Immunity

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Biological characteristics of the pathogen Phoma segeticola *** causing a new tea foliage disease

Biological characteristics of the pathogen Phoma segeticola ...

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中国植物病理学会第十一届全国会员代表大会暨2018年学...

作者： Wei Wei Ren Yafeng Li Dongxue Zhao Xiaozhen Song Baoan Chen Zhuo College of Agriculture Guizhou University State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering Guizhou University Guizhou Institute of Pomology

Tea leaf spot disease caused by Phoma segeticola *** always take place in the tea region with higher altitude of Guizhou Province during the cold *** it is lack of an effective and safe control measure,it led to a very huge loss of tea production and serious decrease in tea *** is a very significance to establish the targeting mode,screen the fungicides with higher activity,and study its action *** this study,we studied the biological characteristics of the representative strain GZSQ-4 of *** *** in *** results indicated that the strain GZSQ-4 can grow on the medium of PDA,OA and MEA,without a significant difference in the growth *** strain GZSQ-4 can produce an amount of conidia on *** was suitable for growth in 25℃ in vitro,and it had a quicker growth rate in neural-than weak acid-or mild *** strain can also grow in PDB medium with the logarithmic phase being from 36 to 120 *** nutrition can influence the growth of the strain and the formation of hyphal *** results would lay a way for the screening the fungicides with a higher activity.

关键词： Camellia sinensis Foliage disease P.segeticola var.camelliae Biological characteristics Screening fungicides

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The transmembrane proteins BcSho1 and BcSln1 regulate vegetative differentiation,pathogenicity and multiple stress tolerance in Botrytis cinerea

The transmembrane proteins BcSho1 and BcSln1 regulate vegeta...

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中国植物病理学会第十一届全国会员代表大会暨2018年学...

作者： Ren Weichao Yang Yalan Li Fengjie Zhou Mingguo Chen Changjun College of Plant Protection Nanjing Agricultural University

The high-osmolarity glycerol(HOG) signaling pathway belongs to the class of mitogenactivated protein kinase(MAPK) cascade that regulates the response of organism to various environmental *** membrane spanning proteins Sho1 and Sln1 serve as signaling sensors of HOG ***,we investigated BcSHOl and BcSLN1 in the gray mold fungus Botrytis *** gene deletion demonstrated that both BcSHO1 and BcSLN1 are important for mycelial growth,conidiation and sclerotial *** tests revealed that the virulence of the BcSHO1 and BcSLN1 double deletion mutant decreased significantly,however,the single deletion mutant of BcSHO1 or BcSLN1 makes no difference with the wild-type *** addition,the BcSHO1 and BcSLN1 double mutant exhibited resistance to osmotic *** of the defects were restored by genetic complementation of the mutants with wild-type BcSHO1 and BcSLN1 *** results indicate that BcSHO1 and BcSLN1 share some functional redundancy and play important roles in the regulation of vegetative differentiation and multiplestress tolerance of ***.

关键词： The transmembrane proteins BcSho1 and BcSln1 regulate vegetative differentiation,pathogenicity and multiple stress tolerance in Botrytis cinerea

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