Host range and molecular analysis of Beet leaf yellowing virus,Beet western yellows virus-JP and Brassica yellows virus in Japan

Beet western yellows virus (BWYV; genus Polerovirus, family Luteoviridae) is one of the most important viruses causing yellowing disease of many field and vegetable crops. This study isolated different poleroviruses from sugar beet, spinach, radish and brassica in Japan, and identified them as BWYV-JP, Beet leaf yellowing virus (BLYV), Brassica yellows virus (BrYV) and BrYV-R (radish strain) based on host range and molecular analysis. Among over 100 plant species from 19 families inoculated with the vector Myzus persicae, about half of the species in 13 families were infected with some of these viruses. BLYV shared a similar host range to Beet mild yellowing virus (BMYV). These had a much more limited host range than BWYV-JP, which resembled BWYV-USA. The host range of BrYV was similar to that of Turnip yellows virus (TuYV). Phylogenetic analyses at the 5′ portion (replication-related gene) of the genome showed that BLYV, BMYV, BWYV (-JP and -USA) and Cucurbit aphid-borne yellows virus (CABYV) formed one large group, whereas BrYV and TuYV were grouped together. BLYV and BWYV were most closely related to each other, and were more closely related to CABYV than to BMYV. However, at the 3′ end (coat protein gene), BLYV and BWYV-JP formed a distinct group, separated from the BrYV group, which in turn was more closely related to BWYV-USA, BMYV, TuYV and Beet chlorosis virus, a group originating from outside Asia. Thus, this study presents host range differences and phylogeographical relationships of BWYV-like poleroviruses that are distributed worldwide.     

北京地区辣椒上黄瓜花叶病毒和甜菜西方黄化病毒复合侵染的分子鉴定

辣椒是我国重要的蔬菜和经济作物,受多种病毒危害。2014年在北京市顺义区调查时发现部分种植的辣椒植株上叶片大面积黄化,边缘症状明显,个别植株叶片轻微上卷。提取典型症状样品的总RNA,反转录得到cDNA,分别用黄瓜花叶病毒(Cucumber mosaic virus,CMV)特异引物和马铃薯卷叶病毒属(Polerovirus)通用引物进行PCR检测,CMV特异引物和马铃薯卷叶病毒属通用引物分别扩增得到约650bp和1 400bp的特异条带。测序和核苷酸序列比对表明,其分别与CMV和甜菜西方黄化病毒(Beet western yellows virus,BWYV)序列同源性最高为99%和96%。这是对我国种植的辣椒上发生的CMV和BWYV复合侵染的首次报道。     

Involvement of Beet western yellows virus, Cauliflower mosaic virus, and Turnip mosaic virus in Internal Disorders of Stored White Cabbage

ABSTRACT Experiments over two growing seasons clearly showed that Turnip mosaic virus (TuMV) infection was associated with internal necrosis (sunken necrotic spots 5 to 10 mm in diameter) and Beet western yellows virus (BWYV) infection was associated with collapse of leaf tissue at the margins (tipburn) in heads of stored white cabbage (Brassica oleracea var. capitata). Virtually no tipburn was seen in cv. Polinius, whereas cv. Impala was affected severely. Internal necrotic spots were seen in both cultivars. BWYV appeared to interact with TuMV. Plants infected with both viruses showed a lower incidence of external symptoms and had less internal necrosis than plants infected with TuMV alone. Cauliflower mosaic virus (CaMV) did not induce significant amounts of internal necrosis or tipburn, but did, in most cases, exacerbate symptoms caused by TuMV and BWYV. BWYV-induced tipburn worsened significantly during storage. Post-transplanting inoculation with TuMV induced more internal necrosis than pre-transplant inoculation. There was a significant association between detection of TuMV just prior to harvest and subsequent development of internal necrotic spots. Individually, all three viruses significantly reduced the yield of cv. Polinius, whereas only BWYV and CaMV treatments reduced the yield of cv. Impala.     

Genetics of resistance to beet western yellows virus in lettuce

Resistance to beet western yellows virus (BWYV) in the lettuce cultivars Burse 17 and Crystal Heart was controlled by a single recessive gene designated bwy. ELISA tests showed that resistant plants were not immune to infection by BWYV, and may develop some mild symptoms. However, the concentration of virus found in resistant plants was less than in susceptible ones. The cultivar Crystal Heart is being used as a source of resistance for breeding BWYV-resistant crisp lettuce (iceberg) cultivars.     

Molecular evidence for the occurrence of beet western yellows virus on chickpea in Morocco

A luteovirus isolate infecting chickpea in Morocco was experimentally transmitted by Myzus persicae to Physalis floridana, on which it produced mild symptoms. When tested in western blots against antisera to known legume luteoviruses, this isolate reacted strongly to beet western yellows virus (BWYV) antiserum, moderately to bean leafroll virus antiserum, while no reaction was recorded with the antiserum against subterranean clover red leaf virus. In PCR, a fragment of ca. 950 bp was amplified, comprising the 3' end of the open reading frame (ORF) 3, the complete coat protein gene (ORF 4), and the non-translated region in between these ORFs. The nucleotide sequence of the amplified fragment showed high similarity with BWYV (approximately 96%), and lower (50–60%) with other luteoviruses reported to infect legumes. On the basis of these data, the Moroccan isolate was identified as BWYV. This is the first molecular evidence for the occurrence of BWYV on chickpea in Morocco, and on food legumes in general in North Africa.     

芜菁花叶病毒对油菜致病力差异及壳蛋白基因序列分析

 2004年春季参试的湖北、安徽2省11个芜菁花叶病毒(Turnip mosaic virus,TuMV)分离物感染4个油菜品种,病情指数幅度为26.2~76.0;秋季参试5个TuMV分离物感染14个油菜品种,病情指数幅度为38.3~55.9,均值方差分析表明,致病力差异分别达到极显著和显著水平。壳蛋白(CP)基因序列分析表明,来源于2省油菜、白菜、红菜薹、芝麻和萝卜的17个TuMV分离物与浙江分离物ZJB3序列同源性在97%以上,同属于MB类群;而另一个萝卜分离物WRS1与ZJR1、CH1和CH2分离物序列同源性在95.4%~98.7%之间,属于MR类群。类群间分离物序列同源性仅为88.0%~92.2%。遗传进化树分析表明,萝卜分离物WRS2在MB类群中单独构成一个分支,可能是MR类群和MB类群发生重组的后代。     

Comparison of phytoplasmas infecting winter oilseed rape in the Czech Republic with Italian Brassica phytoplasmas and their relationship to the aster yellows group

Winter oilseed rape grown in several areas in South Bohemia showed symptoms of stunting, leaf reddening and extensive malformation of floral parts. Phytoplasmas were consistently observed by using electron microscopy only in phloem tissue of symptomatic plants. DNA isolated from infected and healthy control plants was used in PCR experiments. Primer pairs R16F2/R2, P1/P7 and rpF2/R2, amplifying, respectively, 16S rDNA, 16S rDNA plus spacer region and the beginning of the 23S and ribosomal protein gene L22 specific for phytoplasmas, were used. According to RFLP and sequence analyses of PCR products, the phytoplasma from rape was classified in the aster yellows phytoplasma group, subgroup 16SrI-B. The PCR products from the Czech phytoplasma-infected rape also had RFLP profiles identical to those of phytoplasma strains from Italian Brassica . This first molecular characterization of phytoplasmas infecting rape compared with strains from Brassica does not, however, clearly indicate differences among isolates of the same 16SrI-B subgroup. Further studies on other chromosomal DNA portions could help the research on host specificity or on geographical distribution of these phytoplasmas.     

Changes in the glucosinolate content of oilseed rape plants following infection with turnip mosaic virus

Low and high glucosinolate cultivars of oilseed rape were analysed for variation in both individual and total glucosinolate content up to 27 days after inoculation with turnip mosaic virus. Both infected and control plants of the two cultivars differed in the concentration of aliphatic, aromatic and indole glucosinolates. The biggest differences in glucosinolate content between the cultivars were found in the aliphatic and aromatic glucosinolate groups. Infected plants of the high glucosinolate cultivar, Solida, possessed lower amounts of glucosinolates, than did healthy control plants. Similar results were obtained with a low glucosinolate cultivar, S1. No significant difference was found between the two cultivars in resistance to turnip mosaic virus, using qualitative and quantitative ELISA to determine virus content in their tissues.     

Occurrence of yellowing viruses (Beet pseudo-yellows virus, Cucurbit yellow stunting disorder virus and Cucurbit aphid-borne yellows virus) affecting cucurbits in Greece

A survey of the incidence of yellowing viruses in Greek glasshouse (and occasional field) cucumber and melon crops was carried out during 2000–03. In most cases disease incidence ranged from 50 to 80%. Simplex RT-PCR was used for the detection of Beet pseudo-yellows virus (BPYV) and Cucurbit yellow stunting disorder virus (CYSDV), and DAS-ELISA for the detection of Cucurbit aphid-borne yellows virus (CABYV). The results showed that BPYV was the predominant virus in cucumber and melon crops, whereas CYSDV, reported for first time in Greece, was isolated only in three regions of southern Greece: Rhodes, Crete and Arkadia. CABYV was detected only in three cucumber glasshouses in Pella (Macedonia). A simplified triplex RT-PCR method using a simple sample-preparation protocol was developed to allow rapid, sensitive and simultaneous detection of the three viruses. Sequence comparisons of the PCR products of BPYV and CYSDV revealed 98·7 and 100% amino acid identity, respectively, with previously reported sequences. The arable weed species Amaranthus retroflexus , Selosia cristata , Sonchus oleraceus and Sonchus sp. were identified as potential BPYV reservoirs.     

Herbicide sensitivity of transgenic multiple herbicide-tolerant oilseed rape

Glyphosate and glufosinate-ammonium herbicide tolerance traits were combined into both winter and spring lines of Brassica napus L. This allowed the study of possible interactions between these transgenes in two genetic backgrounds when treated with a variety of herbicides. Selective herbicides that are commonly used within Brassica crops showed no adverse effects on the transgenic plants or their null controls. Lines containing both glyphosate and glufosinate transgenes remained tolerant to their respective herbicides, regardless of the presence of the second tolerance transgene. Lines containing only a single transgene retained tolerance to the encoded trait and did not show cross-tolerance to the second. Null lines were killed by either herbicide. All plant lines, regardless of their transgene content, were found to be equally susceptible to three herbicides (paraquat, metsulfuronmethyl and mecoprop), commonly used to remove volunteer B napus from succeeding crops and set-a-side land.     

Latent infection of winter oilseed rape by Leptosphaeria maculans

Plants of oilseed rape, cultivars Primer and Jet Neuf, were grown in a glasshouse and inoculated at G.S. 2.4–2.7 with pycnidiospores or ascospores of Leptosphaeria maculans. The plants were kept for a further 2–4 weeks at 14°C and then transferred, together with uninoculated plants, to a polythene tunnel in winter. The majority of stems of inoculated plants did not have macroscopic symptoms of L. maculans infection 6 weeks after inoculation. Examination of whole mounts of peripheral tissue and transverse sections of fixed and embedded portions of these stems revealed intercellular septate fungal hyphae, often deep in non-necrotic cortical tissue, in symptomless inoculated plants but not in uninoculated plants. L. maculans was recovered following surface sterilization of adjacent portions of the same stems. When symptomless inoculated plants were transferred to a glasshouse at 18–20°C, cankers soon developed. The significance of these latent mycelial infections to canker development in the field is discussed.     

Seed dressings to control slug damage in oilseed rape

Slugs are major pests of oilseed rape that are poorly controlled by conventional bait pellets. A series of laboratory experiments investigated the potential of seed-dressings to control slug damage in this crop. Four compounds: metaldehyde, methiocarb, cinnamamide and 3,5-dimethoxycinnamic acid (DMCA) were tested at a range of doses for phytotoxicity and ability to reduce damage by Deroceras reticulatum (Müller). Metaldehyde and methiocarb were not phytotoxic at any doses, whereas all doses of cinnamamide and DMCA were. All compounds reduced slug damage, but metaldehyde and methiocarb consistently performed better than cinnamamide and DMCA. Metaldehyde and methiocarb seed-dressings were compared with baited pellets containing the same active ingredients at recommended field doses. The seed-dressings protected plants from damage by D reticulatum and Arion subfuscus (Draparnaud) as well as, or better than, baited pellets. We therefore recommend that metaldehyde and methiocarb should be field-tested as seed dressings to control slugs in oilseed rape.     

Prediction of competition between oilseed rape and Stellaria media

Ten experiments have investigated competition between winter oilseed rape and Stellaria media (common chickweed). Yield losses caused by this weed were often high, but differed greatly between experiments, 5% yield loss being calculated to be caused by 1.4–328 plants m^?2. Predictions of yield loss based on relative weed dry weights [weed dry weights/(crop + weed dry weights)] in December were somewhat less variable than those based on weed density, 5% yield loss being caused by 1.4–10.6% relative weed dry weight. The variations in yield loss were related to variations in the competitiveness of the oilseed rape and the S. media, caused by weather differences between years and sites, and the long period between weed assessment and harvest (8–10 months). However, despite the lack of precise relationships, there were indications that the greater the crop dry weights in December, the lower the final yield loss. Delayed sowing of oilseed rape until late September did not clearly increase the competitive effects of the weed compared with late August/early September sowings. Weed competition was not clearly affected by reduced crop density (44–113 plants m^?2), because of the compensatory ability of the lowest density. The results of the experiments are discussed in relation to the prediction of yield loss and, thus, possible adjustment of weed control strategies to meet expected crop losses.     

Biological control methods of pest insects in oilseed rape

Parasitoids, predators, insect pathogenic fungi, nematodes and microsporidia have all shown promise in the biological suppression of the key target pest, the pollen beetle Meligethes aeneus in rapeseed, as well as in the control of other rapeseed pests. Under favourable conditions pollen beetles are locally and temporarily already under efficient natural control by one or several of these agents, but overall the pollen beetle remains the number one pest of rapeseed in Finland, as well as elsewhere in Europe. Conservation biological control to enhance natural control appears the most feasible approach to solve the problem. Field and simulation data are used to demonstrate how a simple change in the tillage practice may be enough to obtain efficient control of the pollen beetle. Several other options are, or will be available, subject to successful registration of microbial products based on the fungi Metarhizium anisopliae and Beauveria bassiana , and the nematode Steinernema feltiae . Strategies and methods of applying these agents have been developed to facilitate either inundative treatments, or ecosystem restoration to (re-)establish the presence of these agents in agricultural fields. Critical components of agricultural practice have been identified for maximising control by biological agents within a functioning IPM system.     

The reaction of two lettuce cultivars to mixed infection by beet western yellows virus, lettuce mosaic virus and cucumber mosaic virus

The interaction of beet western yellows virus (BWYV), lettuce mosaic virus (LMV) and cucumber mosaic virus (CMV) has been studied in lettuce cultivars Little Gem and Saladin. LMV infection alone or in combination with BWYV and or CMV caused the most severe symptoms and yield losses in both cultivars. BWYV caused more severe reactions in Little Gem than in Saladin; the reactions caused by CMV were mild in both cultivars. The interaction between BWYV and CMV infection resulted in a significantly greater yield loss in both cultivars than that caused by BWYV or CMV infection alone.     

The production and release of ascospores of Pyrenopeziza brassicae on oilseed rape

The results of observation for three seasons in oilseed rape crops of the production and release of ascopores of Pyrenopeziza brassicae , the cause of light leaf spot disease of brassicas, are presented. Large apothecia (1–2 mm in diameter) take at least 3 weeks to develop on leaf petioles after infected leaves die, but small apothecia (50–200 um) may form on leaf lamellae after about 15 days. Apothecia were found on all 12 oilseed rape varieties examined. Spore trapping experiments in infected crops show that ascospore release is associated with rain, but most ascospores are released after rainfall when the crop debris bearing apothecia are wet. Ascospores can be released for up to 5 days after rain. Laboratory measurements show that apothecia can continue to release ascospores for up to 3 weeks even when they are subjected to wet and dry cycles. The consequences of the widespread occurrence of the sexual stage of P. brassicae are discussed and possible cycles of disease and spore production in oilseed crops are suggested.