Properties of a new strain of tobacco streak virus from Clematis vitalba (Ranunculaceae)

A distinct strain of tobacco streak virus (TS V/Cle), isolated in Yugoslavia from wild Clematis vitalba showing chlorotic spots or yellow netting of the leaves and from many symptomless shrubs, is described. TSV/Cle was seed transmitted in C. vitalba (70%), and in the experimental hosts Chenopodium quinoa (80%), Nicotiana benthamiana and N. megalosiphon. It was also detected in the pollen of infected C. quinoa. Purified virus preparations mostly contained quasi-spherical particles measuring 24–26 × 28, 26–28 × 28–30 and 28–31 × 32–36 mn, and sedimented in sucrose density gradient and analytical centrifugation as three components with sedimentation coefficients of 76S, 87S and 98S. The virus contained a single polypeptide species of mol. wt of c. 25 000. Unfractionated TSV/Cle preparations contained four RNA species with mol. wts, estimated by gel electrophoresis in agarose, of 1.1 × 10⁶, 0.9 × 10⁶, 0.7 × 10⁶ and 0.3 × 10⁶. In comparative experiments, TSV/Cle differed from four reference strains of TSV (TSV/B, TSV/HF, TSV/RN, and TSV/Ro) in host range and in symptoms induced in some common hosts. In agar gel double diffusion tests it was more closely related to TSV/B and TSV/M (SDI = 5) than to TSV/HF (SDI = 7), TSV/RN (SDI = 7) or TSV/Ro (SDI = 5–8). Immunoelectrophoresis experiments clearly distinguished TSV/Cle from the reference strains. TSV/Cle strain was detected in C. vitalba plants from distant and climatically different regions in Yugoslavia.     

Characterization of a Latent Elongated Virus from Globe Artichoke (Cynara scolymus L.) in Italy

A virus with filamentous particles was isolated from symptomless plants of Cynara scolymus cvs Romanesco and Terom obtained by in vitro meristem culture in northern Italy. The virus was characterized biologically, physico-chemically and serologically. The cytopathology induced by its infection in two artificial hosts (Chenopodium quinoa and Nicotiana benthamiana) was also investigated. The virus has slightly flexuous elongated particles measuring 12 ± 664 nm; its sedimentation coefficient, RNA content, mol. wts. of RNA and coat protein subunits are 150 S, 6 %, 2.2 × 10⁶ and 2.9 × 10⁴, respectively. In microprecipitation tests, it resulted serologically related to poplar mosaic virus (PopMV) (SDI = 4–5). Cellular inclusions and cytopathology observed in both the artificial hosts conform to those of the carlavirus group.     

Artichoke Italian latent virus causes artichoke patchy chlorotic stunting disease

Artichoke patchy chlorotic stunting (APCS) is the most serious disease affecting artichoke in Greece. It is widespread in Argolis, the main artichoke centre in Greece, where the local thornless artichoke, cv. “Argos”, is cultivated. The disease was first noticed in 1982, but to growers it was known since earlier times. Data collected during 1980–1990 revealed its wide distribution in the area, its patchy dispersal in fields, and its annual radial increase of the patches during the years, as well as a centripetal symptomatological severity gradient within each patch. These observations indicated a soil-borne nature for the disease. Field surveys for several viruses indicated a correlation of APCS with artichoke Italian latent nepovirus (AILV), a soil-borne virus known to occur in such patches and transmitted by the vector nematode Longidorus fasciatus Roca et Lamberti. In experiments fulfilling Koch's postulates, it was possible to reproduce the symptoms of APCS, demonstrating that AILV is a cause of the disease, if not the only one. Field surveys also revealed the ubiquitous occurrence of artichoke mottled crinkle virus (AMCV), independently of any particular symptoms. In the same survey, broad bean wilt virus (BBWV), also, was recovered from artichoke in Greece for the first time.     

Further Studies on Cynara Rhabdovirus

Cynara rhabdovirus (CyRV) was isolated from symptomless artichoke plants in southern Italy using Nicotiana langsdorffii as susceptible host and immune serum to artichoke latent virus to eliminate this virus from inoculum. CyRV can infect several solanaceous species, has thermal inactivation point of 40-45°C, dilution end point between 10^?2 and 10^?5 and longevity in vitro at 4 and 20°C of 4-5 days and 6-24 h, respectively. It was purified and used for preparing antisera with homologous titre varying from 1: 16 to 1: 64. In decoration tests, the virus did not react against antisera to eggplant mottled dwarf virus (EMDV) and its antiserum did not decorate ivy vein clearing virus (IVCV). Ultrastructural aspects of CyRV infection in Datura stramonium and Nicotiana glutinosa were very like those described for the same virus in the past except for presence of longer virions which were often encountered in infected cells during this study.     

Viral diversity in Phytophthora cactorum population infecting strawberry

Eighty-eight Phytophthora cactorum strains isolated from crown or leather rot of strawberry in 1971–2019 were screened for viruses using RNA-seq and RT-PCR. Remarkably, all but one isolate were virus-infected, most of them harbouring more than one virus of different genera or species. The most common virus occurring in 94% of the isolates was the Phytophthora cactorum RNA virus 1 (PcRV1) resembling members of Totiviridae. Novel viruses related to members of Endornaviridae, named Phytophthora cactorum alphaendornaviruses 1-3 (PcAEV1-3), were found in 57% of the isolates. Four isolates hosted viruses with affinities to Bunyaviridae, named Phytophthora cactorum bunyaviruses 1-3 (PcBV1-3), and a virus resembling members of the proposed genus ‘Ustivirus’, named Phytophthora cactorum usti-like virus (PcUV1), was found in a single isolate. Most of the virus species were represented by several distinct strains sharing ≥81.4% aa sequence identity. We found no evidence of spatial differentiation but some temporal changes in the P. cactorum virus community were observed. Some isolates harboured two or more closely related strains of the same virus (PcAEV1 or PcRV1) sharing 86.6%–96.4% nt identity in their polymerase sequence. This was surprising as viruses with such a high similarity are typically mutually exclusive.     

Host range and properties of artichoke yellow ringspot virus

The biological, serological and physico-chemical properties of one isolate of artichoke yellow ringspot virus (AYRV) from Greece and another from Italy were compared. Both isolates infected 56 herbaceous species and there were few differences between them in the symptoms they caused. During purification they behaved identically and both tended to aggregate. Virus particles were isometric and measured c. 30 nm in diameter. In CsCl, virus sedimented as mixed aggregates of empty and full particles with buoyant densities varying from 1.20–1.30 g/ml and from 1.40–1.53 g/ml, respectively. The coat protein of AYRV contains a single polypeptide of mol. wt 53000 and the genome consists of two species of single-stranded RNA with mol. wts 2.17 × 10⁶ (RNA-1) and 1.85 × 10⁶ (RNA-2) daltons, estimated under denaturing conditions. The two virus isolates are serologically very closely related but are unrelated to 28 other plant viruses with isometric particles. The characteristics of AYRV suggest that it is a possible member of the nepovirus group.     

Elderberry latent virus: its relationship to Pelargonium ringspot virus and its identification as a distinct member of the genus Carmovirus, family Tombusviridae

The properties of Elderberry latent virus (ELV) and Pelargonium ringspot virus (PelRSV) were compared. The viruses were largely indistinguishable in herbaceous host range and symptomatology, particle morphology, sedimentation coefficient and RNA profiles and size. They were also very closely related serologically with SDI differences in agarose gel double‐diffusion tests of 1 to 3. Purified virus particle preparations of each virus contained isometric particles c. 30 nm in diameter that sedimented as a major component with an s^O_20W of 112–115S. Purified virus particle preparations contained a major and a minor ssRNA species that in polyacrylamide gel electrophoresis (PAGE) had estimated sizes of c. 3.8 kb and c. 1.6 kb respectively. Plants of Chenopodium quinoa infected with ELV or PelRSV each contained three dsRNA species of c. 3.8, 2.6 and 1.8 kbp, although the smallest of these species was not evident in all preparations. Protein from purified virus particle preparations contained a major polypeptide that, in SDS‐PAGE, had an estimated M^r of 40 000 (40K). However, after storage of purified virus particles for 7–10 days, protein preparations from PelRSV particles also contained an additional major polypeptide of estimated M^r of 37 000 that is probably derived by degradation of the 40K protein; this additional component was not observed in freshly prepared preparations of ELV. Neither virus was found to be related serologically to 16 other viruses with isometric particles and similar properties. These data, together with the recent finding by other researchers that the smallest RNA species is a sub‐genomic RNA, suggests that both viruses are members of the genus Carmovirus, and that PelRSV is a minor variant of ELV. However, the taxonomic status of these two viruses is discussed in relation to recent brief reports comparing the nucleotide and amino acid sequences of these two viruses.     

Studies on two Serologically Distinct Raspberry Ringspot Virus Strains from Artichoke

Two raspberry ringspot virus variants, RRV-T and RRV-G, found in artichoke of Turkish and Greek origin, were compared biologically (indexing), biochemically and serologically to two strains of the same virus, RRV-S and RRV-E originating from Scotland and England, respectively. Molecular weight values of protein and nucleic acid of RRV-T and RRV-G were in good agreement with those already known for RRV. RRV-T and RRV-G appeared serologically very similar to each other (serological differentiation index = 1) and well distinguishable from RRV-S and RRV-E (serological differentiation indices varying between 3 and 6). Phaseolus vulgaris cv. La Victoire, Ocimum basilicum cv. Foglia di Lattuga, Cucumis sativus cv. Delicatezza, and Cucurbita pepo cv. Zucchetta striata d'ltalia seemed herbaceous hosts useful for differentiating each strain from the others on symptomatological basis.     

Host range, purification and some properties of rubus Chinese seed-borne virus

A previously undescribed virus, for which the name rubus Chinese seed-borne virus (RCSV) is proposed, was isolated from a single, symptomless plant of an unidentified Rubus species grown from seed collected in the wild in the People's Republic of China, Experimentally RCSV infected 23 out of 39 spp. in six out of eight families. The virus was seed-transmitted in Chenopodium quinoa (100%) and Nicotiana bigelowii (27%). RCSV was not transmitted by the nematodes Xiphinema diversicaudatum or X. index. The particles of RCSV were isometric, c. 30 nm in diameter with some penetrated by negative stains. In thin sections particles were found in double walled tubular structures with an outer membrane enclosing one or more tubules. In crude extracts some particles were found within single-walled tubules. Two virus-associated bands were seen in sucrose density gradients of purified preparations. The upper band was not infective and consisted of penetrated particles apparently devoid of nucleic acid. The lower, infective band was resolved into two components, of density 1.452 and 1.461 g/ml, in caesium chloride isopycnic gradients. There were two polypeptides (mol. wts c. 47 000 and 25 200 daltons) and two nucleic acid species (one of mol. wt c. 1.4 × 10⁶ daltons; the second was poorly defined by the methods used but was of higher molecular weight). RCSV was distantly related serologically (6–7 SDI) to the type isolate of strawberry latent ringspot virus (SLRV) and also reacted with antisera to serologicaly distinct grape and olive isolates of SLRV. It did not react with antisera to 10 other isometric viruses.     

Chemical Composition of the Tuber Essential Oil from Helianthus tuberosus L. (Asteraceae)

Helianthus tuberosus L. (Jerusalem artichoke) is cultivated in Europe and other parts of the world as a food crop and ornamental plant. The volatile oils of the aerial parts of H. tuberosus were investigated more than 30 years ago, but no study could be found to date on the constituents of the tuber essential oil. Herein, the first characterization by GC‐FID, GC/MS, and ¹³C‐NMR analyses of a hydrodistilled essential oil of Jerusalem artichoke tubers was reported. Fresh plant material collected in Serbia (Sample A) and a commercial sample (Sample B) yielded only small amounts of oil (0.0014 and 0.0021% (w/w), resp.). In total, 195 constituents were identified, representing 88.2 and 93.6% of the oil compositions for Samples A and B, respectively. The main constituents identified were β‐bisabolene ( 1 ; 22.9–30.5%), undecanal (0–12.7%), α‐pinene (7.6–0.8%), kauran‐16‐ol ( 2 ; 6.9–9.8%), 2‐pentylfuran (0.0–5.7%), and (E)‐tetradec‐2‐enal (0.0–4.9%). Several rare compounds characteristic for Helianthus ssp. were also detected: helianthol A ( 6 ; 2.1–1.9%), dihydroeuparin ( 10 ; 0.0–2.3%), euparin ( 9 ; 0.0–0.4%), desmethoxyencecalin ( 7 ; traces – 0.2%), desmethylencecalin ( 8 ; 0.0–0.4%), and an isomer of desmethylencecalin (0.0%‐traces). The essential oils isolated from the tuber and the aerial parts share the common major component 1 .     

The Composition of Jerusalem Artichoke (Helianthus tuberosus L.) Spirits Obtained from Fermentation with Bacteria and Yeasts

The composition of spirits distilled from fermentation of Jerusalem artichoke (Helianthus tuberosus L.) tubers was compared by means of gas chromatography. The microorganisms used in the fermentation processes were the bacterium Zymomonas mobilis, strains 3881 and 3883, the distillery yeast Saccharomyces cerevisiae, strains Bc16a and D₂ and the Kluyveromyces fragilis yeast with an active inulinase. The fermentation of mashed tubers was conducted using a single culture of the distillery yeast Saccharomyces cerevisiae and the bacterium Zymomonas mobilis (after acid or enzymatic hydrolysis) as well as Kluyveromyces fragilis (sterilized mashed tubers). The tubers were simultaneously fermented by mixed cultures of the bacterium or the distillery yeast with K. fragilis. The highest ethanol yield was achieved when Z. mobilis 3881 with a yeast demonstrating inulinase activity was applied. The yield reached 94 % of the theoretical value. It was found that the distillates resulting from the fermentation of mixed cultures were characterized by a relatively lower amount of by‐products compared to the distillates resulting from the single species process. Ester production of 0.30–2.93 g/L, responsible for the aromatic quality of the spirits, was noticed when K. fragilis was applied for ethanol fermentation both in a single culture process and also in the mixed fermentation with the bacterium. Yeast applied in this study caused the formation of higher alcohols to concentrations of 7.04 g/L much greater than those obtained with the bacterium. The concentrations of compounds other than ethanol obtained from Jerusalem artichoke mashed tubers, which were fermented by Z. mobilis, were lower than those achieved for yeasts.     

Chemical Composition,Antioxidant Potential and Enzymes Inhibitory Properties of Globe Artichoke By‐Products

In this study, chemical composition and in vitro biological activities of artichoke by‐products (leaves, floral stems and bracts) issued from two Tunisian varieties were evaluated. Analysis was performed by means of high‐performance liquid chromatography with diode array detection coupled to electrospray ionization mass spectrometric (LC/DAD/ESI‐MS). Total phenolic (TPC) and flavonoid (TFC) contents as well as the antioxidant activity conducted by three complementary methods, DPPH, ABTS and FRAP tests, were performed for each sample. Enzyme inhibitory effects against acetylcholinesterase, butyrylcholinesterase and α‐amylase were also studied. Results showed that TPC and TFC varied according to variety as well as the plant part. Bracts presented the highest TPC values (10–15 mg GAE/g DW), while leaves were distinguished by the highest TFC values (52–58 mg EQ/g DW). In vitro assays showed that Violet d'Hyères bracts and Blanc d'Oran leaves present the most antioxidant activities (30.040 and 20.428 mgET/gDW, respectively, by the DPPH method). Leaves demonstrated the highest acetylcholinesterase and butyrylcholinesterase inhibitory effects. Moreover, all organs displayed a noticeable inhibition towards α‐amylase. LC/DAD/MS analysis revealed that artichoke by‐products are a potential source of biopharmaceuticals such as luteolin derivatives from leaves and mono/dicaffeoylquinic acids in the other parts. This research demonstrates that globe artichoke by‐products, unexploited in our country, are a promising source of natural health promoting compounds with potential applications in the food and pharmaceutical industries.     

Metabolism of fructo-oligosaccharides by Bifidobacterium spp.

Summary Bifidobacterium adolescentis ATCC 15703, B. longum ATCC 15707, and B. thermophilum ATCC 25525 were examined for the ability to grow with fructo-oligosaccharides (FOS) as carbohydrate sources. The three species produced cell-associated -fructosidases (inulinases) capable of hydrolysing FOS. Maximum activity was obtained with short-chain FOS with degrees of polymerization (DP) of between three and five (neosugars). The B. thermophilum inulinase was induced by inulin, a long-chain FOS with DP=35, while the enzymes from the other two strains were constitutive. Production of inulinase by all three strains was regulated by catabolite repression. Inulinase activity of the three Bifidobacterium spp. was similar when grown with 0.5% inulin as the carbohydrate source; however, B. thermophilum grew much more rapidly. All three strains utilized crude Jerusalem artichoke flour (JAF) as a carbohydrate source, suggesting that JAF might have commercial application as a food or feed additive to stimulate bifidobacteria in the gut.Contribution no. 802 from the Food Research Centre     

Xylanase production by Aureobasidium pullulans on globe artichoke stem: Bioprocess optimization,enzyme characterization and application in saccharification of lignocellulosic biomass

Statistical optimization of the factors affecting xylanase production by Aureobasidium pullulans NRRL Y-2311-1 on globe artichoke stem was performed for the first time. The optimization strategies used resulted in almost six-fold enhancement of xylanase production (66.48?U/ml). Biochemical and thermal characterization of the crude xylanase preparation was performed to elucidate its feasibility for different industrial applications. The optimum conditions for xylanase activity were pH 4.0 and 30–50°C. The enzyme was very stable over a wide pH range of 3.0–8.0. The thermal stability studies revealed an inactivation energy of 183?kJ/mol. Thermodynamic parameters (enthalpy, entropy, and Gibbs free energy) for thermal inactivation were also determined. Primary application of the crude xylanase preparation in saccharification of corn cob subjected to different pretreatment techniques has been evaluated. The crude xylanase preparation was very promising for saccharification of corn cob pretreated with aqueous ammonia. The maximum yield of reducing sugar was 357?mg/g dry substrate, which revealed that the crude xylanase from A. pullulans could be a very good alternative in saccharification of lignocellulosic biomass for biological fuel generation. This study also provides a basis for further exploitation of globe artichoke by-products in microbial production of several other industrially significant metabolites.     

Studies on the occurrence of tomato bushy stunt virus in English rivers

Tomato bushy stunt virus (TBSV) of unknown source was isolated from water of the River Thames, near Oxford. The isolate designated TBSV-T was mechanically transmissible to several tomato (Lycopersicon esculentum) cvs and to other species including Petunia hybrida, pepper (Capsicum annuum). eggplant (Solanum melongena), Nicotiana clevelandii, Chenopodium amaranticolor and C. quinoa in which it caused systemic symptoms. It caused no infection of globe artichoke (Cynara scolymus) or Pelargonium domesticum. The virus was not adsorbed to soil and could be isolated from leachate of soil in which systemically-infected tomato or C. quinoa plants were grown. Tomato plants became infected when grown in soil watered with virus suspensions. TBSV-T was infective after 10 min at 80°C but not at 90°C and when diluted to 10^-5 but not to 10^-6. Purified virus preparations contained C. 30 nm isometric particles. In gel-diffusion serological tests, TBSV-T reacted with homologous anti-serum and with antiserum to petunia asteroid mosaic virus but not to pelargonium leaf curl virus. Seed-borne infection (50–65%) of TBSV was demonstrated in plants grown from seed of symptomlessly-infected tomato fruit. TBSV was isolated from symptomlessly-infected tomato fruit imported from Morocco during October-April 1981. One of the isolates (TBSV-M) was indistinguishable from TBSV-T in host range, symptomatology and serological reactions. TBSV was also found in tomato plants growing extraneously in primary settlement beds at sewage works; such plants having been derived from undigested seeds in sewage. Because of its ‘alimentary-resistance’ in man, it is possible that one ecological route whereby TBSV enters rivers is by man's consumption of TBSV-infected tomatoes and eventual sewage dispersal into rivers.     

Pathology, soil transmission and characterization of cymbidium ringspot, a virus from cymbidium orchids and white clover (Trifolium repens)

Two strains of a virus, designated cymbidium ringspot virus (CyRSV), were isolated from cymbidium orchids and from Trifolium repens respectively in Britain. Experimentally infected cymbidiums developed slight chlorotic ring-mottle; T. repens developed flecks and mottling in the leaves, and slight stunting. Of 101 plant species tested, the cymbidium strain infected sixty-one (thirteen systemically) in twenty-three of thirty-five families; the clover strain infected sixty-four species (eighteen systemically) in twenty-two families. Both strains were propagated in Nicotiana clevelandii and assayed in Chenopodium quinoa. CyRSV was readily transmitted by inoculation of sap, and by foliage contact between plants, but not by the aphids Myzus persicae or Acyrtho-siphon pisum, nor through seed of T. incarnatum, Phaseolus vulgaris or N. clevelandii. Highly infective virus was released into soil from roots of infected N. clevelandii, and acquired by bait seedlings planted in such soil. Similar transmission occurred when purified virus was applied to the surface of sterilized soil containing bait plants; there was no evidence for any living soil vector. The virus was eliminated from 96 % of small cuttings taken from infected N. clevelandii plants grown at 35–37 °C for 9 wk. CyRSV was still infective in sap of N. clevelandii after dilution to 10?⁵-io–⁶ (only 2 × 10^_1 in cymbidium sap), or after 10min at 85–90 °C. It survived at least 10 months at c. 20 °C and more than 12 yr at 2 °C. Lyophilized sap was highly infective after over 13 yr at laboratory temperatures under high vacuum. Purified preparations made by clarification with n-butanol, followed by differential centrifugation and exclusion chromatography on controlled-pore glass beads, contained isometric particles c. 30 nm diam., with s°_20W= 137 S, and had a buoyant density in caesium chloride of 1–36 g/ml. The A 260/A 280 ratio was 1–55, and A max(26o)/A min(242) was 1–17. The virus contained c. 15 % of single-stranded RNA of mol. wt 1–7 × 10⁶; the nucleotide base ratios were: G27'8; A24/9; C2I-3; U26-I. There was one capsid polypeptide of mol. wt 43600. The virus was a good immunogen and a strongly reacting antigen in vitro; in Immunoelectrophoresis, each strain migrated as a single antigenic component towards the cathode. The cymbidium and clover strains were serologically closely related, although spurs were produced in immunodiffusion. No serological relationship was found to forty-three other isometric viruses, including eighteen tombusvirus isolates; CyRSV nevertheless shares many properties with tombusviruses, and we assign it provisionally to this group. The cryptogram is: R/r:1:7/15:S/S:S/O.     

Partial Characterization of Artichoke Virus M

A virus with filamentous particles 697 nm in length was isolated from artichoke plants in Southern Italy and identified as a new possible member of Carlavirus group, for which the name artichoke virus M (AVM) is suggested. AVM could not be transmitted by sap inoculation to herbaceous hosts and was always present in artichoke in mixed infections with other viruses. Virus particles had a buoyant density in CsCl of 1.31 g × cm^?3 and contained a single species of nucleic acid with an apparent size of 7.5 Kb and a single coat protein species with a mol. wt of 31,000. The virus was distantly related serologically to carnation latent and poplar mosaic carlaviruses but not to other members of the group including the recently described artichoke latent S carlavirus. Cytological alterations consisted of complex cytoplasmic inclusions composed of deranged organelles, lipid droplets and accumulations of membranes.     

MARINICHLORELLA KAISTIAE GEN. ET SP. NOV. (TREBOUXIOPHYCEAE,CHLOROPHYTA) BASED ON POLYPHASIC TAXONOMY1

We describe three coccoid green algal strains belonging to a new genus and species, Marinichlorella kaistiae Z. Aslam, W. Shin, M. K. Kim, W.‐T. Im et S.‐T. Lee, in seawater samples from the South Sea of Korea. These strains were maintained at 25°C–30°C under a 12:12 light:dark (L:D) photoregime in an ASN‐III medium at a pH of 7.5. These strains were tolerant of high salinity (7.5% NaCl) (w/v) and temperature (40°C). Molecular phylogenetic analyses using 18S rRNA gene sequence data resolved these organisms to a clade separate from green coccoid algae with similar morphology. The DNA–DNA hybridization results demonstrated very low relatedness of these organisms to phylogenetically related species of the genera Chlorella and Parachlorella. The molar guanine + cytosine content (G + C mol%) of the genomic DNA of these organisms ranged from 64.7 to 69.1 mol%. Based on molecular phylogeny, DNA–DNA hybridization, and other morphological studies, we propose a new taxon, Marinichlorella kaistiae, to describe these strains and classify them in the family Chlorellaceae. The type strain is KAS007^T (= KCTC AG10303^T = IAM C‐620^T).     

Application of an embryonated chicken egg model to assess the vector competence of Australian Culicoides midges for bluetongue viruses

Culicoides biting midges (Diptera: Ceratopogonidae) are vectors of a number of globally important arboviruses that affect livestock, including bluetongue virus (BTV), African horse sickness virus and the recently emerged Schmallenberg virus. In this study, a model using embryonated chicken eggs (ECEs) was utilized to undertake vector competence studies of Australian Culicoides spp. for 13 laboratory‐adapted or wild‐type virus strains of BTV. A total of 7393 Culicoides brevitarsis were reared from bovine dung, and 3364 Culicoides were induced to feed from ECEs infected with different strains of BTV. Of those, 911 (27%) survived the putative extrinsic incubation period of 9–12 days. In some trials, virus was also transmitted onward to uninfected ECEs, completing the transmission cycle. This model does not rely on the use of colonized midges and has the capacity to assess the vector competence of field‐collected insects with strains of virus that have not previously been passaged in laboratory culture systems. There is also potential for this model to be used in investigations of the competence of Culicoides spp. for other arboviruses.     

Supplemental risk evaluations and status of Puccinia carduorum for biological control of musk thistle

Tests of seven rare and endangered native North American Cirsium species and four modern artichoke lines were requested in response to a proposal for introduction of Puccinia carduorum into the United States for biological control of musk thistle (Carduus nutans ssp. leiophyllus). These tests were supplemental to an earlier extensive host-range study that established P. carduorum from musk thistle as host specific, useful for biological control, and suitable for limited field tests in Virginia. Test plants in the current study were evaluated in support of a proposal to use the rust in the western United States, and particularly, in California. None of the test plants in this study had been evaluated in previous assessments and all were either rare, endangered or threatened in California. Tests were conducted in both field and greenhouse settings. Field tests were run for two seasons, and test plants were inoculated by natural spread of the pathogen from source plants inside rings of test plants. Greenhouse tests involved direct inoculation under optimal conditions of dew and temperature (18–20 °C, 16 h) for infection. None of the seven Cirsium species or subspecies tested became infected by P. carduorum, either in field or greenhouse tests, compared to infection of 98% of the individual musk thistle plants (n = 102) from all the studies. Modern artichoke cultivars were tested only by direct inoculation under optimal greenhouse conditions. All artichoke plants (n = 115) either were immune (no macroscopic symptoms, n = 69) or at most, resistant (n = 46); pustules on all but two of the resistant plants were very small (0.30 mm diam). Despite infections on artichokes, P. carduorum could not be maintained on artichokes under optimal greenhouse conditions. These results confirm earlier findings from host-range tests and risk assessments of P. carduorum. This information suggests that rare, threatened, or endangered Cirsium spp. and modern artichoke cultivars are not likely to be adversely affected by the use of P. carduorum for biological control of musk thistle. These data have been reviewed by grower groups and regulatory agencies in a proposal for permission to use the rust for musk thistle control throughout the United States.