The evolutionary and molecular features of the broad-host-range plant pathogen Sclerotinia sclerotiorum

Sclerotinia sclerotiorum is a pathogenic fungus that infects hundreds of plant species, including many of the world's most important crops. Key features of S. sclerotiorum include its extraordinary host range, preference for dicotyledonous plants, relatively slow evolution, and production of protein effectors that are active in multiple host species. Plant resistance to this pathogen is highly complex, typically involving numerous polymorphisms with infinitesimally small effects, which makes resistance breeding a major challenge. Due to its economic significance, S. sclerotiorum has been subjected to a large amount of molecular and evolutionary research. In this updated pathogen profile, we review the evolutionary and molecular features of S. sclerotiorum and discuss avenues for future research into this important species.     

Xylem structure of four grape varieties and 12 alternative hosts to the xylem-limited bacterium Xylella fastidious

Background and Aims

The bacterium Xylella fastidiosa (Xf), responsible for Pierce''s disease (PD) of grapevine, colonizes the xylem conduits of vines, ultimately killing the plant. However, Vitis vinifera grapevine varieties differ in their susceptibility to Xf and numerous other plant species tolerate Xf populations without showing symptoms. The aim of this study was to examine the xylem structure of grapevines with different susceptibilities to Xf infection, as well as the xylem structure of non-grape plant species that support or limit movement of Xf to determine if anatomical differences might explain some of the differences in susceptibility to Xf.

Methods

Air and paint were introduced into leaves and stems to examine the connectivity between stem and leaves and the length distribution of their vessels. Leaf petiole and stem anatomies were studied to determine the basis for the free or restricted movement of Xf into the plant.

Key Results

There were no obvious differences in stem or petiole vascular anatomy among the grape varieties examined, nor among the other plant species that would explain differences in resistance to Xf. Among grape varieties, the more tolerant ‘Sylvaner’ had smaller stem vessel diameters and 20 % more parenchyma rays than the other three varieties. Alternative hosts supporting Xf movement had slightly longer open xylem conduits within leaves, and more connection between stem and leaves, when compared with alternative hosts that limit Xf movement.

Conclusions

Stem–leaf connectivity via open xylem conduits and vessel length is not responsible for differences in PD tolerance among grape varieties, or for limiting bacterial movement in the tolerant plant species. However, it was found that tolerant host plants had narrower vessels and more parenchyma rays, possibly restricting bacterial movement at the level of the vessels. The implications of xylem structure and connectivity for the means and regulation of bacterial movement are discussed.     

Performance of nymphal and adult Nezara viridula on selected alternate host plants

Laboratory studies with the southern green stink bug, Nezara viridula (L.) (Hemiptera: Pentatomidae), feeding on seeds or fruits of some alternate host plants, demonstrated differences in nymphal survival and development time, and body weight at adult emergence. Lowest mortalities (25%) were observed for nymphs on fruits of dakota mustard [Brassica kaber (DC.)] and siberian motherwort (Leonurus sibiricus L.). On fruits of castor bean (Ricinus communis L.) nymph mortality was ca. 60%, and on mature seeds of L. sibiricus ca. 75%. Total development time from second instar to adult ranged from ca. 26 days on B. kaber to ca. 42 days on R. communis. Fresh body weight at adult emergence was greatest for nymphs on L. sibiricus and lowest on R. communis. Adult survival after 30 days and mean longevity was greater on L. sibiricus than on the remaining foods, being drastically reduced on R. communis. In general, reproductive performance on all foods was similar, except R. communis, on which no female reproduced. Female weight gain of N. viridula was generally greater on L. sibiricus On R. communis, adults lost weight during their first week and showed the lowest weight gain at the end of day 29. Field observations coupled with these laboratory studies suggest that in the Warta area (northern Paraná state, Brazil, 23° 11 S), L. sibiricus and B. kaber are important alternate food plants in the seasonal phenology of N. viridula. R. communis, however, is of poor nutritional quality, serving primarily as a refuge.

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Résumé L'alimentation au laboratoire de N. viridula avec les graines et les fruits de diverses plantes a entraîné différentes mortalités larvaires, différentes vitesses de développement et des différences dans les poids des adultes à l'émergence. Les mortalités les plus faibles (25%) on été observées avec les fruits de Brassica kaber DC. et de Leonurus sibiricus L.Avec les fruits de Ricinus communis L., la mortalité larvaire était de 60%; sur graines mûres de L. sibiricus, elle était de 75%. La durée du développement du second stade à l'adulte a varié de 26 jours sur B. kaber à 42 jours sur R. communis. Le poids frais des adultes à l'émergence était le plus élevé avec L. sibiricus et le plus faible avec de R. communis. La longévité des adultes au-delà de 30 jours était la plus élevée avec L. sibiricus, et était fortement réduite avec R. communis. Dans l'ensemble, la reproduction était de même importance avec les différents aliments, à l'exception de R. communis avec lequel les femelles ne se reproduisaient pas. Le gain de poids des femelles était généralement supérieur avec L. sibiricus; avec R. communis les adultes ont perdu du poids pendant la première semaine et ont présenté le plus faible gain à la fin du 29ème jour.Des observations dans la nature, associées à ces expériences de laboratoire, laissent supposer que dans la zone de Warta (au Nord de l'état de Paraná, 23° 11 S, au Bresil), L. sibiricus et B. kaber sont des plantes hôtes alternatives importantes dans la phénologie saisonnière de N. viridula. Cependant, R. communis, de faible valeur alimentaire, sert avant tout de refuge.

     

Derivation and characterization of cholesterol-independent non-GS NS0 cell lines for production of recombinant antibodies

Presented is an antibody production platform based on the fed-batch culture of recombinant NS0-derived cell lines. NS0 host cells, obtained from the European Collection of Cell Cultures (ECACC, Salisbury, UK, Part No. 85110503), were first adapted to grow in a protein-free, cholesterol-free medium. The resulting host cell line was designated NS0-PFCF (protein-free, cholesterol-free). The five production cell lines presented here were generated using a common protocol consisting of transfection by electroporation and subcloning. The NS0-PFCF host cell line was transfected using a single expression vector containing the Escherichia coli xanthine-guanine phosphoribosyl transferase gene (gpt), and the antibody heavy and light chain genes driven by the CMV promoter. The five cell lines were chosen after one to three rounds of iterative subcloning, which resulted in a 19-64% increase in antibody productivity when four mother-daughter cell pairs were cultured in a fed-batch bioreactor process. The production cell lines were genetically characterized to determine antibody gene integrity, nucleotide sequences, copy number, and the number of insertion sites in the NS0 cell genome. Genetic characterization data indicate that each of the five production cell lines has a single stably integrated copy of the antibody expression vector, and that the antibody genes are correctly expressed. Stability of antibody production was evaluated for three of the five cell lines by comparing the early stage seed bank with the Working Cell Bank (WCB). Antibody productivity was shown to be stable in two of three cell lines evaluated, while one of the cell lines exhibited a 20% drop in productivity after passaging for approximately 4 weeks. These five NS0-derived production cell lines were successfully cultured to produce antibodies with acceptable product quality attributes in a standardized fed-batch bioreactor process, consistently achieving an average specific productivity of 20-60 pg/cell-day, and a volumetric productivity exceeding 120 mg/L-day (Burky et al., 2006). In contrast to the commonly available NS0 host cell line, which requires serum and cholesterol for growth, and the commonly used expression vector system, which uses a proprietary glutamine synthetase selection marker (GS-NS0), these NS0 cells are cholesterol-independent, grow well in a protein-free medium, use a non-proprietary selection marker, and do not require gene amplification for productivity improvement. These characteristics are advantageous for use of this NS0 cell line platform for manufacturing therapeutic antibodies.     

Adaptive Host Preference and the Dynamics of Host–Parasitoid Interactions

Models of two independent host populations and a common parasitoid are investigated. The hosts have density-dependent population growth and only interact indirectly by their effects on parasitoid behavior and population dynamics. The parasitoid is assumed to experience a trade-off in its ability to exploit the two hosts. Three alternative types of parasitoid are investigated: (i) fixed generalists whose consumption rates are those that maximize fitness; (ii) “ideal free” parasitoids, which modify their behavior to maximize their rate of finding unparasitized hosts within a generation; and (iii) “evolving” parasitoids, whose capture rates change between generations based on quantitative genetic determination of the relative attack rates on the two hosts. The primary questions addressed are: (1) Do the different types of adaptive processes stabilize or destabilize the population dynamics? (2) Do the adaptive processes tend to equalize or to magnify differences in host densities? The models show that adaptive behavior and evolution frequently destabilize population dynamics and frequently increase the average difference between host densities.     

Olfactory responses to attractants and repellents in tsetse

The aims of this study were to investigate how antennal olfactory cells of tsetse (Diptera: Glossinidae) code odour quality and how they are able to discriminate between attractive and repellent odours. For Glossina pallidipes Austen, a survey is presented of the cells' responses to attractive (1-octen-3-ol, acetone, 3-methylphenol, carbon dioxide) and repellent stimuli (2-methoxyphenol, acetophenone, lactic acid, naphthalene). In addition, the responses of these cells to binary mixtures and the dose-response curves of 1-octen-3-ol, 3-methylphenol, 2-methoxyphenol and acetophenone are presented. A minority of the cells responded to one attractant or repellent only, whereas the vast majority were excited by more than one of the attractive and/or repellent stimuli. It is proposed that the peripheral olfactory cells of tsetse discriminate between different compounds via an across-fibre pattern coding, in which the cells that specifically code for attractants or repellents may play a substantial role in composing a unique excitation pattern that informs the central nervous system about the specificity of odours.     

Ambrosia beetles (Curculionidae: Scolytinae and Platypodinae) on Fagus crenata Blume: community structure,seasonal population trends and resource utilization patterns

Ambrosia beetles (many Scolytinae and all Platypodinae) are one of the most important insect pests for forestry worldwide, but little is known about the community structure of ambrosia beetles in terms of their vertical variations and resource utilization. We clarified the community structure and seasonal population trends of ambrosia beetles on 11 living and three newly dead Fagus crenata Blume trees using individual tube traps placed up to 10 m high from May to November in 2007 and 2008. We captured seven scolytine species (Ambrosiodmus lewisi (Blandford), Euwallacea validus (Eichhoff), Trypodendron proximum (Niisima), Xyleborinus saxeseni (Ratzeburg), Xyleborus atratus Eichhoff, Xylosandrus brevis (Eichhoff) and Xylosandrus germanus (Blandford)) and three platypodine species (Crossotarsus niponicus Blandford, Platypus hamatus Blandford and Platypus severini Blandford). The ambrosia beetles were suggested to breed at species‐specific height ranges, with the equal host resource use per individual among the species. Of the three major species, C. niponicus, P. hamatus and P. severini, two (C. niponicus and P. hamatus) had male‐biased sex ratios, which is considered a reproductive strategy to increase maternal fitness. Morphological characteristics of the mandibles may play an important role in the difference of sex roles on reproduction in the three major species.     

Infochemical-mediated preference behavior of the parasitoid Microctonus hyperodae when searching for its adult weevil hosts

Infochemicals are the most important cues used by parasitoids for host location. The attractiveness of infochemicals in a tritrophic context is expected to be determined by the degree of specialization of the parasitoid and its host(s). Microctonus hyperodae Loan (Hymenoptera: Braconidae) is an oligophagous parasitoid that attacks adult Curculionidae of the Brachycerinae subfamily, especially Listronotus bonariensis Kuschel, on Gramineae. In 1996, a new host–parasitoid association between the carrot weevil Listronotus oregonensis LeConte and M. hyperodae was created in the laboratory. In this study, the infochemicals used by M. hyperodae when searching for its adult weevil hosts were determined using a Y‐shaped olfactometer. Three curculionid species (L. oregonensis, Listronotus sparsus Say, and Neydus flavicaudis Boheman) and one bruchid species [Callosobruchus maculatus (Fabricius)], and their feces, were tested. It was expected that hosts phylogenetically and ecologically close to L. bonariensis would be more attractive than species less related but in fact, M. hyperodae responded only to L. oregonensis and its feces. When feces and host insects were tested separately, M. hyperodae responded to the odors emitted by L. oregonensis adults but not to their feces, suggesting that most of the kairomones came from the host itself. Host plants were also tested, but M. hyperodae responded neither to Lolium multiflorum Lamark (Gramineae) nor to Daucus carota L. (Umbelliferae) leaves.