Screening for Barley yellow dwarf virus-Resistant Barley Genotypes by Assessment of Virus Content in Inoculated Seedlings

The content of Barley yellow dwarf virus (BYDV) in roots and leaves of barley seedling plants differing in their level of resistance was assessed by quantitative ELISA 1–42 days after inoculation with the strain of BYDV (PAV). High virus accumulation in roots and low concentration in leaves was characteristic of the period 9–15 days after inoculation. In leaves, the differences in virus content between resistant and susceptible genotypes became significant after 15 days and resistance to virus accumulation was better expressed 30–39 days after inoculation. Roots of resistant materials exhibited evident retardation of virus accumulation and the greatest difference in virus content between resistant and susceptible plants was detected 9 days after inoculation. By these criteria, the selected winter and spring barley cultivars and lines (in total 44 materials) fell in to five groups according to field reactions and the presence or absence of the Yd2 resistance gene. There were highly significant and positive relations between ELISA values and 5‐year field data on symptomatic reactions and grain‐yield reductions due to infection. Using the described method, resistant and moderately resistant genotypes (both Yd2 and non‐Yd2) were significantly differentiated from susceptible genotypes. The possible use of this method in screening for BYDV resistance is discussed.     

抗病基因Bdv2抑制大麦黄矮病毒复制和运动的分子证据

小麦-中间偃麦草易位系YW642含有一个源于中间偃麦草7X染色体的抗性基因Bdv2,对大麦黄矮病毒GAV株系具有高度抗性。为有效控制该病毒和阐明抗黄矮病机制,采用半定量RT-PCR的方法,研究了大麦黄矮病毒GAV株系在YW642及其感病姊妹系YW641中积累浓度的差异。分别在接种病毒不同时间、不同部位上取样,用半定量RT-PCR的方法来检测GAV的积累浓度。在接种部位,抗病植株中病毒的浓度远远低于感病植株。在侵染的前5d,抗病植株YW642中病毒会有一定程度的复制和积累,但随后病毒浓度开始下降,接种14—16d时没有检测到病毒;而在感病株系中,病毒积累的浓度远远高于抗病植株,并一直维持一个较高的浓度。在未接种部位．感病植株中可检测到较高浓度的病毒,说明病毒能从接种点很快运动到未接种部位,并大量复制。而在抗病系YW642中,未接种部位始终未检测到病毒。实验结果从分子水平上证明,在抗病植株中BYDV的复制和运动均受到了极大的抑制：这是抗病基因Bdv2与BYDV互作后,激活了一系列防御基因的结果。另外还确定了防御基因诱导表达的时间,为从抗病植株中分离抗病相关基因、研究抗黄矮病机制提供了取样的依据。     

Effect of BYDV Infection on the Cell Surface Glycoproteins in Common Wheat,Wheatgrass and Octoploid Wheat-Wheatgrass

Changes in the cell surface glycoproteins in common wheat 3B-2, Agropyron intermedium and octoploid wheat-wheatgrass Zhong 5 after the inoculation with barley yellow dwarf virus (BYDV) were sdudied using electron microscopy and ruthenium red staining. The results indicated that, after the inoculation with BYDV, different changes in cell surface glycoproreins were observed in the plant species with different levels of resistance. In A. intermediurn which is immune to BYDV, inoculation with BYDV did not cause significant change in cell surface glycoprotein layer. In cotoploid wheat-wheatgrass Zhong 5 which is highly resistent to BYDV, BYDV infection caused significant thickening in most cell surface glycoprotein layer. In common wheat 3B-2 which is susceptible to BYDV, BYDV infection did not cause thickening in cell surface glycoprotein layer, but in most cells, glycoproteins on the cell surface were partially peeled off or disappeared completely. Therefore, it is suggested that the glycoproteins on cell surface play certain roles in BYDV resistance. The phenomenon of the thickening of cell surface glycoprotein layer caused by BYDV infection was possibly a resistant reaction to the virus.     

Differentiation Among Bean Leafroll Virus Susceptible and Resistant Lentil and Faba Bean Genotypes on the Basis of Virus Movement and Multiplication

Systemic movement of Bean leafroll virus (BLRV) in susceptible and resistant lentil and faba bean genotypes was studied using plants grown in a plastic house. All the plants studied were inoculated with BLRV by viruliferous pea aphids (Acyrthosiphon pisum). Five plants/genotype of lentil and faba bean were harvested, respectively, at 3, 6, 9, 12 and 18 days and 1, 2, 3, 4 and 5 weeks after inoculation. Each plant was split into growing point, stem, stem base and root, and each was tested using tissue blot immunoassays (TBIA). Virus concentration in each section was estimated using a 0–3 score and a relative TBIA value was estimated accordingly for each genotype. In susceptible lentil genotypes (ILL 8063 and ILL 2581), BLRV was present in low concentrations in the growing point 3 days after inoculation and in high concentrations in all parts of the plant after 6 days. By contrast, the virus was not detected in the highly resistant genotype (ILL 74) until 18 days after inoculation. In the faba bean genotypes studied, BLRV was detected in high concentrations in all parts of the highly susceptible genotype (Fiord) 1 week after inoculation but only after 3 weeks in resistant genotypes (e.g. BPL 5274), but was not detected in the highly resistant genotypes (BPL 5278 and BPL 5279) 5 weeks after inoculation. The replication and systemic movement of BLRV was thus slower in resistant genotypes than in susceptible genotypes. Moreover, the use of TBIA scores clearly and easily differentiated resistant and susceptible genotypes. Our results suggest that BLRV movement and multiplication can be useful criteria when differentiating resistant from susceptible genotypes. In addition, undertaking the preliminary screening in a plastic house requires less space than direct planting in the field.     

High Levels of Resistance in Agropyron Species to Barley Yellow Dwarf and Wheat Streak Mosaic Viruses

Several Agropyron species were tested for new sources of resistance to barley yellow dwarf virus (Bydv ) and wheat streak mosaic virus (WSMV). With BYDV strain PAV, 11 of the 17 Agropyron species showed no virus transmission when plants were given access feed by viruliferous Rhopalosiphum padi. Similar trials with BYDV strain RMV (vectored by R. maidis) indicated that all plants, except susceptible control plants, remained virus free. Virus status was confirmed by enzyme-linked immunosorbent assays. When plants were mechanically inoculated with WSMV, 11 Agropyron species failed to express symptoms, while five other species showed a segregating response or had some accessions segregating and some resistant. Test results suggest that resistance to BYDV and WSMV in Agropyron species does not appear to be correlated with any specific genome of Agropyron species although most of the Agropyron species containing S genome were resistant to BYDV and WSMV.     

Barley Yellow Dwarf Virus Multiplication and Host Plant Tolerance in Durum Wheat

A Canadian PAV-like isolate of barley yellow dwarf virus (BYDV) was used to infect durum wheat (Triticum durum) cultivars previously identified in field trials involving artificial inoculation as highly sensitive (12 IDSN74), slightly tolerant (La Dulce), and relatively tolerant (Boohai and 12 IDSN227) to BYDV. The cultivars were inoculated in the greenhouse as seedlings, and indexed for virus accumulation by enzyme-linked immunosorbent assay (ELISA) at various intervals between 3 and 60 days thereafter. Mean ELISA values were somewhat consistent with tolerance levels for 4 durum wheat cultivars, but the use of ELISA to screen for BYDV resistance in durum wheat is not practical. The magnitude of the difference between sensitive and tolerant cultivars for the mean ELISA value is not high enough, and it may be necessary to average readings between 3 and 60 days after inoculation to obtain somewhat meaningful ELISA data. The effect of vector aphid numbers on virus titre and aerial biomass in the sensitive durum wheat cv. Karim was also evaluated. There was no significant effect on virus content in a preliminary trial, but a second trial revealed that more viruliferous aphids per plant resulted in higher ELISA values. Infestation with 32 or 50 viruliferous Rhopalosiphum padi per plant depressed biomass yield below the level observed with 1–10 aphids per plant.     

Differential Interaction Between PAV-like Isolates of Barley Yellow Dwarf Virus and Barley (Hordeum vulgare L.) genotypes

Four PAV-like isolates of barley yellow dwarf virus (BYDV) were identified as causing very severe (RG), severe (2t), moderately severe (3b) and mild symptoms (13t) in barley (Hordeum vulgare) cultivar Plaisant in a growth chamber at 25 days after inoculation. These isolates had different effects on a range of barley genotypes. Cultivar Vixen, which contains the Yd2 resistance gene, and 80-81 BQCB10 were not affected by any isolate. Five other genotypes were significantly affected by at, least one of the isolates. Line Ea52 (which is a mutant of the Japanese cultivar Chikurine Ibaraki) was more susceptible to BYDV-PAV than Chikurin Ibaraki 1. No serological differences were detected between the four isolates using monoclonal or polyclonal antibodies. Virus antigen concentration, estimated by enzyme-linked immunosorbent assay (ELISA), was correlated with the decrease in the shoot fresh weight for all isolates and all genotypes except for Vixen and 80-81BQCB10. In field tests, the severity of symptoms induced by the BYDV-PAV isolates was in accordance with that estimated in the growth chamber. However isolate 2t was more severe on cultivar Vixen and overcame the partial resistance of Chikurin Ibaraki 1 to the three other isolates. The results show that virus antigen concentration not only contributes to characterizing the resistance levels of barley genotypes but also the severity of BYDV-PAV isolates.     

Differential Interaction Between PAV-like Isolates of Barley Yellow Dwarf Virus and Barley (Hordeum vulgare L.) genotypes

Four PAV-like isolates of barley yellow dwarf virus (BYDV) were identified as causing very severe (RG), severe (2t), moderately severe (3b) and mild symptoms (13t) in barley (Hordeum vulgare) cultivar Plaisant in a growth chamber at 25 days after inoculation. These isolates had different effects on a range of barley genotypes. Cultivar Vixen, which contains the Yd2 resistance gene, and 80–81BQCB10 were not affected by any isolate. Five other genotypes were significantly affected by at least one of the isolates. Line Ea52 (which is a mutant of the Japanese cultivar Chikurine Ibaraki) was more susceptible to BYDV-PAV than Chikurin Ibaraki 1. No serological differences were detected between the four isolates using monoclonal or polyclonal antibodies. Virus antigen concentration, estimated by enzyme-linked immunosorbent assay (ELISA), was correlated with the decrease in the shoot fresh weight for all isolates and all genotypes except for Vixen and 80–81BQCB10. In field tests, the severity of symptoms induced by the BYDV-PAV isolates was in accordance with that estimated in the growth chamber. However isolate 2t was more severe on cultivar Vixen and overcame the partial resistance of Chikurin Ibaraki 1 to the three other isolates. The results show that virus antigen concentration not only contributes to characterizing the resistance levels of barley genotypes but also the severity of BYDV-PAV isolates.     

Ultrastructural and Immunocytochemical Studies on Effects of Barley Yellow Dwarf Virus – Infection on Fusarium Head Blight, Caused by Fusarium graminearum, in Wheat Plants

The interactions between barley yellow dwarf virus (BYDV) and Fusarium head blight (FHB), caused by Fusarium graminearum, were studied in the two winter wheat cultivars (cvs.), Agent (susceptible to FHB) and Petrus (moderately resistant to FHB), using ultrastructural and immunocytochemical methods. Infections of wheat plants of both cvs. by BYDV increased susceptibility to FHB. BYDV infection caused numerous cytological changes in lemma tissue of both cvs. such as formation of vesicles in the cytoplasm, degradation of fine structures of chloroplasts of both cvs. and accumulation of large starch grains in the chloroplasts. Electron microscopical studies showed that the development of F. graminearum on spike surfaces was not affected in BYDV‐infected plants. After penetration and intercellular growth in lemma tissue, defence responses to Fusarium infections were markedly reduced in BYDV‐diseased plants compared to the tissue of virus‐free plants. At sites of contact of fungal cells with host tissue, depositions of cell wall material were distinctly less pronounced than in tissues of virus‐free plants of cv. Petrus. Detection of β‐1,3‐glucanases and chitinases in lemma tissue of cv. Agent revealed no appreciably increased accumulation of both defence enzymes in F. graminearum‐infected virus‐free and BYDV‐infected tissues compared to the non‐infected control tissue. On the other hand, in cv. Petrus, infection with F. graminearum induced a markedly enhanced activity of both enzymes 3 days after inoculation. The increase of both enzyme activities was less pronounced in BYDV‐infected plants than in tissue exclusively infected with F. graminearum. Cytological studies suggest that in contrast to the susceptible cv. Agent postinfectional defence responses may play still an important role in the resistance of the moderately resistant cv. Petrus to FHB.     

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。     

禾谷多粘菌对小麦梭条斑花叶病毒的传播及其土壤侵染潜力的测定

从大田侵染小麦梭条斑花叶病毒的小麦病根中挑取禾谷多粘菌休眠孢子堆,接种受侵染小麦品种扬麦４号,经砂培养纯化,获得５个禾谷多粘菌分离物,但都为无毒。无毒多粘菌休眠孢子堆接种表现ＷＳＳＭＶ症状的小麦,经培养可饲获病毒,并可经接咱后将病毒传播给无病小麦,供试的４个大小麦禾谷多粘菌分离物都可对大小进行交叉侵染,产生同样数量的游动孢子产量。供试５个病土和２个无病土样品,都具有强大持多粘菌侵染潜力,即使稀释放     

Introgression and characterization of barley yellow dwarf virus resistance from Thinopyrum intermedium into wheat.

Wheatgrasses (species of Agropyron complex) have previously been reported to be resistant to barley yellow dwarf virus (BYDV). To introgress this resistance into wheat, Triticum aestivum x Thinopyrum (Agropyron) intermedium hybrids were advanced through a backcrossing program and reaction to BYDV, as determined by enzyme-linked immunosorbent assay (ELISA), is reported for the first time in backcross populations of wide hybrids between wheat and wheatgrasses. ELISA values revealed highly resistant to highly susceptible segregants in backcrosses. BYDV resistance was expressed in some backcross derivatives. Continued selection, based on cytology and ELISA in each generation, eliminated most of the unwanted wheatgrass chromosomes and produced self-fertile BYDV resistant wheat lines. The BYDV resistant lines with 2n = 42 had normal chromosome pairing similar to wheat, and their F1 hybrids with wheat had two univalents. DNA analyses showed that the source of alien chromatin in these BYDV resistant wheat lines is distinguishable from that in other Th. intermedium derived BYDV resistant wheat lines. Chromosome pairing and restriction fragment length polymorphism analyses indicated that the 42 chromosome resistant Purdue wheat lines are substitution lines in which chromosome 7D was replaced by a chromosome from Th. intermedium that was carrying gene(s) for BYDV resistance.     

The Yd2 resistance to barley yellow dwarf virus is effective in barley plants but not in their leaf protoplasts

The Yd2 gene for "resistance" to barley yellow dwarf virus (BYDV) has been widely used in barley ( Hordeum vulgare ). We have tested Australian isolates of BYDV of varying severity against barley genotypes with and without the Yd2 gene and report here a positive relationship between symptoms and virus levels determined by ELISA. Cultivar Shannon is the result of backcrossing the resistant line CI 3208 to cultivar Proctor, a susceptible line. It appears to be intermediate in reaction to BYDV between Proctor and CI 3208, although it carries the major gene, Yd2. Unlike the whole plant studies, no significant differences were observed with regard to the ability of protoplasts derived from these various genotypes to support BYDV replication. It is therefore demonstrated for the first time that the Yd2 gene is not among the small number of resistance genes which are effective against virus replication in isolated protoplasts.     

Identification and characterization of a novel efficient resistance response to the furoviruses SBWMV and SBCMV in barley

The interaction between the furoviruses Soilborne cereal mosaic virus (SBCMV) and Soilborne wheat mosaic virus (SBWMV) and their main host wheat is well documented; however, to date, only a few reports have addressed the response of other cereal species to these viruses. Here, we show that, in contrast to wheat, barley germplasm is a rich source of resistance to furoviruses. Moreover, we demonstrate that barley genotypes respond differentially to SBCMV and SBWMV, thereby providing an additional biological basis for classification of these viruses as two separate species. Following natural (soil) inoculation, some barley genotypes permitted foliar infection by SBWMV, whereas all 22 genotypes tested were resistant to SBCMV. Resistance is unlikely to be directed toward the virus vector, because Polymyxa graminis DNA was detected in the roots of all tested genotypes. Resistance to SBCMV in some barley genotypes was overcome by artificial virus inoculation onto the leaves, suggesting a block on virus translocation from roots to shoots as in resistant wheat genotypes. However, other genotypes were fully resistant following both inoculation techniques. One barley genotype, 'Dayton,' exhibited extreme resistance to both furoviruses. Further molecular analyses suggested that this novel and highly efficient resistance to furoviruses in barley operates by limiting virus spread from the primary inoculated cells.     

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.     

Interaction Of Barley Yellow Dwarf Virus and Fusarium culmorum (W. G. Sm.) Sacc. in Winter Wheat

The barley yellow dwarf virus (BYDV) epidemics, which occurred predominantly in northern Germany in 1988–90 and caused unusual yield losses of wheat, prompted our study on interactions of BYDV and Fusarium culmorum. At the late stages of plant development (EC 55/65) infections with BYDV resulted in a lower yield reduction of wheat plants than infections with F. culmorum. Combined infections at flowering resulted in severer yield reduction, indicating additive effects of the two pathogens. However, if wheat infected by BYDV at stage EC 25/35 was secondarily inoculated with the fungus at EC 55/65 the yield was less reduced than in combined infections at EC 55/65. Our results proved that the susceptibility of wheat plants to F. culmorum is increased when infection by BYDV takes place during the late stages of growth. These results correspond to observations during 3 years of the epidemic in Germany. In these years BYDV was spread mainly during the late spring, resulting in a severe secondary infection by the fungus. From the results of these investigations it may be concluded that during the years of BYDV epidemic the yield of wheat was reduced to an economically important extent because of the fungal infection, which was favoured by the virus infection.     

Response of wheatgrasses and wheat × wheatgrass hybrids to barley yellow dwarf virus

Summary Resistance to barley yellow dwarf virus (BYDV), manifested by low enzyme-linked immunosorbent assay (ELISA) values in plants exposed to viruliferous aphids, was identified in several wheatgrasses (Agropyron spp.). ELISA results were similar for root and leaf extracts of infested plants. No difference in reaction to BYDV was found between plants grown in the field and those in the growth chamber. Interspecific hybrids were generated using pollen from single resistant plants of Agropyron spp. to pollinate soft red winter wheat spikes. Resistance in hybrids appeared to be at the level of virus replication rather than at the level of vector inoculation. The hybrids varied in their reaction to BYDV. Expression of BYDV resistance in hybrids was influenced not only by wheat genotype and Agropyron species but, in some cases, reaction varied even among hybrids between the same wheat genotype and Agropyron plant. Implications of these results are discussed.Contribution from the Purdue Univ. Agric. Exp. Stn., West Lafayette, IN 47907, and USDA-ARS. The research was supported in part by Public Varieties of Indiana. Purdue Univ. Agric. Exp. Stn. Journal No. 11656     

Haustorium Formation and Cell Wall Appositions in Susceptible and Partially Resistant Wheat and Barley Seedlings Infected with Wheat Leaf Rust

Phase contrast light microscopy observations of wheat and barley seedlings infected with wheat leaf rust spores suggested that cell wall appositions are structural barriers against haustorium formation leading to abortion of infection structures. Nearly equal numbers of cell wall appositions per infection structure were detected in seedlings of susceptible and partially resistant wheat genotypes. Differences between susceptible and partially resistant genotypes became evident after the first haustorium had been formed. This again indicates the presence of a post-haustorial effect of partial resistance. Some factors influencing nutrient uptake are discussed. Wheat leaf rust colonies hardly formed haustoria in barley seedlings, the few not aborted infection structures were accompanied by cell collapse. The mechanisms of partial resistance in wheat and barley to their respective leaf rust fungi seem different, but their non-host reactions appear similar.     

Biological Characterization of an Italian Isolate of Barley Yellow Dwarf Luteovirus from Barley

An isolate of BYDV (BYDV-OC), from barley in Northwest Italy with typical symptoms of yellowing and dwarfing, was transmitted by Rhopalosiphum padi, Sitobion fragariae. S. avenae, Metopolophium festucae, R. maidis and M. dirhodum , but not by Myzus persicae or Schizaphis graminum . It reacted in DAS-ELISA with monoclonal and polyclonal antisera to PAV, but not with antibodies to MAV, RPV and RMV. A polyclonal antiserum prepared to BYDV-OC did not react with MAV-like, RPV-like, or RMV-like isolates of BYDV in ELISA or in Western blots. The concentration of BYDVOC in Avena byzantina plants decreased from weeks 1 to 10 after inoculation, but the total virus content per plant increased up to weeks 7 to 8, following the increase of plant weight.