Genetic aspects of polyacrylic-acid-induced resistance to tobacco mosaic virus and tobacco necrosis virus in Nicotiana plants

Among the Nicotiana tabacum cultivars and hybrids tested, polyacrylic acid (PAA) only induced resistance to tobacco mosaic virus (TMV) and tobacco necrosis virus (TNV) in the cultivars Xanthi-nc (NN) and Xanthi (nn) respectively. This varietal response to the PAA treatment could be sexually transmitted. The complete genetic analysis of the inheritance of the PAA response is reported for the first time, demonstrating that the response is a dominant character with a Mendelian segregation which occurs independently of either the N gene responsible for resistance to TMV or the PRb-protein genes. Possible relationships between the N gene, the gene for the PAA response and the PRb genes are discussed.     

烟草品种对烟草花叶病毒病和黄瓜花叶病毒病的抗性鉴定

由烟草普通花叶病毒(TMV)和烟草黄瓜花叶病毒(CMV)引致的烟草病毒病是世界烟草主产区普遍发生且危害严重的侵染性病害,每年给烟叶生产造成了重大的经济损失。本文采用温室苗期接种鉴定的方法,对16份烟草种质进行了TMV和CMV的抗病性鉴定。结果表明:不同的烟草品种对TMV和CMV的抗病性存在较大差异。在供试种质中,对TMV表现免疫的有‘牛耳烟’、‘8301’、‘台烟7号’、‘三生-NN’共4份材料;表现抗病的有‘吉烟5号’、‘双抗70’、‘大护脖香’、‘秦烟95’共4份材料;表现中抗的有‘铁把子’、‘中烟15’、‘秦烟98’、‘中烟98’共4份材料;表现中感的有‘NC89’、‘翠碧1号’、‘云烟97’共3份材料;表现感病的只有‘秦烟97’。对CMV表现中抗的材料有1份,是‘铁把子’;表现中感的有‘秦烟95’、‘三生-NN’、‘8301’、‘牛耳烟’、‘翠碧1号’共5份材料;表现感病的有‘秦烟98’、‘云烟97’、‘中烟98’、‘NC89’、‘大护脖香’、‘双抗70’、‘秦烟97’、‘中烟15’、‘台烟7号’、‘吉烟5号’共10份材料。研究发现,‘铁把子’是兼抗这两种病毒病的材料。本研究明确了我国16个烟草品种资源的抗病性水平,为抗耐病品种的利用与品种合理布局提供科学依据,同时为烟草抗病毒病育种的亲本选择提供抗源信息。     

Systemic infection of some N-gene-carrying Nicotiana species and cultivars after inoculation with tobacco mosaic virus

Since July 1974Nicotiana tabacum ‘Samsun NN’ plants, inoculated with the common strain of tobacco mosaic virus (TMV), have occasionally been found to develop necrosis on non-inoculated upper leaves 2–7 days after the local necrotic lesions had appeared on the lower leaves. All these plants had been kept in a growth chamber at 17–20°C. Other tobacco species and cultivars carrying theN gene, such asN. glutinosa andN. tabacum ‘Xanthi-nc’, showed the same phenomenon. Substantial amounts of TMV could be recovered from leaves with systemic symptoms. The systemic necrosis somewhat resembled that caused by tobacco rattle virus (TRV). A number of possible causes, such as high concentration of the inoculum, contamination with another strain of TMV or with TRV, change in the genetic composition of the host plants and certain growing conditions (soil, water, pesticides) were investigated. None of these factors could be held fully responsible for the abnormal systemic reaction, although there was evidence that the soil could sometimes play an important rôle.     

激活蛋白PeaT1诱导烟草对TMV的系统抗性

枯斑三生烟草(Samsun-NN)经激活蛋白PeaT1诱导后接种烟草花叶病毒(Tobacco mosaic virus,TMV),对TMV产生了明显的系统获得抗性,枯斑抑制率达54.15%,枯斑大小也受到一定程度的限制。研究结果表明,PeaT1处理烟草植株下位三片叶不同时间后,其上部叶片中PPO、POD和PAL 3种抗病防御酶活性均比对照提高,第4 d酶活性达到最高值。实时荧光定量PCR检测结果显示,经PeaT1诱导4 d后烟草叶片中抗病相关基因PR1a、PR1b、NPR1在转录表达水平上较未诱导对照都有不同程度的上调。由以上结果我们推测PeaT1诱导烟草产生了系统获得抗性,本研究为进一步阐明PeaT1诱导植物抗病的信号传导途径奠定了基础。     

Reactions on Nicotiana species to potato viruses A,X and Y and tobacco mosaic virus in relation to their taxonomy and geographical origin

When testing the response of species of the genusNicotiana to 14 isolates of potato viruses A (PVA), X (PVX) and Y (PVY) and tobacco mosaic virus (TMV), sections and section parts of the genus could be divided into five groups according to the overall reaction of their species. Species from arid regions of australia and belonging to the sectionSuaveolentes (subgenusPetunioides) were most sensitive and least resistant, whereas the sectionsPaniculatae (subgenusRustica),Tomentosae (Tabacum) andNoctiflorae (Petunioides) appeared least sensitive and most resistant. Sixty-one percent of the accessions of the latter sections proved resistant to at least two of the viruses. The most resistant species have their main geographical distribution in the central Andes (southern Peru, Bolivia and north-western Argentina), where the viruses may have originated. One other resistant species wasN. africana (Suaveolentes) indigenous to south-western Africa. The most sensitive American sections, viz.Genuinae (Tabacum) andTrigonophyllae, Alatae, Acuminatae andBigelovianae (Petunioides), were generally more sensitive than species of theSuaveolentes section from outside the arid regions of Australia. The remaining part of the genus, viz. the American sectionsThyrsiflorae andRusticae (Rustica) andUndulatae, Repandea andNudicaules (Petunioides) was intermediate between the latter group ofSuaveolentes species and the resistant group of sections with regard to sensitivity and resistance, but had a high rate of tolerance to PVA, PVX and PVY and of hypersensitivity-associated resistance to TMV. The results indicate that the viral respons in the genusNicotiana is mainly determined geographically and to a lesser extent taxonomically.Samenvatting Na inoculatie van een grote collectieNicotiana-soorten met 14 isolaten van de aardappelvirussen A (PVA), X (PVX) en Y (PVY) en tabaksmozaïekvirus (TMV), konden de secties of sectiedelen van het genus op basis van de gemiddelde reactie van hun soorten in vijf groepen worden verdeeld. Soorten uit Australische woestijngebieden en taxonomisch behorend tot de sectieSuaveolentes (subgenusPetunioides) bleken het meest gevoelig en het minst resistent, terwijl de sectiesPaniculatae (subgenusRustica),Tomentosae (Tabacum) enNoctiflorae (Petunioides) het minst gevoelig en het meest resistent waren. Van de collectienummers van de laatstgenoemde secties bleek 61% resistentie tegen minstens twee van de virussen te bezitten. De meest resistente soorten hebben hun verspreidingsgebied vooral in het centrale deel van het Andesgebergte (het zuidelijk deel van Peru, Bolivia en noord-westelijk Argentinië). Mogelijk is dit het ontstaansgebied van deze virussen. De einge andere resistente soort wasN. africana (Suaveolentes) uit zuid-westelijk Afrika. De meest gevoelige Amerikaanse secties, te wetenGenuinae (Tabacum) enTrigonophyllae, Alatae, Acuminatae enBigelovianae (Petunioides), waren gemiddeld gevoeliger danSuaveolentes-soorten die niet uit de Australische woestijnen afkomstig zijn. Het resterende deel van het genus, bestaande uit de Amerikaanse sectiesThyrsiflorae enRusticae (Rustica) enUndulatae, Repandae enNudicaules (Petunioides), stond met betrekking tot gevoeligheid en resistentie tussen laatstgenoemd deel van de sectieSuaveolentes en de groep resistente secties in, maar vertoonde in hoge mate tolerantie voor PVA, PVX en PVY en met overgevoeligheid samengaande resistentie voor TMV. Uit de resultaten blijkt dat de reacties van het genusNicotiana op virussen vooral geografisch bepaald zijn en in mindere mate taxonomisch.     

植物源病毒抑制剂VFB对烟草抗烟草花叶病毒的诱导作用研究

为揭示植物源病毒抑制剂VFB的抗病毒机理,本研究以普通烟和烟草花叶病毒(TMV)为材料,测定了VFB进行预防和治疗处理后烟草中与抗病性相关的部分生理生化指标变化。结果表明:VFB可明显抑制病毒侵染后导致的细胞膜通透性增大;SOD、CAT、β-1,3-葡聚糖酶和几丁质酶等细胞防御酶活性增强,且预防处理的效果优于治疗处理;与抗病性呈正相关的脯氨酸和总游离氨基酸的含量有显著的提高。表明VFB可诱导植物产生抗病性,增强烟草对TMV侵染的抵抗能力。     

Production of tobacco mosaic virus in and its infectivity from leaves of two Nicotiana species treated with 6-azauracil

A correlation between the amount of virus as determined by infectivity test and that determined by a serological-spectrophotometric method was established in leaf discs ofNicotiana tabacum White Burley which had floated on a solution of 6-azauracil (AzU) before and after inoculation with tobacco mosaic virus. More virus, determined in the two different ways, was present in the AzU-than in the water-treated leaf discs at 24 and 48 h after inoculation. Thereafter, the amount of virus was less than that in the control. The inhibitory effect of the pyrimidine analogue on the amount of infectivity in sap from AzU-treated discs proved to be dependent on the dilution of sap used. The higher the dilution the more inhibition occurred.In inoculated leaf discs ofNicotiana glutinosa the formation of local lesions in the AzU-treated series was strongly inhibited at 48 h after inoculation whereas at that time the amount of virus determined by the serological-spectrophotometric method was higher than that in the water-treated control ones. The latter mentioned stimulating effect disappeared later than 48 h after inoculation and gave way to inhibition.The infectivity of tobacco mosaic virus from AzU-treated leaf discs of White Burley tobacco was 23% of that from water-treated controls when undiluted clarified sap was used.     

Resistance of Capsicum species to tobacco,tomato and pepper strains of tobacco mosaic virus

A number ofCapsicum accessions including nine species were tested for resistance to TMV based on hypersensitivity. The tobacco strain MA and the tomato strain SPS, which were both isolated from tomato, and two pathogenically distinct pepper strains P 11 and P 8, were used. Of the 73Capsicum accessions tested 58 were resistant to MA and SPS, 31 were resistant to P 11 and five were resistant to P 8.Samenvatting Om verschillen in pathogeniteit tussen twee in Nederland voorkomende paprikastammen van het TMV nader vast te stellen, werd een aantalCapsicum-herkomsten, waaronder negen soorten, op resistentie getoetst. Hierbij werden de representatieve paprika-isolaten P 11 en P 8 vergeleken met de uit tomaat afkomstige isolaten MA en SPS als vertegenwoordigers van respectievelijk de tabaks- en tomatestam van het TMV. Bij het beoordelen van de symptomen duidden lokale, necrotische vlekken en afvallen van geïnoculeerde bladeren op resistentie, systemische necrosen of mozaïeksymptomen op vatbaarheid. In deze symptomen kwamen tussen de gebruikte stammen verschillen in virulentie tot uitdrukking. Er werden echter vooral verschillen in agressiviteit waargenomen met betrekking tot zowel afzonderlijke, voor resistentie uitsplitsende,Capsicum-herkomsten als het totale aantal getoetste herkomsten. Van de ruim 73 herkomsten waren er 58 resistent tegen MA en SPS, 31 daarvan tegen P 11, maar slechts vijf daarvan tegen P 8. Deze resistentie tegen P 8 werd gevonden inC. chinense.     

壳寡糖诱导烟草抗烟草花叶病毒的超微结构研究

 电镜检查表明,接种烟草花叶病毒(TMV)表现系统侵染的烟草植株叶肉细胞和筛管内有大量病毒粒子和病毒结晶体,细胞质中出现髓鞘样结构、多泡体和次级囊泡。叶片薄壁细胞内叶绿体、线粒体等细胞器变形解体,膜结构增生。经壳寡糖(50 μg/mL)诱导处理的烟草,系统症状减轻,叶肉细胞和筛管中病毒粒子显著减少,偶见病毒结晶体;叶肉细胞中细胞器基本完好,但叶绿体内出现较大的淀粉粒;液泡内和细胞间隙出现大量电子致密物质。在未接毒和未经壳寡糖处理的植株中未见此种电子致密物,其产生可能与诱导抗病性表达有关。     

防治烟草花叶病的药剂筛选

对目前市场上常见的6种病毒抑制剂进行防治烟草花叶病的田间试验和人工示范，结果表明：2％菌克毒克250倍液防治效果最好。从花叶病发病初期开始每7d喷药1次，共喷3次，第1次喷药后7d调查，防效为76．2％，第3次喷药后7d调查，防效达71．6％，大田示范平均防效为70．1％。其次为病毒必克500倍液和毒消600倍液。3种药剂的防效均显著高于其它几种供试药剂，值得推广应用。为避免抗性的产生，建议在生产中交替使用。     

烟草品种对烟草花叶病毒和黄瓜花叶病毒的抗性鉴定

 2010-2011年采用大田人工接种鉴定的方法,对生产中大面积推广使用的24份烟草品种进行了烟草花叶病毒（TMV）和黄瓜花叶病毒（CMV）的抗性鉴定。结果表明,供试品种对TMV和CMV的抗病性存在较大差异,对TMV表现抗病的有Coker86、吉烟5号、Coker176、CV87、辽烟8号、CV91、中烟90等7份材料;表现中抗的有秦烟98、双抗70、C151等3份材料;表现中感的有秦烟96、G80、金星6007、龙江981、K326、秦烟201、NC89等7份材料;表现感病的有G28、云烟97、净叶黄、红花大金元、RG11、云烟85、云烟87等7份材料。对CMV表现抗病的有Coker86、龙江981、C151、秦烟201、云烟87等5份材料;表现中抗的材料是金星6007;表现中感的有CV91、RG11、Coker176、中烟90、K326、红花大金元、净叶黄、G80、G28等9份材料;表现感病的有秦烟98、云烟85、秦烟96、NC89、双抗70、云烟97、CV87、辽烟8号、吉烟5号等9份材料。其中兼抗TMV和CMV两种病毒病的材料有2份,分别是Coker86和C151。同时研究还发现,抗病性不同的烟草品种在受到病毒危害以后,对烟叶的产量和品质的影响也不同。明确了中国24个烟草品种的抗病性水平,为抗病品种的利用与品种合理布局提供科学依据,同时为烟草抗病毒病育种的亲本选择提供抗源信息。     

电化学酶联免疫分析检测烟草花叶病毒和烟草环斑病毒

以邻苯二胺 (OPD)底物为例 ,用抗原直接包被法 (DAC -ELISA)同线性扫描极谱法 (LSV)相结合 ,检测烟草花叶病毒 (TMV)的提纯液 ,检出限可达 0 2 5ng/ml,TMV粗提液的最高稀释比为 1 :1 0 2 4 0 0 ;检测烟草环斑病毒 (TRSV)提纯液 ,检出限为 1 0ng/ml,粗提液的最高稀释比为 1 :1 0 0 0 0 ,该方法的灵敏度比DAC -ELISA光度法提高了 5倍以上     

Investigating resistance to Barley mild mosaic virus

There are claims that at least 11 genes confer resistance in barley ( Hordeum vulgaris ) to one or more components of the soilborne barley mosaic virus complex but, apart from the immunity conferred by the widely used gene rym4 , little is known about their mode of action. This study used mechanical (sap) and plasmodiophorid vector-inoculation techniques combined with ELISA, RT-PCR, symptom development and virus transmission to investigate the response of different genotypes to Barley mild mosaic virus (BaMMV). Barley genotypes were grown at 20 and 12°C to test for temperature sensitivity. Plants with the genes rym3 or rym6 were fully susceptible to the virus, whereas those with genes rym1 , Rym2 , rym5 or rym11 appeared to be immune, as BaMMV was never detected in any tissue type nor was the virus transmitted from them to susceptible genotypes. The remaining genotypes could all be infected to some extent by BaMMV using one or both inoculation methods, and virus could be transmitted from their roots by the plasmodiophorid vector Polymyxa graminis . Plants with the rym7 gene had delayed symptoms compared to susceptible controls at 12°C. Plants with the rym8 gene could be infected by both inoculation methods, but there was no virus in the leaves at 12°C. Plants with the rym9 gene could be infected only by vector inoculation, and virus remained localized in the roots. Plants with the rym10 gene appeared susceptible by mechanical inoculation at both temperatures, but after vector inoculation virus moved to leaves only at 20°C. This suggests the operation of translocation resistance in plants with the rym8 , rym9 or rym10 genes, which is temperature-sensitive in rym8 and rym10 and perhaps tissue-specific in rym9 . No resistance to P. graminis was observed in any of the genotypes.     

植物源病毒抑制剂VFB诱导烟草抗烟草花叶病毒机理的初步研究

 为揭示植物源病毒抑制剂VFB的抗病毒机理,本研究以VFB对烟草花叶病毒(TMV)进行预防和治疗处理,测定了烟草中与抗病毒性相关的部分生理生化指标的变化。结果表明:VFB的2种处理方式均可使普通烟体内的叶绿素a、叶绿素b、类胡萝卜素、总酚及类黄酮含量升高,使普通烟体内苯丙氨酸解氨酶(PAL)的活性增强,预防处理优于治疗处理。可见,VFB可诱导植物产生抗病性,增强烟草对TMV侵染的抵抗力。