Interaction between the movement protein of barley yellow dwarf virus and the cell nuclear envelope: role of a putative amphiphilic alpha-helix at the N-terminus of the movement protein

The open reading frame 4 (ORF 4) gene product of barley yellow dwarf virus (BYDV) may act as a movement protein (MP) by assisting the transport of viral genomic RNA across the nuclear envelope (NE) of host plant cells. To investigate interactions between BYDV MP and the NE, wild-type and mutant open reading frame (ORF 4)-green fluorescent protein (GFP) fusion cistrons were expressed in insect cells. A fusion protein expressed by the wild-type ORF 4-GFP cistron associated with the NE and caused protrusions from its surface. The fusion protein expressed by the mutant ORF 4-GFP cistron lacked a putative amphiphilic alpha-helix at its N-terminus and although associating with the NE, showed decreased levels of protrusions. A peptide homologue of this putative alpha-helix induced an increase of 7 degrees C in the phase transition temperature of dimyrystoyl phosphatidylserine (DMPS) membranes, accompanied by a decrease in membrane fluidity, but exhibited no significant interaction with either dimyristoyl phosphatidylcholine (DMPC) or dimyristoyl phosphatidylethanolamine (DMPE) membranes. These results strongly support the view that BYDV MP may interact with the NE to help transport viral genomic RNA into the nuclear compartment. This function of BYDV MP appears to involve protrusions on the surface of the NE and may require the presence of an N-terminal amphiphilic alpha-helix, which is speculated to destabilize membranes, thereby assisting the entry of BYDV-GAV into the nuclear compartment.     

The effect of different virus isolates on the expression of tolerance to barley yellow dwarf virus in barley

A barley variety of Ethiopian origin, with a single Mendelian gene con-fering tolerance to barley yellow dwarf virus (BYDV), was equally tolerant to a number of isolates of the virus, whereas a susceptible European barley was more susceptible to isolates transmitted by Rhopalosiphum padi L. than to those transmitted by Macrosiphum (Sitobion) avenae (Fab). However, hybrids between these two varieties homozygous for the Ethiopian tolerance gene were more tolerant to ‘mild’ than to ‘severe’ isolates, irrespective of the vector specificity. The European variety was damaged more severely by all isolates when infected early than when infected late in its development, but the hybrids were damaged more severely by M. awraae-transmitted isolates when infected late. It is suggested that in susceptible plants the concentration, rather than the virulence, of the virus determines disease severity, whereas the reverse is true in plants possessing a gene which reduces virus multiplication. Virus concentration appears to determine the severity of R. padi-transmitted isolates, while virulence determines the severity of M. avenae-transmitted isolates. The latter would also seem to be adapted towards late infection.     

Host plant preference of cereal aphids in the field in relation to the ecology of barley yellow dwarf virus

Observational data obtained by exposing Avena sativa L., Phleum pratense L., Lolium perenne L., Poa pratensis L., and Zea mays L. to the aerial aphid population indicated food plant preference as resulting in a greater number of settlers on the preferred host, on which migrants also remained longer and deposited more nymphs than on the less suitable hosts. This behavioural pattern of alate Rhopalosiphum padi-fitchii is discussed in relation to the epidemiology of barley yellow dwarf virus disease.

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Zusammenfassung Die Besiedlung gewisser Gräserarten durch geflügelte Blattläuse ist verschiedentlich als Ausgangspunkt von Epidemien der Gelbverzwergungsvirose der Gerste (Rotblättrigkeit des Hafers) diskutiert worden. Ziel der vorliegenden Arbeit war es daher, den unterschiedlichen Befall von Hafer, deutschem Weidelgras, Wiesenlieschgras, Wiesenrispengras und Mais durch Blattläuse des Rhopalosiphum padi-fitchii Komplexes täglich oder mehrstündlich in einem Freilandbeet abzulesen. Dabei zeigte es sich, dass der Hafer von den Blattläusen vorgezogen wurde und es auf diesem Wirt zur Ausbildung grösserer Kolonien kam als auf den Gräserarten, während Mais überhaupt nicht besiedelt wurde. Wirtspflanzen-präferenz manifestierte sich ausserdem dadurch, dass die gelandeten Geflügelten längere Zeit auf Hafer verbrachten als auf den offensichtlich weniger geeigneten Gräsern. Die Befunde werden im Zusammenhang mit der Epidemiologie der Gelbverzwergung besprochen.

     

Inheritance and effectiveness of genes in barley that condition tolerance to barley yellow dwarf virus

Tolerance to barley yellow dwarf virus (BYDV) in five Ethiopian barley genotypes was conditioned by major genes which apparently occurred at, or near, the same chromosome locus in each genotype. The genes or alleles differed in effectiveness, one providing a high, three an intermediate and one a low level of tolerance. In plants having a slow rate of growth, the effectiveness of the tolerance genes tended to be masked, but the effectiveness of the gene or allele which provided the highest level of tolerance was masked to a lesser extent than that of genes or alleles providing low or intermediate levels of tolerance.     

Hydroxamic acids affecting barley yellow dwarf virus transmission by the aphid Rhopalosiphum padi

2,4-Dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), a hydroxamic acid (Hx) occurring in wheat, was shown to deter feeding by the aphid Rhopalosiphum padi (L.), and to reduce BYDV transmission to the plant. Dual choice tests with wheat leaves showed the preferential settlement of aphids on leaves with lower levels of DIMBOA. Electric monitoring of aphid feeding behaviour showed that in seedlings with higher DIMBOA levels fewer aphids reached the phloem and they needed longer times to contact a phloem vessel than in those with lower levels. When aphids carrying BYDV were allowed to feed on wheat cultivars with different DIMBOA levels, fewer plants were infected with BYDV in the higher DIMBOA cultivars than in the lower ones. Preliminary field experiments showed a tendency for wheat cultivars with higher Hx levels to be more tolerant to infection by BYDV than lower Hx level ones.     

The relationship between growth rate and the expression of tolerance to barley yellow dwarf virus in barley

Barley varieties were most tolerant to infection with barley yellow dwarf virus (BYDV) when they grew rapidly, whether the rate of growth was determined by manipulation of the environment or by the innate genetic constitution of the host. A specific, incompletely dominant gene conditioning a high level of tolerance to the virus in certain rapidly growing genotypes in which it occurred naturally, failed to do so when transferred to slower growing genotypes. However, reintroduction into genotypes capable of rapid growth led to full restoration of the gene's effectiveness. Virus-free aphids recovered BYDV less readily from quick- than from slow-growing genotypes, all homozygous for the tolerance gene.     

The role of ants in the epidemiology of barley yellow dwarf virus

The effect of ant attendance on the spread of barley yellow dwarf virus by Schizaphis (Toxoptera) graminum (Rondani) was studied under field conditions. Aphids visited by ants multiplied more rapidly than unattended colonies. Secondary spread of virus is caused by apterae which leave overcrowded plants and become established on nearby hosts. Ant-mediated spread of virus is typically confined to the edges of the field. Excavations made by ants around subterranean parts of small grains provide shelter and a suitable feeding site for tht aphid vector.

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Zusammenfassung Die Anregung zur vorliegenden Arbeit wurde dadurch gegeben, daß während einer Schlechtwetterperiode auf unterirdischen Teilen von Hafer- und Gerstenpflanzen von Ameisen besuchte Blattlauskolonien beobachtet wurden. Es handelte sich dabei um die Ameise Lasius neoniger (Emery) und die Blattlaus Schizaphis graminum (Rondani), die als Getreideschädling und als Überträger der Gelbverzwergungsvirose der Gerste in Nordamerika außerordentlich schädlich ist.Die Ergebnisse der Studie bestätigen erneut ältere Beobachtungen, nach denen sich die Populationsdichte der von Ameisen besuchten Blattläuse erheblich erhöht. Verschiedentlich konnte nachgewiesen werden, daß Ameisen Blattlausfeinde, insbesondere Marienkäfer, vertreiben, sobald diese in ihr Territorium eindringen.Die durch Ameisen geförderte Ausbreitung des Virus muß im engen Zusammenhang mit der populationsfördenden Wirkung des Blattlausbesuches gesehen werden. Ungeflügelte Blattläuse verlassen übervölkerte Pflanzen und wandern in die Umgebung der Wirtspflanze ab. Dabei können Entfernungen zurückgelegt werden, die ohne weiteres ausreichen, benachbarte Pflanzen aufzufinden und gegebenenfals mit Virus zu infizieren. Dadurch, daß sich L. neoniger hauptsächlich an den Feldrändern aufhält, bleibt allerdings die Virusausbreitung auf diese Region beschränkt und dringt nicht weiter in den Bestand vor. Selbst unter günstigen Bedingungen, das heißt in der Gegenwart von Ameisen, Blattläusen und Virus während des anfälligen Jugendstadiums des Getreides, bleibt die Ausbreitung des Virus und der dadurch angerichtete Schaden eng lokalisiert.Eine gewisse Bedeutung in der Epidemiologie der Gelbverzwergungsvirose dürfte auch dem Vorkommen von Blattläusen auf unterirdischen Pflanzenteilen zuzuschreiben sein, weil die Läuse hier gegen Witterungs- und Feindeinwirkung geschützt sind.Die von verschiedenen Seiten beobachtete enge Bindung zwischen Ameisen und Blattläusen konnte in dem vorliegenden Fall nicht beobachtet werden. Ein besonderer Instinkt der Ameisen oder Blattläuse scheint mir zur Aufrechterhaltung der Wechselbeziehung nicht notwendig.

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Approved for publication by the Director of the South Dakota Agricultural Experiment Station as Journal Series No. 583.     

Tomato yellow leaf curl virus V3 protein traffics along microfilaments to plasmodesmata to promote virus cell-to-cell movement

Science China Life Sciences -     

Nucleotide sequence of coat protein gene for GPV isolate of barley yellow dwarf virus and construction of expression plasmid for plant

GPV is a Chinese serotype isolate of barley yellow dwarf virus (BYDV) that has no reactionwith antiserum of MAV, PAV, SGV, RPV and RMV. The sequence of the coat protein (CP) of GPV isolate of BYDV was identified and its amino acid sequence was deduced. The coding region for the putative GPV CP is 603 bases nucleotides and encodes a Mr 22218 (22 ku) protein. The same as MAV, PAV and RPV, GPV contained a second ORF within the coat protein coding region. This protein of 17024 Mr (17 ku) is thought to correspond to the Virion protein genome linked (Vpg). Sequence comparisons of the CP coding region between the GPV isolate of BYDV and other isolates of BYDV have been done. The nucleotide and ammo acid sequence homology of GPV has a greater identity to the sequence of RPV than those of PAV and MAV. The GPV CP sequence shared 83.7% of nucleotide similarity and 77.5% of deduced amino add similarity, whereas that of the PAV and MAV shared 56.9%. 53.2% and 44.1%. 43.8% respectively. According to BYDV-GPV CP seque     

Evaluation of oat cultivars and lines under infection with barley yellow dwarf virus

Thirteen domestic and foreign oat cultivars and eight breeding lines bred from the University of Illinois were evaluated for resistance to barley yellow dwarf (BYD) using artificial inoculation with Rhopalosiphum padi viruliferous for an isolate of Barley yellow dwarf virus-PAV endemic to Moscow region origin. Cultivar Blaze and six Illinois lines showed the best grain yields under disease pressure that resembled a BYD epidemic.     

Feeding behavior and transmission of barley yellow dwarf virus by Sitobion avenae on oats

Feeding behavior of Sitobion avenae F. (Homoptera: Aphididae) on oats (Avena sativa cv. Clintland 64) was electronically monitored, and waveforms corresponding to salivation, ingestion, and sieve element penetration described.During 90 min plant access, aphids ingested from phloem for 0–43 min (mean: 8.1 min) and non-phloem for 0–60 min (mean: 19 min). Only 65% of the aphids tested made phloem contact within 90 min, contacting phloem after 18–85 min (mean: 32 min). No significant difference was observed in the feeding behavior of aphids carrying barley yellow dwarf virus (BYDV) from that of non-viruliferous aphids.Penetration of a sieve element was a prerequisite for BYDV transmission but did not insure transmission. Penetration of one sieve element resulted in a 65% chance of transmission independent of the duration of phloem contact. The chance of transmission increased with increasing number of sieve element penetrations.Inoculation of oat seedlings with single, viruliferous aphids for 90 min is estimated to cause 54% of the plants to be infected. Also, it is estimated that no transmission can occur with plant access periods shorter than 17 min.

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Zusammenfassung Das Probeverhalten von Sitobion avenae F. (Homoptera: Aphididae) auf Hafer (Sorte Clintland 64) wurden elektronisch verfolgt. Drei Wellenformen (S=Speichelfluss, X-das Eindringen in ein Siebelement, I=Nahrungsaufname) wurden registriert. In histologischen Untersuchungen wurden diese Wellenformen mit der Position der Stechborsten korreliert. Wenn X-Wellen oder eine X-I-Folge registriert wurden, war die Stechborste immer im Phloëm; bei S-Wellen oder bei einer S-I-Folge, war die Stechborste nie im Phloëm.Die Blattläuse wurden auf gesunden, fünf Tage alten Pflanzen während 90 Min beobachtet. 65% der Blattläuse erreichten nach 18–85 Min (Durchschnitt 32 Min) das Phloëm. Innerhalb 90 Min nahmen die Blattläuse während 0–43 Min (Durchschnitt 8.1 Min) Saft aus dem Phloem und während 0–60 Min (Durchschnitt 19 Min) Saft aus dem Mesophyll auf. Keine signifikanten Unterschiede im Probeverhalten wurden bei Blattläusen mit und ohne Barley Yellow Dwarf Virus (BYDV) festgestellt.Blattläuse, die innerhalb 24 Stunden BYDV übermitteln können, wurden für Übertragungsversuche verwendet. Das Probeverhalten dieser Blattläuser wurde manipuliert, und die Leistungsfähigkeit der Übertragung mit einer immunologischen Technik (ELISA) untersucht. Um BYDV zu übertragen mussten die Blattläuse mit einem Siebelement in Kontakt kommen. Nach der Stechborstenpenetration in ein Siebelement wurden 65% der Pflanzen mit BYDV infiziert. Der Prozentsatz infizierter Pflanzen und der Virusiter in den infizierten Pflanzen waren mit der Dauer der Siebelement-penetration (Anzahl von X-Wellen) nicht proportional. Wenn die Blattläuse mit zwei oder drei Siebelementen in Kontakt kamen, wurde der Prozentsatz infizierter Pflanzen signifikant erhöht, während der Virustiter nich verändert wurde. Infektionsprozente niedriger als 100% nach Siebelement-penetration sind möglicherweise das Resultat von Unterschieden in den Siebelementen.Es wird geschätzt dass 50% Infektion eintritt, wenn Pflanzen während ungefähr 83 Min von einer einzelnen infizierten Blattlaus besogen werden. Keine Übertragung von BYDV kann eintreten, wenn die Probezeit weniger als 17 Minuten beträgt.

     

The role of subterranean aphids in the epidemiology of barley yellow dwarf virus

Survival and dispersal of three root aphid species under greenhouse conditions was greatly enhanced in the presence of Lasius neoniger Emery. the ant was observed carrying three root aphid species but not two above-ground species. All attempts to transmit barley yellow dwarf (BYDV) with Forda olivacea Rohwer, Geoica utricularia (Passerini), Anoecia setariae Gillette & Palmer, or Aphis maidiradicis Forbes failed. However, A. maidiradicis was able to transmit cucumber mosaic virus. Rhopalosiphum padi (L.) could acquire BYDV from barley roots, but it rarely infected plants when feeding on roots.

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Zusammenfassung Die vorliegende Arbeit versucht die Bedeutung unterirdisch lebender Blattläuse für die Gelbverzwergungsvirose der Gerste zu klären. Sie schliesst sich damit früheren Untersuchungen über die Beziehung zwischen Schizaphis graminis (Rond.), einem Vektor des Virus, und der Ameise Lasius neoniger Emery an.Es konnte beobachtet werden, dass L. neoniger drei Wurzellausarten wegtrug, die in einer Versuchsarena freigelassen wurden, während zwei oberirdisch-lebende Blattlausarten nicht transportiert wurden. Die Gegenwart von Ameisen im Versuchsareal förderte auch die Vermehrung von Wurzelläusen auf Getreide und deren Ausbreitung auf benachbarte Pflanzen. Diese ausbreitungsfördende Wirkung des Blattlausbesuchs könnte von epidemiologischer Bedeutung sein, falls sich Wurzelläuse als Vektoren erweisen sollten. Alle Versuche, das Virus mit Forda olivacea Rohwer, Geoica utricularia (Passerini), Anoecia setariae Gillette & Palmer oder Aphis maidiradicis Forbes zu übertragen, schlugen jedoch fehl. Das nichtpersistente Gurkenmosaik konnte allerdings durch A. maidiradicis übertragen werden.Neben den typischen Wurzelläusen wurden unterirdisch lebende Rhopalosiphum padi L. im Freiland angetroffen. Übertragungsversuche ergaben, dass das Gelbverzwergungsvirus von der Blattlaus aus den Wurzeln aufgenommen werden konnte, dagegen gelang es nur selten, Getreidepflanzen durch auf Wurzeln saugende Blattläuse zu infizieren. Da aber im Freiland die Mehrzahl der unterirdisch lebenden R. padi auf den leicht zu infizierenden Stengelteilen angetroffen wurden, dürfte dieser relativen Widerstandsfähigkeit der Wurzeln gegenüber der Virusinfektion keine grössere Bedeutung zukommen.

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Approved for publication by the Director of the South Dakota Agricultural Experiment Station as Journal Series No. 672.     

In vitro interactions of the aphid endosymbiotic SymL chaperonin with barley yellow dwarf virus.

Barley yellow dwarf virus (BYDV)-vector relationships suggest that there are specific interactions between BYDV virions and the aphid's cellular components. However, little is known about vector factors that mediate virion recognition, cellular trafficking, and accumulation within the aphid. Symbionins are molecular chaperonins produced by intracellular endosymbiotic bacteria and are the most abundant proteins found in aphids. To elucidate the potential role of symbionins in BYDV transmission, we have isolated and characterized two new symbionin symL genes encoded by the endosymbionts which are harbored by the BYDV aphid vectors Rhopalosiphum padi and Sitobion avenae. Endosymbiont symL-encoded proteins have extensive homology with the pea aphid SymL and Escherichia coli GroEL chaperonin. Recombinant and native SymL proteins can be assembled into oligomeric complexes which are similar to the GroEL oligomer. R. padi SymL protein demonstrates an in vitro binding affinity for BYDV and its recombinant readthrough polypeptide. In contrast to the R. padi SymL, the closely related GroEL does not exhibit a significant binding affinity either for BYDV or for its recombinant readthrough polypeptide. Comparative sequence analysis between SymL and GroEL was used to identify potential SymL-BYDV binding sites. Affinity binding of SymL to BYDV in vitro suggests a potential involvement of endosymbiotic chaperonins in interactions with virions during their trafficking through the aphid.     

Transmission of barley yellow dwarf virus by field collected aphids (Homoptera: Aphididae) and their relative importance in barley yellow dwarf epidemiology in southwestern Idaho

Populations of cereal aphids were sampled from 1985–1988 and assayed for transmission of barley yellow dwarf virus (BYDV), Rhopalosiphum padi, Rho-palosiphum maidis, Sitobion avenae, Metopolophium dirhodum, Schizaphis graminum and Macrosiphum euphorbiae collected from host plants transmitted BYDV in bioassays. Of the 1028 Diuraphis noxia collected from plants, one may have transmitted BYDV. The isolate involved resembled SGV in serological and biological characteristics, but since it was not recoverable by any of more than 800 D. noxia subsequently tested, we suspect it may have been a contaminant. Among those aphids collected during the autumn from a suction trap adapted for live collection, R. padi transmitted BYDV most frequently. Other trapped species which transmitted BYDV included: R. maidis, Rhopalosiphum insertum, Macrosiphum euphorbiae, Metopolophium dirhodum and Ceruraphis eriophori. An adapted Infectivity Index indicated that R. padi is by far the most important vector of BYDV during the autumn sowing season in southwestern Idaho. Male R. padi consistently transmitted BYDV more frequently than did females collected during the same period.     

兼抗抗白粉病和黄矮病小麦育种材料的创造与鉴定

白粉病和黄矮病是小麦生产上的重要病害,近几年来这两种病害经常在我国一些小麦产区同时发生。为解决该问题,本研究通过杂交、回交方法将抗黄矮病的Bdv2基因（源自于YW642）和抗白粉病的Pm21基因（源自于CB037）聚合在一起,育成了兼抗黄矮病和白粉病的小麦新材料。通过田间抗病性鉴定与分子标记辅助选择相结合,得到聚合了Bdv2基因和Pm21基因的BC1代小麦22株,F2代小麦51株。农艺性状调查显示,这些含Pm21和Bdv2基因的双抗白粉病和黄矮病小麦新材料的农艺性状优于感病植株和原先的亲本,可以在小麦白粉病和黄矮病兼性抗病育种中作为优异种质资源加以利用。     

A single wheatgrass chromosome reduces the concentration of barley yellow dwarf virus in wheat

Seedlings of a series of addition or substitution lines of wheat containing different Thinopyrum intermedium chromosomes were inoculated with the PAV and RPV serotypes of barley yellow dwarf virus (BYDV). Reduced virus titres in infected plants were ascribed to a single pair of homoeologous group 7 chromosomes from Th. intermedium in the disomic addition lines L1 and TAF 2. The group 7 chromosome is associated with red pigmentation of coleoptiles, which was also observed in two lines ditelosomic for the α arm of the chromosome. However, when infected with the PAV serotype of BYDV, the ditelosomic lines had normal virus titres and it is concluded that potential determinants of BYDV resistance are located on the β arm of the Group 7 chromosome.     

Field assessment of the effectiveness of a barley yellow dwarf virus resistance gene following its transference from spring to winter barley

Resistance to infection by barley yellow dwarf virus (BYDV) has been transferred to the winter-hardy barley cv. Vixen. The effects of various sowing and inoculation dates on this resistance have been studied at two levels of BYDV infection. At levels most likely to be encountered in the field cv. Vixen suffered only slight yield losses and even under the most extreme conditions consistently yielded more than the susceptible cv. Igri. Comparison of the range of measurements used to determine the effects of BYDV infection showed that assessment of resistance to BYDV in field-grown winter barleys is most reliably based on yield measurements.