Evaluation of the durability of the <Emphasis Type="Italic">Barley yellow dwarf virus</Emphasis>-resistant <Emphasis Type="Italic">Zhong ZH</Emphasis> and <Emphasis Type="Italic">TC14</Emphasis> wheat lines

Serial passage experiments (SPE) of a Barley yellow dwarf virus-PAV (BYDV-PAV) isolate were performed on Zhong ZH and TC14 wheat lines to evaluate the durability of their resistance to BYDV. At different passage numbers (from the 2nd to the 114th), biological properties of the produced isolates were recorded either by monitoring infection percentages and virus titers of the first 3 weeks of viral infection or by measuring their impact on yield components. Statistical analyses using the area under pathogen progress curves and the area under concentration progress curves demonstrated that these two resistant lines induce, after only a few passages, a selection of variant(s) with significantly modified infection abilities. Isolates resulting from SPE performed on these lines induced important decreases of yield components. These results indicate that the use of Zhong ZH and TC14 lines in BYDV-resistant breeding programmes should be approached with caution.     

Analysis of Accumulation Patterns of <Emphasis Type="Italic">Barley yellow dwarf virus-PAV</Emphasis> (BYDV-PAV) in Two Resistant Wheat Lines

Barley yellow dwarf (BYD) is one of the main viral diseases of small-grain cereals. This disease, reported on numerous plant species of the Poaceae family, is caused by a complex of eight viral species including the species Barley Yellow Dwarf Virus-PAV (BYDV-PAV), frequently found in western Europe. Resistance sources against BYDV-PAV are scarce and only identified in perennial Triticineae. Some BYDV-resistant wheat lines have been obtained by introgressing these resistances into bread wheat germplasms. Genetic and biological characterization of the resulting lines has been undertaken. However, little information on the resistant behaviour of these lines during the early stages of the infection process is available. To evaluate the resistance of two genetically distinct resistant lines (Zhong ZH and TC14), 1740 young plantlets, belonging to susceptible reference hosts (barley cv. Express and wheat cv. Sunstar), Zhong ZH or TC14 wheat lines, were inoculated in controlled conditions with French BYDV-PAV isolates. The infection process was monitored during the first 21 days after inoculation (DAI) using a semi-quantitative ELISA. A standardized protocol including five successive samplings of leaves from all inoculated plants and the collection of plant roots at the end of the monitored period was carried out. This protocol enabled an assessment of the infection percentage and the evolution of the viral load in plants from the 7th DAI to the 21st DAI. Statistical analyses of the BYDV infection kinetics using raw ELISA data, a model of the time-dependent variation of the percentage of infected plants and the area under concentration progress curves (AUCPC) demonstrated that Zhong ZH and TC14 lines (1) reduce the development rate of the BYD disease during the first days of infection, (2) decrease the infection efficiency of BYDV-PAV isolates, in the leaves, from 98.7% for susceptible plant genotypes to 81.9% and 71.7% for Zhong ZH and TC14, respectively, (3) reduce the virus load in the leaves of infected plants and (4) are not spared from BYDV infection, as 95.1% of Zhong ZH and 90.2% of TC14 inoculated plants accumulated viral particles in roots and/or in leaves at 21 DAI. These results confirm the BYDV-partial resistant behaviour of both Zhong ZH and TC14 lines. The development rate of the disease was the single parameter that allowed the distinction between the two resistant sources present in the tested lines.     

Expression of Thinopyrum intermedium-Derived Barley yellow dwarf virus Resistance in Elite Bread Wheat Backgrounds

ABSTRACT Resistance to Barley yellow dwarf virus (BYDV) is not found in wheat but is available in a Thinopyrum intermedium translocation (Ti) carried on chromosome 7DL of bread wheat recombinant lines. We used one of those lines (TC14/2*Spear) to introgress the Ti into bread wheat cultivars and to determine the influence of wheat backgrounds, with and without known tolerance to BYDV, on the expression of resistance. Two single and three backcross populations, segregating for the presence of the alien fragment, were tested under field conditions and artificial inoculation with BYDV isolates MAV-Mex and PAV-Mex. Lines containing the fragment were identified using the microsatellite marker gwm37. Tillering, biomass, grain yield, thousand-kernel weight, and seed quality were evaluated in inoculated and noninoculated plots. Resistance was assessed by enzyme-linked immunosorbent assay. In early generations, the alien fragment followed expected Mendelian segregation, whereas in the advanced ones a slight bias against its transmission was observed. No positive nor negative effects of Ti on agronomic performance and quality were found. A significant optical density reduction in individuals carrying the fragment was observed after PAV infection in crosses with lines Anza and Baviacora but not with Milan. In addition, the fragment was associated with a lower frequency of infected plants for both PAV and MAV isolates. The reduced yield loss associated with the presence of the translocation was due largely to the lower infection rate.     

Virulence spectrum of Puccinia hordei in barley production systems in Ethiopia

Barley leaf rust caused by Puccinia hordei is an important disease of barley in Ethiopia. In the 2003 and 2004 cropping seasons, surveys of P. hordei were conducted on fields in the main rainy, residual moisture and short rainy season-barley production systems. A total of 381 isolates were analysed on 12 barley differential hosts carrying different Rph resistance genes ( Rph1 – Rph12 ). Based on infection phenotypes on leaf rust ( Rph ) resistance genes, seven pathotypes were identified, namely ETPh7611, ETPh7631, ETPh6611, ETPh7651, ETPh7671, ETPh7653 and ETPh7633, with frequencies of 63·0, 21·5, 6·8, 2·9, 2·6, 2·1 and 1·2%, respectively. ETPh7611 and ETPh7631 were the most common pathotypes found in all the surveyed areas of the three production systems. ETPh7653 was found in the small rainy season production system only. ETPh7671, ETPh7653 and ETPh7633 were the most virulent, but the least frequent, pathotypes. All isolates were virulent on resistance genes Rph1 , Rph4 , Rph8 , Rph9 , Rph11 and Rph12 . Virulence against Rph3 and Rph7 was absent. Genes Rph2 , Rph6+2 , Rph5 and Rph10 were effective against 96·3, 88·9, 65 and 2·4% of the rust isolates tested, respectively.     

Genetic Regulation of Polerovirus and Luteovirus Transmission in the Aphid Schizaphis graminum

ABSTRACT Sexual forms of two genotypes of the aphid Schizaphis graminum, one a vector, the other a nonvector of two viruses that cause barley yellow dwarf disease (Barley yellow dwarf virus [BYDV]-SGV, luteovirus and Cereal yellow dwarf virus-RPV, polerovirus), were mated to generate F1 and F2 populations. Segregation of the transmission phenotype for both viruses in the F1 and F2 populations indicated that the transmission phenotype is under genetic control and that the parents are heterozygous for genes involved in transmission. The ability to transmit both viruses was correlated within the F1 and F2 populations, suggesting that a major gene or linked genes regulate the transmission. However, individual hybrid genotypes differed significantly in their ability to transmit each virus, indicating that in addition to a major gene, minor genes can affect the transmission of each virus independently. Gut and salivary gland associated transmission barriers were identified in the nonvector parent and some progeny, while other progeny possessed only a gut barrier or a salivary gland barrier. Hemolymph factors do not appear to be involved in determining the transmission phenotype. These results provide direct evidence that aphid transmission of luteoviruses is genetically regulated in the insect and that the tissue-specific barriers to virus transmission are not genetically linked.     

应用农杆菌介导法获得转大麦黄矮病毒GPV株系复制酶基因ORF1的小麦植株

 以生产用4种基因型小麦为材料,利用农杆菌介导法将大麦黄矮病毒GPV株系复制酶基因ORF1分别转化愈龄40d的幼胚愈伤组织,经过G418筛选后,共再生出15株抗性植株,拓宽了小麦农杆菌转化的受体基因型。PCR及Southern杂交分析证实大麦黄矮病毒复制酶基因ORF1已经整合到小麦基因组中,共获得14株转基因植株。抗病性初步检测结果显示,转基因植株对GPV株系具有抗性。研究结果还表明,对于不同的基因型小麦进行转化时,需选择适合本基因型的菌株。     

小麦条锈病新抗源的抗谱鉴定初析

 利用南京农业大学细胞遗传研究所育成的一套涉及不同簇毛麦染色体的异附加系和代换系以及5个6VS/6AL易位系,经1997、1998、1999连续3年在陕西、北京、四川进行小麦条锈病抗性接种鉴定,结果表明普通小麦-簇毛麦6V异附加系,6V(6A)异代换系和6VS/6AL易位系高抗条锈病菌条中29、条中31、水源11-2、水源11-5、水源11-13和杂46等强毒小种。考虑到含整组V染色体的硬粒小麦-簇毛麦双倍体不抗水源11-13小种,上述普通小麦-簇毛麦6V异附加系、异代换系和6VS/6AL易位系的条锈病抗性可能还与其所涉及的小麦亲本基因的作用有关。     

我国不同地区麦蚜携带BYDV-GAV比率的差异

为了研究我国不同地区麦蚜携带大麦黄矮病毒麦二叉蚜麦长管蚜非专化性株系(BYDV GAV)比率的差异,采用RT-PCR技术,对BYDV-GAV的传毒介体麦蚜带毒情况进行检测.所用方法具有较高的灵敏度和特异性,测定样本用量可少至1/200头蚜虫;对采自我国主要麦区的蚜虫样本进行分子检测,山西、甘肃、青海、陕西11个小麦黄矮病重病区蚜虫样本带毒率为56％～91.5％,而河北、河南两省4个非重病区蚜虫样本带毒率为2.5％～33％.通过试验证实,我国不同地区麦蚜携带BYDV-GAV比率存在差异,小麦黄矮病重病区山西、甘肃、青海、陕西等地的麦蚜带毒率高,而非重病区河北、河南等地的麦蚜带毒率低.     

A simple test for evaluation of transmission efficiency of barley yellow dwarf virus by aphids

Barley yellow dwarf virus (BYDV) transmission test systems involve the use of clip-cages or of whole plants in cages, which are both labor-intensive methods and require large controlled environment units. Employing detached leaves for assessment of the inoculation efficiency of aphids proved reliable for assessing transmission of a BYDV PAV-like isolate byRhopalosiphum padi. One use of the system could be for the rapid determination of the infectivity of field-collected aphids, an essential part of any epidemiological study of BYDV. http://www.phytoparasitica.org posting Aug. 14, 2002.     

应用农杆菌介导法获得转大麦黄矮病毒GPV株系复制酶基因ORF2小麦植株

以生产用5种基因型小麦为材料,利用农杆菌介导法将大麦黄矮病毒GPV株系复制酶基因ORF2转化愈龄40d的幼胚愈伤组织,经过G418筛选后,共再生出9株抗性植株,拓宽了小麦农杆菌转化的受体基因型。PCR及Southern杂交分析证实大麦黄矮病毒复制酶基因ORF2已经整合到小麦基因组中。初步抗病性检测结果显示转基因植株对GPV株系具有抗性。研究结果还表明,受体基因型、外植体的生理状态对农杆菌侵染起着至关重要的作用。     

Phenotype and spectrum of action of the Pvr4 resistance in pepper against potyviruses, and selection for virulent variants

The dominant Pvr4 gene identified in Capsicum annuum cv. Criollo de Morelos 334 (CM334) is frequently used in pepper cultivars because it possesses one of the largest spectra of action among plant virus resistance genes. This gene was previously shown to confer efficient resistance to all known Potato virus Y isolates, to Pepper mottle virus , to Pepper yellow mosaic virus and to Ecuadorian rocoto virus. This study showed that the W4 line, derived from CM334 and carrying Pvr4 , was also resistant to Peru tomato mosaic virus and Pepper severe mosaic virus , but not to Pepper veinal mottle virus , Chilli veinal mottle virus or Tobacco etch virus . It was noticed that the phenotype of the resistance was atypical since, in the W4 line, hypersensitive reaction or extreme resistance could be observed, depending on virus isolates and inoculated organs. Despite the large deployment of Pvr4 in hybrid cultivars, the numerous tests performed in controlled conditions and the use of W4 serial back-inoculations with potyvirus isolates controlled by this line, no virulent variant isolates were obtained. However, it was shown that the use of graft inoculation experiments allow PVY virulent variants to be selected.     

Unterschiedliche Formen der Resistenz gegen bodenbürtige Viren des Weizens

In Germany the furovirus Soil-borne cereal mosaic virus (SBCMV) and the bymovirus Wheat spindle streak mosaic virus (WSSMV) occur often together particularly in several rye production areas. Soil-borne wheat mosaic virus (SBWMV), a wheat infecting furovirus, has so far been found only in one field near Heidelberg. Each of these viruses is transmitted by Polymyxa graminis. The cultivation of resistant varieties is the only promising measure to prevent yield losses caused by soil-borne viruses. Resistance of wheat against the bymovirus WSSMV is comparable to the immunity of barley to the bymoviruses Barley yellow mosaic virus and Barley mild mosaic virus. In case of immunity no virus multiplication is observed in resistant cultivars. In contrast, all wheat cultivars are hosts of the furoviruses. All cultivars – including the resistant ones – can be infected following mechanical inoculation with SBWMV and SBCMV. Resistance to furoviruses is based on reduced levels of virus multiplication in roots and on inhibition of virus movement from roots to leaves. Because of the inhibited virus movement from roots to aerial parts of plants this type of resistance is referred to as translocation resistance. In spite of the different resistance mechanisms the absence of virus symptoms on the leaves is a common selection criterion for both immunity and translocation resistance. Therefore, the symptom free development of plants on uniformly infested fields is the best criterion for selecting wheat lines with resistance to soil-borne viruses. The limited suitability of other selection methods is discussed.     

Extreme Reduction of Disease in Oats Transformed with the 5' Half of the Barley Yellow Dwarf Virus-PAV Genome

ABSTRACT Barley yellow dwarf viruses (BYDVs) are the most serious and widespread viruses of oats, barley, and wheat worldwide. Natural resistance is inadequate. Toward overcoming this limitation, we engineered virus-derived transgenic resistance in oat. Oat plants were transformed with the 5' half of the BYDV strain PAV genome, which includes the RNA-dependent RNA polymerase gene. In experiments on T2- and T3-generation plants descended from the same transformation event, all BYDV-inoculated plants containing the transgene showed disease symptoms initially, but recovered, flowered, and produced seed. In contrast, all but one of the BYDV-PAV-inoculated nontransgenic segregants died before reaching 25 cm in height. Although all of the recovered transgenic plants looked similar, the amount of virus and viral RNA ranged from substantial to undetectable levels. Thus, the transgene may act either by restricting virus accumulation or by a novel transgenic tolerance phenomenon. This work demonstrates a strategy for genetically stable transgenic resistance to BYDVs that should apply to all hosts of the virus.     

大麦黄花叶病抗源筛选

研究比较分析了“青华矮6号”等水稻抗病新品种对我省稻白叶枯病〔Xan-thomonas campestris pv.orvzae(Gshiyama 1923)Dye,1978〕Ⅰ—Ⅳ菌群的抵抗性,它们的抗感性按照品种×菌株的互作反应,可划分为6个类型,以中抗品种“华竹40”为亲本杂交育成的新品种均表现不同程度抗病。其中尤以“青华矮6号”的抗病性最好。“青华矮6号”是采用多系杂交育成的一个纯晚型中迟熟、丰产抗病、米质好的品种,测试证明对广东Ⅰ、Ⅱ、Ⅲ及Ⅳ菌群表现高抗,对优势菌群的抗病等级为1.3～1.4,CK_1(IR_(26)抗病)为0.84～1.2,CK_2(二白矮感病)为6.0,抗性差异达到显著水平,L.S.D=0.77(P=0.05),主要抗性基因为Xa_4。该品种经3年多点大面积验证试验结果,它的丰产抗病性表现一致。     

Disease severity and yield of pure-line wheat cultivars and mixtures in the presence of eyespot, yellow rust, and their combination

Five winter wheat cultivars, six two-component cultivar mixtures, and one four-way mixture were grown in the presence of yellow rust, eyespot, both diseases, and neither disease for three seasons. On average, mixtures reduced the severity of yellow rust relative to their component pure stands by 53%. The four-component mixture provided better yellow rust control than did the two-way mixtures. Eyespot severity was reduced through mixing only in the absence of yellow rust and by only three of the seven mixtures (mean reduction = 13%). Yellow rust was 13% less severe in the presence of eyespot, and eyespot was 10% more severe in the presence of yellow rust. Averaged over all years, the mixtures increased yield relative to the pure stands by 6·2, 1·7, 7·1, and 1·3% in the presence of yellow rust, eyespot, both diseases, and neither disease, respectively. Two mixtures provided significant yield increases over the means of their component pure stands (7% and 9%) in the presence of eyespot even though one of them did not significantly reduce eyespot severity. Accounting for all disease treatments and years, four mixtures provided distinctly higher yield increases than the other three. In mixtures containing a resistant cultivar and a cultivar susceptible to eyespot, yield loss by the susceptible cultivar was not compensated for by increased yield of the resistant cultivar. The mixtures showed improved yield stability relative to the pure stands, with the four-component mixture being particularly stable.     

灰飞虱传播的一种小麦病毒病鉴定

 在实验室中利用灰飞虱接种小麦时出现一种新的病毒病症状,鉴定表明其病原为大麦黄条点花叶病毒(Barley yellow striate mosaic virus, BYSMV)。采用生物学测定、电镜观察、RT-PCR检测和序列分析的方法,明确了该病毒的粒体特性、危害症状及田间发生情况。接种试验表明该病毒通过灰飞虱传播,接种7~10 d后小麦新生叶片出现黄色斑点、斑驳,继而发展成黄色条纹,叶片对生且细而窄,重病株新叶扭曲,叶鞘不能伸长,病株矮化。对小麦病叶超薄切片电镜观察,发现细胞质中存在大量弹状病毒粒子,病毒粒体大小为（315~353）nm×（46~57） nm。利用特异性引物从病株总RNA中扩增出 565 bp基因片段,序列同源性分析显示与大麦黄条点花叶病毒(Barley yellow striate mosaic virus, BYSMV) Zanjan-1分离物聚合酶（L）基因对应序列的一致性为97 %,与北方禾谷花叶病毒(Northern cereal mosaic virus, NCMV) L基因对应序列的一致性为78 %~79 %。对采自河北邯郸、石家庄、保定、唐山的31株样品进行RT-PCR检测,25株检测到BYSMV,7株检测到水稻黑条矮缩病毒(Rice black streaked dwarf virus,RBSDV),其中5株为2种病毒复合侵染,结果表明BYSMV的田间分布较广。系统发育分析表明BYSMV-Lab/TS/ZX/QY 4个分离物与本研究的BYSMV亲缘关系密切。BYSMV是我国小麦上新发现的一种弹状病毒,并已形成危害,暂定名为小麦黄条纹矮缩病,应加强流行动态监测。     

Resistance to Monopartite Begomoviruses Associated with the Bean Leaf Crumple Disease in Phaseolus vulgaris Controlled by a Single Dominant Gene

ABSTRACT Tomato yellow leaf curl virus (TYLCV) and Tomato yellow leaf curl Málaga virus are monopartite begomoviruses (genus Begomovirus, family Geminiviridae) that infect common bean (Phaseolus vulgaris), causing bean leaf crumple disease (BLCD). This disease was found to be widespread in southern Spain and causes stunted growth, flower abortion, and leaf and pod deformation in common bean plants. Commercial yield losses of up to 100% occur. In the present study, we have identified and characterized a resistance trait to BLCD-associated viruses in the common bean breeding line GG12. This resistance resulted in a complete absence of BLCD symptoms under field conditions or after experimental inoculation. Our analysis showed that virus replication was not inhibited. However, a severe restriction to systemic virus accumulation occurred in resistant plants, suggesting that cell-to-cell or long-distance movement were impaired. In addition, recovery from virus infection was observed in resistant plants. The reaction of P. vulgaris lines GG12 (resistant) and GG14 (susceptible), and of F(1), F(2), and backcross populations derived from them, to TYLCV inoculation suggested that a single dominant gene conferred the BLCD resistance described here.     

Emergence of Resistance-breaking Isolates of Rice yellow mottle virus during Serial Inoculations

Two sources of resistance to Rice yellow mottle virus were challenged in host passage experiments. Pronounced changes in pathogenicity occurred over serial passages of virus isolates inoculated to partially or highly resistant cultivars. The changes encompassed the known existing pathogenic variability of field isolates. Ultimately, the high resistance of the Oryza indica cv. Gigante was overcome and the partial resistance of the O. sativa japonica cv. Azucena broke down. The effect was resistance-specific as different isolates overcame partial and high resistance, and may also be allele-specific as different isolates overcame the resistance of cultivars carrying the same resistance gene. The ability of isolates to break resistance was not linked to a high initial pathogenicity of the isolates and did not result in higher virus content in the infected plants. Implications for resistance breeding and deployment are discussed.     

Phenotypic expression, stability, and inheritance of a recessive resistance to monopartite begomoviruses associated with tomato yellow leaf curl disease in tomato

Tomato-infecting begomoviruses comprise a complex of monopartite and bipartite virus species that cause severe yield and quality losses worldwide. Therefore, the availability of wide spectrum resistance for begomovirus control is desirable. However, limited sources of resistance are available. In this study, three tomato inbred lines with resistance to bipartite begomoviruses of Brazil were tested for resistance to monopartite begomoviruses associated with the tomato yellow leaf curl disease (TYLCD). Stable resistance to Tomato yellow leaf curl virus was observed either by inoculation with Bemisia tabaci or with Agrobacterium tumefaciens using an infectious clone. The resistance resulted in a complete absence of TYLCD symptoms and restricted virus accumulation. Further studies performed with the line '468-1-1-12' indicated that the resistance was also effective against three other virus species associated with TYLCD, indicating wide spectrum resistance of this source. Quantitative genetics analyses suggested that a major recessive locus with epistatic interactions is controlling the resistance to TYLCD in '468-1-1-12', which could facilitate introgression of this trait into elite tomato lines. The resistance was stable under field conditions with high TYLCD pressure. Mild symptoms could be observed in these conditions, and recovery from disease and from virus infection suggested an active host antiviral defense mechanism. The differential reaction of '468-1-1-12' against a number of TYLCD-associated viruses and artificial chimeras between them allowed to identify a region of the virus genome that presumably contains a virus determinant for breaking the resistance to infection observed in '468-1-1-12'.