Effect of Metalaxyl Resistance and Cultivar Resistance on Control of Phytophthora nicotianae in Tobacco

Phytophthora nicotianae is a devastating root and stem pathogen of tobacco (Nicotiana tabacum) in South Africa. Growers strive to control the resulting disease, known as black shank, with metalaxyl treatments and resistant cultivars. The aim of this study was to consider whether development of metalaxyl resistance in P. nicotianae has contributed to poor disease control and if recently developed cultivars with high levels of resistance require metalaxyl for effective control. One hundred and thirty-two isolates of P. nicotianae were screened for sensitivity to metalaxyl. P. nicotianae isolates from most tobacco farms were metalaxyl sensitive. Growth of most isolates was inhibited completely at 1.0 μg a.i./ml. However, isolates from the MKTV tobacco producing area showed EC50 values ranging from 1.02 μg a.i./ml to 3.57 μg a.i./ml. Twenty-one tobacco cultivars were planted and treated with and without metalaxyl in two different growing seasons to evaluate their resistance to P. nicotianae and the value of using metalaxyl. Hicks was the most susceptible cultivar. Vuma/3/46, LK30/40/60-1, and LK33/60 exhibited the greatest resistance to P. nicotianae. Use of metalaxyl in combination with moderately resistant cultivars such as NC60 × TL33 and LK10/80/60 effectively reduced black shank in the field. Resistant cultivars were healthy and no significant difference between metalaxyl treated and untreated plants was observed.

A synthetic xylanase as a novel reporter in plants

Transient gene expression assays are often used to screen promoters before stable transformation. Current transient quantification methods have several problems, including a lack of reporter gene stability and expense. Here we report a synthetic, codon-optimised xylanase gene (sXynA) as a reporter gene for quantitative transient analyses in plants. Azurine-crosslinked xylan (AZCL-xylan) was used as a substrate for assaying xylanase activity. The enzymatic nature of the protein allows for sensitive assays at the low levels of transgene protein found in transiently transformed tissue extracts. The xylanase (XYN) protein is stable, activity slopes are linear over long time periods and assays are cost-effective. Coupled with the GUSPlus reporter gene, the XYN reporter allows sensitive and accurate quantification of gene control sequences in transient expression systems.

Modulation of higher-plant NAD(H)-dependent glutamate dehydrogenase activity in transgenic tobacco via alteration of beta subunit levels

Glutamate dehydrogenase (GDH; EC 1.4.1.2-1.4.1.4) catalyses in vitro the reversible amination of 2-oxoglutarate to glutamate. In vascular plants the in vivo direction(s) of the GDH reaction and hence the physiological role(s) of this enzyme remain obscure. A phylogenetic analysis identified two clearly separated groups of higher-plant GDH genes encoding either the alpha- or beta-subunit of the GDH holoenzyme. To help clarify the physiological role(s) of GDH, tobacco (Nicotiana tabacum L.) was transformed with either an antisense or sense copy of a beta-subunit gene, and transgenic plants recovered with between 0.5- and 34-times normal leaf GDH activity. This large modulation of GDH activity (shown to be via alteration of beta-subunit levels) had little effect on leaf ammonium or the leaf free amino acid pool, except that a large increase in GDH activity was associated with a significant decrease in leaf Asp (similar to 51%, P=0.0045). Similarly, plant growth and development were not affected, suggesting that a large modulation of GDH beta-subunit titre does not affect plant viability under the ideal growing conditions employed. Reduction of GDH activity and protein levels in an antisense line was associated with a large increase in transcripts of a beta-subunit gene, suggesting that the reduction in beta-subunit levels might have been due to translational inhibition. In another experiment designed to detect post-translational up-regulation of GDH activity, GDH over-expressing plants were subjected to prolonged dark-stress. GDH activity increased, but this was found to be due more likely to resistance of the GDH protein to stress-induced proteolysis, rather than to post-translational up-regulation.

Host recognition by a polyphagous lepidopteran (Helicoverpa armigera): primary host plants, host produced volatiles and neurosensory stimulation

An important question in the host-finding behaviour of a polyphagous insect is whether the insect recognizes a suite or template of chemicals that are common to many plants? To answer this question, headspace volatiles of a subset of commonly used host plants (pigeon pea, tobacco, cotton and bean) and nonhost plants (lantana and oleander) of Helicoverpa armigera Hubner (Lepidoptera: Noctuidae) are screened by gas chromatography (GC) linked to a mated female H. armigera electroantennograph (EAG). In the present study, pigeon pea is postulated to be a primary host plant of the insect, for comparison of the EAG responses across the test plants. EAG responses for pigeon pea volatiles are also compared between females of different physiological status (virgin and mated females) and the sexes. Eight electrophysiologically active compounds in pigeon pea headspace are identified in relatively high concentrations using GC linked to mass spectrometry (GC-MS). These comprised three green leaf volatiles [(2E)-hexenal, (3Z)-hexenylacetate and (3Z)-hexenyl-2-methylbutyrate] and five monoterpenes (alpha-pinene,beta-myrcene, limonene, E-beta-ocimene and linalool). Other tested host plants have a smaller subset of these electrophysiologically active compounds and even the nonhost plants contain some of these compounds, all at relatively lower concentrations than pigeon pea. The physiological status or sex of the moths has no effect on the responses for these identified compounds. The present study demonstrates how some host plants can be primary targets for moths that are searching for hosts whereas the other host plants are incidental or secondary targets.

A Research Agenda for Assessing the Potential Contribution of Genomic Medicine to Tobacco Control

This paper identifies research priorities in evaluating the ways in which "genomic medicine"-the use of genetic information to prevent and treat disease-may reduce tobacco-related harm by: (1) assisting more smokers to quit; (2) preventing non-smokers from beginning to smoke tobacco; and (3) reducing the harm caused by tobacco smoking. The method proposed to achieve the first aim is pharmacogenetics", the use of genetic information to optimise the selection of smoking-cessation programmes by screening smokers for polymorphisms that predict responses to different methods of smoking cessation. This method competes with the development of more effective forms of smoking cessation that involve vaccinating smokers against the effects of nicotine and using new pharmaceuticals (such as cannabinoid antagonists and nicotine agonists). The second and third aims are more speculative. They include: screening the population for genetic susceptibility to nicotine dependence and intervening (eg, by vaccinating children and adolescents against the effects of nicotine) to prevent smoking uptake, and screening the population for genetic susceptibility to tobacco-related diseases. A framework is described for future research on these policy options. This includes: epidemiological modelling and economic evaluation to specify the conditions under which these strategies are cost-effective; and social psychological research into the effect of providing genetic information on smokers' preparedness to quit, and the general views of the public on tobacco smoking.

The prospects for tobacco harm reduction

Tobacco harm reduction (THR) policies aim to reduce the prevalence of tobacco-related harm by encouraging smokers who are unable or unwilling to quit to adopt less harmful ways of obtaining nicotine, such as pharmaceutical nicotine and oral tobacco snuff. Proponents of THR argue that the effects of tobacco control policies have reduced smoking as much as they reasonably can and that we can best reduce tobacco-related harm by encouraging smokers to use these methods, which substantially reduce the health risks of smoking. Critics argue that THR policies will undermine the two traditional tobacco control goals of preventing the uptake of smoking by young people and encouraging smokers to quit. I assess the main arguments and evidence advanced for and against THR.

Assessment of Swedish snus for tobacco harm reduction: technical appendix

This technical appendix details the methods used in an assessment of the potential of snus for tobacco harm reduction using simulation modelling.

Is YouTube telling or selling you something? Tobacco content on the YouTube video sharing website

The effectiveness of overt tobacco advertising and sponsorship bans is well established. The industry has responded to these bans by implementing “buzz” or “viral” marketing techniques, such as nightclub and dance party promotions. This paper analyses possible tobacco industry content on the burgeoning consumer generated media website, YouTube. Tobacco control efforts need to embrace this new medium in order to counter pro-smoking messages and maximize media advocacy opportunities.

The future of tobacco control: project summary

The National Health and Medical Research Council has funded Professor Wayne Hall (University of Queensland) and Professor Simon Chapman (University of Sydney) for three years 2006-2008, to research aspects of the future of tobacco control, particularly in nations with advanced tobacco control programs like Australia. Dr Coral Gartner (UQ) and Ms Becky Freeman (USyd) are also working on the project. The University of Queensland's eSpace site provides links to papers and data appendices produced by the University of Queensland team on the project. Materials relevant to this project produced by the University of Sydney group are available at the link provided.

Should the health community promote smokeless tobacco (snus) as a harm reduction measure?

Background to the debate: The tobacco control community is divided on whether or not to inform the public that using oral, smokeless tobacco (Swedish snus) is less hazardous to health than smoking tobacco. Proponents of 'harm reduction' point to the Swedish experience. Snus seems to be widely used as an alternative to cigarettes in Sweden, say these proponents, contributing to the low overall prevalence of smoking and smoking-related disease. Harm reduction proponents thus argue that the health community should actively inform inveterate cigarette smokers of the benefits of switching to snus. However, critics of harm reduction say that snus has its own risks, that no form of tobacco should ever be promoted, and that Sweden’s experience is likely to be specific to that culture and not transferable to other settings. Critics also remain deeply suspicious that the tobacco industry will use snus marketing as a 'gateway' to promote cigarettes. In the interests of promoting debate, the authors (who are collaborators on a research project on the future of tobacco control) have agreed to outline the strongest arguments for and against promoting Swedish snus as a form of harm reduction.

The proportion of individual lucerne plants resistant to Phytophthora medicaginis and Colletotrichum trifolii in Australian cultivars

Phytophthora root rot (Phytophthora medicaginis) and colletotrichum crown rot (Colletotrichum trifoli) are the 2 most serious pathogens of lucerne in eastern Australia. Work reported in this paper shows that in glasshouse tests of the 11 most commonly grown Australian lucerne cultivars, the proportion of individual plants with resistance to both pathogens ranges from 0 (Hunter River and Aurora) through to a maximum of 19.8% (Sequel HR). Within 9 of the cultivars, the proportion of individual plants resistant to the 2 pathogens was <7%. Since these 2 diseases are known to cause serious losses in eastern Australia, the results indicate further improvement in lucerne production can be obtained by increasing the proportion of individual plants in a cultivar resistant to both pathogens. This would be best achieved by identifying dominant sources of resistance and incorporating this into on-going lucerne breeding programs.