Glitoria yellow vein virus, a tymovirus from Kenya

Clitoria yellow vein virus (CYW) was found in Clitoria ternatea and Abrus precatorius in coastal districts of Kenya, but was not detected in food legume crops. When transmitted by inoculation of sap, CYW infected many species in the Papilionaceae, commonly causing yellowing of secondary and smaller leaf veins. All the economically important food legumes grown in the area of occurrence were very susceptible, so that CYW is potentially very important. The virus also infected okra (Hibiscus esculentus) and species in the Solanaceae, but none of many species of Cucurbitaceae. CYW is serologically closely related to cocoa yellow mosaic and kennedya yellow mottle viruses, and more distantly to okra mosaic and desmodium yellow mottle viruses. Other properties of CYW^ typical of the tymoviruses include particle morphology (particle diameter c. 28 nm; two components) with sedimentation coefficients of 50S (top) and 109S (bottom); molecular weight of protein sub-units c. 20000; thermal inactivation point c. 72 ^oC; and longevity in vitro c. 3 wk.     

Some new Darwin vascular plant specimens from the Beagle voyage

This paper reports 24 newly discovered specimens of 21 species made by Charles Darwin in Argentina, Australia, Brazil, Chile, Ecuador and Uruguay while on the 1831–1836 voyage of HMS Beagle. They have been found in Cambridge University Herbarium and the herbaria of the Missouri Botanical Garden, Natural History Museum, London, New York Botanical Garden and the Royal Botanic Gardens, Kew, since the earlier publications of Porter. Included are type specimens of Calceolaria darwinii (isotype; = C. uniflora), Cuscuta gymnocarpa (holotype and isotypes), C. sandwichiana var. mimosae (isolectotypes = C. gymnocarpa), Ephedra frustillata (lectotype and isolectotypes), Ourisia breviflora (isolectotype), Polypodium paleaceum (syntype?; = Ctenitis sloanei) and Urera gaudichaudiana (holotype; = Laportea aestuans). © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 12–18.     

Detection of methanogens and methanotrophs in natural environments

The role of methane as a greenhouse gas and the contribution of bacteria to the production (methanogenesis) and destruction (methane oxidation) of methane is described. Using experimental approaches based on DNA sequences identifying either methanogen-specific or methanotroph-specific gene sequences methods were developed to broaden the detection and identification of methane metabolizing bacteria in natural environments. These methods were focused on blanket bog peat but are suitable for other environments. In addition to group specific 16S rRNA DNA sequences, specific functional gene probes based on methane coenzyme reductase sequences for methanogens and methane monooxygenase sequences for methanotrophs, were developed. These sequences were used in PCR-based protocols to detect and amplify specific gene sequences from the total DNA isolated from transverse sections of blanket bog peat. This permitted the analysis of the vertical distribution of methanogen and methanotroph populations, discrimination between different sub-sets of these populations, and the identification of novel organisms not previously detected by culture-based methods.     

Carnation Italian ringspot virus

Carnation Italian ringspot virus (CIRV) was obtained only twice in tests on several thousand carnations in Britain during 15 yr. The two isolates, from cultivars ‘Dusty Sim’ imported from Italy and ‘Orchid Beauty’ from the U.S.A., were indistinguishable serologically and in host reactions. CIRV was cultured in Nicotiana clevelandii and assayed in Chenopodium amaranti-color; it was readily transmitted by leaf-rubbing inoculation to 62 of 104 plant species tested. Virus-free carnations were infected only by injecting purified preparations into the stem, and developed chlorotic spots and oval rings in the younger leaves. CIRV was eliminated from Nicotiana clevelandii plants grown for 8 weeks at 36°C. CIRV presents no threat to carnation growing in Britain. In N. clevelandii sap, CIRV was infective at a dilution of 1/50000 to 1/100000, after heating 10 min at 85 °C (but not 90 °C), and after 16 weeks at 16 °C or 23 weeks at 2 °C. After freeze-drying, the virus survived at least 7 yr storage under vacuum at room temperature. CIRV was still infective and antigenic after treatment for 30 min at 18 °C with ultraviolet radiation (750 μW/cm²), ultrasound, 2% formaldehyde or 0.2% tri-sodium ortho-phosphate (TSP). Infectivity was not wholly abolished in 30 min by 2% TSP. The virus was readily purified by overnight maceration of N. clevelandii leaves extracted in phosphate buffer + butanol, followed by differential centri-fugation. Purified preparations contained abundant isometric particles c. 29 nm diameter, and like other serotypes of the tomato bushy stunt-pelargonium leaf curl group, gave three or four specific bands in density-gradient centri-fugation. The bands corresponded to four Schlieren peaks in analytical centrifugation. Virus from the lower bands was usually less invasive in N. clevelandii than from the upper bands, although the material in the different bands contained similar amounts of nucleic acid. Only one antigenic component was found by Immunoelectrophoresis; different serotypes of the TBSV-PLCV group differed widely in immunoelectrophoretic behaviour. The present cryptogram of CIRV is */*:*/*:S/S:S/*.     

Distribution, host range, properties and purification of cassava latent virus, a geminivirus

Cassava latent virus (CLV) is almost entirely confined in East Africa to upland cassava-growing areas west of the Rift Valley, where it is often associated with cassava mosaic disease (it was isolated from 27 of 38 cassava plants with mosaic, but not from 24 without mosaic). However, it is not the causal agent, because it was not recovered from any of 31 mosaic-diseased plants in coastal districts. All attempts to return CLV to cassava failed. The host range of CLV appears to be limited to Euphorbiaceae (Manihot) and Solanaceae (Nicotiana, Datura, Nicandra, Solanum). N. clevelandii proved the most useful assay and propagation host. The dilution end-point of CLV was about 10^-3, thermal inactivation point about 55°C, and longevity in vitro about 3 days. CLV was purified by clarification of leaf extracts with butanol/chloroform mixtures. Purified preparations (A 260/A 280 ratio c. 16) contained numerous 30 20 nm paired particles with a sedimentation coefficient (s₂₀w) of 76 S. Treatment with RNase and DNase showed that the viral nucleic acid is DNA; CLV closely resembles maize streak virus but is not related to it serologically. The cryptogram for CLV is D/1: 0.8/*: S/S: S/*, geminivirus group.     

A strain of cassava latent virus occurring in coastal districts of Kenya

A strain of cassava latent geminivirus (CLV) was isolated from mosaic-affected cassava plants from coastal districts of Kenya. This virus (CLV-C) did not infect Nicotiana clevelandii, a diagnostic host of the type strain (CLV-T); experimental host range was very restricted and CLV-C only infected N. benthamiana and N. rustica out of several solanaceous hosts readily infected by CLV-T. CLV-C was also isolated from naturally infected Jatropha multifida (Euphorbiaceae) and Hewittia sublobata (Convolvulaceae). CLV-C was propagated in N. benthamiana with difficulty and only those isolates derived from cassava plants infected with severe mosaic symptoms were maintained more or less successfully; these sources usually contained a higher concentration of CLV than plants with mild symptoms. Symptom variants generally remained unchanged when grafted into a highly susceptible South American cassava variety. CLV-C and CLV-T seemed to occur respectively only in coastal and western districts but their ranges overlapped in central Kenya where they could have been introduced in infected material. CLV-C could be purified satisfactorily with the method used for CLV-T but only after modifying the procedure by substituting phosphate for borate in the extraction buffer, n-butanol for n-butanol/chloroform in clarification of extracts, and phosphate for borate buffer when resuspending concentrated virus. A virus serologically indistinguishable from CLV-T was isolated from mosaic- affected material obtained from Nigeria; East African and Nigerian isolates were essentially similar in host range and symptomatology. In gel-diffusion serology tests, pronounced precipitation spurs developed between CLV-T and CLV-C indicating that the isolates were related but not identical serologically. Symptoms typical of cassava mosaic disease appeared in only three of 105 plants in experiments on transmission of CLV-C and CLV-T by whiteflies, when attempted acquisition of either clarified CLV-infective sap or purified CLV was made through ‘Parafilm’ membranes. Because it is possible that the three infections resulted from contamination, they cannot constitute proof of transmission. The presence of CLV in relation to the etiology of cassava mosaic thus remains unresolved.     

Purification and some properties of Tephrosia symptomless virus

Tephrosia symptomless virus (TSV), isolated from Tephrosia villosa, is widely distributed in coastal districts of Kenya. The virus was readily transmitted by inoculation of sap, but not by Aphis craccivora or Apion sp. (Curculionidae) or through soil. Host range was very restricted and it infected only 10 of 70 species tested in one of nine plant families; susceptible species were confined to five genera within the Papilionaceae. The virus was cultured, propagated and assayed in soybean. TSV remained infective after 10 min at 85°C, 3 wk at 20°C and 26 wk at -12°C; crude infective sap of Glycine max retained infectivity when diluted 10^-6 but not 10^-7. Virus was purified from systemically infected soybean by clarifying sap extracted in 0.06 m phosphate buffer containing 0.001 m EDTA and 0.1% thioglycollic acid (pH 7.5) with equal volumes of 1:1 n-butanol/chloroform followed by two cycles of differential and one of sucrose density gradient centrifugation. Purified preparations contained c. 33 nm isometric particles. TSV contained RNA and one protein of molecular weight 1.53. 10⁶ and c. 42 000, respectively. Analytical centrifugation indicated a single component with a sedimentation coefficient (s.20, w) of 127 S; in Cs₂SO₄ and CsCl isopycnic gradients a single virus band formed; buoyant density in CsCl was 1.361. TSV was not related serologically to any of 44 viruses in nine plant virus groups but it resembled the tombusviruses and other ungrouped viruses such as carnation mottle in some of its properties.     

Chronic exposure to increasing background ozone impairs stomatal functioning in grassland species

Two species found in temperate calcareous and mesotrophic grasslands (Dactylis glomerata and Leontodon hispidus) were exposed to eight ozone treatments spanning preindustrial to post‐2100 regimes, and late‐season effects on stomatal functioning were investigated. The plants were grown as a mixed community in 14 L containers and were exposed to ozone in ventilated solardomes (dome‐shaped greenhouses) for 20 weeks from early May to late September 2007. Ozone exposures were based on O₃ concentrations from a nearby upland area, and provided the following seasonal 24 h means: 21.4, 39.9 (simulated ambient), 50.2, 59.4, 74.9, 83.3, 101.3 and 102.5 ppb. In both species, stomatal conductance of undamaged inner canopy leaves developing since a midseason cutback increased linearly with increasing background ozone concentration. Imposition of severe water stress by leaf excision indicated that increasing background ozone concentration decreased the ability of leaves to limit water loss, implying impaired stomatal control. The threshold ozone concentrations for these effects were 15–40 ppb above current ambient in upland UK, and were within the range of ozone concentrations anticipated for much of Europe by the latter part of this century. The potential mechanism behind the impaired stomatal functioning was investigated using a transpiration assay. Unlike for lower ozone treatments, apparently healthy green leaves of L. hispidus that had developed in the 101.3 ppb treatment did not close their stomata in response to 1.5 μm abscisic acid (ABA); indeed stomatal opening initially occurred in this treatment. Thus, ozone appears to be disrupting the ABA‐induced signal transduction pathway for stomatal control thereby reducing the ability of plants to respond to drought. These results have potentially wide‐reaching implications for the functioning of communities under global warming where periods of soil drying and episodes of high vapour pressure deficit are likely to be more severe.     

How specialized is the plant–pollinator association between Eucalyptus globulus ssp. globulus and the swift parrot Lathamus discolor?

Abstract The swift parrot Lathamus discolor (Shaw) (Psittacidae) evolved from granivorous ancestors to become a specialized flower‐feeder in a monotypic genus. Its reproduction is dependent largely on flowers of Eucalyptus globulus Labill. ssp. globulus (Myrtaceae), the birds migrating to breed within the natural distribution of this tree. This paper investigates the extent to which this dependence of L. discolor on E. globulus is mirrored by dependence of the tree on the bird. It was found that L. discolor carried significantly more eucalypt pollen within 22 mm of its bill tip than did the New Holland honeyeater, Phylidonyris novaehollandiae (Latham) (Meliphagidae), and that pollen was concentrated on the regions of the head of L. discolor that consistently contact stigmas. Larger pollen loads on L. discolor can be attributed to it consuming both pollen and nectar, while honeyeaters take nectar only. The short thick bill of L. discolor necessitates regular stigmatic contact while the long slender bills of honeyeaters are unlikely to contact stigmas as often in these bowl‐shaped flowers. These factors suggest that L. discolor has a greater capacity to deposit pollen on stigmas of E. globulus than do honeyeaters. However, the characteristics of L. discolor that make it such an effective pollinator of E. globulus are also exhibited by a lorikeet (Psittacidae) that feeds on flowers of E. globulus. The association between E. globulus and L. discolor is therefore only moderately specialized because the flowers are also adapted to the more recently associated lorikeet and are almost certainly also pollinated by honeyeaters.