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1.
番茄斑萎病毒(tomato spotted wilt orthotospovirus,TSWV)严重危害多种经济作物和园艺植物,其N基因与病毒的侵染力密切相关,将N基因直接构建到VIGS载体上研究其致病功能目前鲜见报道。本研究将TSWV N基因构建到pTRV-PTV00载体上,注射本氏烟,通过qRT-PCR定量分析发现先接种TSWV后注射沉默载体的植株抗TSWV效率达57.60%,先注射沉默载体后接种TSWV的植株抗TSWV效率达99.14%。结果表明先注射载体后接种TSWV的N基因沉默效率更高。TSWV N基因VIGS载体的构建可为研究N基因在病毒致病等方面的功能提供前期材料,并为TSWV抗病育种和田间绿色防控提供一定的理论依据。 相似文献
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番茄斑萎病毒(tomato spotted wilt orthotospovirus,TSWV)严重危害多种经济作物和园艺植物,其N基因与病毒的侵染力密切相关,将N基因直接构建到VIGS载体上研究其致病功能目前鲜见报道。本研究将TSWV N基因构建到pTRV-PTV00载体上,注射本氏烟,通过qRT-PCR定量分析发现先接种TSWV后注射沉默载体的植株抗TSWV效率达57.60%,先注射沉默载体后接种TSWV的植株抗TSWV效率达99.14%。结果表明先注射载体后接种TSWV的N基因沉默效率更高。TSWV N基因VIGS载体的构建可为研究N基因在病毒致病等方面的功能提供前期材料,并为TSWV抗病育种和田间绿色防控提供一定的理论依据。 相似文献
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基于Real-time PCR监测番茄黄化曲叶病毒在番茄植株中的动态变化 总被引：1，自引：0，他引：1
本研究根据番茄黄化曲叶病毒(Tomato yellow leaf curl virus, TYLCV)分离物-SH2基因组序列,设计了一对引物,以番茄25S rRNA基因为内参,建立了番茄黄化曲叶病毒SYBR GreenⅠ实时荧光定量PCR检测方法。该方法可检测到浓度为4.64×10-7 ng/μL的植物DNA中含有TYLCV,其灵敏度是常规PCR的1 000倍。利用该方法,研究了温室条件下以侵染性克隆接种TYLCV后的番茄植株中病毒DNA含量的变化情况。根、茎、叶中病毒DNA定量检测结果表明,TYLCV在3种植物器官中都呈现一个上升、稳定和下降的变化规律;病毒DNA在植株根部最早累积,累积的速度较慢,在叶部和茎部累积较快;叶部和茎部接种18 d后病毒DNA含量达到稳定期;在不同的器官中,病毒的含量不同,在茎部的含量最高,接种33 d后茎部病毒DNA的含量约为根部的100倍。本研究通过对TYLCV含量的动态监测,明确了病毒DNA在番茄植株中的累积和变化规律,为研究TYLCV侵染机制、病毒与寄主互作及病害防治提供了理论依据。 相似文献
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有研究表明烟草花叶病毒(Tobacco mosaic virus, TMV)或黄瓜花叶病毒(Cucumber mosaic virus, CMV)侵染烟草能够激活转录因子NbNAC089,从ER膜移至细胞核。为进一步阐释内质网应激因子NbNAC089对病毒侵染胁迫的响应机制,利用CRISPR/Cas9技术构建敲除载体,经烟草遗传转染获得NbNAC089基因突变植株。植株接种病毒后采用qRT-PCR检测病毒CP基因和寄主UPR基因的表达。结果表明:CRISPR/Cas9系统定点敲除NbNAC089基因后,目的基因靶位点序列有碱基的置换与缺失。正常生长条件下,转基因植株与野生型无差异。植株接种TMV-GFP后24~96 h,突变体中UPR基因(BiP、bZIP28、bZIP60)的表达量显著高于野生型;接种TMV-GFP后2~6 d突变体中病毒的积累量和扩展速度显著高于野生型。表明NbNAC089为UPR的抑制因子,对病毒增殖具有负调控作用。 相似文献
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番茄黄化曲叶病毒(tomato yellow leaf curl virus, TYLCV)是一种由烟粉虱传播的单链环状DNA病毒, 在田间可与多种病毒发生复合侵染, 如番茄褪绿病毒(tomato chlorosis virus, ToCV)等?本文对比了TYLCV单独侵染和TYLCV与ToCV复合侵染对烟粉虱获取和传播TYLCV的影响?结果表明, 与取食TYLCV单独侵染的番茄相比, 取食复合侵染番茄的烟粉虱对TYLCV的传毒率显著提高, 且番茄植株和烟粉虱体内TYLCV的病毒积累量也显著提高?试验结果说明复合侵染会提高烟粉虱的传毒率, 促进TYLCV的发生与流行? 相似文献
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近年来, 番茄病毒病, 特别是番茄黄化曲叶病毒（TYLCV）病在北京地区空前暴发, 给番茄生产造成严重威胁, 使番茄的产量和品质显著降低。2012-2013年在北京周边7个区县, 采集疑似感染病毒的番茄植株样品325份, 分别针对TYLCV、烟草花叶病毒（TMV）、黄瓜花叶病毒（CMV）3种病毒进行了PCR或ELISA 检测。检测结果表明, 目前在北京地区大棚或温室内发生的番茄病毒病以TYLCV为主; 露地番茄病毒复合侵染现象比较普遍, 病毒检出率100%, 其中TYLCV检出率达到75%以上。在TYLCV侵染的样品中, TYLCV和CMV复合侵染占20%左右, TYLCV和TMV复合侵染占15%左右。部分样品检测到TYLCV、CMV 和TMV 3种病毒复合侵染的现象。 相似文献
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钙依赖磷酸酶(Calcineurin)作为Ca2+信号传感器中继因子对受Ca2+调节的生物学过程起重要调控作用。本研究克隆了一个番茄全长cDNA序列,该序列与钙依赖磷酸酶催化亚基(Calcineurin A,CNA)序列同源。推定的蛋白产物由315个氨基酸组成,明显短于非植物来源的该类蛋白。该蛋白含有钙依赖磷酸酶类金属磷酸酶中保守的催化作用活性位点基序,以及两个潜在的钙调蛋白结合位点,故暂时命名为LeCAL1(Lycopersicon esculentum calcineurin A like 1)。对CNA序列的分析结果表明,植物与非植物CNA差异明显,在系统进化树中形成距离很远的两大分支。植物CNA中,LeCAL1与拟南芥CNA同源性较高,而与水稻CNA较低。对LeCAL1在番茄中的表达分析结果显示,Cf/Avr介导的过敏性反应的产生诱导LeCAL1基因的表达,表明钙依赖磷酸酶可能在番茄抗叶霉病中起调节作用。 相似文献
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为明确烟粉虱传播的番茄褪绿病毒(Tomato chlorosis virus,ToCV)与番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)对不同番茄品种的复合侵染情况,于2015年11月在山东省寿光市温室内采集13个番茄品种共390份疑似发病植株叶片,对不同番茄品种的TYLCV抗性和2种病毒的复合侵染以及温室内发病番茄植株上烟粉虱成虫的带毒率进行检测。结果表明,采集的13个番茄品种经分子标记检测鉴定均为TYLCV杂合抗性;不同番茄品种ToCV与TYLCV的复合侵染率存在明显差异,大果番茄粉宴和贝瑞上复合侵染率最高可达73.3%,而樱桃番茄八喜上未检测到这2种病毒的复合侵染。此外,在发病番茄植株上采集的烟粉虱成虫体内可检测到2种病毒,其中烟粉虱ToCV带毒率为90.7%,TYLCV带毒率为80.0%,同时检测到ToCV与TYLCV的概率为71.3%。表明ToCV和TYLCV的复合侵染在山东省番茄生产中普遍发生,烟粉虱可同时携带这2种病毒并广泛传播。 相似文献
9.
番茄褪绿病毒Tomato chlorosis virus(ToCV)是一种由烟粉虱Bemisia tabaci传播的正义单链RNA病毒,在田间常与番茄黄化曲叶病毒Tomato yellow leaf curl virus(TYLCV)复合侵染而造成番茄生产上重大的经济损失。为了明确ToCV与TYLCV的复合侵染对烟粉虱传播ToCV所造成的影响,本文采用RT-PCR以及qRT-PCR检测了复合侵染的番茄对烟粉虱获取和传播ToCV的影响。研究表明,烟粉虱取食复合侵染的番茄后对ToCV的传播效率显著提高,仅25头烟粉虱的传毒率即可达到100%,ToCV在烟粉虱以及番茄体内的累积量均显著提高。说明这种复合侵染促进了烟粉虱对ToCV的传播,在田间应当及时防控烟粉虱,警惕病毒与烟粉虱的蔓延。 相似文献
10.
尖孢镰刀菌番茄专化型中SNARE蛋白FolSso1的功能分析 总被引：1，自引：0，他引：1
SNARE(Soluble N-ethylmaleimide-sensitive factor attachment protein receptor)蛋白保守存在于丝状真菌中,在膜泡转运的过程中起着关键的作用。番茄枯萎病是由尖孢镰刀菌番茄专化型(Fusarium oxysporum f. sp. lycopersici,Fol)引起的,严重威胁着番茄的生产。我们使用反向遗传学的方法来研究番茄枯萎病菌中SNARE蛋白FolSso1的功能,实验结果发现FolSSO1的基因缺失突变体菌丝生长速率降低,且产孢数量减少。另外,FolSSO1基因的缺失导致突变体相较于野生型菌株对细胞壁压力与细胞膜压力更加敏感。然后,在番茄果实和番茄植株的致病性实验中,我们发现FolSSO1的缺失并没有引起Fol致病性显著的变化。综上所述,本研究发现FolSSO1可以调控Fol营养生长,繁殖和对环境压力的响应过程,然而对Fol的致病过程并没有显著的调控作用。 相似文献
11.
广东番茄上检测到Tospovirus病毒 总被引：1，自引：0，他引：1
Some tomato samples possibly infected by tospovirus in Guangdong were detected with indirect ELISA and RT-PCR. The results showed that the virus infected tomato did not react with the antiserum of Tomato spotted wilt virus (TSWV), but about 500 bp fragment of RT-PCR shared 83%-84% nucleotide identities with N gene of those reported tospoviruses. The phylogenetic tree of the N gene fragment compared with those of other tospoviruses indicated that the virus infected tomato was belonged to Tospovirus. 相似文献
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Gil Atzmon Hadassa van Oss Henryk Czosnek 《European journal of plant pathology / European Foundation for Plant Pathology》1998,104(2):189-194
DNA of tomato yellow leaf curl virus (TYLCV), a geminivirus transmitted by the whitefly Bemisia tabaci, was amplified from squashes of infected tomato plants and of viruliferous vectors using the polymerase chain reaction (PCR). Samples of infected tissues as small as 1 mm2 were squashed onto a nylon membrane. A 1 × 2 mm strip containing the squash was introduced into a 25 µl PCR reaction mix. The reaction products were subjected to gel electrophoresis, blotted and hybridized with a radiolabeled virus-specific DNA probe. TYLCV DNA was amplified from squashes of leaves, roots, and stem of infected tomato and from individual viruliferous whiteflies. The same squash could be used several times to amplify different virus DNA fragments with various sets of primers. Thus plant and insect squashes can be used as templates for the amplification of geminiviral DNA with no need to prepare tissue extracts or purify nucleic acids. The squash-PCR procedure was applied to study whitefly transmission of TYLCV. Tomato plants were inoculated by placing a single viruliferous insect in the center of a young leaflet. In some plants TYLCV DNA was detected at the site of inoculation as early as 5 min after the beginning of the access feeding and in all plants after 30 min. The squash-PCR procedure also was applied to the study of TYLCV acquisition by the insect vector. TYLCV DNA was detected in the head of whiteflies as early as 5 min after the beginning of the access feeding on infected tomato plants. Viral DNA was detected in the thorax after 10 min and in the abdomen after 25 min. 相似文献
13.
M. R. HAJIMORAD A. KHEYR-POUR V. ALAVI A. AHOONMANESH M. BAHAR M. A. REZAIAN & B. GRONENBORN 《Plant pathology》1995,45(3):418-425
A virus causing a disease of tomato, prevalent in the southern provinces of Iran, with symptoms of leaf-curling, stunting, reduction of leaf size, leaf corrugation, shortening of internodes and severe reduction in fruit yield, was shown to be transmissible to healthy tomato plants by grafting and by whiteflies ( Bemisia tabaci ), but not by sap inoculation. Geminivirus DNA was detected in extracts of diseased tomato plants by dot-blot hybridization assays using as probes full-length cloned DNA of Australian, Italian (Sardinian) or Jordanian strains of tomato yellow leaf curl virus (TYLCV). Geminivirus coat protein was detected in whitefly inoculated plants by dot immunobinding assay using polyclonal antibody raised against Jordanian TYLCV. A limited survey using the dot-blot hybridization assay for virus detection indicated the presence of the virus in tomato-growing provinces of southern but not northern Iran. Whitefly transmission experiments to tomato under controlled greenhouse conditions showed that some isolates of TYLCV-like geminiviruses from different parts of Iran differ in symptomatology. 相似文献
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In recent years since 2018, the disease of tomato fruit rot has been often noted in Jiangxi province. In order to ascertain the causal agent, common tissue isolation method was used to isolate the pathogen collected from 8 counties and cities of Jiangxi province. A total of 17 isolates was obtained, which exhibited similar phenotype on V8 agar plates with production of antheridia, oogonia and oospore indicating the characteristics of Phytophthora spp.. The pathogenicity test for the isolates showed the similar disease symptoms with that in the field and the pathogen was reisolated from the infected tomato tissues, which fulfilled the Koch’s postulate. BLAST search with rDNA-ITS, partial Ypt1 and β-tubulin gene sequences for 17 isolates showed 99%-100% of identities to Phytophthora capsici that in correspond with the clustering result of phylogenetic analysis for two represented strains. Combined with morphologic characteristic observation, pathogenicity test and sequence ana-lysis, the pathogen causing tomato fruit rot was identified as Phytophthora capsici. This is the first report of P. capsici causing fruit rot on tomato in Jiangxi province, China. 相似文献
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2012年,江苏南京菜豆上出现了一种新的病毒病害,病株表现明显的叶片皱缩、植株矮化等症状。根据其症状及介体发生状况,对其伴随的病毒种类进行了研究,结果从中检测到一种粉虱传双生病毒,对其基因组DNA-A组分克隆测序后发现其全长2 781 bp,编码6个ORF,BLAST及聚类分析结果显示该病毒与番茄黄化曲叶病毒同源性最高(99%),是番茄黄化曲叶病毒的一个分离物。这是江苏省番茄黄化曲叶病毒侵染菜豆的首次报道,暗示番茄黄化曲叶病毒可能是我国菜豆种植的重要潜在威胁。 相似文献
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M. Lapidot O. Goldray R. Ben-Joseph S. Cohen M. Friedmann A. Shlomo S. Nahon L. Chen M. Pilowsky 《EPPO Bulletin》2000,30(2):317-321
Tomato yellow leaf curl begomovirus (TYLCV) can be devastating to tomato crops in tropical and subtropical regions. The development of resistant cultivars is the best option for the control of TYLCV. The TYLCV-resistance level of a new breeding line, TY172, alongside that of commercial cultivars known to be resistant to the virus, was evaluated in a field test by comparing the yield performance of inoculated plants with that of uninoculated plants of the same line or cultivar. There were substantial differences among the different entries tested in the extent of yield loss relative to the corresponding uninoculated control plants. This comparison between inoculated and uninoculated plants of the same entry provides a quantitative assay for resistance level. All resistant commercial cultivars tested developed different levels of disease symptoms. Only line TY172 showed no symptoms of the disease. A low level of viral DNA was detected in infected TY172, showing that it is a symptomless carrier of TYLCV. When TY172 was crossed with susceptible lines, the hybrids exhibited milder symptoms than the susceptible parent, yet higher than that of TY172, suggesting a partial dominance for TY172 resistance. Upon inoculation of F2 populations, the amount of symptomless individuals appeared in a ratio approximating 7:64. This suggests that at least three genes appear to account for the resistance. 相似文献