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二重RT-PCR快速检测马铃薯病毒的方法 总被引：6，自引：0，他引：6
本研究采用传统的蛋白酶K法和病毒RNA简易浸提法,从马铃薯块茎、茎干、叶梗、叶片中提取马铃薯X病毒,马铃薯Y病毒,马铃薯A病毒及马铃薯卷叶病毒RNA,并设计了4种马铃薯病毒引物,优化了二重RT-PCR反应条件,可以同步扩增出上述4种病毒,扩增产生的靶带分别为562bp(PVX)、480bp(PVY)、336bp(PLRV)、255bp(PVA).应用病毒RNA简易制样技术和优化的二重RT-PCR反应条件,可以同步快速检测田间自然感染的马铃薯病毒,此研究还可适合于检测马铃薯脱毒种薯及试管苗,对马铃薯病毒病早期监测有一定的作用. 相似文献
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马铃薯X病毒和马铃薯Y病毒胶体金免疫层析试纸条的研制 总被引：8，自引：1，他引：8
采用柠檬酸三钠还原法制备胶体金颗粒,标记马铃薯X病毒和马铃薯Y病毒的兔多克隆抗体。用微定量喷头在硝酸纤维素膜上喷好病毒检测线和羊抗兔抗体质控线,制成免疫层析检测试纸条。马铃薯X病毒试纸条检测烟草病汁液可稀释10000倍(重量/体积)马铃薯Y病毒试纸条检测烟草病汁液可稀释1000倍。10min内即可出检测结果。两种试纸条相互测试,未出现交叉反应。用健康和缓冲液对照测试,两种试纸条结果都为阴性。田间采集的马铃薯叶片用试纸条检测的结果和酶联结果相吻合。在密封、干燥、低温的条件下保存,有利于维持试纸条的稳定性。 相似文献
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在马铃薯植株上接种Y病毒,往往经过很久时间也不出现症状。偶尔发现茎部折断的植株在新生枝上表现明显的皱缩花叶症状。 相似文献
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1986年自安徽省全椒县药菊产区显黄化病叶症状的药菊上分离到马铃薯Y病毒(Potato Virus Y简称PVY)。由于药菊含有抑制病毒体外传染的挥发性油类物质,所以该病毒极难用一般常见的接种方法传染,仅能以0.2%Na2SO3和0。2%Vc(1:1)混合液为缓冲液,活性碳为磨料来接种传染。 相似文献
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2017年调查福建福清地区马铃薯病毒病的发生情况,以明确该地区马铃薯主要病毒病原。共采集了46份疑似感染病毒的马铃薯植株,提取总RNA,利用RT-PCR技术进行分子检测,结果表明,福清地区危害马铃薯的病毒有马铃薯Y病毒Potato virus Y(PVY)、马铃薯卷叶病毒Potato leaf roll virus(PLRV)、马铃薯S病毒Potato virus S(PVS),检出率分别为56.52%、17.39%和10.87%,以PVY检出率最高,说明PVY是危害该地区马铃薯样品的主要病毒病原。通过病毒复合侵染进行分析,发现该地区存在病毒复合侵染马铃薯现象。研究结果可为福清地区马铃薯种薯的引进和病毒病害防治提供参考依据。 相似文献
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测定了自然感染花叶病毒(x y)的马铃薯男爵品种在不同地理(北京和克山)、季节(春播和秋播)和人工控制的土温(25°及15℃)三类条件下形成的退化及未退化块茎中的总氮(TN)、非蛋白质氮(NPN)、蛋白质氮(PN)、氨态氮、醯胺氮(AN)和氨基氮(NH_2—N)的变化。各种条件下形成的退化及未退化块茎中,TN 都没有显著差别,而 NPN 和PN 却有明显变化,退化块茎中 NPN 高于 PN.(NPN)/(PN)值为1.2—1.6;而未退化块茎中 PN 高于 NPN,(NPN)/(PN)值为0.6—1.0。此比值的大小与块茎退化程度成正相关。在进一步的测定中证明 NPN 的升高,主要是 NH_2—N 和 AN 的增加。 相似文献
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基于小RNA深度测序和RT-PCR检测侵染广东省冬种马铃薯的病毒 总被引：2，自引：1，他引：1
为明确侵染广东省冬种马铃薯的病毒种类及优势病毒,结合小RNA深度测序技术及RTPCR检测方法,对采集于广东省冬种马铃薯7个主产区的189份疑似病样进行检测分析。结果表明,经小RNA深度测序技术检测马铃薯病毒病混合样,发现存在马铃薯Y病毒(Potato virus Y,PVY)、马铃薯S病毒(Potato virus S,PVS)和马铃薯卷叶病毒(Potato leaf-roll virus,PLRV)3种病毒。进一步设计3种病毒的特异性引物并利用国内已报道的其它5种马铃薯病毒的特异性引物进行RT-PCR检测,发现189份马铃薯病毒病疑似病样中仅检测到PVY、PVS和PLRV这3种病毒,检出率依次为75.13%、10.05%和4.76%,且3种病毒在马铃薯上还存在复合侵染,复合侵染率为14.19%,其中PVY在各马铃薯产区均可检测到。表明侵染广东省冬种马铃薯的病毒为PVY、PVS和PLRV,其中PVY是优势病毒。 相似文献
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ABSTRACT Following regeneration, putative virus-resistant transgenic plants are usually transferred from tissue culture to a greenhouse or growth chamber to screen for resistance to infection and disease development using mechanical, graft, or insect vector inoculation methods. To reduce initial screening costs and time, we developed mechanical and graft inoculation methods suitable for tissue culture use. The in vitro methods were validated by comparing them with similar greenhouse screens using putative potato virus Y strain o (PVY degrees ) replicase-mediated resistant regenerants of the potato cultivar Atlantic. Five transgenic lines were tested, with similar results obtained from in vitro and greenhouse experiments. Two of the transgenic lines, A1 and A3, showed the greatest resistance to PVY degrees infection, as indicated by low enzyme-linked immunosorbent assay values and infection rates. In vitro mechanical inoculation methods were also used to infect wild-type tomato and tobacco plants with cucumber mosaic virus and potato virus Y. Potato plants were also infected with the phloem-restricted potato leafroll virus, a low-titer virus, using in vitro graft inoculation methods. These results suggest the potential usefulness of these simple, effective, and economical techniques for screening large numbers of putative virus-resistant plants. 相似文献
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Frédéric Boulard Gallien Rohaut David Dupré Florian Barbe Barrailh Amélie Person Melen Leclerc Emmanuel Jacquot Laurent Glais 《Plant pathology》2022,71(6):1323-1334
Aged plants are more difficult to infect than young plantlets. This modification of susceptibility is described as mature plant resistance (MPR). For potato virus Y (PVY), MPR is known to lead to low infection rates of plants inoculated at the postflowering stage and a decrease in the number of infected daughter tubers. However, the impact of inoculation date on the capacity of PVY to accumulate in daughter tubers has not been studied so far. Field and greenhouse experiments were carried out to better understand PVY epidemiology and to help potato growers to evaluate consequences of early/late infections on the quality of their crops. In field trials, potato plants (cv. Bintje) were covered by insectproof nets from planting to harvest except for a 14-day period to expose plants to natural PVY infections. Under controlled conditions, potato plants were mechanically inoculated with PVY at different dates from preflowering stages (early inoculations) to postflowering stage (late inoculations). At harvest, daughter tubers were individually collected and analysed to define proportions and viral load of infected tubers according to the time between virus inoculation and harvest. Our results showed that although the age of plants at the time of inoculation can modify their susceptibility to PVY infection, in return, early and late PVY inoculations lead to similar rates of infected tubers at the plant scale and equivalent viral accumulation in infected tubers. All together, these data revealed that both early/late infections are high risks for the sanitary quality of potato tubers. 相似文献
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ABSTRACT Soilborne wheat mosaic virus (SBWMV) is an agronomically important pathogen of wheat that is transmitted by the soilborne plasmodiophorid vector Polymyxa graminis. In the laboratory, attempts to generate SBWMV-infected plants are often hampered by poor infectivity of the virus. To analyze the mechanism for virus resistance in wheat cultivars, we developed novel inoculation techniques. A new technique for foliar inoculation of SBWMV was developed that eliminated wound-induced necrosis normally associated with rub inoculating virus to wheat leaves. This new technique is important because we can now uniformly inoculate plants in the laboratory for studies of host resistance mechanisms in the inoculated leaf. Additionally, wheat plants were grown hydroponically in seed germination pouches and their roots were inoculated with SBWMV either by placing P. graminis-infested root material in the pouch or by mechanically inoculating the roots with purified virus. The susceptibility of one SBWMV susceptible and three field resistant wheat cultivars were analyzed following inoculation of plants using these novel inoculation techniques or the conventional inoculation technique of growing plants in P. graminis-infested soil. The results presented in this study suggest that virus resistance in wheat likely functions in the roots to block virus infection. 相似文献
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J. A. T. Woodford 《European journal of plant pathology / European Foundation for Plant Pathology》1992,98(Z2):47-54
This paper reviews the contribution of vector activity and plant age to virus spread in potato crops. Determining which aphid species are vectors is particularly important for timing haulm destruction to minimize tuber infection by potato virus Y (PVY). Alate aphids of more than 30 species transmit PVY, and aphids such asRhopalosiphum padi, that migrate in large numbers before flights of the more efficient vector,Myzus persicae, appear to be important vectors. Differences in methodology, aphid biotypes and virus strains prevent direct comparisons between estimates of vector efficiencies obtained for aphids in different countries in north western Europe.
M. persicae is also the most efficient vector of potato leafroll virus (PLRV), but some clones ofMacrosiphum euphorbiae transmit PLRV efficiently toNicotiana clevelandii and potato test plants. The removal of infected plants early in the season prevents the spread of PLRV in cool regions with limited vector activity. The proportion of aphids acquiring PLRV from infected potato plants decreases with plant age, and healthy potato plants are more resistant to infection later in the season. Severe symptoms of secondary leafroll developed on progeny plants of cv. Maris Piper derived from mother plants inoculated with PLRV in June or July of the previous year. Progeny plants derived from mother plants inoculated in August showed only mild symptoms, but the concentration of PLRV in these plants was as high as that in the plants with severe symptoms. 相似文献
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J. F. J. M. van den Heuvel R. A. A. van der Vlugt M. Verbeek P. T. de Haan H. Huttinga 《European journal of plant pathology / European Foundation for Plant Pathology》1994,100(5):347-356
An Austrian isolate of potato virus YNTN, the causal agent of potato tuber necrotic ringspot disease (PTNRD), was serologically compared with seven Dutch PVYN isolates. Using polyclonal and monoclonal antibodies, it was found indistinguishable from PVYN. Determination of the nucleotide sequence of the coat protein cistron and comparison of the deduced amino acid sequence with coat protein sequences of other potyviruses revealed a high level of homology with PVYN coat protein sequences. This confirmed the close taxonomic relationship of PVYNTN with the PVYN subgroup of potato virus Y. PVYNTN is able to overcome all resistance genes known so far in commercial potato cultivars. Remarkably, transgenic PVY-protected tobacco plants are also resistant to PVYNTN infection upon mechanical and aphid-mediated inoculation. These experiments indicate that genetically engineered resistance offers great potential in protection of potato to new aggressive strains of PVYN. 相似文献
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Studies of a wilt disease of the potato plant in Israel caused by Erwinia chrysanthemi 总被引：5，自引：3，他引：2
In Israel field infections of potato plants by Erwinia chrysanthemi are characterized by wilting of the leaves followed by total desiccation of the plants. These symptoms are indistinguishable from those caused by Verticillium dahliae or those that develop during the normal process of plant senescence. Diagnosis of E. chrysanthemi in the spring-sown (February) crop in Israel is difficult because all three conditions often appear at approximately the same time, late in the growing season in May when the air temperature exceeds 25°C. The symptoms of E. chrysanthemi infection were reproduced in the field when potato seed tubers, tested and found to be contaminated at a low level with E. carotovora pv. carotovora , were inoculated with a strain of E. chrysanthemi isolated from a diseased potato plant. When plants in a growth cabinet at 30°C were stem-inoculated with E. chrysanthemi , similar symptoms developed when the relative humidity was low ( c . 80%). Presence of the disease only on plants grown from seed contaminated with E. chrysanthemi and not from uncontaminated seed suggests that the bacterium is seed borne, as is E. carotovora pv. atroseptica , the blackleg pathogen. 相似文献
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J. A. T. Woodford 《European journal of plant pathology / European Foundation for Plant Pathology》1992,98(2):47-54
This paper reviews the contribution of vector activity and plant age to virus spread in potato crops. Determining which aphid species are vectors is particularly important for timing haulm destruction to minimize tuber infection by potato virus Y (PVY). Alate aphids of more than 30 species transmit PVY, and aphids such asRhopalosiphum padi, that migrate in large numbers before flights of the more efficient vector,Myzus persicae, appear to be important vectors. Differences in methodology, aphid biotypes and virus strains prevent direct comparisons between estimates of vector efficiencies obtained for aphids in different countries in north western Europe.M. persicae is also the most efficient vector of potato leafroll virus (PLRV), but some clones ofMacrosiphum euphorbiae transmit PLRV efficiently toNicotiana clevelandii and potato test plants. The removal of infected plants early in the season prevents the spread of PLRV in cool regions with limited vector activity. The proportion of aphids acquiring PLRV from infected potato plants decreases with plant age, and healthy potato plants are more resistant to infection later in the season. Severe symptoms of secondary leafroll developed on progeny plants of cv. Maris Piper derived from mother plants inoculated with PLRV in June or July of the previous year. Progeny plants derived from mother plants inoculated in August showed only mild symptoms, but the concentration of PLRV in these plants was as high as that in the plants with severe symptoms. 相似文献
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M. J. Foxe 《European journal of plant pathology / European Foundation for Plant Pathology》1992,98(2):13-20
Conventional breeding methods based primarily on the principles of Mendelian genetics have provided the basis for the majority of resistance to viruses in potato cultivars. This development of resistant cultivars has resulted from sexual hybridisation followed by selection of the best genetic recombinant. Selection is one of the most powerful tools available for crop improvement. Successful selection depends on reproducible genetic variability, ability to identify genetically superior individual plants or families, populations large enough to ensure the likely occurrence of rare genetic combinations, and the combination of resistance expression with commercial utility. Resistance to the major potato viruses namely potato virus A, potato virus M, potato virus S, potato virus X, potato virus Y and potato leafroll virus has been studied and selected for. A number of different types of resistance have been identified including (a) major gene resistance; (b) minor gene resistance; and (c) immunity. With the exception of potato leafroll virus, stable resistance to the other viruses has been achieved either singly or in combination in a range of potato cultivars. In addition for both potato leafroll virus and potato virus Y, the focus has been on the development of resistance to aphid transmission of the viruses. 相似文献
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L.?Tsror O.?Erlich S.?Lebiush M.?Hazanovsky U.?Zig M.?Slawiak G.?Grabe J.?M.?van der?Wolf J.?J.?van de?Haar 《European journal of plant pathology / European Foundation for Plant Pathology》2009,123(3):311-320
Suspected Dickeya sp. strains were obtained from potato plants and tubers collected from commercial plots. The disease was observed on crops
of various cultivars grown from seed tubers imported from the Netherlands during the spring seasons of 2004–2006, with disease
incidence of 2–30% (10% in average). In addition to typical wilting symptoms on the foliage, in cases of severe infection,
progeny tubers were rotten in the soil. Six strains were characterised by biochemical, serological and PCR-amplification.
All tests verified the strains as Dickeya sp. The rep-PCR and the biochemical assays showed that the strains isolated from blackleg diseased plants in Israel were
very similar, if not identical to strains isolated from Dutch seed potatoes, suggesting that the infection in Israel originated
from the Dutch seed. The strains were distantly related to D. dianthicola strains, typically found in potatoes in Western Europe, and were similar to biovar 3 D. dadanti or D. zeae. This is the first time that the presence of biovar 3 strains in potato in the Netherlands is described. One of the strains
was used for pathogenicity assays on potato cvs Nicola and Mondial. Symptoms appeared 2 to 3 days after stem inoculation,
and 7 to 10 days after soil inoculation. The control plants treated with water, or plants inoculated with Pectobacterium carotovorum, did not develop any symptoms with either method of inoculation. The identity of Dickeya sp. and P. carotovorum re-isolated from inoculated plants was confirmed by PCR and ELISA. 相似文献