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1.
Transgenic tomato plants containing the coat protein (CP) gene of Cucumber mosaic virus (CMV) of subgroup IB were developed through Agrobacterium-mediated transformations. The progenies of transgenic plants showed the presence of transgene, its expression and translation of 26 KDa CP. The T1 and T2 generation plants were evaluated for resistance against challenge inoculations by a homologous strain of CMV. Visual observations of challenged transgenic plants categorized them into resistant, tolerant and susceptible as compared with untransformed control plants. Out of 33 plants of the T1 generation, 36.3% showed resistance and remained symptomless throughout their life, 48.4% showed tolerance which developed delayed symptoms of mild mosaic, and 15.1% showed susceptibility to CMV which developed severe systemic mosaic and leaf distortion symptoms after 30?days of virus challenge. Out of 120 plants of the T2 generation, 60% showed resistance, 26.6% were tolerant and only 13.3% were found susceptible to challenge inoculations of CMV. Resistant transgenic plants also showed less CP accumulation in systemic upper leaves as compared with challenged untransformed plants. In this study, CP of a CMV subgroup IB strain has demonstrated a significant level of resistance in transgenic tomato plants against the CMV strain. The strategy may be applied for better quality and productivity of tomato crops.  相似文献   

2.
Gunduz I  Buss GR  Chen P  Tolin SA 《Phytopathology》2004,94(7):687-692
ABSTRACT Resistance to Soybean mosaic virus (SMV) was identified in PI 88788 soybean, a germ plasm accession from China that is used widely as a source of resistance to soybean cyst nematode. Strains SMV-G1 through -G7 infected the inoculated leaves of PI 88788 but were not detected in upper, noninoculated trifoliolate leaves. Inheritance of resistance was determined by inoculating progenies of crosses of PI 88788 with susceptible cvs. Essex and Lee 68 with SMV strains G1 and G7. Allelomorphic relationships with known genes for resistance to SMV were tested in crosses with the resistant genotypes PI 96983, L29, and V94-5152, possessing Rsv1, Rsv3, and Rsv4 genes, respectively. Data analyses showed that resistance in PI 88788 to SMV-G1 is controlled by a single, partially dominant gene; however, to SMV-G7, the same gene was completely dominant. The PI 88788 gene was independent of the Rsv1 and Rsv3 loci, but allelic to Rsv4 in V94-5152. Expression of the Rsv4 gene in PI 88788 resulted in a reduced number of infection sites and restricted short- and long-distance movement of virus, rather than hypersensitivity. A unique late susceptible phenotype was strongly associated with heterozygosity. This gene has potential value for use in gene pyramiding to achieve resistance to several SMV strains, as well as for rate-reducing resistance.  相似文献   

3.
Gal-On A  Wolf D  Wang Y  Faure JE  Pilowsky M  Zelcer A 《Phytopathology》1998,88(10):1101-1107
ABSTRACT Tomato breeding lines were transformed with a defective replicase gene from RNA 2 of cucumber mosaic virus (CMV). A total of 63 transformants from five tomato genotypes were evaluated for resistance to CMV strains. The responses of R1 transgenic offspring fit into three categories: fully susceptible lines (44%), fully resistant lines (8%), and an intermediate-type mixture of susceptible and resistant seedlings in variable proportions (48%). Further characterization of the response of two highly resistant lines was performed by mechanical inoculation, aphid transmission, or grafting experiments. No virus was detected in noninoculated leaves from these lines, although a low level of virus accumulated initially in the inoculated leaf. The homozygous R2 plants and further generations that were evaluated (up to R5) showed resistance to the Fny-CMV strain, two Israeli isolates tentatively classified as subgroup IA, and K-CMV (a representative of subgroup IB). These lines were partially resistant to LS-CMV (a representative of subgroup II) when a high-virus-titer inoculum was used. Expression of the viral transgene was verified in these lines; however, the expected translation product was not detectable. In grafting experiments, we demonstrated that CMV virions were blocked in their ability to move from infected rootstocks of nontransformed tomato or tobacco into the transgenic scions. Interestingly, virions could not move through a transgenic intersection into the upper scion. These results provide an additional indication that replicase-mediated resistance affects long-distance movement.  相似文献   

4.
Kobori T  Osaki T  Ohki ST 《Phytopathology》2003,93(11):1445-1451
ABSTRACT A potential regulatory site for Cucumber mosaic virus (CMV, pepo strain) movement necessary to establish systemic infection was identified through immunological and hybridization studies on Tetragonia expansa, which was systemically infected by CMV at 36 degrees C but not at 24 degrees C. In inoculated leaves, cell-to-cell movement of CMV was enhanced at 36 degrees C compared with that observed at 24 degrees C. CMV was distributed in the phloem cells of minor veins as well as epidermal and mesophyll cells at both 36 and 24 degrees C. CMV was detected in the petioles of inoculated leaves, stems, and petioles of uninoculated upper leaves at 36 degrees C, whereas CMV was detected only in the petioles of inoculated leaves and in stems at 24 degrees C. CMV moved into the phloem and was transported to the stem within 24 h postinoculation (hpi) at 36 degrees C. However, it did not accumulate in the petioles of the upper leaves until 36 hpi. In petioles of inoculated leaves at 24 degrees C, CMV was detected in the external phloem but not in the internal phloem. From these results, we conclude that systemic infection is established after viral entrance into the phloem pathway in T. expansa at 36 degrees C.  相似文献   

5.
西瓜抗小西葫芦黄花叶病毒基因的连锁分子标记研究   总被引:10,自引:0,他引:10  
 小西葫芦黄花叶病毒中国株系(Zucchini yellow mosaic virus Chinese strain,ZYMV-CH)是危害我国西瓜的主要病毒。本实验以抗病毒病西瓜野生种质P.I.595203与感病的普通西瓜自交系98R为亲本,采用单粒传方式得到109个E代株系,分别对亲本、F1及109个F3代株系群体进行了苗期抗ZYMV-CH接种鉴定,通过F3代群体的抗感分离情况,推测得到F2代各单株的基因型,采用集团分离分析法(bulked segregant analysis,BSA)在F2代建立抗感基因池,以亲本、F1和抗感基因池为模板,对640条RAPD引物进行PCR扩增筛选,其中引物AK13在亲本、F1和抗感基因池之间扩增出一条多态性片段(644bp),在F2代群体上验证该多态性条带与ZYMV-CH的抗性基因呈现连锁关系,遗传连锁距离为8cM,定名为AK13-644,该连锁标记在ZYMV-CH抗性转育后代自交系上得到了验证。最终将此RAPD标记成功转化成SCAR标记SCAK13-644,该标记可以作为西瓜抗病毒病辅助选择的分子标记。  相似文献   

6.
7.
ABSTRACT Begomoviruses (the family Geminiviridae) are transmitted by the whitefly Bemisia tabaci and contain monopartite or bipartite circular single-stranded (ss)DNA genomes. They have emerged as severe problems in the production of agricultural and horticultural crops worldwide. Here, we report the identification of a tomato breeding line, FLA653, that confers a high level of resistance to Tomato leaf curl virus (TLCV, monopartite). Genetic analysis indicated that the resistance is controlled by a single recessive allele named tgr-1, which is in contrast to previous reports that multiple genetic factors are involved in tomato resistance to begomoviruses. Particle bombardment of an infectious TLCV DNA construct into the detached leaves of FLA653 resulted in the viral replication, but the viral ssDNA accumulated at a much lower level than that in susceptible controls. In situ localization of TLCV in the bombarded leaves suggests that tgr-1 impaired TLCV movement, raising the possibility that it may specify a host factor essential for viral systematic infection. This makes tgr-1 a strong candidate for developing resistance in major crops carrying the gene homologue.  相似文献   

8.
Zea mays has been historically imported to Japan via two independent geographical routes: into southern Japan by trading with Europe in the 16th century and into northern Japan by import from North America in the 19th century. Breeding to genetically improve on quality traits and high yields has led to the current domestic landraces in each region. In a survey of 82 domestic landraces, nine out of 38 landraces originating from southern Japan showed complete immunity to cucumber mosaic virus yellow strain (CMV(Y)) without the formation of necrotic local lesions (NLLs). In contrast, three out of 44 landraces originating from northern Japan developed NLLs, but revealed no systemic spread of the virus. Due to the absence of good documentation on NLL formation in Z. mays, the response of domestic landraces Aso‐1 and Aso‐3, originating from Ibaraki in northern Japan, to a challenge with CMV(Y) and CMV(Ma‐1) was further analysed. Aso‐3 only formed NLL in response to CMV(Y) but not to CMV(Ma‐1). Moreover, in CMV(Y)‐inoculated Aso‐3, virus spread was restricted to the primary infection site and the expression of defence‐related genes was up‐regulated, whereas Aso‐1 became systemically infected with either CMV(Y) or CMV(Ma‐1). The response of Aso‐3 to CMV(Y) was inherited as a single dominant trait. Together, these results pointed towards the induction of hypersensitive response (HR)‐mediated resistance to CMV(Y) in Aso‐3. Although HR‐mediated resistance to viruses has been studied mainly in dicots, the pathosystem CMV–Z. mays may provide a model to investigate HR‐mediated resistance to viruses in monocot plants.  相似文献   

9.
转CP基因线辣椒对CMV和CMV-RNA的抗病性比较   总被引:7,自引:0,他引:7  
 本试验以转化CMV-CP和TMV-CP基因线辣椒纯合系作试材,比较了接种CMV粒体和CMV-RNA后的发病特点和叶片中的病毒含量。结果表明:转化线辣椒不仅能抵抗CMV粒体的侵染,而且还能抵抗CMV-RNA的侵染。不论接种CMV粒体或CMV-RNA,CP(+)线辣椒的系统症状都延迟出现,显症株率和病害严重度级别大幅度降低,病毒增殖和运转受到抑制,接种叶片与新生叶片中的病毒含量明显减低。这一结果证实CMV-RNA不能克服线辣椒由CP基因介导的抗病性。  相似文献   

10.
ABSTRACT Tobacco cultivar Virgin A Mutant (VAM) is reported to have the recessive potyvirus resistance gene va. Varied levels of resistance were observed in VAM plants inoculated with Japanese potato virus Y (PVY) isolates. VAM was highly resistant to most of the PVY isolates tested and tolerant to three necrotic strain isolates of PVY-T. Based on data obtained from tissue printing and press blotting, the resistance appeared to be mainly at the level of cell-to-cell movement. PVY replicated in VAM proto-plasts, but the replication was 30% lower than in susceptible tobacco, suggesting that impairment of replication also contributes to resistance. To identify the viral gene product or products involved in VAM resistance, we isolated spontaneous resistance-breaking mutants by passing vein-banding (O strain) isolates several times through VAM plants. By comparing the amino acid sequences of the mutants with their original isolates, we identified a single amino acid substitution in the viral genome-associated protein (VPg) domain that is correlated with VAM resistance breaking. Together, these results suggest that, in addition to its role in replication, VPg plays an important role in the cell-to-cell movement of PVY.  相似文献   

11.
 本文以抗病自交系黄野四-3、感病自交系8112和Mo17、耐病自交系478为材料,采用直接组织斑免疫测定法(IDDTB)和酶联免疫吸附测定法(ELISA)结合生物学症状研究了玉米矮花叶病毒对玉米的侵染及其运转规律,试验表明玉米抗玉米矮花叶病毒主要是抗系统传导,其次抗病毒增殖和细胞间扩展,但不抗侵入。  相似文献   

12.
系统侵染的番茄植株中黄瓜花叶病毒的时序变化   总被引:1,自引:0,他引:1  
 采用实时荧光定量PCR (FQ-PCR)和DAS-ELISA方法,研究了22~26℃温室条件下番茄幼苗中黄瓜花叶病毒CNA株系(CMV-CNA)各基因组RNA组分及其外壳蛋白(CP)含量的动态变化,同时结合同期感病植株症状发展和病情指数,分析并探讨CMV各基因组RNA、CP以及病症显示程度之间的时间效应及其相关性。以18S rRNA为内参照,FQ-PCR相对定量分析结果显示:接种后5~30 d,CMV三分体基因组RNA在系统侵染的番茄组织中负荷量变化趋势大体一致,但是不同时期含量差异显著,均经历对数增长期、稳定期和回落期。其中,以RNA2负荷量变化情况最为平缓。DAS-ELISA检测结果显示:CP含量随接种时间延长而持续升高,但其对数增长趋势相对滞后于基因组RNA。番茄幼苗发病症状与CMV基因组RNA及CP负荷量的变化趋势大体一致,但症状表现时间相对滞后。CMV-CNA株系在番茄幼苗中以基因组RNA、CP以及病症显示先后次序出现高峰期,显示病毒基因组RNA及其CP在植物组织内负荷量的变化与植株症状表现并不同步。其动态变化规律将为研究CMV侵染机制,病毒与寄主互作及防病控病提供量化依据。  相似文献   

13.
14.
ABSTRACT Transformation of plants with the nucleocapsid (N) gene of Tomato spotted wilt tospovirus (TSWV) provides resistance to disease development; however, information is lacking on the response of plants to natural inoculum in the field. Three tobacco cultivars were transformed with the N gene of a dahlia isolate of TSWV (TSWV-D), and plants were evaluated over several generations in the greenhouse. The resistant phenotype was more frequently observed in 'Burley 21' than in 'KY-14' or 'K-326', but highly resistant 'Burley 21' transgenic lines were resistant to only 44% of the heterologous TSWV isolates tested. Advanced generation (R(3) and R(4)) transgenic resistant lines of 'Burley 21' and a 'K-326' F(1) hybrid containing the N genes of two TSWV isolates were evaluated in the field near Tifton, GA, where TSWV is endemic. Disease development was monitored by symptom expression and enzyme-linked immunosorbent assay (ELISA) analysis. Whereas incidence of TSWV infection in 'Burley 21' susceptible controls was 20% in 1996 and 62% in 1997, the mean incidence in transgenic lines was reduced to 4 and 31%, respectively. Three transgenic 'Burley 21' lines were identified that had significantly lower incidence of disease than susceptible controls over the two years of the study. In addition, the rate of disease increase at the onset of the 1997 epidemic was reduced for all the 'Burley 21' transgenic lines compared with the susceptible controls. The 'K-326' F(1) hybrid was as susceptible as the 'K-326' nontransformed control. ELISA analysis demonstrated that symptomless plants from the most resistant 'Burley 21' transgenic lines accumulated detectable nucleocapsid protein, whereas symptomless plants from more susceptible lines did not. We conclude that transgenic resistance to TSWV is effective in reducing incidence of the disease in the field, and that accumulation of transgene protein may be important in broad-spectrum resistance.  相似文献   

15.
16.
A variant of Cucumber mosaic virus, CMV(Y/GM2), was isolated from a tobacco plant with mild green mosaic symptoms that was regenerated in vitro from a yellow strain of CMV [CMV(Y)]-infected tobacco leaves by tissue culture. CMV(Y/GM2) has two amino acid substitutions at 36 and 111 positions in the coat protein encoded on RNA3. CMV, assembled by mixing in vitro transcribed CMV(Y) RNA1 and RNA2 plus infectious RNA3 transcribed in vitro from cDNA to RNA3 of CMV(Y/GM2), was prepared and designated as CMV(Y/GM2)tr. When tobacco (Nicotiana tabacum cv. Xanthi nc) plants were inoculated with CMV(Y/GM2)tr, large necrotic local lesions in which the virus was localized, developed on the inoculated leaves. This host response unique to CMV(Y/GM2)tr was similar to the hypersensitive response (HR), which is a common resistance response to avirulent pathogens and was observed in five cultivars of Nicotiana tabacum and eight Nicotiana species. The revertant virus, however, accumulated to quite different levels in the various hosts. CMV(Y/GM2)tr induced pathogenesis-related 1 (PR-1) protein accumulation and systemic acquired resistance (SAR) which were generally observed in the HR. However, when tobaccos were inoculated with CMV(S36P)tr and CMV(V111I)tr, which have an amino acid substitution at either the 36 or 111 position in the coat protein of CMV(Y), respectively, CMV(S36P)tr was restricted to the primary infection site without necrotic local lesion formation and PR-1 protein and SAR induction. CMV(V111I)tr, however, systemically spread and induced mild green mosaic symptoms, while the host had the HR to CMV(Y/GM2)tr. The localization of CMV(Y/GM2)tr at the primary infection site may not only be caused by the HR, but also by the restriction of virus systemic movement resulting from the amino acid substitution at position 36 in the coat protein of CMV(Y). Received 15 December 1999/ Accepted in revised form 18 April 2000  相似文献   

17.
A strong recovery response occurs in cantaloupe (Cucumis melo) and watermelon (Citrullus lanatus) infected with the bipartite begomovirus Cucurbit leaf crumple virus (CuLCrV). This response is characterized by initially severe symptoms, which gradually become attenuated (almost symptomless). An inverse relationship was detected between viral DNA levels and recovery, indicating that recovered tissues had reduced viral titers. Recovered tissues also were resistant to reinfection with CuLCrV; i.e., recovered leaves reinoculated with the virus did not develop symptoms or have an increased level of viral DNA. In contrast, infection of CuLCrV-recovered leaves with the RNA virus, Cucumber mosaic virus (CMV), disrupted recovery, resulting in the development of severe disease symptoms (more severe than those induced by CMV or CuLCrV alone) and increased CuLCrV DNA levels. Small RNAs with homology to CuLCrV DNA were detected in recovered and nonrecovered tissues; as well as in phloem exudates from infected, but not uninfected plants. Levels of these small RNAs were positively correlated with viral titer; thus, recovered tissues had lower levels than symptomatic tissues. In addition, viral DNA from a host that undergoes strong recovery (watermelon) was more highly methylated compared with that from a host that undergoes limited recovery (zucchini). Furthermore, inoculation of CuLCrV-infected zucchini with a construct expressing an inverted repeat of the CuLCrV common region enhanced recovery and reduced viral symptoms and viral DNA levels in newly emerged leaves. Taken together, these results suggest that recovery from CuLCrV infection is an adaptive antiviral defense mechanism, most likely mediated by gene silencing.  相似文献   

18.
ABSTRACT Arabidopsis thaliana ecotypes vary in their responses to viruses. In this study, we analyzed the variation in response of A. thaliana ecotype Tsu-0 to Cauliflower mosaic virus (CaMV). This ecotype was previously reported to be resistant to two CaMV isolates (CM1841 and CM4-184), but susceptible to W260. In this study, we show that Tsu-0 is resistant to four additional CaMV isolates. CaMV propagated within the rosette leaves of Tsu-0 plants, but did not appear to spread systemically into the inflorescence. However, virus viability in rosette leaves of Tsu-0 plants apparently was not compromised because infectious CaMV could be recovered from these organs. W260 overcomes Tsu-0 resistance by a passive mechanism (i.e., this virus avoids activating plant defenses). The portion of the viral genome responsible for W260 resistance breakage was mapped to the 5' third of gene VI, which we have termed RBR-1. This region is also responsible for controlling the ability of CaMV to infect different types of solanaceous plants. Hence, the pathways by which plants of different families interact with CaMV may be conserved through evolution.  相似文献   

19.
ABSTRACT A quantitative method to screen common bean (Phaseolus vulgaris) plants for resistance to Bean common mosaic necrosis virus (BCMNV) is described. Four parameters were assessed in developing the quantitative method: symptoms associated with systemic virus movement, plant vigor, virus titer, and plant dry weight. Based on these parameters, two rating systems (V and VV rating) were established. Plants from 21 recombinant inbred lines (RILs) from a Sierra (susceptible) x Olathe (partially resistant) cross inoculated with the BCMNV-NL-3 K strain were used to evaluate this quantitative approach. In all, 11 RILs exhibited very susceptible reactions and 10 RILs expressed partially resistant reactions, thus fitting a 1:1 susceptible/partially resistant ratio (chi(2) = 0.048, P = 0.827) and suggesting that the response is mediated by a single gene. Using the classical qualitative approach based only on symptom expression, the RILs were difficult to separate into phenotypic groups because of a continuum of responses. By plotting mean percent reduction in either V (based on visual symptoms) or VV (based on visual symptoms and vigor) rating versus enzyme-linked immunosorbent assay (ELISA) absorbance values, RILs could be separated clearly into different phenotypic groups. The utility of this quantitative approach also was evaluated on plants from 12 cultivars or pure lines inoculated with one of three strains of BCMNV. Using the mean VV rating and ELISA absorbance values, significant differences were established not only in cultivar and pure line comparisons but also in virus strain comparisons. This quantitative system should be particularly useful for the evaluation of the independent action of bc genes, the discovery of new genes associated with partial resistance, and assessing virulence of virus strains.  相似文献   

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