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1.
Antibodies were prepared against two synthetic peptides, P19 and P11, derived from the coat protein N-terminal region of two pepper isolates of Potato virus Y from Tunisia (PVY-P21 and PVY-P2, respectively). The peptides were selected by comparing the predicted amino acid sequences of three pepper and four potato PVY isolates on the basis of their polymorphism and hydrophilicity. Sera with high titres were only obtained against P19. Three MAbs, raised in response to P19, reacted with the homologous virus (PVY-P21) in TAS-ELISA. When tested against a broad range of PVY isolates and related viruses, MAb 3C5 proved to be PVY species specific, whereas MAbs 8A4 and 1D6 reacted specifically with standard isolates of PVYO, PVYC and PVYN-W strains, but not with other PVY isolates. Consequently, epitope(s) recognized by 8A4 and 1D6 MAbs may be specific to a PVY group comprising all serologically PVYnon–N isolates. Surprisingly, and unlike isolate PVY-P21, many Tunisian field pepper isolates did not carry this epitope(s), thus revealing serological heterogeneity within the PVY pepper group. As PVY is one of the most economically important plant pathogens in a range of crops, including pepper, these MAbs will provide a useful tool for practical diagnosis and strain identification of PVY.  相似文献   

2.
Detailed characterization of a number of isolates of PVYSYR, a novel recombinant strain of Potato virus Y (PVY) from Syria, was conducted to elucidate their origin, assess their significance and achieve a final classification of PVYSYR. Recombination analysis grouped isolates of PVYSYR into three recombination patterns, SYR-I, SYR-II and SYR-III, which varied in the first 700 nucleotides of their genomes, with the second recombination pattern, SYR-II, the most frequent. PVYSYR isolates shared highest genomic identity and close phylogenetic relationships with PVYNTN and PVYNW isolates from Syria, suggesting a common origin and local emergence of these isolates in Syria. All PVYSYR isolates (total of 20) induced tobacco veinal necrosis, but reacted to a PVYO monoclonal antibody, typical characteristics of the previously reported PVYNW (or PVYN:O). In potato, however, four isolates tested (one of SYR-I and three of SYR-II) induced potato tuber necrotic ringspot disease (PTNRD), which is the characteristic phenotype of PVYNTN. Given the shared properties of SYR-I and SYR-II isolates with PVYNTN and PVYNW, it was decided that they represent a new recombinant strain of the PVYN strain group, with the proposed name PVYNTN-NW. The classification of SYR-III will be possible only after testing the phenotype in potato. The high prevalence of PVYNTN-NW in potatoes and weeds, as well as its ability to induce PTNRD, demonstrates its importance and the necessity for its control.  相似文献   

3.
一株PVYNTN-NW黑龙江马铃薯分离物的检测鉴定   总被引:1,自引:0,他引:1  
 马铃薯Y病毒(Potato virus Y,PVY)是马铃薯、烟草等茄科作物上的重要病毒,在与寄主共同进化过程中产生了许多株系。本文从黑龙江马铃薯样品中得到PVY分离物A12。ELISA结果表明A12被PVYO的单克隆抗体特异性识别。A12开放阅读框为9 186个核苷酸,编码3 061个氨基酸,与SYR-II-Be1分离物的核苷酸和氨基酸序列一致率均最高,分别为98.3%和99.2%。系统发育分析发现A12与PVYNTN-NW株系SYR-II型的分离物聚类到一起;重组分析表明,A12是N-605和Oz的重组体,重组类型与SYR-II-Be1相同。综合以上结果表明,A12属于PVYNTN-NW株系SYR-II型。但与常见PVYNTN-NW株系分离物在珊西烟引起叶脉坏死不同,A12产生花叶症状。A12辅助成分-蛋白酶在182位和245位的氨基酸均为精氨酸,而其它PVYNTN-NW株系分离物为赖氨酸。本研究结果可为黑龙江马铃薯PVY的早期检测和有效防控提供理论指导。  相似文献   

4.
 马铃薯Y病毒(Potato virus Y,PVY)是马铃薯、烟草等茄科作物上的重要病毒,在与寄主共同进化过程中产生了许多株系。本文从黑龙江马铃薯样品中得到PVY分离物A12。ELISA结果表明A12被PVYO的单克隆抗体特异性识别。A12开放阅读框为9 186个核苷酸,编码3 061个氨基酸,与SYR-II-Be1分离物的核苷酸和氨基酸序列一致率均最高,分别为98.3%和99.2%。系统发育分析发现A12与PVYNTN-NW株系SYR-II型的分离物聚类到一起;重组分析表明,A12是N-605和Oz的重组体,重组类型与SYR-II-Be1相同。综合以上结果表明,A12属于PVYNTN-NW株系SYR-II型。但与常见PVYNTN-NW株系分离物在珊西烟引起叶脉坏死不同,A12产生花叶症状。A12辅助成分-蛋白酶在182位和245位的氨基酸均为精氨酸,而其它PVYNTN-NW株系分离物为赖氨酸。本研究结果可为黑龙江马铃薯PVY的早期检测和有效防控提供理论指导。  相似文献   

5.
 马铃薯是我国重要粮食和经济作物。马铃薯Y病毒(potato virus Y,PVY)是危害马铃薯生产的重要病害。种植脱毒种薯是防治PVY最有效的途径。马铃薯种薯携带PVY问题严重,但种薯中PVY株系还不清楚。本研究利用PVY特异性抗体检测了7个马铃薯品种362个种薯,发现不同品种种薯带毒率差异较大,最高达12%。通过RT-PCR方法扩增获得了7个PVY分离物编码区全序列。重组分析发现7个分离物基因组均为重组型,根据重组位点的差异可以分为PVYNTN-NW(SYR-II型)、Rec-1、Rec-2和Rec-3等4种重组类型,后3种为新重组类型。系统进化分析发现,分离物HQH18G3-10与PVYNTN-NW(SYR-II型)处于同一个大的分支,但与中国PVY大田分离物聚集在一起形成一个相对独立的组,命名为PVYNTN-NW(CN型);其余6个分离物与数据库中的中国分离物聚集在PVYN-Wi组。这暗示PVY中国分离物具有相对独立的进化过程,PVY马铃薯大田分离物和种薯分离物进化上相近。所有分离物均能在珊西烟上引起典型叶脉坏死症状,HQH18G3-10引起的坏死症状最为严重。本研究首次报道了我国种薯内PVY发生情况,对分析病毒发生发展规律和防控具有借鉴作用。  相似文献   

6.
 马铃薯是我国重要粮食和经济作物。马铃薯Y病毒(potato virus Y,PVY)是危害马铃薯生产的重要病害。种植脱毒种薯是防治PVY最有效的途径。马铃薯种薯携带PVY问题严重,但种薯中PVY株系还不清楚。本研究利用PVY特异性抗体检测了7个马铃薯品种362个种薯,发现不同品种种薯带毒率差异较大,最高达12%。通过RT-PCR方法扩增获得了7个PVY分离物编码区全序列。重组分析发现7个分离物基因组均为重组型,根据重组位点的差异可以分为PVYNTN-NW(SYR-II型)、Rec-1、Rec-2和Rec-3等4种重组类型,后3种为新重组类型。系统进化分析发现,分离物HQH18G3-10与PVYNTN-NW(SYR-II型)处于同一个大的分支,但与中国PVY大田分离物聚集在一起形成一个相对独立的组,命名为PVYNTN-NW(CN型);其余6个分离物与数据库中的中国分离物聚集在PVYN-Wi组。这暗示PVY中国分离物具有相对独立的进化过程,PVY马铃薯大田分离物和种薯分离物进化上相近。所有分离物均能在珊西烟上引起典型叶脉坏死症状,HQH18G3-10引起的坏死症状最为严重。本研究首次报道了我国种薯内PVY发生情况,对分析病毒发生发展规律和防控具有借鉴作用。  相似文献   

7.
A range of selected PVY isolates that induce superficial necrosis on potato tubers, originating from several countries, were compared with standard strains of PVA, PVV and PVY. Biological properties (e.g., host range, aphid transmissibility and relationships based on cross-protection between virus isolates) were studied. PVYNN isolates differ from the normal PVYC, PVYN and PVO strains by their ability to infect Capsicum annuum but not Chenopodium amaranticolor and C. quinoa. All PVYNN isolates are transmissible by Myzus persicae , without any significant differences from one standard strain. These additional data confirm that these tuber-necrosing isolates belong to PVY. However, they could be ranged in a homogeneous and distinct group inside the PVYN group, based on the differences revealed in the host range, in addition to the specific ability naturally to induce necrosis on tubers.  相似文献   

8.
Potato virus Y (PVY) strains were originally defined by interactions with different resistance genes in standard potato cultivars. Five distinct strain groups are defined that cause local or systemic hypersensitive responses (HRs) in genetic background with a corresponding N gene: PVY(O), PVY(N), PVY(C), PVY(Z), and PVY(E). The nucleotide sequences of multiple isolates of PVY(O) and PVY(N) differ from each other by ≈8% along their genomes. Additionally, complete genome sequences of multiple recombinant isolates are composed of segments of parental PVY(O) and PVY(N) sequences. Here, we report that recombinant isolate PVY-L26 induces an HR in potato 'Maris Bard' carrying the putative Nz gene, and is not recognized by two other resistance genes, Nc and Ny(tbr). These genetic responses in potato, combined with the inability of PVY-L26 to induce vein necrosis in tobacco, clearly define it as an isolate from the PVY(Z) strain group and provide the first information on genome structure and sequence of PVY(Z). The genome of PVY-L26 displays typical features of European NTN-type isolates with three recombinant junctions (PVY(EU-NTN)), and the PVY-L26 is named PVY(Z)-NTN. Three typical PVY(NTN) isolates and two PVY(N) isolates, all inducing vein necrosis in tobacco, were compared with PVY-L26. One PVY(NTN) isolate elicited HR reactions in Maris Bard, similar to PVY-L26, while two induced a severe systemic HR-like reaction quite different from the quasi-symptomless reaction induced by two PVY(N) isolates. 'Yukon Gold' potato from North America produced HR against several PVY(NTN) isolates, including PVY-L26, but only late and limited systemic necrosis against one PVY(N) isolate. Consequently, according to symptoms in potato indicators, both PVY(Z) and PVY(NTN) isolates appeared biologically very close and clearly distinct from PVY(O) and PVY(N) strain groups.  相似文献   

9.
A survey of Potato virus Y (PVY) was carried out on weeds growing in and around potato fields in Syria during the autumn growing seasons of 2002, 2004 and 2006. A total of 59 samples of eight weed species and three tobacco ( Nicotiana tabacum ) samples were tested by ELISA using PVY antisera. Among them 15 samples belonged to Solanum nigrum (7 samples), Physalis sp. (6 samples) and tobacco (2 samples) were PVY infected. This suggests that weed hosts and neighbouring tobacco fields are natural reservoirs of PVY in Syria. According to their biological, serological and molecular characteristics, the majority of weed PVY isolates belonged to the PVYSYR variant indicating a possible correlation between the high incidence of PVYSYR in potato and weed hosts. This is the first report on the occurrence and characterization of weed PVY isolates from Syria.  相似文献   

10.
Bhat  Varma  Pappu  Rajamannar  Jain  & Praveen 《Plant pathology》1999,48(5):648-654
A potyvirus (eggplant mottle virus, EMoV) causing mosaic mottling in eggplant ( Solanum melongena ) was characterized on the basis of biological, serological and partial nucleotide sequence properties. EMoV infected Chenopodium amaranticolor and members of the Solanaceae. Polyclonal antiserum against EMoV showed antigenic relationship with henbane mosaic potyvirus (HMV) and potato Y potyvirus (PVY). Virus-specific antibodies directed to the N-terminal region of EMoV cross-reacted only with PVY. Determination and comparison of nucleotide sequence of the coat protein (CP) and the 3'-untranslated region (UTR) of EMoV with other potyviruses showed that the level of homology was highest with PVY isolates. Comparative sequence analyses of the CP amino acid and 3'-UTR sequences with distinct PVY isolates placed EMoV within the PVYO subgroup.  相似文献   

11.
The biological and molecular relationships between a large number of Potato virus Y (PVY) isolates were examined, concentrating mainly on isolates associated with potato tuber necrotic ringspot disease (PTNRD). Following detailed analysis of the coat-protein gene, four main groups were identified which broadly corresponded to the phenotype of the different isolates. The groups comprised the ordinary strain (PVYO), the necrotic strain (PVYN), the C strain (PVYC) and a group of recombinant (between ordinary and necrotic) isolates. In the latter group, all members were associated with PTNRD. However, four nonrecombinant isolates were also identified which were associated with PTNRD or tuber necrosis. Three were from tubers showing PTNRD symptoms in the field, while the fourth originated from symptomless tubers, but could cause necrotic rings on tubers under glasshouse conditions. The results show that although coat-protein recombination is always found associated with the PTNRD phenotype, some nonrecombinant isolates have very similar biological properties.  相似文献   

12.
13.
A potyvirus known to be an important agent involved in causing a disease of trailing petunias, was identified as being a member of the necrotic strain of potato virus Y (PVY) using a number of monoclonal antibodies. The sequence of the coat protein gene for the PVY isolate was determined and when compared with sequences for other PVY strains it was shown to cluster closely with isolates of PVYNTN and to have a recombination point present within the coat protein common with other isolates of PVYNTN. When inoculated onto potato tuber necrotic ringspot disease (PTNRD) susceptible potato cultivars the petunia isolate was found to be capable of causing necrotic tuber symptoms, consistent with those caused by other isolates of PVYNTN. Due to the number of similarities it is thought the petunia isolate belongs to the PVYNTN group of isolates. Out of 24 species of bedding and pot plant crops tested, 19 were shown by mechanical inoculation to be susceptible to PVY, highlighting not only a clear risk to a number of commercially important plant species from PVYNTN infected trailing petunias, but also other susceptible crops grown in these areas.  相似文献   

14.
Infection with potato virus S Andean (PVSA) and ordinary (PVSo) strains was found in potato breeder's selection No. 8163-511 imported from West Germany; the two PVS strains were differentiated by their reactions on Chenopodium quinoa Tests on potato leaf samples using enzyme-linked immunosorbent assay followed by inoculation to C quinoa were subsequently used to detect PVSA and PVSo in a large-scale survey of imported and domestic potato material. PVSA was detected in breeders' selections and cultivars imported from the Netherlands and West Germany, but not in domestic certified seed potato stocks or farmers' once-grown stocks. PVSo was found in both imported and domestic certified stocks, but infection was commoner in the imported ones. When plants of C. quinoa, C. amaranticolor, C. murale and Nicotiana debneyi were inoculated with four isolates of PVSA, one induced mild symptoms while the reactions of the others ranged from moderate to severe. When plants of different potato cultivars were inoculated with three isolates, the plants were mostly infected without symptoms. However, when tubers from some were grown on, the progeny plants of most of the different combinations of cultivar and isolate of PVSA developed one or more of the following symptoms: vein deepening, rugosity, interveinal chlorosis, premature senescence and early loss of lower leaves.  相似文献   

15.
Two Potato virus Y (PVY) isolates collected in Brazil, PVY‐AGA and PVY‐MON, were identified as recombinants between two parent genomes, PVYNTN and PVY‐NE‐11, with a novel type of genomic pattern. The new recombinants had an ordinary PVYNTN genome structure for approximately 6·7‐kb from the 5′‐end of the genome whereas the 3′‐terminal 3·0‐kb segment had two fragments of NE‐11‐like sequence separated by another small PVYNTN‐like fragment. PVY strains are defined based on the hypersensitive resistance (HR) response in potato indicators. Both PVY‐AGA and PVY‐MON isolates did not induce the HR in potato cultivars carrying Ny, Nc, or (putative) Nz genes and thus were able to overcome all known resistance genes to PVY. Only one of the two isolates, PVY‐AGA, induced a vein necrosis reaction in tobacco. The biological responses of the potato indicators and tobacco defined PVY‐MON as an isolate of the PVYE strain. To distinguish PVY‐AGA and PVY‐MON from other PVYNTN isolates, an RT‐PCR test was developed utilizing new specific primers from the capsid protein gene area and producing a characteristic 955‐bp band. Serological profiling of these PVY isolates with three monoclonal antibodies revealed an unusual reactivity, where one of the two commercial PVYN‐specific monoclonal antibodies did not recognize PVY‐AGA. The ability of these new PVY recombinants to overcome resistance genes in potato producing mild or no symptoms, combined with the lack of serological reactivity towards at least one PVYN‐specific antibody may present a significant threat posed by these isolates to seed potato production areas.  相似文献   

16.
Surveys were conducted of symptomatic potato plants in late season crops, from the major potato production regions in Northern Tunisia, for infection with six common potato viruses. The presence of Potato leafroll virus (PLRV), Potato virus Y (PVY), Potato virus X (PVX), Potato virus A (PVA), Potato virus S (PVS) and Potato virus M (PVM) was confirmed serologically with virus infection levels up to 5.4, 90.2, 4.3, 3.8, 7.1 and 4.8%, respectively. As PVY was prevalent in all seven surveyed regions, further biological, serological and molecular typing of 32 PVY isolates was undertaken. Only one isolate was shown to induce PVYO-type symptoms following transmission to tobacco and to react only against anti-PVYO-C antibodies. Typical vein necrosis symptoms were obtained from 31 samples, six of which reacted against both anti-PVYN and anti-PVYO-C antibodies showing they contained mixed isolates, while 25 of them reacted only with anti-PVYN antibodies. An immunocapture RT-PCR molecular test using a PVYNTN specific primer pair set in the 5’NTR/P1 genomic region and examination of recombinant points in three genomic regions (HC-Pro/P3, CI/NIa and CP/3’NTR) showed that all 25 serotype-N PVY isolates were PVYNTN variants with similar recombinations to the standard PVYNTN-H isolate. This is the first report of the occurrence of the PVYNTN variant and its high incidence in late season potatoes in Tunisia.  相似文献   

17.
Endogenous levels of free and conjugated salicylic (SA) and gentisic (GA) acids, both putative signal molecules in plant defence, were analysed in order to investigate their involvement in the resistance of four potato ( Solanum tuberosum ) genotypes with different susceptibilities to Potato virus YNTN (PVYNTN) infection: the highly susceptible cv. Igor and its extremely resistant transgenic line, the extremely resistant cv. Sante and the tolerant cv. Pentland Squire. The lowest levels of free and conjugated SA were observed in the extremely resistant cv. Sante, while free GA, which was detected in all the other varieties, was absent. The extremely resistant transgenic cv. Igor contained the highest basal total SA level and the lowest level of total GA of all four cultivars. In susceptible cv. Igor, but not in resistant transgenic cv. Igor, a systemic increase of free SA was measured 1 day postinfection (dpi). Even more significant increases of free and conjugated SA and GA were detected 11 dpi when systemic symptoms appeared. In inoculated but not in upper noninoculated leaves of resistant transgenic cv. Igor, significant increase of SA conjugates occurred, but not before 11 dpi. The increase of SA and GA in susceptible cv. Igor could contribute to the general elevated levels of phenolic compounds as a response to stress caused by virus infection. It appears that basal levels of SA and GA do not correlate with resistance to PVYNTN in potato plants.  相似文献   

18.
ABSTRACT Four monoclonal antibodies (MAbs) were prepared against an isolate of soilborne wheat mosaic furovirus from Oklahoma (SBWMV Okl-7). Three MAbs had different reactivities in tests on SBWMV isolates from Nebraska (Lab1), France, and Japan. One MAb (SCR 133) also reacted with oat golden stripe furovirus. None of the MAbs cross-reacted with other rod-shaped viruses including beet necrotic yellow vein furovirus, potato mop-top furovirus, and tobacco rattle tobravirus. Sequence analysis of nucleotides between 334 and 1,000 of RNA 2, the region that encodes the coat protein (CP) and the first 44 amino acids of a readthrough protein, of the four SBWMV isolates revealed up to 27 base changes from the published sequence of a Nebraska field isolate of SBWMV. Most changes were translationally silent, but some caused differences of one to three amino acids in residues located near either the N- or C-terminus of the CPs of the different isolates. Two further single amino acid changes were found at the beginning of the readthrough domain of the CP-readthrough protein. Some of these amino acid changes could be discriminated by MAbs SCR 132, SCR 133, and SCR 134. Peptide scanning (Pepscan) analysis indicated that the epitope recognized by SCR 134 is located near the N-terminus of the CP. SCR 132 was deduced to react with a discontinuous CP epitope near the C-terminus, and SCR 133 reacted with a surface-located continuous epitope also near the C-terminus. Predictions of CP structure from computer-assisted three-dimensional model building, by comparison with the X-ray fiber diffraction structure of tobacco mosaic virus, suggested that the three CP amino acids found to differ between isolates of SBWMV were located near the viral surface and were in regions predicted to be antigenic.  相似文献   

19.
 马铃薯Y病毒(potato virus Y,PVY)主要侵染马铃薯和烟草等茄科作物,给世界农业造成巨大经济损失。本文对测定的23个及GenBank中注册的52个中国PVY分离物ORF序列进行了系统发育、重组和选择压等分析。系统发育分析表明,根据ORF序列可把我国75个PVY分离物和国外30个参比分离物分成O、C、E、NTN-NW(SYR-I型)、NTN-NW(SYR-II型)、NTN(NTN-a型)、NTN(NTN-b型)、NA-N/NTN、Eu-N、N-Wi(N:O型)和N-Wi(N-Wi型)等11个分子株系,其中中国PVY分离物属于除E和C株系外的9个分子株系。除ME162、guiyang、PVYzu、SD-G、WA-13和CN:JL-1:17等 6个分离物基因组中未检测到重组,其余69个分离物均存在明显重组。根据重组位点的不同,中国PVY可分为11种重组类型,其中5种为新的重组类型。选择压分析表明,中国PVY分离物的11个基因均处于负选择,其中核内含体b基因受到的选择压最大,PIPO受到的选择压最小。基因流分析表明,黑龙江、河南和山东PVY分离物间基因交流频繁,马铃薯与烟草PVY分离物之间基因交流频繁。本研究的结果明确了中国PVY分离物的分子株系组成,对指导PVY的检测和防控具有积极作用。  相似文献   

20.
A virus, isolated from faba bean (Vicia faba) obtained from Algeria, was readily recognized as a tobravirus by its particle sizes and morphology. Pea (Pisum sativum) and French bean (Phaseolus vulgaris) characteristically reacted to the isolate like pea early-browning virus (PEBV), but faba bean,Antirrhinum majus, Nicotiana rustica, andN. tabacum reacted with line-pattern symptoms which were unusually brilliant on theNicotiana species. In electronmicroscope decoration tests, the isolate did not react with an antiserum to the Dutch type strain of PEBV, but with one to the broad bean yellow band (BBYB) serotype from Italy. It resembles this serotype in reaction on faba bean, but seems to differ appreciably onN. rustica, N. tabacum, andPetunia hybrida. It is described as a deviant isolate of the BBYB serotype of PEBV.All thirteen faba-bean genotypes tested were found to be susceptible to the Algerian isolate and two Dutch type strain isolates of the virus, and to react with erratic line-pattern symptoms to the Algerian isolate only. All ten genotypes of chickpea (Cicer arietinum) tested reacted hypersensitively, and four out of ten genotypes of lentil (Lens culinaris) were susceptible to the virus but reacted differentially to the three isolates. Seed transmission of PEBV, including the new isolate, in faba bean is confirmed (9% for the Algerian isolate, and over 45% for one of the Dutch type strain isolates), and seed transmission of the virus in a non-legume (N. rustica, 4%) is herewith first reported. This is the first report on the occurrence of the BBYB serotype of PEBV outside Italy, and of PEBV outside Morocco in North Africa.  相似文献   

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